Previous studies in our laboratory have shown that C-ASWS, an alkali-soluble, water-soluble cell wall extract of Coccidioides immitis mycelia (C-ASWS-M) and spherules (C-ASWS-S) is biologically active in: (i) detecting in vivo and in vitro cellular immune responses in experimentally-infected animals and patients with various stages of coccidioidomycosis; (ii) detecting IgM tube precipitin antibody to C. immitis; and (iii) protecting mice against challenge with viable Coccidioides arthroconidia. These studies have established that C-ASWS-M and C-ASWS-S exhibit the spectrum of biological activities that are associated with coccidioidin (CDN), and autolysate of C. immitis mycelia, and spherulin, an autolysate of spherules, with the single exception of reactivity with complement-fixing IgG antibody. More recent studies provide evidence that the C-ASWS extracts are antigenically similar, if not identical, to an antigen (designated Antigen 2) present in both CDN and spherulin. This evidence is based upon our finding that the C-ASWS extracts fuse with Antigen 2 in tandem two-dimensional immunoelectrophoresis (2D-IEP) of CDN against burro anti-CDN and that solid-phase immunoadsorption (SPIA) of CDN on a column containing goat antiserum to C-ASWS-M or C-ASWS-S results in the adsorption of Antigen 2. Desorption of Antigen 2 from SPIA columns yields a preparation which is homogeneous by 2D-IEP and biologically active in eliciting skin test responses in Coccidioides-infected guinea pigs and in detecting tube precipitin antibody in patients with active coccidioidomycosis. On the basis of these results, we propose a study with the following specific aims: (i) isolate Antigen 2 from CDN and spherulin by SPIA using goat antisera to the C-ASWS extracts; (ii) assess the homogeneity of Antigen 2 preparations using the techniques of 2D-IEP and electroblotting; (iii) utilize conventional methods for purifying Antigen 2 preparations; (iv) establish the sensitivity and specificity of Antigen 2 in humoral and cellular immune assays of Coccidioides-, Histoplasma-, and Blastomyces-infected animals; and (v) characterize the chemical composition of Antigen 2 and assess whether the biological activities are attributed to a protein moiety, polysaccharride moiety or both.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021431-03
Application #
3131556
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1984-08-01
Project End
1987-07-31
Budget Start
1986-08-01
Budget End
1987-07-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Texas Center for Infectious Disease
Department
Type
DUNS #
City
San Antonio
State
TX
Country
United States
Zip Code
78223
Jiang, C; Magee, D M; Cox, R A (1999) Coadministration of interleukin 12 expression vector with antigen 2 cDNA enhances induction of protective immunity against Coccidioides immitis. Infect Immun 67:5848-53
Jiang, C; Magee, D M; Quitugua, T N et al. (1999) Genetic vaccination against Coccidioides immitis: comparison of vaccine efficacy of recombinant antigen 2 and antigen 2 cDNA. Infect Immun 67:630-5
Zhu, Y; Tryon, V; Magee, D M et al. (1997) Identification of a Coccidioides immitis antigen 2 domain that expresses B-cell-reactive epitopes. Infect Immun 65:3376-80
Cox, R A; Dolan, M J; Magee, D M et al. (1993) Production of a murine monoclonal antibody that recognizes an epitope specific to Coccidioides immitis antigen 2. Infect Immun 61:1895-9
Cox, R A; Sun, S H; Dolan, M J et al. (1992) Localization of the tube precipitin and complement fixation antigens of Coccidioides immitis by immunoelectron microscopy with murine monoclonal antibodies. Infect Immun 60:3315-24
Dolan, M J; Cox, R A (1991) Production and characterization of a monoclonal antibody to the complement fixation antigen of Coccidioides immitis. Infect Immun 59:2175-80
Dolan, M J; Cox, R A; Williams, V et al. (1989) Development and characterization of a monoclonal antibody against the tube precipitin antigen of Coccidioides immitis. Infect Immun 57:1035-9
Cox, R A (1989) Antigenic structure of Coccidioides immitis. Immunol Ser 47:133-70
Cox, R A; Britt, L A; Michael, R A (1987) Isolation of Coccidioides immitis F antigen by immunoaffinity chromatography with monospecific antiserum. Infect Immun 55:227-32
Cox, R A; Britt, L A (1987) Antigenic identity of biologically active antigens in coccidioidin and spherulin. Infect Immun 55:2590-6

Showing the most recent 10 out of 13 publications