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 ant igenic integrity of A. marginale grown in vitro in bovine erythrocytes co-c ultured with endothelial cells. Both the mAbs reacted in the indirect immun ofluorescent antibody test with A. marginale grown in vitro and also detect ed the antigens in Western immunoblots of SDS-PAGE separated antigens made from A, marginale infected erythrocytes from the cultures. Furthermore, act ive replication was evident as [S-35]-methionine is incorporated by A. marg inale present in the second passage of a culture maintained for six weeks a s shown by immunoprecipitation of labeled antigens by the mAbs. This indica tes that A. marginale grown in the in vitro culture system described previo usly [Waghela et al., Vet. Parasitol. 73 (1997) 43] maintain antigenic char acter, and with further development the system can be used for preparing im munogens or diagnostic antigens. (C) 2000 Elsevier Science B.V. All rights reserved.