Using a monoclonal antibody specific for IgM and IgA and immunohi

Using a monoclonal antibody specific for IgM and IgA and immunohistological techniques, we found that IgM and IgA were more abundant in lesions from patients with lepromatous leprosy, those with

the disseminated form of the disease – accounting for 8% of the cells BMS-354825 mouse in the infiltrate compared with < 2% of the cells in lesions from patients with T-lep (Fig. 5). These results correlate the expression of IgM and IgA in leprosy with the clinical form of the disease – being greatest in those patients in whom the disease is disseminated – and, by inference, also correlate with the T helper type 2 immunity to the pathogen. We reasoned that immunoglobulins should be expressed on mature B cells or plasma cells so we examined the expression of CD138, a specific marker for plasma cells in leprosy tissue, using immunoperoxidase. Plasma cells were more abundant in L-lep patients, accounting for approximately 15% of the cells in the infiltrate. In contrast, CD138-expressing cells were rare or absent in T-lep lesions (Fig. 6a). To identify the phenotype of the cells containing IgM at the site of disease in leprosy, we performed two-colour immunofluorescence labelling using a monoclonal antibody that detected mature B cells followed by confocal laser

scanning microscopy. Double immunofluorescence labelling showed that cells containing IgM in L-lep lesions were plasma cells (Fig. 6b). We hypothesized that increased IL-5 in addition to the mycobacteria at the site of disease may play a role in increasing the production of antibodies by B cells. B cells were purified INK 128 manufacturer from healthy donors and stimulated with M. leprae sonicate in the presence or absence of IL-5. Neither individually nor in combination did these stimuli enhance the production of total IgM, IgG or IgA. However, when added to PBMC cultures, M. leprae-stimulated cells produced almost 20 times more IgM in the presence of IL-5 whereas Rebamipide there was no significant difference in IgA or IgG

production (Fig. 7). While PBMC with added IL-5 showed a trend toward increased IgA and IgG, these increases were not statistically significant. These studies suggest a role for IL-5 and T cells in the ability of M. leprae to stimulate IgM production from B cells. To investigate the pathways and functional gene sets that are differentially expressed across the spectrum of leprosy, we performed pathways analysis of gene expression profiles comparing leprosy lesions from patients with progressive (L-lep) versus self-limited (T-lep) infections.10 Using analysis of canonical pathways and functional groups, we found that B-cell pathways recurred in the top gene sets (P < 0·005) in the progressive L-lep form of the disease where antibody levels, including those of IgM, are high. Pathways analysis of IgM production at the site of disease suggested a role for IL-5.

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