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Ravkov EV, Ventura MF, Gudipaty S, Ng D, Delgado JC, Lin L. Converting an HLA-B27 flow assay from the BD FACSCanto to the BD FACSLyric. CYTOMETRY. PART B, CLINICAL CYTOMETRY 2025; 108:67-76. [PMID: 39287115 DOI: 10.1002/cyto.b.22206] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/07/2024] [Revised: 08/02/2024] [Accepted: 08/28/2024] [Indexed: 09/19/2024]
Abstract
HLA-B27 is a major histocompatibility complex (MHC) class I antigen which exhibits strong association (90%) with ankylosing spondylitis. HLA-B27 detection in patients by flow cytometry is a widely used clinical test, performed on many different flow cytometer models. We sought to develop and validate a test conversion protocol for the HLA-B27 test performed on the BD FACSCanto to BD's newer FACSLyric flow cytometers. The development and validation experiments were performed using anti-HLA-B27*FITC/CD3*PE antibody-stained whole blood patient specimens. The anti-HLA-B27*FITC logarithmic median fluorescence (LMF) results on the BD FACSCanto were converted to median fluorescence intensity (MFI) values on the BD FACSLyric. Clustering of the HLA-B27 positive and negative values, using a 3rd order polynomial equation, resulted in a conversion of the BD FACSCanto cutoff values, negative (<150 LMF) and positive (≥160 LMF), to negative (<4530 MFI) and positive (≥6950 MFI) on the BD FACSLyric. Accuracy was assessed by comparing the flow results obtained on the BD FACSCanto and BD FACSLyric to a molecular PCR based assay. Additional validation parameters (compensation verification, intra- and inter-assay precision, and instrument comparison) were performed per the recommendations outlined in the Clinical and Laboratory Standards Institute (CLSI) H62 guidelines for validation of flow cytometry assays.
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Affiliation(s)
- Eugene V Ravkov
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
| | - Miguel F Ventura
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
| | - Swapna Gudipaty
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
| | - David Ng
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
- Department of Pathology, University of Utah School of Medicine, Salt Lake, Utah, USA
| | - Julio C Delgado
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
- Department of Pathology, University of Utah School of Medicine, Salt Lake, Utah, USA
| | - Leo Lin
- APUP Institute for Clinical and Experimental Pathology, Salt Lake, Utah, USA
- Department of Pathology, University of Utah School of Medicine, Salt Lake, Utah, USA
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Giri PS, Bhimani R, Laddha NC, Dwivedi M. HLAB27 may confer protection to COVID-19 in generalized vitiligo patients from South Gujarat population. NUCLEOSIDES, NUCLEOTIDES & NUCLEIC ACIDS 2024; 44:1-15. [PMID: 38240306 DOI: 10.1080/15257770.2024.2303710] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/30/2023] [Revised: 01/03/2024] [Accepted: 01/05/2024] [Indexed: 12/06/2024]
Abstract
Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), while generalized vitiligo(GV) is an autoimmune disease that causes the loss of functional melanocytes, resulting in white patches all over the body. Human Leukocyte Antigen (HLA) plays a crucial role in immune response to pathogens. Studies assessing the link between GV and COVID-19 are lacking; therefore, our current study was aimed to establish the association between GV and HLAB27 by genotyping the HLAB27 allele in 150 GV patients and 150 controls from South Gujarat population through polymerase chain reaction-sequence-specific primers (PCR-SSP) method. Additionally, we assessed the correlation of GV with COVID-19 and the influence of HLAB27 on COVID-19 development. Interestingly, our study suggested that the HLAB27 allele was prevalent in GV patients as compared to controls (52% vs 35.33%; p = 0.0051). Moreover, the occurrence of COVID-19 was significantly lower in GV patients than in controls (10% vs 32.66%; p < 0.0001). Disease activity-based analysis suggested that COVID-19 occurrence was significantly lower in active vitiligo (AV) patients as compared to stable vitiligo (SV) patients(6.87% vs 31.57%; p = 0.0045). Furthermore, COVID-19 development was significantly reduced in HLAB27 positive individuals as compared to HLAB27 negative individuals (p = 0.0025). Overall, our study suggests, for the first time, that HLAB27 allele might be a genetic risk factor for GV susceptibility, and an ongoing immune response in GV patients, more specifically in AV patients, might protect against COVID-19 infection in South Gujarat population. Additionally, our study highlighted the likely role of HLAB27 in protection against COVID-19 development.
