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Serum-specific antibodies in rabbits naturally infected with Trichophyton mentagrophytes
1Clinic for Reproduction and Horses, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia. petra.zrimsek@vf.uni-lj.si
Abstract:
In the present study, we investigated the humoral immune response of rabbits to Trichophyton mentagrophytes (sensu lato) proteins obtained from keratin-rich media in vitro. The test rabbits were naturally infected with T. mentagrophytes. The production of keratinolytic enzymes in T. mentagrophytes was stimulated by growing cultures with keratin as a sole nitrogen source. The proteins were isolated from a protein extract prepared from the fungal mat. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed three bands. Bands with Mr 20 and 30 kDa were glycosylated, whereas a band of 18 kDa was not. The rabbits' humoral immune responses to the total protein extract of T. mentagrophytes and to the proteins with keratinolytic activity was studied by immunoblotting. IgG from infected rabbits' sera revealed eight dominant bands with apparent molecular weights between 20 and 75 kDa. Bands of 20, 30 and 33 kDa appeared with a frequency rate of 76% only on immunoblots of infected rabbit serum. Using an indirect enzyme-linked immunosorbent assay (ELISA), we observed a significant increase in specific antibodies in a group of infected rabbits compared to a control group (P < 0.001). The ELISA exhibited 95% sensitivity and 83% specificity at the optimal cut-off value, with 90% predictive values of a positive and a negative result. Under these conditions, it could be used in the accurate detection of specific antibodies in sera of infected rabbits.
Insights
This study explored rabbit immune responses to Trichophyton mentagrophytes proteins, identifying key antigens. An enzyme-linked immunosorbent assay (ELISA) shows promise for detecting specific antibodies in infected rabbits.
Area of Science:
- Mycology
- Immunology
- Veterinary Dermatology
Background:
- Trichophyton mentagrophytes (sensu lato) is a common dermatophyte causing infections.
- Understanding the host immune response is crucial for diagnosis and treatment strategies.
Purpose of the Study:
- To investigate the humoral immune response in rabbits naturally infected with T. mentagrophytes.
- To identify specific proteins from T. mentagrophytes that elicit an immune response.
- To evaluate the diagnostic potential of an indirect enzyme-linked immunosorbent assay (ELISA) for detecting antibodies.
Main Methods:
- Proteins were extracted from T. mentagrophytes cultured on keratin-rich media.
- Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze protein components.
- Immunoblotting and indirect ELISA were employed to assess rabbit immune responses to fungal proteins.
Main Results:
- SDS-PAGE revealed three protein bands, with two being glycosylated.
- Immunoblotting identified eight dominant IgG-binding bands in infected rabbit sera, with specific bands at 20, 30, and 33 kDa appearing frequently.
- ELISA demonstrated a significant increase in specific antibodies in infected rabbits compared to controls, with high sensitivity (95%) and specificity (83%).
Conclusions:
- Specific proteins of T. mentagrophytes, particularly those with keratinolytic activity, elicit a humoral immune response in infected rabbits.
- The developed indirect ELISA is a sensitive and specific method for detecting antibodies against T. mentagrophytes in rabbit sera.
- This assay can aid in the accurate diagnosis of T. mentagrophytes infections in rabbits.