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Fayyaz A, Cugia L, Noli M, Jasemi S, Simula ER, Sechi LA. A Characterization of the Humoral Immune Response to Human Endogenous Retroviruses and Mycobacterium paratuberculosis in Crohn's Disease. Pathogens 2025; 14:361. [PMID: 40333136 PMCID: PMC12030244 DOI: 10.3390/pathogens14040361] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2025] [Revised: 04/03/2025] [Accepted: 04/05/2025] [Indexed: 05/09/2025] Open
Abstract
Crohn's disease (CD) is a multifactorial polygenic inflammatory bowel disease linked to aberrant immune response. Mycobacterium paratuberculosis (MAP) has been associated with CD; however, detecting MAP in CD tissues remains highly challenging. Recently, Human Endogenous Retroviruses (HERVs) differential gene expression has been reported in CD, but little is known about the involvement of MAP and HERVs in CD pathology. This study aimed to characterize the humoral response against HERV-K, HERV-W, and MAP antigens using an indirect ELISA in plasma samples from CD patients and age- and gender-matched healthy controls (HCs). We observed a significant antibody response against HERV-K and HERV-W epitopes in CD patients in comparison to MAP epitopes, as well as a higher overall antibody response in patients compared to HCs. This study is the first to report the presence of humoral immune response against HERVs antigens in CD. Considering the pro-inflammatory nature of CD, HERVs may contribute to the development or progression of disease in genetically predisposed individuals. However, further research is needed to better understand the complex role of HERVs in CD.
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Affiliation(s)
- Alishba Fayyaz
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy; (A.F.); (M.N.); (S.J.)
- Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy
| | - Luigi Cugia
- Gastroenterology and Digestive Endoscopy Department, Azienda Ospedaliera Universitaria di Sassari, 07100 Sassari, Italy;
| | - Marta Noli
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy; (A.F.); (M.N.); (S.J.)
| | - Somaye Jasemi
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy; (A.F.); (M.N.); (S.J.)
| | - Elena Rita Simula
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy; (A.F.); (M.N.); (S.J.)
| | - Leonardo A. Sechi
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy; (A.F.); (M.N.); (S.J.)
- SC Microbiologia e Virologia, Azienda Ospedaliera Universitaria, 07100 Sassari, Italy
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Ashraf H, Dikarlo P, Masia A, Zarbo IR, Solla P, Ijaz UZ, Sechi LA. Mycobacterium avium subspecies paratuberculosis (MAP) infection, and its impact on gut microbiome of individuals with multiple sclerosis. Sci Rep 2024; 14:24027. [PMID: 39402079 PMCID: PMC11479286 DOI: 10.1038/s41598-024-74975-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2024] [Accepted: 09/30/2024] [Indexed: 10/17/2024] Open
Abstract
The microbial ecology of Mycobacterium avium subspecies paratuberculosis infections (MAP) within the context of Multiple Sclerosis (MS) is largely an unexplored topic in the literature. Thus, we have characterized the compositional and predicted functional differences of the gut microbiome between MS patients with MAP (MAP+) and without (MAP-) infection. This was done in the context of exposome differences (through self-reported filled questionnaires), principally in anthropometric and sociodemographic patterns to gain an understanding of the gut microbiome dynamics. 16S rRNA microbiome profiling of faecal samples (n = 69) was performed for four groups, which differed by disease and MAP infection: healthy cohort (HC) MAP-; HC MAP+ ; MS MAP-; and MS MAP+ . Using a dynamic strategy, with MAP infection and time of sampling as occupancy models, we have recovered the core microbiome for both HC and MS individuals. Additional application of neutral modeling suggests key genera that are under selection pressure by the hosts. These include members of the phyla Actinobacteriota, Bacteroidota, and Firmicutes. As several subjects provided multiple samples, a Quasi Conditional Association Test that incorporates paired-nature of samples found major differences in Archaea. To consolidate treatment groups, confounders, microbiome, and the disease outcome parameters, a mediation analysis is performed for MS cohort. This highlighted certain genera i.e., Sutterella, Akkermansia, Bacteriodes, Gastranaerophilales, Alistipes, Balutia, Faecalibacterium, Lachnospiraceae, Anaerostipes, Ruminococcaceae, Eggerthellaceae and Clostridia-UCG-014 having mediatory effect using disease duration as an outcome and MAP infection as a treatment group. Our analyses indicate that the gut microbiome may be an important target for dietary and lifestyle intervention in MS patients with and without MAP infection.
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Affiliation(s)
- Hajra Ashraf
- Department of Biomedical Sciences, University of Sassari, Sassari, Italy
- Water & Environment Research Group, Mazumdar-Shaw Advanced Research Centre, University of Glasgow, Glasgow, UK
| | - Plamena Dikarlo
- BIOMES NGS GmbH, Schwartzkopffstraße 1, Halle 21, 15745, Wildau, Germany
| | - Aurora Masia
- Department of Medicine and Pharmacy, Neurology, University of Sassari, Sassari, Italy
| | - Ignazio R Zarbo
- Department of Medicine and Pharmacy, Neurology, University of Sassari, Sassari, Italy
| | - Paolo Solla
- Department of Medicine and Pharmacy, Neurology, University of Sassari, Sassari, Italy
| | - Umer Zeeshan Ijaz
- Water & Environment Research Group, Mazumdar-Shaw Advanced Research Centre, University of Glasgow, Glasgow, UK.
- National University of Ireland, University Road, Galway, Ireland.
- Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK.
| | - Leonardo A Sechi
- Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
- Complex Structure of Microbiology and Virology, AOU Sassari, Sassari, Italy.
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Aitken JM, Aitken JE, Agrawal G. Mycobacterium avium ssp. paratuberculosis and Crohn's Disease-Diagnostic Microbiological Investigations Can Inform New Therapeutic Approaches. Antibiotics (Basel) 2024; 13:158. [PMID: 38391544 PMCID: PMC10886072 DOI: 10.3390/antibiotics13020158] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2023] [Revised: 01/24/2024] [Accepted: 02/03/2024] [Indexed: 02/24/2024] Open
Abstract
Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of Johne's disease (JD), which is a chronic infectious gastrointestinal disease of ruminants and is often fatal. In humans, MAP has been associated with Crohn's disease (CD) for over a century, without conclusive evidence of pathogenicity. Numerous researchers have contributed to the subject, but there is still a need for evidence of the causation of CD by MAP. An infectious aetiology in CD that is attributable to MAP can only be proven by bacteriological investigations. There is an urgency in resolving this question due to the rising global incidence rates of CD. Recent papers have indicated the "therapeutic ceiling" may be close in the development of new biologics. Clinical trial outcomes have demonstrated mild or inconsistent improvements in therapeutic interventions over the last decades when compared with placebo. The necessity to revisit therapeutic options for CD is becoming more urgent and a renewed focus on causation is essential for progress in identifying new treatment options. This manuscript discusses newer interventions, such as vaccination, FMT, dietary remediation and gut microbiome regulation, that will become more relevant as existing therapeutic options expire. Revisiting the MAP theory as a potential infectious cause of CD, rather than the prevailing concept of an "aberrant immune response" will require expanding the current therapeutic programme to include potential new alternatives, and combinations of existing treatments. To advance research on MAP in humans, it is essential for microbiologists and medical scientists to microscopically detect CWDM and to biologically amplify the growth by directed culture.
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Affiliation(s)
- John M Aitken
- Otakaro Pathways Ltd., Innovation Park, Christchurch 7675, New Zealand
| | - Jack E Aitken
- Otakaro Pathways Ltd., Innovation Park, Christchurch 7675, New Zealand
| | - Gaurav Agrawal
- Division of Diabetes & Nutritional Sciences, Franklin-Wilkins Building, King's College London, London SE1 9NH, UK
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Ekundayo TC, Olasehinde TA, Falade AO, Adewoyin MA, Iwu CD, Igere BE, Ijabadeniyi OA. Systematic review and meta-analysis of Mycobacterium avium subsp. paratuberculosis as environmental trigger of multiple sclerosis. Mult Scler Relat Disord 2022; 59:103671. [DOI: 10.1016/j.msard.2022.103671] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2021] [Revised: 01/27/2022] [Accepted: 02/05/2022] [Indexed: 12/24/2022]
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An Escherichia coli carrier vaccine with surface-displayed protein MAP3061c elicits protective immunity against Mycobacterium paratuberculosis in mice. Res Vet Sci 2021; 141:180-189. [PMID: 34763254 DOI: 10.1016/j.rvsc.2021.10.019] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/13/2021] [Revised: 10/06/2021] [Accepted: 10/28/2021] [Indexed: 11/24/2022]
Abstract
Johne's disease, or paratuberculosis, is a chronic granulomatous enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). This disease occurs worldwide and results in considerable economic losses in the livestock industry. There are no effective treatments for Johne's disease, so there is an urgent need to develop an efficient, economical, and stable vaccine for MAP control. Here, a live Escherichia coli (E. coli) surface display vaccine harboring the MAP3061c gene was developed through an ice nucleation protein (INP) surface display system. The experimental data demonstrated that MAP3061c has strong immunogenicity and that the surface displayed vaccine can stimulate mice to produce high levels of antibodies. Both CD4+ and CD8+ T cell counts as well as several cytokines - including IFN-γ, IL-4, IL-10, IL-17A and IL-23 - were significantly increased in the display vaccine group. Post-vaccination challenge with MAP in mice resulted in improved fitness of the mice as demonstrated by a lack of weight loss. Pathological results revealed that the surface display vaccine could reduce the degree of pathological damage and slowed the course of disease. Taken together, our data suggests that the E. coli carrier vaccine with surface-displayed MAP3061c elicits protective immunity against MAP, providing new insights into the development of a MAP vaccine.
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Mallikarjunappa S, Brito LF, Pant SD, Schenkel FS, Meade KG, Karrow NA. Johne's Disease in Dairy Cattle: An Immunogenetic Perspective. Front Vet Sci 2021; 8:718987. [PMID: 34513975 PMCID: PMC8426623 DOI: 10.3389/fvets.2021.718987] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/01/2021] [Accepted: 07/19/2021] [Indexed: 12/25/2022] Open
Abstract
Johne's disease (JD), also known as paratuberculosis, is a severe production-limiting disease with significant economic and welfare implications for the global cattle industry. Caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP), JD manifests as chronic enteritis in infected cattle. In addition to the economic losses and animal welfare issues associated with JD, MAP has attracted public health concerns with potential association with Crohn's disease, a human inflammatory bowel disease. The lack of effective treatment options, such as a vaccine, has hampered JD control resulting in its increasing global prevalence. The disease was first reported in 1895, but in recognition of its growing economic impact, extensive recent research facilitated by a revolution in technological approaches has led to significantly enhanced understanding of the immunological, genetic, and pathogen factors influencing disease pathogenesis. This knowledge has been derived from a variety of diverse models to elucidate host-pathogen interactions including in vivo and in vitro experimental infection models, studies measuring immune parameters in naturally-infected animals, and by studies conducted at the population level to enable the estimation of genetic parameters, and the identification of genetic markers and quantitative trait loci (QTL) putatively associated with susceptibility or resistance to JD. The main objectives of this review are to summarize these recent developments from an immunogenetics perspective and attempt to extract the principal and common findings emerging from this wealth of recent information. Based on these analyses, and in light of emerging technologies such as gene-editing, we conclude by discussing potential future avenues for effectively mitigating JD in cattle.
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Affiliation(s)
- Sanjay Mallikarjunappa
- Department of Animal Biosciences, Centre for Genetic Improvement of Livestock, University of Guelph, Guelph, ON, Canada
| | - Luiz F Brito
- Department of Animal Sciences, Purdue University, West Lafayette, IN, United States
| | - Sameer D Pant
- Graham Centre for Agricultural Innovation, Charles Sturt University, Wagga Wagga, NSW, Australia
| | - Flavio S Schenkel
- Department of Animal Biosciences, Centre for Genetic Improvement of Livestock, University of Guelph, Guelph, ON, Canada
| | - Kieran G Meade
- School of Agriculture and Food Science, University College Dublin, Dublin, Ireland
| | - Niel A Karrow
- Department of Animal Biosciences, Centre for Genetic Improvement of Livestock, University of Guelph, Guelph, ON, Canada
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Hosseiniporgham S, Biet F, Ganneau C, Bannantine JP, Bay S, Sechi LA. A Comparative Study on the Efficiency of Two Mycobacterium avium subsp. paratuberculosis (MAP)-Derived Lipopeptides of L3P and L5P as Capture Antigens in an In-House Milk ELISA Test. Vaccines (Basel) 2021; 9:vaccines9090997. [PMID: 34579234 PMCID: PMC8471605 DOI: 10.3390/vaccines9090997] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2021] [Revised: 09/01/2021] [Accepted: 09/02/2021] [Indexed: 02/07/2023] Open
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) surface-exposed lipopeptides could be specific capture-antigen molecules targeting antibodies against MAP, in milk, through ELISA. Previous studies have revealed that MAP strains, isolated from sheep (S) or cow (C), could produce specific lipopeptides, L3P or L5P, respectively. In this study, we used L3P and L5P as capture antigens in an in-house milk ELISA (H-MELISA) to assess how these antigens perform, in comparison with other ELISA tests, on well-defined milk samples from MAP-infected sheep. The overall positivity rates of H-MELISA via L3P and L5P varied by the source of milk samples, in which, at bulk-tank-milk (BTM) level, the majority of positive cases (63.83%) reacted more against L5P, whereas a predominant number (69.14%) of milk samples were more responsive against L3P at the individual level. To clarify whether the positivity status of milk samples in H-MELISA L3P/L5P were predictive of MAP strain-types (S/C), strain-typing was carried out using PCR IS1311-restriction enzyme analysis. Although the presence of three MAP strains (S/C/bison types) was detected among the milk samples, the C-type (46.67%) and S-type (75%) MAP strains were detected with higher incidence among BTMs and individual milk samples, respectively. However, further examination on the H-MELISA L3P/L5P-positivity pattern of each C/S-type-MAP sample revealed that some samples had a reverse reactivity against both L3P and L5P. These results could be the consequence of either cross-reactivity between L3P and L5P (due to the similarity in the structures of the two epitopes) or simply a within-herd mixed infection with MAP strains of C and S types. These findings suggest that lipopeptide antigens could contribute a diagnostic test with optimal performance, considering the diversity of MAP strains.