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Affiliation(s)
- Prashant S Giri
- C. G. Bhakta Institute of Biotechnology, Uka Tarsadia University, Surat, Gujarat, India
| | - Radhika Bhimani
- C. G. Bhakta Institute of Biotechnology, Uka Tarsadia University, Surat, Gujarat, India
| | | | - Mitesh Dwivedi
- C. G. Bhakta Institute of Biotechnology, Uka Tarsadia University, Surat, Gujarat, India
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Analysis of choroidal and central foveal thicknesses in acute anterior uveitis by enhanced-depth imaging optical coherence tomography. BMC Ophthalmol 2017; 17:225. [PMID: 29191218 PMCID: PMC5709927 DOI: 10.1186/s12886-017-0628-7] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2017] [Accepted: 11/22/2017] [Indexed: 11/25/2022] Open
Abstract
Background Currently, there are a limited number of reports of structural changes in the retina and choroid in acute anterior uveitis (AAU). The aim of this study was to evaluate choroidal and central foveal thicknesses during episodes of AAU. Methods The medical records of 120 patients with AAU and 120 healthy subjects matched for age, sex, and spherical equivalent of refractive error were reviewed. Subjects were divided into group 1 (AAU-affected eyes), 2 (unaffected fellow eyes), and 3 (healthy control eyes). Results In the uveitis group, etiologic diagnoses included human leukocyte antigen (HLA)-B27-associated (n = 71) and idiopathic (n = 49) AAU. The mean subfoveal choroidal thicknesses (SFCTs) in groups 1–3 were 326.7 ± 64.2, 296.1 ± 66.6, and 294.9 ± 41.7 μm, respectively. The corresponding mean central foveal thicknesses (CFTs) were 273.5 ± 29.3, 264.4 ± 24.6, and 263.0 ± 30.8 μm, respectively. The AAU group exhibited a significantly greater SFCT than the control groups (P < .001). Relative to the control group, while eyes with idiopathic AAU exhibited a significantly greater CFT, those with HLA-B27-associated AAU exhibited no such significant difference. Anterior chamber cell grade was not associated with SFCT or CFT. Conclusions The SFCT increased significantly during AAU. This indicates the importance of OCT examination for detection of subclinical choroidal and retinal changes in all types of AAU.
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D’Ambrosio EM, La Cava M, Tortorella P, Gharbiya M, Campanella M, Iannetti L. Clinical Features and Complications of the HLA-B27-associated Acute Anterior Uveitis: A Metanalysis. Semin Ophthalmol 2016; 32:689-701. [DOI: 10.3109/08820538.2016.1170158] [Citation(s) in RCA: 36] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Affiliation(s)
| | - Maurizio La Cava
- Department of Ophthalmology, “Sapienza” University of Rome, Rome, Italy
| | - Paolo Tortorella
- Department of Ophthalmology, “Sapienza” University of Rome, Rome, Italy
| | - Magda Gharbiya
- Department of Ophthalmology, “Sapienza” University of Rome, Rome, Italy
| | - Michelangelo Campanella
- RVC Department of Comparative Biomedical Sciences and UCL Consortium for Mitochondrial Research, University of London, London, UK
| | - Ludovico Iannetti
- Department of Ophthalmology, “Sapienza” University of Rome, Rome, Italy
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Optic neuritis with positive HLA-B27: Characteristic phenotype in the Chinese population. J Neurol Sci 2016; 362:100-5. [DOI: 10.1016/j.jns.2016.01.027] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2015] [Revised: 01/11/2016] [Accepted: 01/15/2016] [Indexed: 01/18/2023]
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Louwen R, Hays JP. Is there an unrecognised role for Campylobacter infections in (chronic) inflammatory diseases? World J Clin Infect Dis 2013; 3:58-69. [DOI: 10.5495/wjcid.v3.i4.58] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/07/2013] [Revised: 10/30/2013] [Accepted: 11/16/2013] [Indexed: 02/06/2023] Open
Abstract
Campylobacter species are one of the major causes of global bacterial-related diarrheal disease worldwide. The disease is most frequently associated with the ingestion of contaminated meat, raw milk, pets, contaminated water, and the organism may be frequently cultured from the faeces of chicken and other domesticated farm animals. Of the 17 established Campylobacter species, the most important pathogens for humans are Campylobacter jejuni (C. jejuni), Campylobacter coli (C. coli) and Campylobacter fetus (C. fetus), which are all associated with diarrheal disease. Further, C. jejuni and C. coli are also associated with the neuroparalytic diseases Guillain-Barré syndrome and Miller Fischer syndrome, respectively, whereas C. fetus is linked with psoriatic arthritis. The