Abstract
Dog Erythrocyte Antigens (DEA) have thus far been found by sensitizing dogs with canine allogeneic blood and are clinically important regarding blood transfusion incompatibilities, but remain poorly defined. The goals of this study were to discover and characterize two DEAs, named as Kai 1 and Kai 2. The monoclonal antibodies were produced by mouse hybridoma techniques and examined by ELISA isotyping, immunoblotting, and affinity chromatography. Canine blood samples were typed and the development of alloantibodies was examined in transfused dogs. The monoclonal Kai 1 and Kai 2 antibodies were isotyped as IgM kappa and IgG3 lamda, respectively, and identified two different erythrocyte membrane proteins of 200 kDa and 80 kDa in molecular weights, respectively. Either Kai 1 or Kai 2 can be expressed but not both in an individual dog. There were no naturally occurring anti-Kai 1 or Kai 2 alloantibodies. In addition, Kai 1- and/or Kai 2- dogs developed Kai 1 and Kai 2 alloantibodies, respectively, when transfused with mismatched blood. This is the first discovery of canine blood types by screening monoclonal antibodies. Kai 1 and Kai 2 are novel blood types which can induce anti-Kai 1 or anti-Kai 2 alloantibodies when Kai 1- and/or Kai 2- dogs are transfused with Kai 1+ or Kai 2+ blood. These canine blood types may explain some of the blood incompatibilities and transfusion reactions observed in dogs in clinical practice.
Highlights
Dogs have been used in early xenotransfusions to humans as well as animal models to characterize transfusion reactions [1]
We isolated two hybridoma cell lines which produced different, but not cross-reacting, antibodies against the red blood cell (RBC) from the two dogs typed as Dog Erythrocyte Antigens (DEA) 1.1 and DEA 1.2, respectively
While based upon experimental and clinically mismatched transfusions more than a dozen blood groups have been described in dogs, the nature of these canine alloantibodies and RBC antigens have remained mostly elusive
Summary
Dogs have been used in early xenotransfusions to humans as well as animal models to characterize transfusion reactions [1]. In veterinary clinical practice, when anemic or bleeding dogs are transfused, blood type incompatibilities have been documented based upon hemolytic transfusion reactions and incompatible agglutination crossmatch test results [2,3]. Based upon experimentally sensitizing dogs with canine blood transfusions, eight Dog Erythrocyte Antigens (DEA) were classified with polyclonal alloantibodies by an international committee in 1974, but currently only reagents for DEA 1, DEA 3, DEA 4, and DEA 7 are commercially available. Kai dog erythrocyte antigens group system where dogs were classified to be DEA 1.1+ or DEA 1.1- and those DEA 1.1- could be DEA 1.2+ or DEA 1.2- [3,4]. Recent flow cytometry and strip kit typing studies with monoclonal antibodies reveal that dogs are either DEA 1- or weakly to strongly DEA 1+ [5], but the biochemical and molecular basis remains elusive for DEA 1 and other DEAs
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