Biased immunoglobulin G1 isotype responses induced in cattle with DNA expressing msp1a of Anaplasma marginale

Immunization with the native major surface protein 1 (MSP1) (a heterodimer containing disulfide and noncovalently banded polypeptides designated MSP1a and MSP1b) of the erythrocytic stage of Anaplasma marginale conferred prot ection against homologous challenge (G. H. Palmer, A. F. Barbet, W. C. Davi s, and T. C. McGuire, Science 231:1299-1302, 1986). The MSP1a polypeptide p ossesses a conserved neutralization-sensitive epitope. In the present study , the immune response to DNA-mediated immunization using msp1a was studied. The plasmid pVCL/MSP1a, which encodes the complete msp1a gene of A. margin ale under the control of human cytomegalovirus immediate-early enhancer/pro moter and intron A, was constructed. The immune responses elicited by immun ization with pVCL/MSP1a into cardiotoxin-induced regenerating muscle were e valuated in mice and cattle. Antibody reactive,vith native MSP1a was detect ed in pooled sera of immunized BALB/c mice 3 weeks following primary immuni zation. Two calves seronegative for A. marginale were immunized four times, at weeks 0, 3, 7, and 13, with pVCL/MSP1a. By 8 weeks, both calves respond ed to MSP1a with an antibody titer of 1:100, which peaked at 1:1,600 and 1: 800 by 16 weeks after the initial immunization. Interestingly, immunoblotti ng with anti-immunoglobulin G1 (anti-IgG1) and anti-IgG2 specific monoclona l antibodies revealed a restricted IgG1 anti-MSP1a response in both animals . T-lymphocyte lines, established after the fourth immunization, proliferat ed specifically against A. marginale homogenate and purified MSP1 in a dose dependent manner. These data provide a basis for an immunization strategy to direct bovine immune responses by using DNA vaccine vectors containing s ingle or multiple genes encoding major surface proteins of A. marginale.