discovery of both “molecular mimicry” and translocation-related virulence in the pathogenesis of C. jejuni-induced disease, indicates that Campylobacter-related gastrointestinal infections may not only generate localized, acute intestinal infection in the human host, but may also be involved in the establishment of chronic inflammatory diseases. Indeed, pathogenicity studies on several Campylobacter species now suggest that molecular mimicry and translocation-related virulence is not only related to C. jejuni, but may play a role in human disease caused by other Campylobacter spp. In this review, the authors provide a review based on the current literature describing the potential links between Campylobacter spp. and (chronic) inflammatory diseases, and provide their opinions on the likely role of Campylobacter in such diseases.
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The role of Klebsiella in Crohn's disease with a potential for the use of antimicrobial measures. Int J Rheumatol 2013; 2013:610393. [PMID: 24223596 PMCID: PMC3810322 DOI: 10.1155/2013/610393] [Citation(s) in RCA: 35] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2013] [Accepted: 09/04/2013] [Indexed: 12/23/2022] Open
Abstract
There is a general consensus that Crohn's disease (CD) develops as the result of immune-mediated tissue damage triggered by infections with intestinal microbial agents. Based on the results of existing microbiological, molecular, and immunological studies, Klebsiella microbe seems to have a key role in the initiation and perpetuation of the pathological damage involving the gut and joint tissues in patients with CD. Six different gastroenterology centres in the UK have reported elevated levels of antibodies to Klebsiella in CD patients. There is a relationship between high intake of starch-containing diet, enhanced growth of gut microbes, and the production of pullulanases by Klebsiella. It is proposed that eradication of these microbes by the use of antibiotics and low starch diet, in addition to the currently used treatment, could help in alleviating or halting the disease process in CD.
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Sinigaglia M, Antunes DA, Rigo MM, Chies JAB, Vieira GF. CrossTope: a curate repository of 3D structures of immunogenic peptide: MHC complexes. DATABASE-THE JOURNAL OF BIOLOGICAL DATABASES AND CURATION 2013; 2013:bat002. [PMID: 23396301 PMCID: PMC3567486 DOI: 10.1093/database/bat002] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 11/14/2022]
Abstract
The CrossTope is a highly curate repository of three-dimensional structures of peptide:major histocompatibility complex (MHC) class I complexes (pMHC-I). The complexes hosted by this databank were obtained in protein databases and by large-scale in silico construction of pMHC-I structures, using a new approach developed by our group. At this moment, the database contains 182 ‘non-redundant’ pMHC-I complexes from two human and two murine alleles. A web server provides interface for database query. The user can download (i) structure coordinate files and (ii) topological and charges distribution maps images from the T-cell receptor-interacting surface of pMHC-I complexes. The retrieved structures and maps can be used to cluster similar epitopes in cross-reactivity approaches, to analyse viral escape mutations in a structural level or even to improve the immunogenicity of tumour antigens. Database URL: http://www.crosstope.com.br
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Affiliation(s)
- Marialva Sinigaglia
- NBLI - Núcleo de Bioinformática do Laboratório de Imunogenética, Department of Genetics, Universidade Federal do Rio Grande do Sul, 9500 Bento Gonçalves Avenue, Bldg 43323, Rm 225, 91501-970 Porto Alegre, RS, Brazil
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Boldt ABW, Goeldner I, de Messias-Reason IJT. Relevance of the lectin pathway of complement in rheumatic diseases. Adv Clin Chem 2012; 56:105-53. [PMID: 22397030 DOI: 10.1016/b978-0-12-394317-0.00012-1] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Due to its importance both in the clearance of pathogens that contribute as rheumatic etiological agents and in the disposal of apoptotic bodies and potential autoimmune initiators, deficiencies of the components of the lectin pathway of complement have been found to increase susceptibility and modulate the severity of most rheumatic disorders. This chapter introduces the general aspects of the structure, function, and genetics of lectin pathway components and summarizes current knowledge of the field regarding rheumatic diseases predisposition and modulation.