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Affiliation(s)
| | - Franck Biet
- UMR1282, Infectiologie et Santé Publique (ISP-311), INRAE Centre Val de Loire, 37380 Nouzilly, France;
| | - Christelle Ganneau
- Unité de Chimie Des Biomolécules, Département de Biologie Structurale et Chimie, Institut Pasteur, 75015 Paris, France; (C.G.); (S.B.)
- CNRS UMR 3523, 75015 Paris, France
| | - John P. Bannantine
- USDA-Agricultural Research Service, National Animal Disease Center, Ames, IA 50010, USA;
| | - Sylvie Bay
- Unité de Chimie Des Biomolécules, Département de Biologie Structurale et Chimie, Institut Pasteur, 75015 Paris, France; (C.G.); (S.B.)
- CNRS UMR 3523, 75015 Paris, France
| | - Leonardo A. Sechi
- Dipartimento di Scienze Biomediche, Università di Sassari, 07100 Sassari, Italy;
- SC Microbiologia e Virologia, Azienda Ospedaliera Universitaria, 07100 Sassari, Italy
- Mediterraneam Center for Disease Control, 07100 Sassari, Italy
- Correspondence:
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Crohn’s Disease: The infectious Disease Incorporated’s Perspective. GASTROINTESTINAL DISORDERS 2021. [DOI: 10.3390/gidisord3030015] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/17/2022] Open
Abstract
Infectious Diseases Incorporated (IDI) is an infectious disease think-tank, established in 1973. Crohn’s disease (CD) is a chronic, recurrent disease of the gastrointestinal tract that has reached epidemic proportions within industrialized nations. CD is said to be without cure. Since 2003, therapeutic interventions have focused on disruption of the pro-inflammatory Th1 response against an unknown antigen. In 2015, the Hruska Postulate was introduced and, in so doing, explained how, in the absence of acquired immunity, newborn infection by Mycobacterium avium subspecies paratuberculosis could cause fixation of the immune system’s Th1 response against the organism. The Hruska Postulate was utilized to answer all the documented epidemiological facts embedded in the natural history of Crohn’s disease and, in particular, why breastfeeding confers protection against the future development of Crohn’s disease. It is Infectious Diseases Incorporated’s (IDI) stated opinion that Crohn’s disease is both preventable and curable if treated appropriately in its early stages.
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Kleinwort KJH, Hobmaier BF, Mayer R, Hölzel C, Degroote RL, Märtlbauer E, Hauck SM, Deeg CA. Mycobacterium avium subsp. paratuberculosis Proteome Changes Profoundly in Milk. Metabolites 2021; 11:metabo11080549. [PMID: 34436489 PMCID: PMC8399727 DOI: 10.3390/metabo11080549] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2021] [Revised: 08/10/2021] [Accepted: 08/18/2021] [Indexed: 11/16/2022] Open
Abstract
Mycobacterium avium subspecies paratuberculosis (MAP) are detectable viable in milk and other dairy products. The molecular mechanisms allowing the adaptation of MAP in these products are still poorly understood. To obtain information about respective adaptation of MAP in milk, we differentially analyzed the proteomes of MAP cultivated for 48 h in either milk at 37 °C or 4 °C or Middlebrook 7H9 broth as a control. From a total of 2197 MAP proteins identified, 242 proteins were at least fivefold higher in abundance in milk. MAP responded to the nutritional shortage in milk with upregulation of 32% of proteins with function in metabolism and 17% in fatty acid metabolism/synthesis. Additionally, MAP upregulated clusters of 19% proteins with roles in stress responses and immune evasion, 19% in transcription/translation, and 13% in bacterial cell wall synthesis. Dut, MmpL4_1, and RecA were only detected in MAP incubated in milk, pointing to very important roles of these proteins for MAP coping with a stressful environment. Dut is essential and plays an exclusive role for growth, MmpL4_1 for virulence through secretion of specific lipids, and RecA for SOS response of mycobacteria. Further, 35 candidates with stable expression in all conditions were detected, which could serve as targets for detection. Data are available via ProteomeXchange with identifier PXD027444.
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Affiliation(s)
- Kristina J. H. Kleinwort
- Chair of Physiology, Department of Veterinary Sciences, LMU Munich, D-82152 Martinsried, Germany; (K.J.H.K.); (B.F.H.); (R.L.D.)
| | - Bernhard F. Hobmaier
- Chair of Physiology, Department of Veterinary Sciences, LMU Munich, D-82152 Martinsried, Germany; (K.J.H.K.); (B.F.H.); (R.L.D.)
| | - Ricarda Mayer
- Chair of Hygiene and Technology of Milk, Department of Veterinary Sciences, LMU Munich, D-85764 Oberschleißheim, Germany; (R.M.); (C.H.); (E.M.)
| | - Christina Hölzel
- Chair of Hygiene and Technology of Milk, Department of Veterinary Sciences, LMU Munich, D-85764 Oberschleißheim, Germany; (R.M.); (C.H.); (E.M.)
- Institute of Animal Breeding and Husbandry, Faculty of Agricultural and Nutritional Sciences, CAU Kiel, D-24098 Kiel, Germany
| | - Roxane L. Degroote
- Chair of Physiology, Department of Veterinary Sciences, LMU Munich, D-82152 Martinsried, Germany; (K.J.H.K.); (B.F.H.); (R.L.D.)
| | - Erwin Märtlbauer
- Chair of Hygiene and Technology of Milk, Department of Veterinary Sciences, LMU Munich, D-85764 Oberschleißheim, Germany; (R.M.); (C.H.); (E.M.)
| | - Stefanie M. Hauck
- Research Unit Protein Science, Helmholtz Center Munich, German Research Center for Environmental Health, D-80939 Munich, Germany;
| | - Cornelia A. Deeg
- Chair of Physiology, Department of Veterinary Sciences, LMU Munich, D-82152 Martinsried, Germany; (K.J.H.K.); (B.F.H.); (R.L.D.)
- Correspondence:
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Yaman Y, Aymaz R, Keleş M, Bay V, Ün C, Heaton MP. Association of TLR2 haplotypes encoding Q650 with reduced susceptibility to ovine Johne's disease in Turkish sheep. Sci Rep 2021; 11:7088. [PMID: 33782507 PMCID: PMC8007707 DOI: 10.1038/s41598-021-86605-4] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/12/2021] [Accepted: 03/18/2021] [Indexed: 02/06/2023] Open
Abstract
Ovine Johne’s disease (OJD) is caused by Mycobacterium avium subsp. paratuberculosis (MAP) and carries a potential zoonotic risk for humans. Selective breeding strategies for reduced OJD susceptibility would be welcome tools in disease eradication efforts, if available. The Toll-like receptor 2 gene (TLR2) plays an important signaling role in immune response to MAP, and missense variants are associated with mycobacterial infections in mammals. Our aim was to identify and evaluate ovine TLR2 missense variants for association with OJD in Turkish sheep. Eleven TLR2 missense variants and 17 haplotype configurations were identified in genomic sequences of 221 sheep from 61 globally-distributed breeds. The five most frequent haplotypes were tested for OJD association in 102 matched pairs of infected and uninfected ewes identified in 2257 Turkish sheep. Ewes with one or two copies of TLR2 haplotypes encoding glutamine (Q) at position 650 (Q650) in the Tir domain were 6.6-fold more likely to be uninfected compared to ewes with arginine (R650) at that position (CI95 = 2.6 to 16.9, p-value = 3.7 × 10–6). The protective TLR2 Q650 allele was present in at least 25% of breeds tested and thus may facilitate selective breeding for sheep with reduced susceptibility to OJD.
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Affiliation(s)
- Yalçın Yaman
- Department of Biometry and Genetics, Sheep Breeding and Research Institute, 10200, Bandırma, Balıkesir, Turkey.
| | - Ramazan Aymaz
- Department of Biometry and Genetics, Sheep Breeding and Research Institute, 10200, Bandırma, Balıkesir, Turkey
| | - Murat Keleş
- Department of Biometry and Genetics, Sheep Breeding and Research Institute, 10200, Bandırma, Balıkesir, Turkey
| | - Veysel Bay
- Department of Biometry and Genetics, Sheep Breeding and Research Institute, 10200, Bandırma, Balıkesir, Turkey
| | - Cemal Ün
- Department of Biology, Faculty of Science, Ege University, 35000, İzmir, Turkey
| | - Michael P Heaton
- USDA, ARS, U.S. Meat Animal Research Center, Clay Center, NE, 68933, USA
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Mallikarjunappa S, Shandilya UK, Sharma A, Lamers K, Bissonnette N, Karrow NA, Meade KG. Functional analysis of bovine interleukin-10 receptor alpha in response to Mycobacterium avium subsp. paratuberculosis lysate using CRISPR/Cas9. BMC Genet 2020; 21:121. [PMID: 33138773 PMCID: PMC7607837 DOI: 10.1186/s12863-020-00925-4] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/23/2019] [Accepted: 10/20/2020] [Indexed: 12/21/2022] Open
Abstract
Background The interleukin-10 receptor alpha (IL10RA) gene codes for the alpha chain of the IL-10 receptor which binds the cytokine IL-10. IL-10 is an anti-inflammatory cytokine with immunoregulatory function during the pathogenesis of many inflammatory disorders in livestock, including Johne’s disease (JD). JD is a chronic enteritis in cattle caused by Mycobacterium avium subsp. paratuberculosis (MAP) and is responsible for significant economic losses to the dairy industry. Several candidate genes including IL10RA have been found to be associated with JD. The aim of this study was to better understand the functional significance of IL10RA in the context of immune stimulation with MAP cell wall lysate. Results An IL10RA knock out (KO) bovine mammary epithelial cell (MAC-T) line was generated using the CRISPR/cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9) gene editing system. These IL10RA KO cells were stimulated with the immune stimulant MAP lysate +/− IL-10, or with LPS as a positive control. In comparison to unedited cells, relative quantification of immune-related genes after stimulation revealed that knocking out IL10RA resulted in upregulation of pro-inflammatory cytokine gene expression (TNFA, IL1A, IL1B and IL6) and downregulation of suppressor of cytokine signaling 3 (SOCS3), a negative regulator of pro-inflammatory cytokine signaling. At the protein level knocking out IL10RA also resulted in upregulation of inflammatory cytokines - TNF-α and IL-6 and chemokines - IL-8, CCL2 and CCL4, relative to unedited cells. Conclusions The findings of this study illustrate the broad and significant effects of knocking out the IL10RA gene in enhancing pro-inflammatory cytokine expression and further support the immunoregulatory role of IL10RA in eliciting an anti-inflammatory response as well as its potential functional involvement during the immune response associated with JD.
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Affiliation(s)
- Sanjay Mallikarjunappa
- Animal and Bioscience Research Department, Animal and Grassland Research and Innovation Centre, Teagasc, Grange, Co. Meath, Ireland.,Centre for Genetic Improvement of Livestock, Department of Animal Biosciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
| | - Umesh K Shandilya
- Centre for Genetic Improvement of Livestock, Department of Animal Biosciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
| | - Ankita Sharma
- Centre for Genetic Improvement of Livestock, Department of Animal Biosciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
| | - Kristen Lamers
- Centre for Genetic Improvement of Livestock, Department of Animal Biosciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
| | - Nathalie Bissonnette
- Agriculture and Agri-Food Canada, Sherbrooke Research and Development Centre, Sherbrooke, QC, J1M 0C8, Canada
| | - Niel A Karrow
- Centre for Genetic Improvement of Livestock, Department of Animal Biosciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
| | - Kieran G Meade
- Animal and Bioscience Research Department, Animal and Grassland Research and Innovation Centre, Teagasc, Grange, Co. Meath, Ireland.
