Abstract

An immunochemical approach to the characterization of the capsular polysaccharides of Cryptococcus neoformans is proposed. Heteroglycans from several chains of the four recognized serotypes and mutants partially blocked in capsule synthesis will be fractionated based on differential complexation with cetyltrimethylammonium bromide, ion exchange chromatography, lectin affinity, and immunoaffinity. The native polysaccharides, polysaccharide derivatives and oligosaccharides derived by special chemical treatments, e.g., lithium: ethylenediamine, anhydrous hydrofluoric acid and periodate oxidation, will be examined by various analytical methods, including gas liquid chromatography, high field nuclear magnetic resonance spectrometry, gas liquid chromatography-mass spectrometry, and glycosidase susceptibility. The data obtained will be used to make linkage and sequence assignments for the native polysaccharides. Purified and immunochemically defined capsular polysaccharides and their derivatives will be used in the development of an enzyme immunoassay (EIA). This assay in conjunction with monoclonal antibodies will be used to delineate the chemical nature of the type specific and other immunodominant epitopes of C. neoformans. Structurally defined cryptoccal polysaccharides, their derivatives and model compounds will be studied by both direct and hapten inhibition variations of EIA.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI018877-03
Application #
3128273
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1985-09-01
Project End
1989-02-28
Budget Start
1987-09-01
Budget End
1989-02-28
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
Georgia State University
Department
Type
Schools of Arts and Sciences
DUNS #
837322494
City
Atlanta
State
GA
Country
United States
Zip Code
30302

  Publications

Cherniak, R; Sundstrom, J B (1994) Polysaccharide antigens of the capsule of Cryptococcus neoformans. Infect Immun 62:1507-12
Turner, S H; Cherniak, R (1991) Glucuronoxylomannan of Cryptococcus neoformans serotype B: structural analysis by gas-liquid chromatography-mass spectrometry and 13C-nuclear magnetic resonance spectroscopy. Carbohydr Res 211:103-16
Stortz, C A; Cherniak, R; Jones, R G et al. (1990) Polysaccharides from Peptostreptococcus anaerobius and structure of the species-specific antigen. Carbohydr Res 207:101-20
James, P G; Cherniak, R; Jones, R G et al. (1990) Cell-wall glucans of Cryptococcus neoformans Cap 67. Carbohydr Res 198:23-38
James, P G; Cherniak, R (1990) 4-Methylmorpholine N-oxide-methyl sulfoxide soluble glucan of Piptoporus betulinus. Carbohydr Res 206:167-72
Todaro-Luck, F; Reiss, E; Cherniak, R et al. (1989) Characterization of Cryptococcus neoformans capsular glucuronoxylomannan polysaccharide with monoclonal antibodies. Infect Immun 57:3882-7
Todaro-Luck, F; White, E H; Reiss, E et al. (1989) Immunoelectronmicroscopic characterization of monoclonal antibodies (MAbs) against Cryptococcus neoformans. Mol Cell Probes 3:345-61
Cherniak, R; Jones, R G; Reiss, E (1988) Structure determination of Cryptococcus neoformans serotype A-variant glucuronoxylomannan by 13C-n.m.r. spectroscopy. Carbohydr Res 172:113-38
Cherniak, R; Cheeseman, M M; Reyes, G H et al. (1988) Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay. Diagn Clin Immunol 5:344-8
Murphy, J W; Mosley, R L; Cherniak, R et al. (1988) Serological, electrophoretic, and biological properties of Cryptococcus neoformans antigens. Infect Immun 56:424-31

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