ISOLATION AND PURIFICATION OF AG 85 COMPLEX FROM MYCOBADVRIUM BOVIS (BCG) AND ASSESSMENT OF ITS CELL PROLIFERATION RESPONSE IN WHOLE BLOOD

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NIKOUKAR I.; KAJBAF M.J.; MAK VANDI M.; FARAJZADEH A.; KAMALI E.; MOSTAFAEI ALI; HUYGEN K.

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Journal: Cell Journal (Yakhteh) Year:2004 | Volume:6 | Issue:23 Page(s): 144-151

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Title

ISOLATION AND PURIFICATION OF AG 85 COMPLEX FROM MYCOBADVRIUM BOVIS (BCG) AND ASSESSMENT OF ITS CELL PROLIFERATION RESPONSE IN WHOLE BLOOD

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Keywords

MYCOBACTERIUM BOVIS (BCG)Q3

AG 85 COMPLEXQ3

CELL PROLIFERATIONQ3

WHOLE BLOOD ASSAYQ3

Abstract

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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APA: Copy

NIKOUKAR, I., KAJBAF, M.J., MAK VANDI, M., FARAJZADEH, A., KAMALI, E., MOSTAFAEI, ALI, & HUYGEN, K.. (2004). ISOLATION AND PURIFICATION OF AG 85 COMPLEX FROM MYCOBADVRIUM BOVIS (BCG) AND ASSESSMENT OF ITS CELL PROLIFERATION RESPONSE IN WHOLE BLOOD. CELL JOURNAL (YAKHTEH), 6(23), 144-151. SID. https://sid.ir/paper/16429/en

Vancouver: Copy

NIKOUKAR I., KAJBAF M.J., MAK VANDI M., FARAJZADEH A., KAMALI E., MOSTAFAEI ALI, HUYGEN K.. ISOLATION AND PURIFICATION OF AG 85 COMPLEX FROM MYCOBADVRIUM BOVIS (BCG) AND ASSESSMENT OF ITS CELL PROLIFERATION RESPONSE IN WHOLE BLOOD. CELL JOURNAL (YAKHTEH)[Internet]. 2004;6(23):144-151. Available from: https://sid.ir/paper/16429/en

IEEE: Copy

I. NIKOUKAR, M.J. KAJBAF, M. MAK VANDI, A. FARAJZADEH, E. KAMALI, ALI MOSTAFAEI, and K. HUYGEN, “ISOLATION AND PURIFICATION OF AG 85 COMPLEX FROM MYCOBADVRIUM BOVIS (BCG) AND ASSESSMENT OF ITS CELL PROLIFERATION RESPONSE IN WHOLE BLOOD,” CELL JOURNAL (YAKHTEH), vol. 6, no. 23, pp. 144–151, 2004, [Online]. Available: https://sid.ir/paper/16429/en

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