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Liaskos C, Gkoutzourelas A, Spyrou V, Koutsoumpas A, Athanasiou LV, Amiridis GS, Billinis C, Bogdanos DP. Pancreatic anti-GP2 and anti-Saccharomyces cerevisiae antibodies in ruminants with paratuberculosis: A better understanding of the immunopathogenesis of Crohn's disease. Clin Res Hepatol Gastroenterol 2020; 44:778-785. [PMID: 32035824 DOI: 10.1016/j.clinre.2019.12.013] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/04/2019] [Revised: 12/12/2019] [Accepted: 12/27/2019] [Indexed: 02/04/2023]
Abstract
INTRODUCTION Ruminants (cattle and sheep) with Mycobacterium avium (MAP)-induced paratuberculosis (ptb), the ruminant model of Crohn's disease (CD), exhibit pancreatic specific autoantibodies (PAB) against GP2 but not against CUZD1. Since anti-Saccharomyces cerevisiae antibodies (ASCAs) is a CD marker, we tested MAP-infected ptb ruminants for ASCA, and compared them with ruminants lacking evidence of anti-MAP serology or with ruminants, which were positive for anti-GP2 antibodies. MATERIAL AND METHODS A total of 98 samples from ruminants (48 cattle and 50 sheep) were studied. IgG anti-MAP antibodies, and CD-related ASCA and anti-GP2 antibodies were tested by modified ELISAs. RESULTS Nine cattle (18.75%) and 20 sheep (40%) were suffered from ptb. ASCA antibodies were present in 21/48 (43.7%) cattle and 10/50 (20%) sheep while anti-GP2 antibodies were present in 14/48 (29.2%) cattle, and 8/50 (16%) sheep. ASCA antibodies were more prevalent in anti-MAP antibody positive (14/29, 48.3%) than in anti-MAP negative ruminants (17/69, 24.6%, P=0.022) and also in anti-GP2 antibody positive (13/23, 56.5%) than in anti-GP2 negative ruminants (18/75, 24%, P=0.003). No association between ASCA and anti-MAP antibody concentrations were found (r=0.159, P=0.117). A significant association between ASCA and anti-GP2 antibody concentration were observed (r=0.211 and P=0.037). CONCLUSION ASCA are present in a significant proportion of ruminants with ptb and correlate with anti-GP2 antibody positivity, a finding further supporting the notion that Crohn's disease and ptb share common immunological mechanisms of antigen-driven loss of self-tolerance.
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Affiliation(s)
- Christos Liaskos
- Department of Rheumatology and Clinical Immunology, Faculty of Medicine, School of Health Sciences, University of Thessaly, 41110 Larissa, Greece.
| | - Athanasios Gkoutzourelas
- Department of Rheumatology and Clinical Immunology, Faculty of Medicine, School of Health Sciences, University of Thessaly, 41110 Larissa, Greece
| | - Vasiliki Spyrou
- Department of Animal Production, University of Thessaly, 41110 Larissa, Greece
| | - Andreas Koutsoumpas
- Department of Rheumatology and Clinical Immunology, Faculty of Medicine, School of Health Sciences, University of Thessaly, 41110 Larissa, Greece
| | - Labrini V Athanasiou
- Department of Medicine, Faculty of Veterinary Medicine, University of Thessaly, Karditsa, Greece
| | - Georgios S Amiridis
- Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Thessaly, Karditsa, Greece
| | - Charalambos Billinis
- Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Thessaly, Karditsa, Greece
| | - Dimitrios P Bogdanos
- Department of Rheumatology and Clinical Immunology, Faculty of Medicine, School of Health Sciences, University of Thessaly, 41110 Larissa, Greece
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Garg A, Singhal N, Kumar M. Discerning novel drug targets for treating Mycobacterium avium ss. paratuberculosis-associated autoimmune disorders: an in silico approach. Brief Bioinform 2020; 22:5902595. [PMID: 32895696 DOI: 10.1093/bib/bbaa195] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/13/2020] [Revised: 06/24/2020] [Accepted: 07/30/2020] [Indexed: 11/13/2022] Open
Abstract
Mycobacterium avium subspecies paratuberculosis (MAP) exhibits 'molecular mimicry' with the human host resulting in several autoimmune diseases such as multiple sclerosis, type 1 diabetes mellitus (T1DM), Hashimoto's thyroiditis, Crohn's disease (CD), etc. The conventional therapy for autoimmune diseases includes immunosuppressants or immunomodulators that treat the symptoms rather than the etiology and/or causative mechanism(s). Eliminating MAP-the etiopathological agent might be a better strategy to treat MAP-associated autoimmune diseases. In this case study, we conducted a systematic in silico analysis to identify the metabolic chokepoints of MAP's mimicry proteins and their interacting partners. The probable inhibitors of chokepoint proteins were identified using DrugBank. DrugBank molecules were stringently screened and molecular interactions were analyzed by molecular docking and 'off-target' binding. Thus, we identified 18 metabolic chokepoints of MAP mimicry proteins and 13 DrugBank molecules that could inhibit three chokepoint proteins viz. katG, rpoB and narH. On the basis of molecular interaction between drug and target proteins finally eight DrugBank molecules, viz. DB00609, DB00951, DB00615, DB01220, DB08638, DB08226, DB08266 and DB07349 were selected and are proposed for treatment of three MAP-associated autoimmune diseases namely, T1DM, CD and multiple sclerosis. Because these molecules are either approved by the Food and Drug Administration or these are experimental drugs that can be easily incorporated in clinical studies or tested in vitro. The proposed strategy may be used to repurpose drugs to treat autoimmune diseases induced by other pathogens.
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14
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Honap S, Johnston E, Agrawal G, Al-Hakim B, Hermon-Taylor J, Sanderson J. Anti- Mycobacterium paratuberculosis (MAP) therapy for Crohn's disease: an overview and update. Frontline Gastroenterol 2020; 12:397-403. [PMID: 35401965 PMCID: PMC8989010 DOI: 10.1136/flgastro-2020-101471] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/16/2020] [Revised: 05/28/2020] [Accepted: 06/14/2020] [Indexed: 02/06/2023] Open
Abstract
The role of Mycobacterium avium subspecies paratuberculosis (MAP) in the pathogenesis of Crohn's disease (CD) has been strongly debated for many years. MAP is the known aetiological agent of Johne's disease, a chronic enteritis affecting livestock. At present, due to the paucity of high-quality data, anti-MAP therapy (AMT) is not featured in international guidelines as a treatment for CD. Although the much-quoted randomised trial of AMT did not show sustained benefits over placebo, questions have been raised regarding trial design, antibiotic dosing and the formulation used. There are several lines of evidence supporting the CD and MAP association with uncontrolled and controlled trials demonstrating effectiveness, including a retrospective review of cases treated at our own institution. Here, we provide an overview of the evidence supporting and refuting AMT in CD before focussing on updates of the current research in the field, including the ongoing trials with the novel RHB-104 formulation and the MAP vaccine trial. While controversial, gastroenterologists are often asked about long-term combination antibiotic therapy for CD. There has been broadcast and social media coverage surrounding this, particularly with regard to current trials. Although patients should not be deterred from treatments of proven effectiveness, this review aims to help with commonly asked questions and highlights our own approach for the use of anti-MAP in specific circumstances.
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Affiliation(s)
- Sailish Honap
- IBD Centre, Guy's and Saint Thomas' NHS Foundation Trust, London, UK
| | - Emma Johnston
- Department of Gastroenterology, Chelsea and Westminster Hospital NHS Foundation Trust, London, UK
| | - Gaurav Agrawal
- IBD Centre, Guy's and Saint Thomas' NHS Foundation Trust, London, UK,Department of Nutritional Sciences, King's College London, London, UK
| | - Bahij Al-Hakim
- IBD Centre, Guy's and Saint Thomas' NHS Foundation Trust, London, UK
| | | | - Jeremy Sanderson
- IBD Centre, Guy's and Saint Thomas' NHS Foundation Trust, London, UK,Department of Nutritional Sciences, King's College London, London, UK
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15
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Dow CT, Sechi LA. Cows Get Crohn's Disease and They're Giving Us Diabetes. Microorganisms 2019; 7:microorganisms7100466. [PMID: 31627347 PMCID: PMC6843388 DOI: 10.3390/microorganisms7100466] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2019] [Revised: 10/14/2019] [Accepted: 10/15/2019] [Indexed: 12/20/2022] Open
Abstract
Increasingly, Johne's disease of ruminants and human Crohn's disease are regarded as the same infectious disease: paratuberculosis. Mycobacterium avium ss. paratuberculosis (MAP) is the cause of Johne's and is the most commonly linked infectious cause of Crohn's disease. Humans are broadly exposed to MAP in dairy products and in the environment. MAP has been found within granulomas such as Crohn's disease and can stimulate autoantibodies in diseases such as type 1 diabetes (T1D) and Hashimoto's thyroiditis. Moreover, beyond Crohn's and T1D, MAP is increasingly associated with a host of autoimmune diseases. This article suggests near equivalency between paucibacillary Johne's disease of ruminant animals and human Crohn's disease and implicates MAP zoonosis beyond Crohn's disease to include T1D.
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Affiliation(s)
- Coad Thomas Dow
- McPherson Eye Research Institute, University of Wisconsin, 9431 WIMR, 1111 Highland Avenue, Madison, WI 53705, USA.
| | - Leonardo A Sechi
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43b, 07100 Sassari, Italy.
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16
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Fawzy A, Prince A, Hassan A, Fayed A, Zschöck M, Naga M, Omar M, Salem M, El-Sayed A. Epidemiological studies on Johne’s disease in ruminants and Crohn’s disease in humans in Egypt. Int J Vet Sci Med 2019. [DOI: 10.1016/j.ijvsm.2013.09.004] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023] Open
Affiliation(s)
- A. Fawzy
- Laboratory of Molecular Epidemiology, Faculty of Veterinary Medicine, Cairo University, Egypt
- Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Egypt
| | - A. Prince
- Laboratory of Molecular Epidemiology, Faculty of Veterinary Medicine, Cairo University, Egypt
- Agricultural Research Service, USDA, Beltsville, MD, USA
| | - A.A. Hassan
- Department of Veterinary Public Health, College of Veterinary Medicine, University of Mosul, Mosul, Iraq
| | - A. Fayed
- Laboratory of Molecular Epidemiology, Faculty of Veterinary Medicine, Cairo University, Egypt
- Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Egypt
| | - M. Zschöck
- Landesbetrieb Hessisches Landeslabor, Giessen, Germany
| | - M. Naga
- Endoscopy Unit, Department of Internal Medicine, Faculty of Medicine Kasralainy, Cairo University, Egypt
| | - M. Omar
- Department of Microbiology and Immunology, Faculty of Medicine, King Abdul Aziz University, Saudi Arabia
- Department of Microbiology and Immunology, Faculty of Medicine, Ain Shams University, Egypt
| | - M. Salem
- Laboratory of Molecular Epidemiology, Faculty of Veterinary Medicine, Cairo University, Egypt
- Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Egypt
| | - A. El-Sayed
- Laboratory of Molecular Epidemiology, Faculty of Veterinary Medicine, Cairo University, Egypt
- Landesbetrieb Hessisches Landeslabor, Giessen, Germany
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17
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Savarino E, Bertani L, Ceccarelli L, Bodini G, Zingone F, Buda A, Facchin S, Lorenzon G, Marchi S, Marabotto E, De Bortoli N, Savarino V, Costa F, Blandizzi C. Antimicrobial treatment with the fixed-dose antibiotic combination RHB-104 for Mycobacterium avium subspecies paratuberculosis in Crohn's disease: pharmacological and clinical implications. Expert Opin Biol Ther 2019; 19:79-88. [PMID: 30574820 DOI: 10.1080/14712598.2019.1561852] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
INTRODUCTION Crohn's disease (CD) is an inflammatory bowel disease of unknown etiology. However, increasing evidence suggests Mycobacterium avium subspecies paratuberculosis (MAP) as a putative causative agent: 1) MAP is the etiological agent of Johne's disease, a granulomatous enteritis affecting ruminants, which shares clinical and pathological features with CD; 2) MAP has been detected in tissues and blood samples from CD patients; 3) case reports have documented a favorable therapeutic response to anti-MAP antibiotics. Area covered: This review provides an appraisal of current information on MAP characteristics, diagnostic methodologies and emerging drug treatments. The authors focus on RHB-104, a novel oral formulation containing a fixed-dose combination of clarithromycin, clofazimine and rifabutin, endowed with synergistic inhibitory activity on MAP strains isolated from CD patients. Expert opinion: Based on encouraging in vitro data, RHB-104 has entered recently the clinical phase of its development, and is being investigated in a randomized, placebo-controlled phase III trial aimed at evaluating its efficacy and safety in CD. Provided that the overall clinical development will support the suitability of RHB-104 for inducing disease remission in CD patients with documented MAP infection, this novel antibiotic combination will likely take a relevant position in the therapeutic armamentarium for CD management.
