Leprosy is a disease caused by Mycobacterium leprae (M. leprae), an intracellular pathogen that has tropism and affects skin and nervous system cells. The disease has two forms of presentation: Paucibacillary and multibacillary, with different clinical and immunological manifestations. Unlike what occurs in the multibacillary form , the diagnostic tests for the paucibacillary form are nonspecific and not very sensitive, allowing the existence of infected individuals without treatment, which contributes to the spread of the pathogen in the population. To mitigate this contamination, more sensitive diagnostic tests capable of detecting paucibacillary patients are needed.

To predict the three-dimensional structure models of M. leprae antigens with serodiagnostic potential for leprosy.

In this in silico study, satisfactory templates were selected in the Protein Data Bank (PDB) using Basic Local Alignment Search Tool to predict the structural templates of ML2038, ML0286, ML0050, and 85B antigens by comparative modeling. The templates were selected according to general criteria such as sequence identity, coverage, X-ray resolution, Global Model Quality Estimate value and phylogenetic relationship; Clustal X 2.1 software was used in this analysis. Molecular modeling was completed using the software Modeller 9v13. Visualization of the models was made using ViewerLite 4.2 and PyMol software, and analysis of the quality of the predicted models was performed using the QMEAN score and Z-score. Finally, the three-dimensional moels were validated using the MolProbity and Verify 3D platforms.

The three-dimensional structure models of ML2038, ML0286, ML0050, and 85B antigens of M. leprae were predicted using the templates PDB: 3UOI (90.51% identity), PDB: 3EKL (87.46% identity), PDB: 3FAV (40.00% identity), and PDB: 1F0N (85.21% identity), respectively. The QMEAN and Z-score values indicated the good quality of the structure models. These data refer to the monomeric units of antigens, since some of these antigens have quaternary structure. The validation of the models was performed with the final three-dimensional structure - monomer (ML0050 and 85B antigens) and quaternary structures (ML2038 and ML0286). The majority of amino acid residues were observed in favorable and allowed regions in the Ramachandran plot, indicating correct positioning of the side chain and absence of steric impediment. The MolProbity score value and Verify 3D results of all models indicated a satisfactory prediction.

The polarized immune response against M. leprae creates a problem in leprosy detection. The selection of immunodominant epitopes is essential for the development of more sensitive serodiagnostic tests, for this it is important to know the three-dimensional structure of the antigens, which can be predicted with bioinformatics tools.

• Antigens
• Leprosy diagnosis
• Mycobacterium leprae
• Molecular modelling
• Serological test
• In silico study

Leprosy is caused by Mycobacterium leprae (M. leprae) and M. lepromatosis, an obligate intracellular organism, and over 200,000 new cases occur every year. M. leprae parasitizes histiocytes (skin macrophages) and Schwann cells in the peripheral nerves. Although leprosy can be treated by multidrug therapy, some patients relapse or have a prolonged clinical course and/or experience leprosy reaction. These varying outcomes depend on host factors such as immune responses against bacterial components that determine a range of symptoms. To understand these host responses, knowledge of the mechanisms by which M. leprae parasitizes host cells is important. This article describes the characteristics of leprosy through bacteriology, genetics, epidemiology, immunology, animal models, routes of infection, and clinical findings. It also discusses recent diagnostic methods, treatment, and measures according to the World Health Organization (WHO), including prevention. Recently, the antibacterial activities of anti-hyperlipidaemia agents against other pathogens, such as M. tuberculosis and Staphylococcus aureus have been investigated. Our laboratory has been focused on the metabolism of lipids which constitute the cell wall of M. leprae. Our findings may be useful for the development of future treatments.

