Comparison of selected protocols for the generation of IgA isotype monoclonal antibodies against the gut parasite, Eimeria tenella

Abstract

Investigations into the role of mucosal IgA in the protective immune response to alimentary tract parasites could be facilitated by the production of specific IgA monoclonal antibodies. To this end, we sought to generate IgA hybridomas against sporozoites of the protozoan, Eimeria tenella using different protocols. Of the methods investigated, intra-enteric priming followed by intravenous boosting of germ-free mice, using splenic lymphocytes in fusions, optimised the yield of IgA hybridomas. Using this protocol, 7 27 specific anti- Eimeria hybridoma antibodies isolated were of IgA isotype. When lymphocytes from mesenteric lymph nodes (MLN) were used in fusions more non-specific IgA secretors were produced than when splenic lymphocytes were used, but the yield of specific anti- Eimeria IgA secreting hybridomas was not improved. By all protocols, a total of nine IgA secreting hybridomas were identified of which eight have been cloned for further studies.