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Affiliation(s)
- Edoardo Savarino
- a Gastrointestinal Unit, Department of Surgery, Oncology and Gastroenterology , University of Padua , Padua , Italy
| | - Lorenzo Bertani
- b Gastrointestinal Unit, Division of Gastroenterology, Department of Translational Research and New Technologies in Medicine and Surgery , University of Pisa , Pisa , Italy
| | - Linda Ceccarelli
- b Gastrointestinal Unit, Division of Gastroenterology, Department of Translational Research and New Technologies in Medicine and Surgery , University of Pisa , Pisa , Italy
| | - Giorgia Bodini
- c Gastrointestinal Unit, Department of Internal Medicine and Medical Specialties , University of Genoa , Genoa , Italy
| | - Fabiana Zingone
- a Gastrointestinal Unit, Department of Surgery, Oncology and Gastroenterology , University of Padua , Padua , Italy
| | - Andrea Buda
- a Gastrointestinal Unit, Department of Surgery, Oncology and Gastroenterology , University of Padua , Padua , Italy
| | - Sonia Facchin
- a Gastrointestinal Unit, Department of Surgery, Oncology and Gastroenterology , University of Padua , Padua , Italy
| | - Greta Lorenzon
- a Gastrointestinal Unit, Department of Surgery, Oncology and Gastroenterology , University of Padua , Padua , Italy
| | - Santino Marchi
- b Gastrointestinal Unit, Division of Gastroenterology, Department of Translational Research and New Technologies in Medicine and Surgery , University of Pisa , Pisa , Italy
| | - Elisa Marabotto
- c Gastrointestinal Unit, Department of Internal Medicine and Medical Specialties , University of Genoa , Genoa , Italy
| | - Nicola De Bortoli
- b Gastrointestinal Unit, Division of Gastroenterology, Department of Translational Research and New Technologies in Medicine and Surgery , University of Pisa , Pisa , Italy
| | - Vincenzo Savarino
- c Gastrointestinal Unit, Department of Internal Medicine and Medical Specialties , University of Genoa , Genoa , Italy
| | - Francesco Costa
- b Gastrointestinal Unit, Division of Gastroenterology, Department of Translational Research and New Technologies in Medicine and Surgery , University of Pisa , Pisa , Italy
| | - Corrado Blandizzi
- d Unit of Pharmacology and Pharmacovigilance, Department of Clinical and Experimental Medicine , University of Pisa , Pisa , Italy
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18
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Salem M, El-Deeb W, Abdel-Moein K, El-Sayed A, Fayed A, Housawi F, Al-Naeem A, Zschöck M. Detection of Mycobacterium avium subsp. paratuberculosis in an Egyptian mixed breeding farm and comparative molecular characterisation of isolates from cattle, camels and cats – a case report. BULGARIAN JOURNAL OF VETERINARY MEDICINE 2019. [DOI: 10.15547/bjvm.2033] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022] Open
Abstract
The present study records and investigates an outbreak of Johne’s Disease in a mixed breeding camel – cattle farm and the possible role of non-domestic non-ruminants animals in the epidemiology of Mycobacterium avium subspecies paratuberculosis in Egypt. For this reason, faecal samples were collected from 24 dairy cattle and from 15 one humped Arabian camels suffering from diarrhoea. Moreover, intestinal tissue samples were provided from 7 cats and 2 rats that were caught from the same farm and were euthanized before necropsy. Samples were examined using traditional culture and IS900 PCR techniques together with the application of BstEII-IS900 RFLP for typing of obtained isolates. Interestingly, MAP was recovered from cattle (n=8) and from camels (n=3) and non-domestic cats (n=3) reared under local conditions in this farm in Egypt. The obtained results highlight the potential role of cats in the epidemiology of MAP, a subject which needs further investigation and might have a public health importance, catsbeing common members of many families.
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19
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Balseiro A, Perez V, Juste RA. Chronic regional intestinal inflammatory disease: A trans-species slow infection? Comp Immunol Microbiol Infect Dis 2018; 62:88-100. [PMID: 30711052 DOI: 10.1016/j.cimid.2018.12.001] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2018] [Revised: 11/27/2018] [Accepted: 12/07/2018] [Indexed: 01/08/2023]
Abstract
Crohn's disease and ulcerative colitis in humans and paratuberculosis in domestic and wild ruminants can be defined as chronic regional intestinal inflammatory diseases (CRIID). This review is a literature overview on these diseases in humans, non-human primates, dogs, cats, rabbits, equids and ruminants with a focus on pathological and microbiological features aimed identifying common characteristics that could lead to a unified pathological classification for a better understanding of their mechanisms and causes. The result is a framework of inflammatory forms throughout the different species indicative of common mechanisms of the slow infection type characterized by a time course varying from weeks to months or even years, and where the inflammatory component would be more prominent in the intestinal interphase between host and environment and be morphologically characterized by an infiltrate ranging from lymphoplasmacytic to histiocytic. This should provide new insights for causation demonstration and therapeutic approaches in human IBD.
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Affiliation(s)
- Ana Balseiro
- Centro de Biotecnología Animal, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), 33394 Gijón, Asturias, Spain
| | - Valentin Perez
- Departamento de Sanidad Animal, Instituto de Ganadería de Montaña (CSIC-ULE), Facultad de Veterinaria, Universidad de León, 24071 León, Spain
| | - Ramon A Juste
- Direccion. Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), 33300 Villaviciosa, Asturias, Spain.
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20
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Kim WS, Shin MK, Shin SJ. MAP1981c, a Putative Nucleic Acid-Binding Protein, Produced by Mycobacterium avium subsp. paratuberculosis, Induces Maturation of Dendritic Cells and Th1-Polarization. Front Cell Infect Microbiol 2018; 8:206. [PMID: 29977867 PMCID: PMC6021526 DOI: 10.3389/fcimb.2018.00206] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2018] [Accepted: 06/05/2018] [Indexed: 12/23/2022] Open
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) is the causative pathogen of chronic granulomatous enteropathy (Johne's disease) in animals, and has been focused on its association with various autoimmune diseases in humans, including Crohn's disease. The discovery of novel mycobacterial antigens and exploring their role in host immunity can contribute to the advancement of effective defense strategies including vaccines and diagnostic tools. In a preliminary study, we identified cellular extract proteins of MAP that strongly react with the blood of patients with Crohn's disease. In particular, MAP1981c, a putative nucleic acid-binding protein, showed high expression levels and strong reactivity to IgG and IgM in the sera of patients. Here, we investigated the immunological features of MAP1981c and focused on its interaction with dendritic cells (DCs), confirming its immunomodulatory ability. MAP1981c was shown to recognize Toll-like receptor (TLR) 4, and induce DC maturation and activation by increasing the expression of co-stimulatory (CD80 and CD86) and MHC class I/II molecules and the secretion of pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) in DCs. This DC activation by MAP1981c was mediated by downstream signaling of TLR4 via MyD88- and TRIF-, MAP kinase-, and NF-κB-dependent signaling pathways. In addition, MAP1981c-treated DCs activated naïve T cells and induced the differentiation of CD4+ and CD8+ T cells to express T-bet, IFN-γ, and/or IL-2, but not GATA-3 and IL-4, thus indicating that MAP1981c contributes to Th1-type immune responses both in vitro and in vivo. Taken together, these results suggest that MAP1981c is a novel immunocompetent antigen that induces DC maturation and a Th1-biased response upon DC activation, suggesting that MAP1981c can be an effective vaccine and diagnostic target.
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Affiliation(s)
- Woo Sik Kim
- Department of Microbiology and Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea.,Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup, South Korea
| | - Min-Kyoung Shin
- Department of Microbiology, Institute of Health Sciences, College of Medicine, Gyeongsang National University, Jinju, South Korea
| | - Sung Jae Shin
- Department of Microbiology and Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea
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21
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Garvey M. Mycobacterium avium subspecies paratuberculosis: A possible causative agent in human morbidity and risk to public health safety. Open Vet J 2018; 8:172-181. [PMID: 29911021 PMCID: PMC5987349 DOI: 10.4314/ovj.v8i2.10] [Citation(s) in RCA: 49] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2017] [Accepted: 05/04/2018] [Indexed: 12/14/2022] Open
Abstract
Mycobacterium avium subspecies paratuberculosis is a bacterial parasite and the causative agent of paratuberculosis, a disease predominately found in cattle and sheep. Infection with this microorganism results in substantial farming economic losses and animal morbidity. The link between infection with this pathogen and human disease has been theorised for many years with Crohn's disease being one of many suspected resultant conditions. Mycobacterium avium may be spread from animal to human hosts by water and foodborne transmission routes, where the foodborne route of exposure represents a significant risk for susceptible populations, namely children and the immune-compromised. Following colonisation of the host, the parasitic organism evades the host immune system by use of molecular mimicry, displaying peptide sequences similar to that of the host cells causing a disruption of self-verses non self-recognition. Theoretically, this failure to recognise the invading organism as distinct from host cells may result in numerous autoimmune conditions. Here, the author presents current information assessing the link between numerous diseases states in humans such inflammatory bowel disease, Type 1 diabetes, rheumatoid arthritis, Hashimoto\'s thyroiditis, multiple sclerosis and autism following infection with Mycobacterium avium paratuberculosis. The possibility of zoonotic transmission of the organism and its significant risk to public health safety as a consequence is also discussed.
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Affiliation(s)
- Mary Garvey
- Cellular Health and Toxicology Research Group, Institute of Technology, Sligo, Ash Lane, Sligo, Ireland
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22
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Kim WS, Kim JS, Shin MK, Shin SJ. A novel Th1-type T-cell immunity-biasing effect of malate dehydrogenase derived from Mycobacterium avium subspecies paratuberculosis via the activation of dendritic cells. Cytokine 2018; 104:14-22. [PMID: 29414321 DOI: 10.1016/j.cyto.2018.01.022] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/25/2017] [Revised: 12/30/2017] [Accepted: 01/25/2018] [Indexed: 01/13/2023]
Abstract
Mycobacterium avium subspecies paratuberculosis (MAP) is the causative pathogen of Johne's disease in ruminants, characterized by chronic granulomatous enteritis; it also has zoonotic potential and is associated with Crohn's disease in humans. A better understanding of the mycobacterial antigens and their roles in the host immune response may facilitate the rational design of control strategies, including the development of effective vaccines and diagnostic tools. However, the functional roles of a large proportion of MAP antigens involved in modulating the host immune response remain unknown. In this study, an immunological role of MAP malate dehydrogenase (MDH, MAP2541c), an antigen that is upregulated in stress culture conditions, such as nutrient starvation and hypoxia, in polarizing naïve CD4+/CD8+ T cells toward Th1-biased T-cell immunity via the activation of dendritic cells (DCs) was identified. DCs treated with MAP MDH displayed characteristics of the activated and mature immune status, with augmented expression of cell surface molecules and pro-inflammatory cytokines, including TNF-α, IL-1β, IL-6, and IL-12p70, but not IL-10, along with a dose-dependent decrease in the antigen uptake capacity. A mechanistic investigation revealed that the observed DC maturation is mediated by the activation of JNK, ERK, and p38 MAP kinases, and the NF-κB signaling pathway. Notably, DCs activated by MAP MDH treatment promoted naïve CD4+/CD8+ T cell proliferation; in particular, they effectively polarized naïve CD4+ T cells to secrete IFN-γ and IL-2 and activate T-bet, but, unlike the LPS control, did not influence IL-5 and GATA-3. These results indicated that MAP MDH has the potential to induce the Th1 cell response via DC activation. Collectively, our data demonstrated that MAP MDH is a novel immunostimulatory antigen that drives Th1-biased T cell polarization via interactions with DCs, suggesting that MDP MDH has the potential to be an effective MAP vaccine antigen target and diagnostic marker.
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Affiliation(s)
- Woo Sik Kim
- Department of Microbiology and Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185, Republic of Korea
| | - Jong-Seok Kim
- Department of Microbiology and Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea
| | - Min-Kyoung Shin
- Department of Microbiology, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 52727, Republic of Korea.
| | - Sung Jae Shin
- Department of Microbiology and Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
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Chaubey KK, Singh SV, Gupta S, Singh M, Sohal JS, Kumar N, Singh MK, Bhatia AK, Dhama K. Mycobacterium avium subspecies paratuberculosis - an important food borne pathogen of high public health significance with special reference to India: an update. Vet Q 2018; 37:282-299. [PMID: 29090657 DOI: 10.1080/01652176.2017.1397301] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
This review underlines the public health significance of 'Indian Bison Type' of Mycobacterium avium subspecies paratuberculosis (MAP) and also its potential as 'zoonotic infection'. In the absence of control programs, bio-load of MAP is increasing and if we take total population of animals (500 million plus) and human beings (1.23 billion plus) into account, the number of infected animals and human beings will run into millions in India. Our research on screening of over 26,000 domestic livestock for MAP infection using 4 different diagnostic tests (microscopy, culture, ELISA and PCR), during last 31 years has shown that the average bio-load of MAP in the livestock population of India is very high (cattle 43%, buffaloes 36%, goats 23% and sheep 41%). 'Mass screening' of 28,291 human samples between 2008-2016 revealed also high bio-load of MAP. It has been proved that MAP is not in-activated during pasteurization and therefore live bacilli are continuously reaching human population by consumption of even pasteurized milk and other milk products. Live bacilli have also been recovered from meat products and the environment thus illustrating the potential of MAP as pathogen of public health concern. However, at present, there is inadequate scientific evidence to confirm a conclusive link between MAP infection and Johne's disease in ruminants and some cases of Crohn's disease in human beings.