• Leprosy
• Mycobacterium leprae
• Schwann cell
• lipids metabolism
• macrophage
• pseudogene

Mycobacterium leprae, the causative agent of leprosy, is an obligate intracellular pathogen primarily residing within host macrophages and Schwann cells. Whole genome sequencing predicts a highly degraded genome with approximately one third of the coding capacity resulting in the loss of many catabolic pathways. Therefore, it can be assumed that M. leprae obtains many of the necessary metabolites for intracellular survival and growth from the host cells. In this study, global transcriptomic analyses were done on freshly harvested M. leprae growing in athymic mouse footpads for five months (MFP5) and compared to those held in axenic medium for 48 (ML48) and 96 (ML96) hours. Results show that all of the genes and pseudogenes were transcribed under both in vivo and in vitro conditions. 24% and 33% of gene transcript levels were significantly altered in ML48 and ML96 respectively, compared to MFP5. Approximately 45% (39/86) of lipid metabolism genes were significantly downregulated in ML96 compared to MFP5, majority of which are in the β-oxidation pathway. Cholesterol oxidase, acyl-CoA dehydrogenase, and coenzyme F420-dependent oxidoreductase, were significantly upregulated in both ML48 and ML96 compared to MFP5. 30% of cell wall and cell processes functional category genes had altered gene transcription at 96hr compared to MFP5. 40% of 57 genes associated with mycobacterial virulence showed significantly altered transcript levels with 52% significantly downregulated in ML96, including most of the Pro-Glu/Pro-Pro-Glu genes. All 111 hypothetical protein genes with unknown function were expressed. Adenosine triphosphate (ATP) synthesis in M. leprae appears to be significantly downregulated under ex vivo conditions. This is the first study comparing M. leprae global gene expression during in vivo growth and ex vivo stationery phase in axenic medium confirming that during the growth phase in the footpads of experimentally infected mice, M. leprae is metabolically active and its primary source of energy production is probably lipids.

• Mycobacterium leprae
• RNA-Seq
• metabolic pathways
• mouse footpad
• transcriptome

• Humans
• 1-Acylglycerol-3-Phosphate O-Acyltransferase/metabolism
• 1-Acylglycerol-3-Phosphate O-Acyltransferase/genetics
• Leprosy/microbiology
• Leprosy/metabolism
• Lipid Metabolism
• Macrophages/metabolism
• Macrophages/microbiology
• Mycobacterium leprae
• THP-1 Cells
• Triglycerides/metabolism
• Triglycerides/biosynthesis

• Ministry of Education, Culture, Sports, Science and Technology
• Japan Society for the Promotion of Science
• Japan Agency for Medical Research and Development
• Sasakawa Scientific Research

In leprosy, classic diagnostic tools based on bacillary counts and histopathology have been facing hurdles, especially in distinguishing latent infection from active disease and diagnosing paucibacillary clinical forms. Serological tests and IFN-gamma releasing assays (IGRA) that employ humoral and cellular immune parameters, respectively, are also being used, but recent results indicate that quantitative PCR (qPCR) is a key technique due to its higher sensitivity and specificity. In fact, advances concerning the structure and function of the Mycobacterium leprae genome led to the development of specific PCR-based gene amplification assays for leprosy diagnosis and monitoring of household contacts. Also, based on the validation of point-of-care technologies for M. tuberculosis DNA detection, it is clear that the same advantages of rapid DNA detection could be observed in respect to leprosy. So far, PCR has proven useful in the determination of transmission routes, M. leprae viability, and drug resistance in leprosy. However, PCR has been ascertained to be especially valuable in diagnosing difficult cases like pure neural leprosy (PNL), paucibacillary (PB), and patients with atypical clinical presentation and histopathological features compatible with leprosy. Also, the detection of M. leprae DNA in different samples of the household contacts of leprosy patients is very promising. Although a positive PCR result is not sufficient to establish a causal relationship with disease outcome, quantitation provided by qPCR is clearly capable of indicating increased risk of developing the disease and could alert clinicians to follow these contacts more closely or even define rules for chemoprophylaxis.