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Affiliation(s)
- Angelica B W Boldt
- Molecular Immunopathology Laboratory, Department of Medical Pathology, Federal University of Paraná, Curitiba, Brazil
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Abstract
Autoimmune and inflammatory uveitis are a group of potentially blinding intraocular inflammatory diseases that arise without a known infectious trigger and are often associated with immunological responses to unique retinal proteins. In the United States, about 10% of the cases of severe visual handicap are attributed to this group of disorders. As I discuss here, experimental models of ocular autoimmunity targeting retinal proteins have brought about a better understanding of the basic immunological mechanisms involved in the pathogenesis of uveitis and are serving as templates for the development of novel therapies.
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Affiliation(s)
- Rachel R Caspi
- Laboratory of Immunology, National Eye Institute, NIH, Bethesda, Maryland 20892, USA.
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Accorinti M, Iannetti L, Liverani M, Caggiano C, Gilardi M. Clinical Features and Prognosis of HLA B27-associated Acute Anterior Uveitis in an Italian Patient Population. Ocul Immunol Inflamm 2010; 18:91-6. [DOI: 10.3109/09273941003597268] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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Darke C, Coates E. One-tube HLA-B27/B2708 typing by flow cytometry using two "Anti-HLA-B27" monoclonal antibody reagents. CYTOMETRY PART B-CLINICAL CYTOMETRY 2009; 78:21-30. [PMID: 19693889 DOI: 10.1002/cyto.b.20490] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
BACKGROUND Flow cytometry-based methods are widely used to detect the ankylosing spondylitis-associated HLA-B27/B2708 antigens. However, the generally used "HLA-B27" monoclonal antibodies (moabs) cross-react with many HLA specificities, including the common HLA-B7 antigen. Thus, using two "B27" moabs is highly recommended. METHODS The assay used two "HLA-B27" reagents, FITC and PE conjugated, respectively and a PE-Cy5 anti-CD3 antibody. Assay verification used 51 reference subjects possessing B*2705, B*2702, and B*2708 and a range of cross-reactive HLA antigens. A total of 1,006 consecutive patients' samples, referred for "HLA-B27 typing", were assayed alongside our standard flow cytometry method. A further 12 low frequency HLA-B*27 specificities were tested. Samples reacting with one "B27" moab only were B*27 allele typed by PCR using sequence-specific primers. RESULTS All patient B27/B2708 positives (28.3%) were identified by our one-tube method which detected B*2705, B*2702, B*2708, and 8/12 other B*27 specificities. It was unaffected by HLA-B7 and other cross-reactive antigens but required a minor adjustment, a reduction in the volume of one of the "B27" moabs used, to avoid detecting a minority of HLA-B57 subjects. CONCLUSIONS Our one-tube B27/B2708 assay is simple, robust, uses two "B27" moabs for typing precision and security, does not suffer from interference by HLA-B7 or other cross-reactive antigens and has the obvious advantage of using a single tube per typing.
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Affiliation(s)
- Chris Darke
- Welsh Transplantation and Immunogenetics Laboratory, Welsh Blood Service, Wales, United Kingdom.
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Feng M, Yin B, Shen T, Ma Q, Liu L, Zheng J, Zhao Y, Qian K, Liu D. TAP1 and TAP2 polymorphisms associated with ankylosing spondylitis in genetically homogenous Chinese Han population. Hum Immunol 2009; 70:257-61. [DOI: 10.1016/j.humimm.2009.01.028] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2008] [Revised: 01/16/2009] [Accepted: 01/28/2009] [Indexed: 12/31/2022]
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