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Affiliation(s)
- Kundan Kumar Chaubey
- a Animal Health Division, Central Institute for Research on Goats (CIRG) , Mathura , UP , India.,b Department of Microbiology and Immunology , GLA University , Mathura , UP , India
| | - Shoor Vir Singh
- a Animal Health Division, Central Institute for Research on Goats (CIRG) , Mathura , UP , India
| | - Saurabh Gupta
- a Animal Health Division, Central Institute for Research on Goats (CIRG) , Mathura , UP , India.,b Department of Microbiology and Immunology , GLA University , Mathura , UP , India
| | - Manju Singh
- a Animal Health Division, Central Institute for Research on Goats (CIRG) , Mathura , UP , India
| | - Jagdip Singh Sohal
- c Amity Institutes of Microbial Technology, Amity University , Jaipur , India
| | - Naveen Kumar
- d Veterinary Type Culture Collection, NRC On Equines , Indian Council of Agricultural Research , Hisar , India
| | - Manoj Kumar Singh
- a Animal Health Division, Central Institute for Research on Goats (CIRG) , Mathura , UP , India
| | - Ashok Kumar Bhatia
- b Department of Microbiology and Immunology , GLA University , Mathura , UP , India
| | - Kuldeep Dhama
- e Pathology Division , Indian Veterinary Research Institute (IVRI) , Bareilly , UP , India
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Shen ZH, Zhu CX, Quan YS, Yang ZY, Wu S, Luo WW, Tan B, Wang XY. Relationship between intestinal microbiota and ulcerative colitis: Mechanisms and clinical application of probiotics and fecal microbiota transplantation. World J Gastroenterol 2018; 24:5-14. [PMID: 29358877 PMCID: PMC5757125 DOI: 10.3748/wjg.v24.i1.5] [Citation(s) in RCA: 450] [Impact Index Per Article: 64.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/01/2017] [Revised: 11/07/2017] [Accepted: 11/21/2017] [Indexed: 02/06/2023] Open
Abstract
Ulcerative colitis (UC) is an inflammatory disease that mainly affects the colon and rectum. It is believed that genetic factors, host immune system disorders, intestinal microbiota dysbiosis, and environmental factors contribute to the pathogenesis of UC. However, studies on the role of intestinal microbiota in the pathogenesis of UC have been inconclusive. Studies have shown that probiotics improve intestinal mucosa barrier function and immune system function and promote secretion of anti-inflammatory factors, thereby inhibiting the growth of harmful bacteria in the intestine. Fecal microbiota transplantation (FMT) can reduce bowel permeability and thus the severity of disease by increasing the production of short-chain fatty acids, especially butyrate, which help maintain the integrity of the epithelial barrier. FMT can also restore immune dysbiosis by inhibiting Th1 differentiation, activity of T cells, leukocyte adhesion, and production of inflammatory factors. Probiotics and FMT are being increasingly used to treat UC, but their use is controversial because of uncertain efficacy. Here, we briefly review the role of intestinal microbiota in the pathogenesis and treatment of UC.
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Affiliation(s)
- Zhao-Hua Shen
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Chang-Xin Zhu
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Yong-Sheng Quan
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Zhen-Yu Yang
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Shuai Wu
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Wei-Wei Luo
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Bei Tan
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
| | - Xiao-Yan Wang
- Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
- Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Changsha 410008, Hunan Province, China
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Cecchi F, Russo C, Fratini F, Turchi B, Preziuso G, Cantile C. Preliminary association analysis of microsatellites and Mycobacterium avium subspecies paratuberculosis infection in the native Garfagnina goats. JOURNAL OF APPLIED ANIMAL RESEARCH 2017. [DOI: 10.1080/09712119.2017.1417129] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Affiliation(s)
- Francesca Cecchi
- Department of Veterinary Science, University of Pisa, Pisa, Italy
| | - Claudia Russo
- Department of Veterinary Science, University of Pisa, Pisa, Italy
| | - Filippo Fratini
- Department of Veterinary Science, University of Pisa, Pisa, Italy
| | - Barbara Turchi
- Department of Veterinary Science, University of Pisa, Pisa, Italy
| | | | - Carlo Cantile
- Department of Veterinary Science, University of Pisa, Pisa, Italy
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26
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Marino R, Capoferri R, Panelli S, Minozzi G, Strozzi F, Trevisi E, Snel GGM, Ajmone-Marsan P, Williams JL. Johne's disease in cattle: an in vitro model to study early response to infection of Mycobacterium avium subsp. paratuberculosis using RNA-seq. Mol Immunol 2017; 91:259-271. [PMID: 28988040 DOI: 10.1016/j.molimm.2017.08.017] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/19/2017] [Revised: 07/07/2017] [Accepted: 08/21/2017] [Indexed: 02/07/2023]
Abstract
Johne's disease is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratubercolosis (MAP) which affects ruminants worldwide and has a significant economic impact. MAP has also been associated with human Crohn's disease, although this connection is not well established. MAP is highly adapted for survival within host macrophages and prevents macrophage activation, blocks phagosome acidification and maturation, and attenuates presentation of antigens to the immune system. The consequence is a very long silent infection before clinical signs are observed. The present work examined the transcriptome of bovine monocyte-derived macrophages (MDM) infected with the L1 strain of MAP at 2h, 6h and 24h post infection using RNA-seq. Pathway over-representation analysis of genes differentially expressed between infected vs. control MDM identified that immune related pathways were affected. Genes belonging to the cytokine-cytokine receptor interaction pathway and members of the JAK-STAT pathway, which is involved in the regulation of immune response, were up-regulated. However, in parallel inhibitors of immune functions were activated, including suppressor of cytokine signaling (SOCS) and cytokine-inducible SH2-containing protein (CISH), which most likely suppresses IFNγ and the JAK/STAT signaling cascade in infected MDM, which may favour MAP survival. After exposure, macrophages phagocytise pathogens, activate the complement cascade and the adaptive immune system through the antigen presentation process. However, data presented here suggest that genes related to phagocytosis and lysosome function are down regulated in MAP infected MDM. Genes of MHC class II and complement pathway were also down-regulated. This study therefore shows that MAP infection is associated with changes in expression of genes related to the host immune response that may affect its ability to survive and multiply inside the host cell.
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Affiliation(s)
- Rosanna Marino
- CREA Research Centre for Animal Production and Aquaculture, Via Antonio Lombardo 11, 26900 Lodi, Italy; Istituto Sperimentale Italiano "Lazzaro Spallanzani", 26027, Rivolta d'Adda, Cremona, Italy; Institute of Zootechnics, Università Cattolica del S. Cuore, Via Emilia Parmense 84, 29122 Piacenza, Italy.
| | - Rossana Capoferri
- Istituto Sperimentale Italiano "Lazzaro Spallanzani", 26027, Rivolta d'Adda, Cremona, Italy.
| | - Simona Panelli
- Parco Tecnologico Padano, via Einstein, 26900 Lodi, Italy.
| | | | | | - Erminio Trevisi
- Institute of Zootechnics, Università Cattolica del S. Cuore, Via Emilia Parmense 84, 29122 Piacenza, Italy; Nutrigenomics and Proteomic Research Center - PRONUTRIGEN, Università Cattolica del S. Cuore, via Emilia Parmense 84, 29122 Piacenza, Italy.
| | | | - Paolo Ajmone-Marsan
- Institute of Zootechnics, Università Cattolica del S. Cuore, Via Emilia Parmense 84, 29122 Piacenza, Italy; Nutrigenomics and Proteomic Research Center - PRONUTRIGEN, Università Cattolica del S. Cuore, via Emilia Parmense 84, 29122 Piacenza, Italy.
| | - John L Williams
- Davies Research Centre, School of Animal and Veterinary Sciences, University of Adelaide, Roseworthy, SA 5371, Australia.
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Kiser JN, White SN, Johnson KA, Hoff JL, Taylor JF, Neibergs HL. Identification of loci associated with susceptibility to subspecies () tissue infection in cattle. J Anim Sci 2017; 95:1080-1091. [PMID: 28380509 DOI: 10.2527/jas.2016.1152] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Johne's disease is a contagious bacterial infection of cattle caused by ssp. (). A previous genome-wide association analysis (GWAA) in Holstein cattle identified QTL on BTA3 and BTA9 that were highly associated (P < 5 × 10) and on BTA1, BTA16, and BTA21 that were moderately associated (P < 5 × 10) with Map tissue infection. The objectives of this study were to validate previous GWAA results in Jersey cattle ( = 57), Holstein cattle from the Pacific Northwest (PNW, = 205) and a combined Holstein population from the PNW and the Northeast (PNW + NE, = 423), and also identify new loci associated with tissue infection. DNA was genotyped using the Illumina BovineSNP50 BeadChip, and the PNW + NE data was also imputed to whole genome sequence level using Run4 of the 1000 Bull Genomes project with Beagle v 4.1 and FImpute. Cases were ileocecal node positive and controls were negative for by quantitative PCR (qPCR). Individuals were removed for SNP call rate < 90%, and SNP were removed for genotype call rate < 90% or minor allele frequency < 1%. For the Jersey, PNW, and PNW + NE, GWAA were conducted using an allelic dosage model. For the PNW and the PNW + NE, an additional efficient mixed-model association eXpedited (EMMAX) analysis was performed using additive, dominance and recessive models. Seven QTL on BTA22 were identified in the Jersey population with the most significant ( = 4.45 × 10) located at 21.7 megabases (Mb). Six QTL were associated in the PNW and the PNW + NE analyses, including a QTL previously identified on BTA16 in the NE population. The most significant locus for the PNW was located on BTA21 at 61 Mb ( = 8.61 × 10) while the most significant locus for the PNW + NE was on BTA12 at 90 Mb ( = 2.33 × 10). No additional QTL were identified with the imputed GWAA. Putative positional candidate genes were identified within 50 kb 5' and 3' of each QTL. Two positional candidate genes were identified in Jersey cattle, 1 identified in the PNW and 8 in the PNW + NE populations. Many identified positional candidate genes are involved in signal transduction, have immunological functions, or have putative functional relevance in entry into host cells. This study supported 2 previously identified SNP within a QTL on BTA16 and identified 16 new QTL, including 2 found in the PNW and the PNW+NE, associated with tissue infection.
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Parrish N, Vadlamudi A, Goldberg N. Anaerobic adaptation of Mycobacterium avium subspecies paratuberculosis in vitro: similarities to M. tuberculosis and differential susceptibility to antibiotics. Gut Pathog 2017; 9:34. [PMID: 28616081 PMCID: PMC5466712 DOI: 10.1186/s13099-017-0183-z] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/25/2017] [Accepted: 06/01/2017] [Indexed: 01/29/2023] Open
Abstract
Background Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of Johne’s disease in ruminants and is associated with Crohn’s disease (CD) in humans, although the latter remains controversial. In this study, we investigated the ability of MAP to adapt to anaerobic growth using the “Wayne” model of non-replicating persistence (NRP) developed for M. tuberculosis. Results All strains adapted to anaerobiosis over time in a manner similar to that seen with MTB. Susceptibility to 12 antibiotics varied widely between strains under aerobic conditions. Under anaerobic conditions, no drugs caused significant growth inhibition (>0.5 log) except metronidazole, resulting in an average decrease of ~2 logs. Conclusions These results demonstrate that MAP is capable of adaptation to NRP similar to that observed for MTB with differential susceptibility to antibiotics under aerobic versus anaerobic conditions. Such findings have significant implications for our understanding of the pathogenesis of MAP in vivo and the treatment of CD should this organism be established as the causative agent.
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Affiliation(s)
- Nicole Parrish
- The Johns Hopkins Medical Institutions, 600 North Wolfe Street, Meyer B1-193, Baltimore, Maryland USA
| | - Aravinda Vadlamudi
- The Johns Hopkins Medical Institutions, 600 North Wolfe Street, Meyer B1-193, Baltimore, Maryland USA
| | - Neil Goldberg
- Saint Joseph Medical Center, University of Maryland, Towson, Maryland USA
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Superior Protection from Live-Attenuated Vaccines Directed against Johne's Disease. CLINICAL AND VACCINE IMMUNOLOGY : CVI 2017; 24:CVI.00478-16. [PMID: 27806993 DOI: 10.1128/cvi.00478-16] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/11/2016] [Accepted: 10/27/2016] [Indexed: 12/27/2022]
Abstract
Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) is the etiological agent of Johne's disease in ruminants. Johne's disease is an important enteric infection causing large economic losses associated with infected herds. In an attempt to fight this infection, we created two novel live-attenuated vaccine candidates with mutations in sigH and lipN (pgsH and pgsN, respectively). Earlier reports in mice suggested these vaccines are promising candidates to fight Johne's disease in ruminants. In this study, we tested the performances of the two constructs as vaccine candidates using the goat model of Johne's disease. Both vaccines appeared to provide significant immunity to goats against challenge from wild-type M. paratuberculosis The pgsH and pgsN constructs showed a significant reduction in histopathological lesions and tissue colonization compared to nonvaccinated goats and those vaccinated with an inactivated vaccine. Unlike the inactivated vaccine, the pgsN construct was able to eliminate fecal shedding from challenged animals, a feature that is highly desirable to control Johne's disease in infected herds. Furthermore, strong initial cell-mediated immune responses were elicited in goats vaccinated with pgsN that were not demonstrated in other vaccine groups. Overall, the results indicate the potential use of live-attenuated vaccines to control intracellular pathogens, including M. paratuberculosis, and warrant further testing in cattle, the main target for Johne's disease control programs.
