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Cell Journal، جلد ۶، شماره ۳، صفحات ۱۴۴-۱۵۱
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عنوان انگلیسی Isolation and Purification of Ag 85 Complex from Mycobacterium bovis (BCG) and assessment of its cell proliferation response in whole blood
چکیده انگلیسی مقاله Introduction: Tuberculosis (TB) caused by Mycobacterium tuberculosis remains a major worldwide health problem. The only TB vaccine currently available is anattenuated strain of mycobacterium bovis termed Bacillus Calmette -Guerin (BCG).The efficacy of BCG remains controversial. Mycobacterium secretory proteins are generally considered important antigens for protection against TB. A major protein component of mycobacterial culture filtrate is Antigen 85 complex which is apromising potential vaccine candidate. Material and Methods: Antigen 85 complex was purified from Mycobacterium bovis (BCG) culture filtrate by sequential chromatography on phenyl Sepharose 4B, DEAE-Sephacel and Superdex G75. Purification was confirmed by SDS-PAGE and immunoblotting with a specific monoclonal antibody. The in vitro ability of Ag 85 complex to stimulate cell proliferation was compared with that of Purified Protein Derivative (PPD) and the polyclonal T cell mitogen PHA in a whole blood assay in which the target cells were derived from 25 healthy PPD-positive and 25 healthy PPD-negative subjects. Results: Antigen 85 complex was found to have a molecular weight of 30-32 KDa by SDS-polyacrylamide gel electrophoresis and reacted strongly by immunoblotting with the monoclonal antibody specified against Ag 85. The responses to Ag 85 and PPD were significantly higher in cells of PPD- positive than in cells of PPD-negative donors. Eighty eight percent (22/25) of the PPD- positive cells responded to Ag 85 whereas only 16% (4/25) of the PPD-negative cells responded. Conclusion: The cell proliferation response to Ag 85 complex is significantly different between cells of skin-test positive and skin- test negative subjects and may suggest an immuno-protective role for Ag 85 complex against M. tuberculosis infection.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/3508
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