Antigenic analysis of Anaplasma marginale grown in bovine erythrocytes co-cultured with bovine endothelial cells.
Two monoclonal antibodies (mAbs) for A. marginale were used to test the antigenic integrity of A. marginale grown in vitro in bovine erythrocytes co-cultured with endothelial cells. Both the mAbs reacted in the indirect immunofluorescent antibody test with A. marginale grown in vitro and also detected the antigens in Western immunoblots of SDS-PAGE separated antigens made from A. marginale infected erythrocytes from the cultures. Furthermore, active replication was evident as [35S]-methionine is incorporated by A. marginale present in the second passage of a culture maintained for six weeks as shown by immunoprecipitation of labeled antigens by the mAbs. This indicates that A. marginale grown in the in vitro culture system described previously [Waghela et al., Vet. Parasitol. 73 (1997) 43] maintain antigenic character, and with further development the system can be used for preparing immunogens or diagnostic antigens.
Waghela
S D
SD
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843-4467, USA. swaghela@cvm.tamu.edu
Melendy
D
D
Cruz
D
D
Wagner
G G
GG
D000709:Anaplasma / Q000276:immunology*; D000712:Anaplasmosis / Q000276:immunology / Q000469:parasitology; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology / Q000469:parasitology; D018920:Coculture Techniques; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004727:Endothelium / Q000166:cytology / Q000469:parasitology; D004912:Erythrocytes / Q000469:parasitology; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D015151:Immunoblotting / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests / Q000662:veterinary; D013462:Sulfur Radioisotopes / Q000737:chemistry
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D013462:Sulfur Radioisotopes