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Zamani S, Zali MR, Aghdaei HA, Sechi LA, Niegowska M, Caggiu E, Keshavarz R, Mosavari N, Feizabadi MM. Mycobacterium avium subsp. paratuberculosis and associated risk factors for inflammatory bowel disease in Iranian patients. Gut Pathog 2017; 9:1. [PMID: 28053669 PMCID: PMC5209900 DOI: 10.1186/s13099-016-0151-z] [Citation(s) in RCA: 40] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/28/2016] [Accepted: 12/14/2016] [Indexed: 12/11/2022] Open
Abstract
BACKGROUND Inflammatory bowel disease (IBD) is described as a relapsing condition with high morbidity and uncertain complex pathogenesis. The association of Mycobacterium avium ssp. paratuberculosis (MAP) with Crohn's disease (CD) in human has been debated for decades, however there is no confirmed data to verify such relations in Iran. The aim of this study was to investigate risk factors and a possible role of MAP in Iranian patients with CD. METHODS Anti-MAP antibodies were detected in serum of IBD patients and subjects without IBD (nIBD) through ELISA; MAP DNA and viable MAP cells were identified in patients' biopsies through nested PCR and direct culture methods, respectively. Principal component analysis (PCA) was used to investigate the risk factors in relation to IBD and MAP infection. RESULTS Positivity for IS900 PCR was detected in 64% (n = 18) of CD, 33% (n = 10) of ulcerative colitis (UC) and 9.7% (n = 6) of nIBD samples. Live MAP cells were isolated from biopsies of 2 CD patients only. Among 28 patients with CD, 46% (n = 13) and 39% (n = 11) were positive for antibodies against MAP3865c133-141 and MAP3865c125-133 peptides, respectively, whereas much lower seroreactivity was detected in UC subjects accounting for 3% (n = 1) for MAP3865c133-141 and 16.7% (n = 5) for MAP3865c125-133. A high immune reactivity to MAP epitopes among CD patients was positively correlated with consumption of fast food meals and IBD familiarity. For both CD and UC, breastfeeding period and consumption of fruit/vegetables presented negative correlation with the presence of anti-MAP antibodies. CONCLUSIONS This study provided evidences that high prevalence of MAP DNA and anti-MAP antibodies in CD patients is significantly associated with the development of CD. Despite the role of several factors contributing to IBD, the presence of MAP DNA and anti-MAP antibodies in Iranian CD patients highlights a possible transmission of MAP from animal-derived products to humans.
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Affiliation(s)
- Samin Zamani
- Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Mohammad Reza Zali
- Gastroenterology and Liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Hamid Asadzadeh Aghdaei
- Gastroenterology and Liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Leonardo Antonio Sechi
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43b, 07100 Sassari, Italy
| | - Magdalena Niegowska
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43b, 07100 Sassari, Italy
| | - Elisa Caggiu
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43b, 07100 Sassari, Italy
| | - Rouhollah Keshavarz
- PPD Tuberculin Department, Razi Vaccine & Serum Research Institute, Karaj, Iran
- Reference Laboratory for Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran
| | - Nader Mosavari
- PPD Tuberculin Department, Razi Vaccine & Serum Research Institute, Karaj, Iran
- Reference Laboratory for Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran
| | - Mohammad Mehdi Feizabadi
- Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
- Thoracic Diseases Research Center, Tehran University of Medical Sciences, Tehran, Iran
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Fiebiger U, Bereswill S, Heimesaat MM. Dissecting the Interplay Between Intestinal Microbiota and Host Immunity in Health and Disease: Lessons Learned from Germfree and Gnotobiotic Animal Models. Eur J Microbiol Immunol (Bp) 2016; 6:253-271. [PMID: 27980855 PMCID: PMC5146645 DOI: 10.1556/1886.2016.00036] [Citation(s) in RCA: 106] [Impact Index Per Article: 11.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/14/2016] [Accepted: 11/21/2016] [Indexed: 02/06/2023] Open
Abstract
This review elaborates the development of germfree and gnotobiotic animal models and their application in the scientific field to unravel mechanisms underlying host-microbe interactions and distinct diseases. Strictly germfree animals are raised in isolators and not colonized by any organism at all. The germfree state is continuously maintained by birth, raising, housing and breeding under strict sterile conditions. However, isolator raised germfree mice are exposed to a stressful environment and exert an underdeveloped immune system. To circumvent these physiological disadvantages depletion of the bacterial microbiota in conventionally raised and housed mice by antibiotic treatment has become an alternative approach. While fungi and parasites are not affected by antibiosis, the bacterial microbiota in these "secondary abiotic mice" have been shown to be virtually eradicated. Recolonization of isolator raised germfree animals or secondary abiotic mice results in a gnotobiotic state. Both, germfree and gnotobiotic mice have been successfully used to investigate biological functions of the conventional microbiota in health and disease. Particularly for the development of novel clinical applications germfree mice are widely used tools, as summarized in this review further focusing on the modulation of bacterial microbiota in laboratory mice to better mimic conditions in the human host.
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Affiliation(s)
| | | | - Markus M. Heimesaat
- Gastrointestinal Microbiology Research Group, Institute of Microbiology and Hygiene, Charité – University Medicine Berlin, Campus Benjamin Franklin
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Singh AV, Chauhan DS, Singh SV, Kumar V, Singh A, Yadav A, Yadav VS. Current status of Mycobacterium avium subspecies paratuberculosis infection in animals & humans in India: What needs to be done? Indian J Med Res 2016; 144:661-671. [PMID: 28361818 PMCID: PMC5393076 DOI: 10.4103/ijmr.ijmr_1401_14] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/18/2014] [Indexed: 12/13/2022] Open
Abstract
Mycobacterium avium subspecies paratuberculosis (MAP) has emerged as a major health problem for domestic livestock and human beings. Reduced per animal productivity of domestic livestock seriously impacts the economics of dairy farming globally. High to very high bioload of MAP in domestic livestock and also in the human population has been reported from north India. Presence of live MAP bacilli in commercial supplies of raw and pasteurized milk and milk products indicates its public health significance. MAP is not inactivated during pasteurization, therefore, entering into human food chain daily. Recovery of MAP from patients with inflammatory bowel disease or Crohn's disease and animal healthcare workers suffering with chronic gastrointestinal problems indicate a close association of MAP with a number of chronic and other diseases affecting human health. Higher bioload of MAP in the animals increases the risk of exposure to the human population with MAP. This review summarizes the current status of MAP infection in animals as well as in human beings and also highlights the prospects of effective management and control of disease in animals to reduce the risk of exposure to human population.
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Affiliation(s)
- Ajay Vir Singh
- Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases, Agra, India
| | - Devendra Singh Chauhan
- Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases, Agra, India
| | - Shoor Vir Singh
- Animal Health Division, Central Institute for Research on Goats, Mathura, India
| | - Vijay Kumar
- Department of Biotechnology, Institute of Biomedical Education & Research, Mangalayatan University, Aligarh, India
| | - Abhinendra Singh
- Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases, Agra, India
| | - Anjali Yadav
- Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases, Agra, India
| | - Virendra Singh Yadav
- Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases, Agra, India
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Evaluation of the association between the lactation stage and serum and milk ELISA results in the diagnosis of ovine Paratuberculosis. Small Rumin Res 2016. [DOI: 10.1016/j.smallrumres.2016.04.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
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Verin R, Perroni M, Rossi G, De Grossi L, Botta R, De Sanctis B, Rocca S, Cubeddu T, Crosby-Durrani H, Taccini E. Paratuberculosis in sheep: Histochemical, immunohistochemical and in situ hybridization evidence of in utero and milk transmission. Res Vet Sci 2016; 106:173-9. [PMID: 27234557 DOI: 10.1016/j.rvsc.2016.04.006] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/27/2015] [Revised: 02/05/2016] [Accepted: 04/25/2016] [Indexed: 11/16/2022]
Abstract
To investigate in utero and milk transmission of Mycobacterium avium subsp. paratuberculosis (MAP), tissues from thirteen pregnant sheep, naturally infected and serologically positive to MAP, were examined by means of histochemistry, immunohistochemistry and in situ hybridization. Soon after parturition, ewes were euthanized and tissues samples were collected and prepared. The offspring (18 lambs) were divided into three groups to investigate different routes of MAP transmission. Lambs were sacrificed at three months old and the tissue samples collected, formalin-fixed and paraffin embedded. Hematoxylin and eosin and Ziehl-Neelsen staining methods were performed on fixed tissues for general examination and for detection of acid-fast bacteria. Additionally, immunohistochemical and in situ hybridization techniques were used to detect MAP antigen and MAP DNA respectively. This study of a flock of MAP-infected sheep indicates both in utero and milk transmission of MAP from dams to their offspring. Importantly, this study detected the presence of MAP in the mammary gland and mammary lymph nodes of adult ewes therefore indicating a significant route for the potential exposure to humans from this bacterial infection.
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Affiliation(s)
- Ranieri Verin
- Department of Animal Pathology, School of Veterinary Medicine, University of Pisa, Italy; Department of Veterinary Pathology, School of Veterinary Science, University of Liverpool, UK.
| | - Marco Perroni
- Department of Animal Pathology, School of Veterinary Medicine, University of Pisa, Italy
| | - Giacomo Rossi
- School of Biosciences and Veterinary Medicine, University of Camerino, Italy
| | - Luigi De Grossi
- Istituto Zooprofilattico Sperimentale Lazio and Toscana, Viterbo, Italy
| | | | - Bruno De Sanctis
- Istituto Zooprofilattico Sperimentale Lazio and Toscana, Viterbo, Italy
| | - Stefano Rocca
- Department of Veterinary Medicine, University of Sassari, Italy
| | - Tiziana Cubeddu
- Department of Veterinary Medicine, University of Sassari, Italy
| | - Hayley Crosby-Durrani
- Department of Veterinary Pathology, School of Veterinary Science, University of Liverpool, UK
| | - Ennio Taccini
- Department of Animal Pathology, School of Veterinary Medicine, University of Pisa, Italy
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Khan IA, Pilli S, A S, Rampal R, Chauhan SK, Tiwari V, Mouli VP, Kedia S, Nayak B, Das P, Makharia GK, Ahuja V. Prevalence and Association of Mycobacterium avium subspecies paratuberculosis with Disease Course in Patients with Ulcero-Constrictive Ileocolonic Disease. PLoS One 2016; 11:e0152063. [PMID: 27019109 PMCID: PMC4809507 DOI: 10.1371/journal.pone.0152063] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2015] [Accepted: 03/08/2016] [Indexed: 12/30/2022] Open
Abstract
BACKGROUND Association of Mycobacterium avium subspecies paratuberculosis (MAP) and Crohn's disease (CD) has been controversial due to contradictory reports. Therefore, we determined the prevalence of MAP in patients with CD and intestinal tuberculosis (ITB) and its association with clinical course. METHODOLOGY Blood and intestinal biopsies were taken from 69 CD, 32 ITB patients and 41 patients with haemorrhoidal bleed who served as controls. qPCR targeting of MAP-specific IS900 gene was used to detect the presence of MAP DNA. qPCR results were further validated by sequencing. Immunohistochemistry (IHC) was used to detect the presence of MAP antigen in biopsy specimens. CD and ITB patients were followed-up for disease course and response to therapy. PRINCIPAL FINDINGS The frequency of MAP-specific DNA in biopsies by qPCR was significantly higher in CD patients (23.2%, p = 0.03) as compared to controls (7.3%). No significant difference in intestinal MAP presence was observed between ITB patients (12.5%, p = 0.6) and controls (7.3%). MAP presence in blood of CD patients was 10.1% as compared to 4.9% in controls while no patients with ITB were found to be positive (p = 0.1). Using IHC for detection of MAP antigen, the prevalence of MAP in CD was 2.9%, 12.5% in ITB patients and 2.4% in controls. However, long-term follow-up of the patients revealed no significant associations between clinical characteristics and treatment outcomes with MAP positivity. CONCLUSION We report significantly high prevalence of MAP in intestinal biopsies of CD patients. However, the presence of MAP does not affect the disease course and treatment outcomes in either CD or ITB patients.
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Affiliation(s)
- Imteyaz Ahmad Khan
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Sucharita Pilli
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Surendranath A
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Ritika Rampal
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Sudhir Kumar Chauhan
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Veena Tiwari
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Venigalla Pratap Mouli
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Saurabh Kedia
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Baibaswata Nayak
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Prasenjit Das
- Department of Pathology, All India Institute of Medical Sciences, New Delhi, India
| | - Govind K. Makharia
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
| | - Vineet Ahuja
- Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, New Delhi, India
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36
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Niegowska M, Paccagnini D, Mannu C, Targhetta C, Songini M, Sechi LA. Recognition of ZnT8, Proinsulin, and Homologous MAP Peptides in Sardinian Children at Risk of T1D Precedes Detection of Classical Islet Antibodies. J Diabetes Res 2016; 2016:5842701. [PMID: 26824044 PMCID: PMC4707333 DOI: 10.1155/2016/5842701] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/17/2015] [Revised: 11/09/2015] [Accepted: 11/10/2015] [Indexed: 12/12/2022] Open
Abstract
As numerous studies put in evidence the increasing incidence of type 1 diabetes (T1D) in children, an early diagnosis is of great importance to define correct treatment and diet. Currently, the identification of classical islet autoantibodies is the primary biomarker for diagnosis in subjects at risk, especially in pediatric patients. Recent studies suggest that detection of antibodies against ZnT8 protein in preclinical phase can predict the development of T1D. We previously demonstrated a significant association of Mycobacterium avium subspecies paratuberculosis (MAP) with T1D in adult Sardinian patients. To enforce this finding, we investigated the presence of antibodies against ZnT8 and proinsulin (PI) with respective homologous epitopes: MAP3865c133-141/ZnT8186-194, MAP3865c125-133/ZnT8178-186, MAP2404c70-85/PI46-61, and MAP1,4αgbp157-173/PI64-80, in 23 children at risk for T1D, formerly involved in the TRIGR study, and 22 healthy controls (HCs). Positivity to anti-MAP and homologous human peptides was detected in 48% of at-risk subjects compared to 5,85% HCs, preceding appearance of islet autoantibodies. Being MAP easily transmitted to humans with infected cow's milk and detected in retail infant formulas, MAP epitopes could be present in extensively hydrolyzed formula and act as antigens stimulating β-cell autoimmunity.
