Immunologic characterization of a 35-kilodalton recombinant antigen of Mycobacterium tuberculosis.

Journal of Clinical Microbiology
H S RumschlagR C Good

Abstract

A 35-kilodalton (kDa) recombinant antigen (35-kDa antigen) produced by Escherichia coli JM107 carrying DNA from Mycobacterium tuberculosis was purified and immunologically examined by in vivo and in vitro methods. A monoclonal antibody (2B2) was produced against the 35-kDa antigen. The protein was purified from the insoluble fraction of the recombinant E. coli strain by either affinity chromatography with the 2B2 monoclonal antibody or preparative isoelectric focusing. In enzyme-linked immunosorbent assay and Western blot (immunoblot) analyses, antibody to 2B2 reacted with whole-cell sonic extracts of M. tuberculosis and other slowly growing mycobacteria but not with two rapid growers, M. chelonae and M. fortuitum. An injection series totaling less than 1 mg of purified protein without adjuvant elicited a humoral response in guinea pigs. In one guinea pig, 10 micrograms of purified protein injected intradermally elicited both a humoral and a cell-mediated response. Results of these studies suggest that the 35-kDa antigen is a membrane-associated protein that stimulates both humoral and cell-mediated immune responses and should be evaluated as a vaccine candidate.

References

Jan 15, 1989·Annals of Internal Medicine·R C Good
Dec 1, 1985·Infection and Immunity·J E TholeJ D van Embden
Dec 27, 1985·Journal of Immunological Methods·J G KennaR S Williams
Jan 1, 1983·Infection and Immunity·M AchtmanR P Silver

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Citations

Jun 30, 2000·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·M L Gennaro
May 16, 2018·Pathogens and Disease·Julia PuffalYasu S Morita
Jun 1, 1994·Comparative Biochemistry and Physiology. Biochemistry and Molecular Biology·L F MontañoE Zenteno

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