Isolation of perchloric acid soluble, heat stable, ethanol extractable protein from Mycobacterium tuberculosis.
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. bovis 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.
Kim
D S
DS
Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea.
Han
B H
BH
Lee
K Y
KY
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology / Q000302:isolation & purification*; D016207:Cytokines / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006968:Hypersensitivity, Delayed; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D010472:Perchlorates; D012867:Skin / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology
D000942:Antigens, Bacterial; D016207:Cytokines; D010472:Perchlorates