Mycobacterium leprae (M. leprae) lives and replicates within macrophages in a foamy, lipid-laden phagosome. The lipids provide essential nutrition for the mycobacteria, and M. leprae infection modulates expression of important host proteins related to lipid metabolism. Thus, M. leprae infection increases the expression of adipophilin/adipose differentiation-related protein (ADRP) and decreases hormone-sensitive lipase (HSL), facilitating the accumulation and maintenance of lipid-rich environments suitable for the intracellular survival of M. leprae. HSL levels are not detectable in skin smear specimens taken from leprosy patients, but re-appear shortly after multidrug therapy (MDT). This study examined the effect of MDT components on host lipid metabolism in vitro, and the outcome of rifampicin, dapsone and clofazimine treatment on ADRP and HSL expression in THP-1 cells. Clofazimine attenuated the mRNA and protein levels of ADRP in M. leprae-infected cells, while those of HSL were increased. Rifampicin and dapsone did not show any significant effects on ADRP and HSL expression levels. A transient increase of interferon (IFN)-β and IFN-γ mRNA was also observed in cells infected with M. leprae and treated with clofazimine. Lipid droplets accumulated by M. leprae-infection were significantly decreased 48 h after clofazimine treatment. Such effects were not evident in cells without M. leprae infection. In clinical samples, ADRP expression was decreased and HSL expression was increased after treatment. These results suggest that clofazimine modulates lipid metabolism in M. leprae-infected macrophages by modulating the expression of ADRP and HSL. It also induces IFN production in M. leprae-infected cells. The resultant decrease in lipid accumulation, increase in lipolysis, and activation of innate immunity may be some of the key actions of clofazimine.

Leprosy, caused by Mycobacterium leprae (M. leprae), is an ancient infectious disease that remains the leading infectious cause of disability. After infection, M. leprae lives inside host macrophages that contain a large amount of lipids, which is thought to be an essential microenvironment for M. leprae to survive in host cells. M. leprae infection increases lipid accumulation in macrophages and decreases the metabolic breakdown of lipids (catabolism). In addition, the treatment of leprosy with multidrug therapy (MDT) reverses the effect of infection on the modulation of lipid metabolism. We therefore aimed to use cultured human macrophage cells to determine which of the three MDT drugs (clofazimine, dapsone, or rifampicin) is responsible for this effect. We found that only clofazimine affects lipid accumulation and catabolism in M. leprae-infected cells in vitro. The amounts of lipids accumulated in the cells decreased when clofazimine was added to the cell culture medium. Clofazimine also activated immune responses in M. leprae-infected cells. These results suggest that the effectiveness of clofazimine against leprosy is due to the modulation of lipid metabolism and activation of immune reactions in M. leprae-infected host cells.

• Animals
• Blotting, Western
• Clofazimine/pharmacology
• Dapsone/pharmacology
• Gene Expression Profiling
• Humans
• Interferons/biosynthesis
• Leprostatic Agents/pharmacology
• Lipid Metabolism/drug effects
• Macrophages/drug effects
• Macrophages/microbiology
• Mycobacterium leprae/drug effects
• Rats
• Rats, Nude
• Real-Time Polymerase Chain Reaction
• Rifampin/pharmacology

Leprosy is a chronic infectious disease and is a major source of morbidity in developing countries. Leprosy is caused by the obligate intracellular bacterium Mycobacterium leprae, which infects as primary target Schwann cells. Lepromatous leprosy exhibits multiple lesions of the skin, eyes, nerves, and lymph nodes. The sites of infection are characterized by the presence of foamy macrophages, fully packed with lipid droplets (LDs), which are induced by M. leprae. In the last years, it has become evident that M. tuberculosis imports lipids from foamy macrophages and is dependent on fatty acids for growth in infected macrophages. M. leprae seems to have similar mechanisms for scavenging lipids from the host. But due to the inability to culture M. leprae on laboratory media, research progresses only slowly. However, in the last years, substantial progress has been made in the field of lipid metabolism in M. leprae. Herein, we will present and summarize the lipid droplets formation and the metabolism of lipids during M. leprae infection.