Characterization of hemagglutinating components on the Anaplasma marginale initial body surface and identification of possible adhesins.
Interaction of Anaplasma marginale initial bodies with the bovine erythrocyte surface was examined by a direct hemagglutination assay. Purified initial bodies were shown to specifically hemagglutinate bovine erythrocytes but not erythrocytes from nonhost animal species. Hemagglutination was inhibited by treatment of purified initial bodies with trypsin, alpha-chymotrypsin, or proteinase K but not by treatment with neuraminidase or sodium periodate. Treatment of bovine erythrocytes with alpha-chymotrypsin or neuraminidase partially inhibited hemagglutination of the treated cells by initial bodies. In contrast, no inhibition occurred after treatment of erythrocytes with trypsin, phospholipases, or sodium periodate or when monosaccharides and disaccharides were used as potential competitive inhibitors. Thus, the initial body receptor is probably a surface protein, whereas the bovine receptor may comprise both protein and carbohydrate. Hemagglutination was unaffected by treatment of initial bodies with monoclonal or polyclonal antibodies raised against the A. marginale 31-kDa (MSP4) major surface polypeptide or non-A. marginale proteins or by treatment with a monoclonal antibody to the A. marginale MSP1a neutralization-sensitive epitope. In contrast, antiserum raised against whole A. marginale initial bodies or monospecific antibodies raised against purified A. marginale major surface polypeptides with molecular sizes of 105 (MSP1a), 100 (MSP1b), 61, and 36 (MSP2) kDa completely or partially inhibited hemagglutination. These data confirm the proposed surface location of the proteins susceptible to inhibition and suggest that they mediate hemagglutination of bovine erythrocytes. We propose that these surface proteins are possible adhesins.
McGarey
D J
DJ
Department of Infectious Diseases, University of Florida, Gainesville 32611.
Allred
D R
DR
D000709:Anaplasma / Q000472:pathogenicity*; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000502:physiology; D002417:Cattle; D005260:Female; D006384:Hemagglutination; D006385:Hemagglutination Inhibition Tests
D001426:Bacterial Proteins