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KMID : 0357319920270030253
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Journal of the Korean Society for Microbiology
1992 Volume.27 No. 3 p.253 ~ p.268
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Lymphocyte Proliferation and Antibody Response against 30-kDa Protein Antigen of Mycobacterium tuberculosis
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백태현
이봉규/김화중/조은경/최대경
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Abstract
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The 30-kDa Protein antigen(also known as antigen 85B, or alpha antigen) is a major and biologically important constituent of M. bovis BCG and M. tuberculosis culture fluds.
We purified the 30-kDa antiten from the unheated culture filtrated of M. tuberculosis H37Rv by 50% ammonium sulfate precipitation, hydroxylapatite chromatography and Sephadex G-75 gel filtration and examined the immunological activities of the
30-kDa
antigen in mice by splenocyte proliferation, ELISA and Western blot after different immunization.
The spllen index was increased in all immunized mice, higher value was observed in the 30-kDa antigen immunized moce primed with live BCG(/30K group) but the splenocyte proliferaion and antibody respones to the 30-kDa antigen were lower than
other
immunized groups without BCG-priming. Mice immunized with the 30-kDa antigen in incomplete freund's adjuvant(IFA/30K group) showed relatively low spleen index but the highest increases in splenocyte proliferation and IgG antibody response to the
30-kDa
antigen.
Sequential IgG levels of immunized mice with the 30-kDa antigen in complete Freund's adjuvant (CFA/30K group) or IFA/30K group were showed marked increases after 4 weeks and reaches the highest level and showed plateau pattern after 6 weeks but
IgM
levels showd a little decrease at 4 weeks and rose again after 6 weeks. The final IgG antibody level was rather higher in IFA/30K group than CFA/30K group nd approximately 100 fold groater than the final IgM level in both groups. BCG/30K group
showed
and initially slow and depressed IgG antibody response but followed a rapid rise 8 weeks after BCG-priming. This result suggests that live BCG suppress the early phase of immune response to the 30-kDa antigen.
On the Western blot r4esults, all sera from immunized mice except BCG/30K group intensely reacted with the molecular weight of 30-kDa as early as 2 weeks after immunization and the reactivities become gradually larger and stronger. Many other
minor
bands were also appeared during the course of boost immunization but the BCG/30K mice showed very slow reactivities after 8 weeks.
From the above results, it is suggested that the 30-kDa antigen of M. tubericulosis is highly immunogenic but its immunological activity in mice may be suppressed by live M. bovis BCG and has many cross-reactive epitopes on the molecule.
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