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United States Department of Agriculture

Agricultural Research Service

Title: Generation of Chicken-Chicken Monoclonal Antibody Against Eimeria Acervulina Antigen Associated with Cd8+t Cell: Effect on Ultrastructure Using Immunofluorescent Scanning Microscopy and Cellular Invasion

item Sasai Kazumi, - UNIV. OF OSAKA PREF.
item Lillehoj, Hyun
item Matsuda H, - HIROSHIMA UNIV
item Miyamoto T, - HIROSHIMA UNIV
item Fukata T, - HIROSHIMA UNIV
item Arakawa A, - HIROSHIMA UNIV

Submitted to: Japanese German Cooperative Symposium on Protozoan Diseases
Publication Type: Proceedings
Publication Acceptance Date: September 1, 1994
Publication Date: N/A

Technical Abstract: Biochemical characterization of eimerian parasite antigens is important to assess the target antigens of the host immune response. Many mice monoclonal antibodies (mAbs) have been used to characterize coccidial antigens and to identify cDNAs encoding proteins of eimerian parasites. However, the relevance of using mouse mAbs to define the epitopes important tin the chicken's immune response to Eimeria and to identify potential Eimeria vaccine antigens are debatable. Furthermore, differences in the target antigens recognized by immune sera from chickens, rabbits and mice have been reported. For these reasons, it is important to develop chicken- chicken B cell hybridoma to identify coccidial antigens that are recognized by the natural host during coccidial infection. Recently we have reported that sporozoites entered CD8+ T cells in the intestinal epithelium shortly following oral infection and were transported dby CD8+ T cell to crypt epithelium where sporozoites developed. In the present study, we have fused TK-chicken B cell line with spleen cells from chickens that were immunized with CD8+ T cells incubated with Ea sporozoite antigens to identify sporozoite antigens that interact with surface receptor on CD8+ T cell. A stable hybridoma (6D-12-G10) that produced monoclonal antibody against eimerian parasites was developed and characterized using immunofluorescent microscopy. This monoclonal antibody was used to inhibition assay of invasion of sporozoites into CD8+ T cells in vitro. The results showed that immunofluorescent staining by this monoclonal antibody is localized to apical portion of sporozoite and statistically significant inhibition of invasion of Ea sporozoites can be achieved by using a single anti-sporozoite monoclonal antibody.

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