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The coccidioidal complement fixation and immunodiffusion-complement fixation antigen is a chitinase
1Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis 95616.
Insights
Researchers identified a chitinase from Coccidioides immitis as a key antigen for diagnosing coccidioidomycosis. This enzyme is crucial for serodiagnosis and may play a role in fungal development.
Area of Science:
- Mycology
- Immunology
- Biochemistry
Background:
- Culture filtrates and autolysates of Coccidioides immitis are used as crude antigens for coccidioidomycosis serodiagnosis.
- A specific heat-labile antigen, involved in immunodiffusion and complement fixation reactions (IDCF), was previously characterized as a 110-kDa protein yielding a 48-kDa component under reducing and heat conditions.
Purpose of the Study:
- To provide serologic and biochemical evidence identifying the heat-labile IDCF antigen as a chitinase.
- To investigate the enzymatic and antigenic properties of this C. immitis chitinase.
Main Methods:
- Isolation of the antigen from a 48-h culture filtrate of the spherule-endospore-phase C. immitis using affinity adsorption to chitin.
- Serologic testing including immunodiffusion and complement fixation reactions with human serum.
- Biochemical analysis of enzymatic and antigenic activity following heat treatment.
Main Results:
- The isolated chitinase formed a line of identity with the IDCF reference antigen in immunodiffusion.
- The chitinase participated in the complement fixation reaction with human serum.
- Enzymatic and antigenic activity were lost upon heating, but enzymatic activity was retained when not heated, even after precipitation with coccidiodal antibody.
Conclusions:
- The identified chitinase is a significant serodiagnostic substance for coccidioidomycosis.
- This chitinase may play a role in the morphogenesis of Coccidioides immitis.
Abstract:
Culture filtrates and autolysates of Coccidioides immitis have provided suitable crude antigens for the serodiagnosis and prognosis of coccidioidomycosis. One of these, a heat-labile antigen which participates in the immunodiffusion reaction corresponding to the complement fixation reaction (IDCF), has been characterized as a 110-kDa native protein that, when subjected to reducing conditions and heat, yields a 48-kDa component. The present report provides serologic and biochemical evidence that this antigen is a chitinase. This chitinase, isolated from 48-h culture filtrate of the spherule-endospore-phase C. immitis by affinity adsorption to chitin, formed a line of identity with the IDCF reference antigen and participated in the complement fixation reaction with human serum. It lost its enzymatic as well as antigenic activity when heated, but when not heated it retained its enzymatic activity even when precipitated with coccidiodal antibody present in human serum. This chitinase represents a significant serodiagnostic substance and may be important in the morphogenesis of C. immitis.
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