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Purification and immunoreactivity of three components from the 30/32-kilodalton antigen 85 complex in Mycobacterium
1Department of Microbiology, College of Medicine, Chungnam National University, Taejeon 301-131, Korea.
Abstract:
The three proteins of the antigen 85 complex (85A, 85B, and 85C), which are major secretory products of Mycobacterium tuberculosis, were purified to homogeneity in large amounts by a combination of chromatography on hydroxylapatite, DEAE-Sepharose, and DEAE-Sephacel and gel filtration from M. tuberculosis culture filtrate. Then we examined the immunological reactivity of the three proteins in tuberculosis patients and healthy controls. Antibody responses to the 85B and 85A proteins in patients were significantly greater than responses to the 85C protein. In contrast, all three antigens induced significant lymphoproliferation and gamma interferon production in peripheral blood mononuclear cells from healthy tuberculin reactors.
Insights
Researchers purified three key Mycobacterium tuberculosis proteins (antigen 85A, 85B, 85C). Antibody responses were higher to 85A and 85B in tuberculosis patients, while all antigens triggered immune responses in healthy individuals.
Area of Science:
- Immunology
- Microbiology
- Biochemistry
Background:
- The antigen 85 complex (85A, 85B, 85C) are major secretory proteins of Mycobacterium tuberculosis.
- These proteins play a role in the pathogenesis of tuberculosis.
Purpose of the Study:
- To purify the three antigen 85 proteins (85A, 85B, 85C) from Mycobacterium tuberculosis.
- To investigate the immunological reactivity of these purified proteins in tuberculosis patients and healthy controls.
Main Methods:
- Purification of antigen 85A, 85B, and 85C using hydroxylapatite, DEAE-Sepharose, DEAE-Sephacel chromatography, and gel filtration.
- Assessment of antibody responses in tuberculosis patients and healthy controls.
- Evaluation of lymphoproliferation and gamma interferon production in peripheral blood mononuclear cells from healthy tuberculin reactors.
Main Results:
- Antibody responses to antigen 85B and 85A were significantly higher in tuberculosis patients compared to antigen 85C.
- All three antigens (85A, 85B, 85C) induced significant lymphoproliferation.
- All three antigens stimulated gamma interferon production in peripheral blood mononuclear cells from healthy tuberculin reactors.
Conclusions:
- Antigen 85B and 85A are more immunogenic in tuberculosis patients than antigen 85C.
- The antigen 85 complex proteins elicit a cellular immune response in healthy individuals with a history of tuberculosis exposure.