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Isolation of perchloric acid soluble, heat stable, ethanol extractable protein from Mycobacterium tuberculosis

 Dong Soo Kim  ;  Bu Hyun Han  ;  Ki Young Lee 
 YONSEI MEDICAL JOURNAL, Vol.37(5) : 339-349, 1996-10 
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Animals ; Antigens, Bacterial / immunology ; Antigens, Bacterial / isolation & purification* ; Cytokines / biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Humans ; Hypersensitivity, Delayed ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis / immunology* ; Perchlorates ; Skin / pathology ; T-Lymphocytes / immunology
The perchloric acid soluble, heat stable, and ethanol insoluble antigen of M. tuberculosis (TB-PBE) was prepared, and antigenicity of this antigen was studied in vivo and in vitro. TB-PBE showed a single band of 60 kDa by SDS-PAGE. Sera from the patients with active pulmonary tuberculosis did not react with this antigen by ELISA. A delayed hypersensitivity skin reaction was induced with this antigen and was correlated with the reaction with PPD. Skin biopsy was performed in this skin lesion induced by TB-PBE and stained by H-E and immunohistochemical methods. TB-PBE induced an inflammatory lesion similar to a lesion induced by PPD. Blastogenic activity of the peripheral blood mononuclear cells stimulated by TB-PBE increased, and showed a peak reaction at 7 days after stimulation. The blastogenic activity changed in a dose-dependent manner. After stimulation with TB-PBE, mononuclear cells were analyzed by FACS. DR+ T cells and CD4/CD8 ratio increased after stimulation by TB-PBE. These cells secreted IL-2, not IL-4 after stimulation with TB-PBE. In the immunofluorescence test, mouse antiserum against TB-PBE showed a positive reaction with M. tuberculosis and showed cross-reactivity with M. bovive and other atypical mycobacteria, but not with S. aureus. With these results, it is evident that TB-PBE is an antigen which can induce cell mediated immunity in vivo and in vitro.
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1. College of Medicine (의과대학) > Dept. of Pediatrics (소아과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Dong Soo(김동수)
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