Antigen specificity in canine autoimmune haemolytic anaemia

Autoimmune haemolytic anaemia (AIHA) is the most common clinical manifestation of autoimmunity in the dog and generally presents as a profound, regenerative, Coombs' positive anaemia of acute or chronic onset. The disease pathogenesis involves formation of erythrocyte-specific autoantibodies of the IgG and IgM class that may fix complement resulting in intra- or extravascular haemolysis. Western blotting and immunoprecipitation studies using autoantibody eluted from the erythrocytes of dogs with AIHA have demonstrated specificity for erythrocyte glycophorins and the membrane anion-exchange molecule (band 3). Autoantibodies specific for the cytoskeletal molecule spectrin have been identified in serum by ELISA. The specificity of autoreactive T-cells has been examined in vitro using bulk cultures stimulated with a panel of autoantigens including intact erythrocyte membranes, purified glycophorin and spectrin fractions and a panel of overlapping 15-mer glycophorin peptides. Control responses to ConA and recall (vaccine antigens) and non-recall (KLH) antigens were measured in the same system. PBMC obtained from dogs that had recovered from AIHA consistently proliferated in response to erythrocyte membranes, with occasional responses to spectrin or glycophorin. PBMC from sone clinically normal dogs also responded to erythrocyte membranes. PBMC obtained from dogs closely related to AIHA cases gave the most consistent responses, including proliferation when stimulated by the glycophorin peptides. These data suggest that normal dogs harbour erythrocyte autoreactive lymphocytes, and that these cells may be primed in dogs recovered from AIHA or having genetic susceptibility to the disease.