S A Staroverov1,2, A S Fomin1, S V Kozlov2, A A Volkov2, E S Kozlov2, K P Gabalov1, L A Dykman3. 1. Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 13 Prospekt Entuziastov, Saratov, Russia, 410049. 2. Saratov State Vavilov Agrarian University, 1 Teatralnaya Ploshchad, Saratov, Russia, 410012. 3. Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 13 Prospekt Entuziastov, Saratov, Russia, 410049. dykman_l@ibppm.ru.
Abstract
PURPOSE: The aim of this study was to isolate Babesia canis soluble antigens and to investigate the effect of their conjugates with gold nanoparticles on the immunogenicity in laboratory animals. METHODS: A procedure was developed for isolating and purifying B. canis antigens. The isolated culture antigen of B. canis 495 was coupled to gold nanoparticles, and the conjugate was used to immunize laboratory mice. RESULTS: Western blotting showed that the resultant antiserum specifically recognized the proteins of the B. canis strains isolated from naturally infected dogs. The antibody titer, the respiratory activity of peritoneal macrophages, the proliferative activity of splenocytes, and the production of cytokines were maximal when the animals were immunized with the antigen-nanoparticle conjugate emulsified in complete Freund's adjuvant. Without adjuvant, the babesial antigen was weakly immunogenic. CONCLUSION: Therefore, the use of gold nanoparticles as an antigen carrier induced a broad immune response involving both cellular and humoral responses. The antibodies raised by the proposed procedure are potentially effective at immunodetection of Babesia canis infections in dogs.
PURPOSE: The aim of this study was to isolate Babesiacanis soluble antigens and to investigate the effect of their conjugates with gold nanoparticles on the immunogenicity in laboratory animals. METHODS: A procedure was developed for isolating and purifying B. canis antigens. The isolated culture antigen of B. canis 495 was coupled to gold nanoparticles, and the conjugate was used to immunize laboratory mice. RESULTS: Western blotting showed that the resultant antiserum specifically recognized the proteins of the B. canis strains isolated from naturally infecteddogs. The antibody titer, the respiratory activity of peritoneal macrophages, the proliferative activity of splenocytes, and the production of cytokines were maximal when the animals were immunized with the antigen-nanoparticle conjugate emulsified in complete Freund's adjuvant. Without adjuvant, the babesial antigen was weakly immunogenic. CONCLUSION: Therefore, the use of gold nanoparticles as an antigen carrier induced a broad immune response involving both cellular and humoral responses. The antibodies raised by the proposed procedure are potentially effective at immunodetection of Babesia canis infections in dogs.
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