A major, immunodominant, 47,000 (47K) MW surface immunogen of Treponema pallidum was identified using functionally-defined anti-T. pallidum monoclonal antibodies. It may represent the major immunogen of T. pallidum.
The aim of this proposal is to further examine the biological activities of both the """"""""native"""""""" 47K immunogen as well as antibodies elicited against it, and to assess their overall role(s) in the pathogenesis and immunology of treponemal infection. The 47K surface immunogen will be affinity purified from in vivo- or in vitro-cultivated T. pallidum using monoclonal antibodies, and animals will be immunized. The antibody response in animals immunized with 47K immunogen will be assessed using radioimmunoassay, microhemagglutination assay for T. pallidum, indirect immunofluorescence, radioimmunoprecipitation, Western blotting, T. pallidum immobilization assay, blockage of T. pallidum attachment to host cells in vitro, and in the in vitro-in vivo neutralization test of Bishop and Miller. Animals exhibiting strong humoral responses will be challenged with viable T. pallidum to assess the vaccinogenic potential of the purified 47K immunogen. Simultaneously, anti-47K monoclonal antibodies will be used in passive immunization studies with rabbits or hamsters. The complete primary amino acid sequence for the 47K immunogen will be deduced from studies combining monoclonal antibody immunoaffinity purification, N-terminal amino acid microsequencing, and recombinant DNA techniques. Polyclonal or monoclonal anti-idiotypic antibodies, initially raised against the idiotype of anti-47K monoclonal antibody 11E3, will be used in attempt to elicit an anti-T. palldium response in animals never exposed to T. pallidum. In ongoing studies, monoclonal antibody affinity purification of the 102K, 84K, 54K, 53K, 52K, 32K, 29K, and 24K immunogens of T. pallidum will also be attempted. Production of anti-T. pallidum monoclonal antibodies for the isolation of monoclonal antibodies directed against the 36K, 34K, and 13K immunogens will continue. The study attempts to narrow the information gap regarding the role of the prospective immunogens of T. pallidum, and the antibodies they elicit, in the immunopathogenesis of syphilis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017366-05
Application #
3127183
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1983-04-01
Project End
1990-08-31
Budget Start
1987-09-01
Budget End
1988-08-31
Support Year
5
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Type
Overall Medical
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
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Burstain, J M; Grimprel, E; Lukehart, S A et al. (1991) Sensitive detection of Treponema pallidum by using the polymerase chain reaction. J Clin Microbiol 29:62-9
Brandt, M E; Riley, B S; Radolf, J D et al. (1990) Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins. Infect Immun 58:983-91
Swancutt, M A; Radolf, J D; Norgard, M V (1990) The 34-kilodalton membrane immunogen of Treponema pallidum is a lipoprotein. Infect Immun 58:384-92
Purcell, B K; Chamberlain, N R; Goldberg, M S et al. (1989) Molecular cloning and characterization of the 15-kilodalton major immunogen of Treponema pallidum. Infect Immun 57:3708-14
Swancutt, M A; Riley, B S; Radolf, J D et al. (1989) Molecular characterization of the pathogen-specific, 34-kilodalton membrane immunogen of Treponema pallidum. Infect Immun 57:3314-23
Chamberlain, N R; Brandt, M E; Erwin, A L et al. (1989) Major integral membrane protein immunogens of Treponema pallidum are proteolipids. Infect Immun 57:2872-7
Isaacs, R D; Hanke, J H; Guzman-Verduzco, L M et al. (1989) Molecular cloning and DNA sequence analysis of the 37-kilodalton endoflagellar sheath protein gene of Treponema pallidum. Infect Immun 57:3403-11
Chamberlain, N R; DeOgny, L; Slaughter, C et al. (1989) Acylation of the 47-kilodalton major membrane immunogen of Treponema pallidum determines its hydrophobicity. Infect Immun 57:2878-85
Sanchez, P J; McCracken Jr, G H; Wendel, G D et al. (1989) Molecular analysis of the fetal IgM response to Treponema pallidum antigens: implications for improved serodiagnosis of congenital syphilis. J Infect Dis 159:508-17

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