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Affiliation(s)
- Magdalena Niegowska
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy
| | - Daniela Paccagnini
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy
| | - Carla Mannu
- Centre for the Treatment of Complications of Diabetes, Hospital “G. Brotzu”, 09134 Cagliari, Italy
| | - Clara Targhetta
- Centre for the Treatment of Complications of Diabetes, Hospital “G. Brotzu”, 09134 Cagliari, Italy
| | - Marco Songini
- Centre for the Treatment of Complications of Diabetes, Hospital “G. Brotzu”, 09134 Cagliari, Italy
| | - Leonardo A. Sechi
- Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy
- *Leonardo A. Sechi:
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Host Transcriptional Profiles and Immunopathologic Response following Mycobacterium avium subsp. paratuberculosis Infection in Mice. PLoS One 2015; 10:e0138770. [PMID: 26439498 PMCID: PMC4595071 DOI: 10.1371/journal.pone.0138770] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2015] [Accepted: 09/03/2015] [Indexed: 01/11/2023] Open
Abstract
Paratuberculosis or Johne’s disease is a chronic granulomatous enteropathy in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP) infection. In the present study, we examined the host response to MAP infection in spleens of mice in order to investigate the host immunopathology accompanying host-pathogen interaction. Transcriptional profiles of the MAP-infected mice at 3 and 6 weeks p.i. showed severe histopathological changes, whereas those at 12 weeks p.i. displayed reduced lesion severity in the spleen and liver. MAP-infected mice at 3 and 6 weeks p.i. showed up-regulation of interferon-related genes, scavenger receptor, and complement components, suggesting an initial innate immune reaction, such as macrophage activation, bactericidal activity, and macrophage invasion of MAP. Concurrently, MAP-infected mice at 3 and 6 weeks p.i. were also suggested to express M2 macrophage phenotype with up-regulation of Mrc1, and Marco and down-regulation of MHC class II, Ccr7, and Irf5, and canonical pathways related to the T cell response including ICOS-ICOSL signaling in T helper cells, calcium-induced T lymphocyte apoptosis, and CD28 signaling in T helper cell. These results provide information which furthers the understanding of the immunopathologic response to MAP infection in mice, thereby providing insights valuable for research into the pathogenesis for MAP infection.
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38
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Monif GRG. The Hruska postulate of Crohn's disease. Med Hypotheses 2015; 85:878-81. [PMID: 26432629 DOI: 10.1016/j.mehy.2015.09.019] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2015] [Revised: 09/09/2015] [Accepted: 09/20/2015] [Indexed: 02/07/2023]
Abstract
Crohn's disease is due to the loss of immunological tolerance within the gastrointestinal tract to the antigenic array of Mycobacterium avium subspecies paratuberculosis (MAP) and closely related polymorphic variants. The loss of immune tolerance results in an effector cytokine responsive upon re-exposure to MAP. For immune tolerance to MAP to be induced, infection must occur when acquired immunity is markedly underdeveloped.
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Affiliation(s)
- Gilles R G Monif
- Infectious Diseases Incorporated, Bellevue, NE, United States; University of Florida, College of Veterinary Medicine, Gainesville, FL, United States.
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Otsubo S, Cossu D, Eda S, Otsubo Y, Sechi LA, Suzuki T, Iwao Y, Yamamoto S, Kuribayashi T, Momotani E. Seroprevalence of IgG1 and IgG4 class antibodies against Mycobacterium avium subsp. paratuberculosis in Japanese population. Foodborne Pathog Dis 2015; 12:851-6. [PMID: 26267654 DOI: 10.1089/fpd.2015.1956] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023] Open
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) is the established causative agent of Johne's disease in cattle and other ruminants, and it has also been speculated to be a putative etiological agent of several human autoimmune diseases. It is acknowledged that dairy products deriving from infected animals play a role (could be vehicles) in exposing humans to MAP. MAP could stimulate the human immune system by means of their complex antigen (in the case of lipids, multivalent antigens) and may modulate it, acting as adjuvant molecules such as Freund's complete adjuvant. The immune system might be abnormally stimulated by the constant presence of MAP antigens (for example, in the dairy products), and this might be particularly relevant in genetically predisposed individuals. However, there is limited understanding about the current human exposure to MAP. The present study analyzed the antibody recognition profile of MAP lipophilic antigens in a cohort of 126 healthy Japanese. We measured the serum levels of total immunoglobulin G (IgG) and subclasses targeting MAP surface antigens through ethanol vortex indirect enzyme-linked immunosorbent assay (EVELISA) by using serum absorbed with Mycobacterium phlei. Elevated IgG (especially IgG1 and IgG4) responses were observed in 14% of the sera. To assess the specificity of EVELISA, the same samples were analyzed by means of a commercially available Johnelisa II kit. It was noteworthy that a high degree of correlation was observed when comparing the two methodologies (rs=0.7, p<0.0001). Moreover, in order to investigate the specificity of the binding, inhibition assay experiments were carried out also searching for antibodies against Bacillus Calmette-Guérin antigens, but no cross-reaction was observed. The result obtained represents the first evidence implying that the Japanese population is exposed to MAP, and additionally the existence of a foodborne chain of exposure that transmits MAP antigens to humans.
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Affiliation(s)
| | - Davide Cossu
- 2 Department of Biomedical Sciences, Division of Microbiology and Virology, University of Sassari , Viale San Pietro, Italy .,3 Department of Human-Care, Tohto College of Health Sciences (TCHS) , Saitama, Japan
| | - Shigetoshi Eda
- 4 Center for Wildlife Health, Department of Forestry, Wildlife, and Fisheries, University of Tennessee , Knoxville, Tennessee
| | | | - Leonardo Antonio Sechi
- 2 Department of Biomedical Sciences, Division of Microbiology and Virology, University of Sassari , Viale San Pietro, Italy
| | - Tsuyoshi Suzuki
- 3 Department of Human-Care, Tohto College of Health Sciences (TCHS) , Saitama, Japan
| | - Yumiko Iwao
- 3 Department of Human-Care, Tohto College of Health Sciences (TCHS) , Saitama, Japan
| | - Shizuo Yamamoto
- 5 Laboratories of Immunology, School of Life and Environmental Science, Azabu University , Kanagawa, Japan
| | - Takashi Kuribayashi
- 5 Laboratories of Immunology, School of Life and Environmental Science, Azabu University , Kanagawa, Japan
| | - Eiichi Momotani
- 3 Department of Human-Care, Tohto College of Health Sciences (TCHS) , Saitama, Japan
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Liaskos C, Spyrou V, Athanasiou LV, Orfanidou T, Mavropoulos A, Rigopoulou EI, Amiridis GS, Shoenfeld Y, Billinis C, Bogdanos DP. Crohn's disease-specific anti-CUZD1 pancreatic antibodies are absent in ruminants with paratuberculosis. Clin Res Hepatol Gastroenterol 2015; 39:384-390. [PMID: 25575461 DOI: 10.1016/j.clinre.2014.12.001] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/28/2014] [Revised: 11/23/2014] [Accepted: 12/01/2014] [Indexed: 02/07/2023]
Abstract
BACKGROUND Pancreatic autoantibodies (PABs) specifically recognizing GP2 and/or CUZD1 are present in more than 35% of patients with Crohn's disease (CrD). We have recently provided evidence of the presence of GP2-specific PABs in ruminants with paratuberculosis (ptb), a Mycobacterium avium paratuberculosis (MAP)-induced disease resembling CrD. OBJECTIVE To assess whether anti-CUZD1 antibodies are also present in ruminants with ptb. METHODS A total of 110 samples (73 cattle/37 sheep) were studied including 40 with ptb (24 cattle/16 sheep; 20 anti-GP2 antibody pos) and 70 without ptb (49 cattle/21 sheep; 10 anti-GP2 antibody pos). The samples were pre-characterized for anti-MAP and anti-GP2 antibodies by ELISA. Evidence of MAP was confirmed by PCR. Anti-CUZD1 antibody testing was performed by indirect immunofluorescence (IIF) based on transfected HEK293 cells expressing CUZD1. Anti-sheep or anti-cattle specific antisera were used as revealing antibodies. RESULTS None of the ruminant sera had anti-CUZD1 antibodies by IIF testing at dilutions varying from 1/10 to 1/160. Methodological flaws were prevented by a series of tests. Control sera from anti-CUZD1 positive CrD samples have shown anti-CUZD1 antibody reactivity at various concentrations. Antibody reactivity to GP2-expressing HEK293 cells has confirmed the reactivity to GP2 in ruminant sera found positive for anti-GP2 antibodies by ELISA. CONCLUSION The present study has found no evidence of anti-CUZD1 PABs in MAP-induced ptb. Our findings indicate that the induction of CUZD1-specific PABs is unrelated to MAP infection and that the mechanisms responsible for the loss of tolerance to GP2 and CUZD1 are probably quite distinct.
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Affiliation(s)
- Christos Liaskos
- Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece; Department of Rheumatology, Faculty of Medicine, School of Health Sciences, University of Thessaly, Biopolis, 45000 Larissa, Greece.
| | - Vassiliki Spyrou
- Department of Animal Production, Technological Educational Institute of Thessaly, Larissa 41110, Greece.
| | - Labrini V Athanasiou
- Department of Medicine, Faculty of Veterinary Medicine, University of Thessaly, Karditsa 43100, Greece.
| | - Timoklia Orfanidou
- Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece.
| | - Athanasios Mavropoulos
- Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece.
| | - Eirini I Rigopoulou
- Division of Transplantation Immunology and Mucosal Biology, King's College London School of Medicine at King's College Hospital, London SE5 9RJ, UK.
| | - Georgios S Amiridis
- Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Thessaly, Karditsa 43100, Greece.
| | - Yehuda Shoenfeld
- The Zabludowicz Center for Autoimmune Diseases, Sheba Medical Center, Tel-Hashomer, Israel.
| | - Charalambos Billinis
- Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Thessaly, 43100 Karditsa, Greece.
| | - Dimitrios P Bogdanos
- Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece; Department of Rheumatology, Faculty of Medicine, School of Health Sciences, University of Thessaly, Biopolis, 45000 Larissa, Greece; Division of Transplantation Immunology and Mucosal Biology, King's College London School of Medicine at King's College Hospital, London SE5 9RJ, UK.
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Nazareth N, Magro F, Machado E, Ribeiro TG, Martinho A, Rodrigues P, Alves R, Macedo GN, Gracio D, Coelho R, Abreu C, Appelberg R, Dias C, Macedo G, Bull T, Sarmento A. Prevalence of Mycobacterium avium subsp. paratuberculosis and Escherichia coli in blood samples from patients with inflammatory bowel disease. Med Microbiol Immunol 2015; 204:681-92. [PMID: 25994082 DOI: 10.1007/s00430-015-0420-3] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2015] [Accepted: 05/13/2015] [Indexed: 12/31/2022]
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) and adherent-invasive Escherichia coli (AIEC) have been implicated as primary triggers in Crohn's disease (CD). In this study, we evaluated the prevalence of MAP and E. coli (EC) DNA in peripheral blood from 202 inflammatory bowel disease (IBD) patients at various disease periods and compared against 24 cirrhotic patients with ascites (CIR) (non-IBD controls) and 29 healthy controls (HC). MAP DNA was detected by IS900-specific nested PCR, EC DNA by malB-specific nested PCR and AIEC identity, in selected samples, by sequencing of fimH gene. CD patients with active disease showed the highest MAP DNA prevalence among IBD patients (68 %). Infliximab treatment resulted in decreased MAP detection. CIR patients had high individual and coinfection rates (75 % MAP, 88 % EC and 67 % MAP and EC), whilst HC controls had lower MAP prevalence (38 %) and EC was undetectable in this control group. EC DNA prevalence in IBD patients was highly associated with CD, and 80 % of EC from the selected samples of CD patients analyzed carried the fimH30 allele, with a mutation strongly associated with AIEC. Our results show that coinfection with MAP and AIEC is common and persistent in CD, although the high MAP and EC detection in CIR patients suggested that colonization is, at least, partially dependent on increased gut permeability. Nevertheless, facilitative mechanisms between a susceptible host and these two potential human pathogens may allow their implication in CD pathogenesis.
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Affiliation(s)
- Nair Nazareth
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
| | - Fernando Magro
- Institute of Pharmacology and Therapeutics, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
- Gastroenterology Department, Centro Hospitalar São João, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
- MedInUP -Center for Drug Discovery and Innovative Medicines, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Elisabete Machado
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
- REQUIMTE, Laboratory of Microbiology, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal
| | - Teresa Gonçalves Ribeiro
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
- REQUIMTE, Laboratory of Microbiology, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal
| | - António Martinho
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
| | - Pedro Rodrigues
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
- Infection and Immunity Unit, IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal
| | - Rita Alves
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
| | - Gonçalo Nuno Macedo
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal
| | - Daniela Gracio
- Institute of Pharmacology and Therapeutics, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
- MedInUP -Center for Drug Discovery and Innovative Medicines, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Rosa Coelho
- Gastroenterology Department, Centro Hospitalar São João, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Candida Abreu
- Department of Infectious Diseases, Centro Hospitalar S. João, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
- Nephrology Research and Development Unit, Faculdade de Medicina da Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Rui Appelberg
- Infection and Immunity Unit, IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal
| | - Camila Dias
- Department of Biostatistics and Medical Informatics, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Guilherme Macedo
- Gastroenterology Department, Centro Hospitalar São João, Alameda Professor Hernâni Monteiro, 4200-319, Porto, Portugal
| | - Tim Bull
- Institute for Infection and Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK
| | - Amélia Sarmento
- FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), University Fernando Pessoa, Rua Carlos da Maia, 296, 4200-150, Porto, Portugal.
