Production and preliminary characterization of monoclonal antibodies to Actinobacillus sp isolated from epididymitis lesions in a ram.
A panel of 55 monoclonal antibodies to an Actinobacillus sp isolate (As8C strain) cultured from the epididymides of an infected ram was produced. Cell lines producing 5 of these antibodies were cloned and expanded by hybridoma tumor production in Balb/c mice. An isotype profile revealed that 1 cloned antibody belonged to the immunoglobulin (Ig) M class and the 4 remaining antibodies belonged to the IgG class. Within the IgG class, 1 clone produced IgG1, 1 clone produced IgG2a, and 2 clones produced IgG2b. Ascites fluid antibody titers from the cloned hybridomas ranged from 6,400 to 51,200, as determined by enzyme-linked immunosorbent assay. Specificity of the antibodies to target As8C antigens could be demonstrated by enzyme-linked immunosorbent assay inhibition. Ascites fluid from 2 clones contained antibodies that agglutinated As8C. Two additional clones produced antibodies capable of only partial agglutination, whereas 1 clone produced antibody that did not agglutinate As8C. The indirect fluorescent antibody test revealed that target antigens for at least 4 of the 5 monoclonal antibodies were most likely located on the bacterial cell surface. Antigens were extracted from As8C, using 5 surface active chemicals. An attempt to immunoprecipitate these antigens in agarose by reacting individual extracts with each of the antibodies was unsuccessful.
D000188:Actinobacillus / Q000276:immunology*; D000372:Agglutination Tests; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D000937:Antigen-Antibody Reactions; D001202:Ascitic Fluid / Q000276:immunology; D004823:Epididymitis / Q000662:veterinary*; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology*; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M