Abstract
In the current study, fractions of culture filtrate proteins isolated at different time periods from M. tuberculosis sputum culture were evaluated for T cell activity (ADA, IFN-γ, TNF-α, & IL-12) using in vitro PBMC model. These isolated fractions were later on partially characterized by antibody detection assay using panel of six M. tuberculosis H37RV antigens (Ag85B, ESAT-6, CFP-10, GroES, 45 KD, and Hsp 16). Our results suggested us that PBMCs induced with culture filtrate proteins particularly those secreted towards later phase of growth curve of M. tuberculosis culture have good potential T cell activity as compared to BCG vaccine. On partial characterization we found levels of all secretary antigens increased towards later phase fractions (fraction C). Moreover on further evaluating individual purified M. tuberculosis H37RV antigens for T cell activity, we found that PBMCs induced with these antigens induces good T-cell response but not as consistent as fraction C. Thus to conclude, Culture Filtrate Proteins of M. tuberculosis sputum culture are important T-cell targets, and thus potential of such culture filtrate proteins may be further explored in near future for development of effective vaccination strategies for improving efficacy of currently available TB vaccine.
Highlights
Tuberculosis (TB) caused by the intracellular bacterium Mycobacterium tuberculosis (MTB) remains a major worldwide health problem responsible for approximately three million deaths annually [1]
Our current study focused on testing vaccine potential of CFPs isolated from different phases of MTB sputum culture using in vitro Peripheral blood mononuclear cells (PBMC) model
Cell culture supernatants induced with different culture filtrate fractions showed good Adenosine Deaminase (ADA) activity, which was statistically significant as compared Bacillus Calmette Guerin (BCG) vaccine
Summary
Tuberculosis (TB) caused by the intracellular bacterium Mycobacterium tuberculosis (MTB) remains a major worldwide health problem responsible for approximately three million deaths annually [1]. The demonstration that non-living vaccines based on secreted proteins could effectively protect against subsequent MTB infection in animal models, has led to the initiation of extensive antigen discovery programs which aimed to identify crucial antigenic molecules in culture filtrates [2]. Recent data demonstrates that some antigen are expressed in culture filtrates of MTB but absent in Mycobacterium bovis BCG and most environmental mycobacterial species investigated [7]. These findings have increased our interest in these molecules both as potential vaccine candidate and a novel specific diagnostic reagent
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