- Infection and Immunity Unit, IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal.
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Masala S, Cossu D, Piccinini S, Rapini N, Mameli G, Manca Bitti ML, Sechi LA. Proinsulin and MAP3865c homologous epitopes are a target of antibody response in new-onset type 1 diabetes children from continental Italy. Pediatr Diabetes 2015; 16:189-95. [PMID: 25720593 DOI: 10.1111/pedi.12269] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/25/2014] [Revised: 01/12/2015] [Accepted: 02/03/2015] [Indexed: 12/31/2022] Open
Abstract
Mycobacterium avium subspecies paratuberculosis (MAP) asymptomatic infection is speculated to play a role in type 1 diabetes (T1D) among Sardinian subjects. Data obtained analyzing a pediatric population from mainland Italy lends support to the hypothesis, which envisions MAP as an environmental factor at play in T1D pathogenesis. Aiming to investigate the likelihood of cross-recognition between linear determinants shared by self (proinsulin) and non-self (MAP) proteins, 59 children with new onset T1D and 60 healthy controls (HCs) from continental Italy were enrolled in the study. Serum samples were subjected to indirect enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies (Abs) toward four homologues MAP/proinsulin epitopes. The rate of MAP infection (42.4% in T1D children and 5% in HCs; p < 0.0001) was estimated searching for Abs against MAP specific protein MptD. The homologous MAP2404c70-85 and proinsulin (PI)46-61 peptides were recognized by 42.4 and 39% of new-onset T1D children and only in 5% of HCs (AUC = 0.76, AUC = 0.7, p < 0.0001); whereas the prevalence of Abs against MAP 1,4-α-gbp157-173 and PI64-80 peptides was 45.7 and 49.1% in new-onset T1D children, respectively, compared with 3.3% of HCs (AUC = 0.74 and p < 0.0001 in both). Pre-incubation of MAP Ab-positive sera with proinsulin peptides was able to block the binding to the correspondent MAP epitopes, thus showing that Abs against these homologous peptides are cross-reactive. MAP/Proinsulin Ab mediated cross-recognition, most likely via molecular mimicry, maybe a factor in accelerating and/or initiating T1D in MAP-infected children. Indeed, it is known that anti-proinsulin and anti-Insulin autoantibodies are the earliest to appear.
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Affiliation(s)
- Speranza Masala
- Department of Biomedical Sciences, Section of Experimental and Clinical Microbiology, University of Sassari, Sassari, 07100, Italy
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Shin MK, Shin SW, Jung M, Park H, Park HE, Yoo HS. Host gene expression for Mycobacterium avium subsp. paratuberculosis infection in human THP-1 macrophages. Pathog Dis 2015; 73:ftv031. [PMID: 25877879 DOI: 10.1093/femspd/ftv031] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 04/06/2015] [Indexed: 11/13/2022] Open
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, which causes considerable economic loss in the dairy industry and has a possible relationship to Crohn's disease (CD) in humans. As MAP has been detected in retail pasteurized milk samples, its transmission via milk is of concern. Despite its possible role in the etiology of CD, there have been few studies examining the interactions between MAP and human cells. In the current study, we applied Ingenuity Pathway Analysis to the transcription profiles generated from a murine model with MAP infection as part of a previously conducted study. Twenty-one genes were selected as potential host immune responses, compared with the transcriptional profiles in naturally MAP-infected cattle, and validated in MAP-infected human monocyte-derived macrophage THP-1 cells. Of these, the potential host responses included up-regulation of genes related to immune response (CD14, S100A8, S100A9, LTF, HP and CHCIL3), up-regulation of Th1-polarizing factor (CCL4, CCL5, CXCL9 and CXCL10), down-regulation of genes related to metabolism (ELANE, IGF1, TCF7L2 and MPO) and no significant response of other genes (GADD45a, GPNMB, HMOX1, IFNG and NQO1) in THP-1 cells infected with MAP.
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Affiliation(s)
- Min-Kyoung Shin
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea Dairy and Swine Research and Development Centre, Agriculture and Agri-Food Canada, Sherbrooke, QC J1M 1Z3, Canada
| | - Seung Won Shin
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea
| | - Myunghwan Jung
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea
| | - Hongtae Park
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea
| | - Hyun-Eui Park
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea
| | - Han Sang Yoo
- Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 151-742 Korea Institute of Green Bio Science and Technology, Seoul National University, Pyeongchang, 232-916, Korea
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Comparative Evaluation of Different Test Combinations for Diagnosis of Mycobacterium avium Subspecies paratuberculosis Infecting Dairy Herds in India. BIOMED RESEARCH INTERNATIONAL 2015; 2015:983978. [PMID: 25945351 PMCID: PMC4402184 DOI: 10.1155/2015/983978] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Subscribe] [Scholar Register] [Received: 11/25/2014] [Revised: 02/24/2015] [Accepted: 03/09/2015] [Indexed: 11/30/2022]
Abstract
A total of 355 cows were sampled (serum, n = 315; faeces, n = 355; milk, n = 209) from dairy farms located in the Punjab state of India. Faeces and serum/milk samples were screened by acid fast staining and “indigenous ELISA,” respectively. IS900 PCR was used to screen faeces and milk samples. Bio-load of MAP in dairy cows was 36.9, 15.6, 16.3, and 14.4%, using microscopy, serum ELISA, milk ELISA and milk PCR, respectively. Estimated kappa values between different test combinations: serum and milk ELISA, faecal microscopy and faecal PCR, milk ELISA and milk PCR, faecal PCR and serum ELISA were 0.325, 0.241, 0.682, and 0.677, respectively. Estimation of the relative sensitivity and specificity of different tests in the present study indicated that “serum ELISA” and “milk ELISA” were good screening tests, add “milk PCR” was “confirmatory test” for MAP infection. Combination of milk ELISA with milk PCR may be adopted as a model strategy for screening and diagnosis of JD in lactating/dairy cattle herds in Indian conditions.
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Increased viability but decreased culturability of Mycobacterium avium subsp. paratuberculosis in macrophages from inflammatory bowel disease patients under Infliximab treatment. Med Microbiol Immunol 2015; 204:647-56. [PMID: 25702170 DOI: 10.1007/s00430-015-0393-2] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/24/2014] [Accepted: 02/10/2015] [Indexed: 12/26/2022]
Abstract
Mycobacterium avium subsp. paratuberculosis (MAP) has long been implicated as a triggering agent in Crohn's disease (CD). In this study, we investigated the growth/persistence of both M. avium subsp. hominissuis (MAH) and MAP, in macrophages from healthy controls (HC), CD and ulcerative colitis patients. For viability assessment, both CFU counts and a pre16SrRNA RNA/DNA ratio assay (for MAP) were used. Phagolysosome fusion was evaluated by immunofluorescence, through analysis of LAMP-1 colocalization with MAP. IBD macrophages were more permissive to MAP survival than HC macrophages (a finding not evident with MAH), but did not support MAP active growth. The lower MAP CFU counts in macrophage cultures associated with Infliximab treatment were not due to increased killing, but possibly to elevation in the proportion of intracellular dormant non-culturable MAP forms, as MAP showed higher viability in those macrophages. Increased MAP viability was not related to lack of phagolysosome maturation. The predominant induction of MAP dormant forms by Infliximab treatment may explain the lack of MAP reactivation during anti-TNF therapy of CD but does not exclude the possibility of MAP recrudescence after termination of therapy.
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McMullen L, 1 School of Women's and Children's Health, University of New South Wales, Sydney, Australia;, T Leach S, A Lemberg D, S Day A. Current roles of specific bacteria in the pathogenesis of inflammatory bowel disease. AIMS Microbiol 2015. [DOI: 10.3934/microbiol.2015.1.82] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022] Open
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Performance of interferon-gamma release assay for tuberculosis screening in inflammatory bowel disease patients. Inflamm Bowel Dis 2014; 20:2067-72. [PMID: 25159454 DOI: 10.1097/mib.0000000000000147] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
BACKGROUND Screening for latent tuberculosis (TB) is mandatory in inflammatory bowel disease (IBD) before starting anti-tumor necrosis factor therapy. Data on the utility of screening tests in populations with moderate background risk of TB are limited. This study aims to evaluate the performance of interferon-gamma release assay (IGRA) with QuantiFERON-TB Gold in IBD patients in a TB endemic region. METHODS Two hundred sixty-eight consecutive adult IBD patients and 234 healthy controls were prospectively recruited. Detailed clinical history, chest x-ray findings, and IGRA results were documented for all individuals. The IGRA positive rates between IBD patients, with or without immunosuppressant, and healthy controls were compared. RESULTS The IGRA result was positive in 21.9% of IBD patients and 19.2% of healthy controls (P = 0.535). IBD patients on immunosuppressive therapy had a significantly lower IGRA positive rate (13.0% versus 29.6%; P = 0.002) compared with immunosuppressant-naive IBD patients. This difference seemed to be most prominent for patients taking azathioprine (11.8% versus 27.3%, P = 0.006). CONCLUSIONS IGRA results are negatively impacted by immunosuppressive therapy. Current guidelines suggesting TB screening before anti-tumor necrosis factor therapy may be inadequate in patients already on immunosuppressive drugs. Latent TB testing seems best performed before the initiation of immunosuppressive therapies in IBD patients.
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Wang ZK, Yang YS, Chen Y, Yuan J, Sun G, Peng LH. Intestinal microbiota pathogenesis and fecal microbiota transplantation for inflammatory bowel disease. World J Gastroenterol 2014; 20:14805-14820. [PMID: 25356041 PMCID: PMC4209544 DOI: 10.3748/wjg.v20.i40.14805] [Citation(s) in RCA: 88] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/26/2014] [Revised: 06/16/2014] [Accepted: 07/16/2014] [Indexed: 02/06/2023] Open
Abstract
The intestinal microbiota plays an important role in inflammatory bowel disease (IBD). The pathogenesis of IBD involves inappropriate ongoing activation of the mucosal immune system driven by abnormal intestinal microbiota in genetically predisposed individuals. However, there are still no definitive microbial pathogens linked to the onset of IBD. The composition and function of the intestinal microbiota and their metabolites are indeed disturbed in IBD patients. The special alterations of gut microbiota associated with IBD remain to be evaluated. The microbial interactions and host-microbe immune interactions are still not clarified. Limitations of present probiotic products in IBD are mainly due to modest clinical efficacy, few available strains and no standardized administration. Fecal microbiota transplantation (FMT) may restore intestinal microbial homeostasis, and preliminary data have shown the clinical efficacy of FMT on refractory IBD or IBD combined with Clostridium difficile infection. Additionally, synthetic microbiota transplantation with the defined composition of fecal microbiota is also a promising therapeutic approach for IBD. However, FMT-related barriers, including the mechanism of restoring gut microbiota, standardized donor screening, fecal material preparation and administration, and long-term safety should be resolved. The role of intestinal microbiota and FMT in IBD should be further investigated by metagenomic and metatranscriptomic analyses combined with germ-free/human flora-associated animals and chemostat gut models.
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Okafor C, Grooms D, Alocilja E, Bolin S. Comparison between a conductometric biosensor and ELISA in the evaluation of Johne's disease. SENSORS (BASEL, SWITZERLAND) 2014; 14:19128-37. [PMID: 25320903 PMCID: PMC4239859 DOI: 10.3390/s141019128] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/11/2014] [Revised: 10/08/2014] [Accepted: 10/09/2014] [Indexed: 11/24/2022]
Abstract
Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is an important gastrointestinal disease of cattle worldwide because of the economic losses encountered in JD-affected herds. These losses include reduction in milk yield in cows, premature culling and reduced carcass weight of culled diseased animals. In the U.S. dairy industry, economic losses from reduced productivity associated with JD are estimated to cost between $200 and $250 million annually. The development of non-laboratory-based assays would support more frequent testing of animals for JD and could improve its control. Conductometric biosensors combine immunomigration technology with electronic signal detection and have been adapted for the detection of IgG antibody against MAP. In the present study, a capture membrane with limited variability in the immunomigration channel and an optimal concentration of the secondary anti-bovine antibody used in a previously developed conductometric biosensor were compared with a commercially available antibody detection ELISA in their evaluation of JD, using samples of serum from cattle whose JD status where unknown. There was a moderate strength of agreement (kappa = 0.41) between the two assays. Findings from this preliminary study support the continued development of conductometric biosensors for use in the diagnosis of JD.
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Affiliation(s)
- Chika Okafor
- Large Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA.
| | - Daniel Grooms
- Large Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA.
| | - Evangelyn Alocilja
- Biosystems and Agricultural Engineering, Michigan State University, East Lansing, MI 48824, USA.
| | - Steven Bolin
- Diagnostic Center for Population and Animal Health, Michigan State University, East Lansing, MI 48824, USA.
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Liverani E, Scaioli E, Cardamone C, Monte PD, Belluzzi A. Mycobacterium avium subspecies paratuberculosis in the etiology of Crohn’s disease, cause or epiphenomenon? World J Gastroenterol 2014; 20:13060-13070. [PMID: 25278700 PMCID: PMC4177485 DOI: 10.3748/wjg.v20.i36.13060] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/20/2014] [Revised: 04/30/2014] [Accepted: 05/26/2014] [Indexed: 02/06/2023] Open
Abstract
The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne’s disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis (MAP) and shows some analogies with Crohn’s disease (CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the “MAP-hypothesis”, analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven.
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