Summary Mycobacteria including Mycobacterium tuberculosis and Mycobacterium leprae possess multiple antigens some of which inhibit other anti-mycobacterial immune responses. Whole cell vaccines are not free from these suppressive molecules and may adversely affect the immunogenic response(s). Purified protein components having only immunogenic properties should prove to be superior vaccine(s). Mycobacterium habana, a candidate vaccine for mycobacterial infections has been dissected for analysing its antigenic myriad. A 65 kDa protein of this mycobacterium has been isolated and characterized for its protective and celJ mediated immune responses. The protein was isolated in pure form using an isotachophoresis (SDS-PAGE filtration) technique and identified with low molecular weight markers along with mAb using the immunoblot technique. Mab IIH9 has identified a 65 kDa protein in M. habana. This protein has been found to be immunoprotective in mice against M. tuberculosis H37RV infection. It generates high levels of DTH responses in mice against M. tuberculosis and M. leprae antigens and inhibits migration of sensitized cells under the antigenic influence of homologous and heterologous origin. Possibilities of developing this protein as a subunit vaccine are discussed in this report.
A method to assay anti-HCG activity was developed. Polystyrene micro-ELISA plates were coated with HCG and uterine and tubal washings were incubated in the wells. Sera were diluted to equate the protein content with that in the washings and were included in the assay to serve as internal reference standards. Anti-rabbit-IgG (sheep) conjugated to horse-radish peroxidase was added to each well and enzyme activity was monitored using ortho-phenyl-diamine as chromogen agent. Enzyme activity was a direct measure of anti-HCG activity. This method was used to compare anti-HCG activity in the genital tract fluid of rabbits immunized with HCG with that of serum from the same animal. The results of the present study show that at equal protein concentrations, the anti-HCG activity was only 2.64% to 18.73% of the activity present in serum. Thus, it seems that antibody activity in the genital tract of the female rabbits immunized systemically may not be sufficient enough to neutralize the biological function of the antigen needs for contraceptive protection.
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