Pre- and Post-Transfusion Alloimmunization in Dogs Characterized by 2 Antiglobulin-Enhanced Cross-match Tests

Abstract

Background: When dogs are transfused, blood compatibility testing varies widely but may include dog erythrocyte antigen (DEA) 1 typing and rarely cross-matching.

Objectives: Prospective study to examine naturally occurring alloantibodies against red blood cells (RBCs) and alloimmunization by transfusion using 2 antiglobulin-enhanced cross-match tests.

Animals: Eighty client-owned anemic, 72 donor, and 7 control dogs.

Methods: All dogs were typed for DEA 1 and some also for DEA 4 and DEA 7. Major cross-match tests with canine antiglobulin-enhanced immunochromatographic strip and gel columns were performed 26-129 days post-transfusion (median, 39 days); some dogs had an additional early evaluation 11-22 days post-transfusion (median, 16 days). Plasma from alloimmunized recipients was cross-matched against RBCs from 34 donor and control dogs.

Results: The 2 cross-match methods gave entirely concordant results. All 126 pretransfusion cross-match results for the 80 anemic recipients were compatible, but 54 dogs died or were lost to follow up. Among the 26 recipients with follow-up, 1 dog accidently received DEA 1-mismatched blood and became cross-match-incompatible post-transfusion. Eleven of the 25 DEA 1-matched recipients (44%) became incompatible against other RBC antigens. No naturally occurring anti-DEA 7 alloantibodies were detected in DEA 7- dogs.

Conclusions and clinical importance: The antiglobulin-enhanced immunochromatographic strip cross-match and laboratory gel column techniques identified no naturally occurring alloantibodies against RBC antigens, but a high degree of post-transfusion alloimmunization in dogs. Cross-matching is warranted in any dog that has been previously transfused independent of initial DEA 1 typing and cross-matching results before the first transfusion event.

Keywords: Alloantibodies; Blood compatibility; Canine; Dog erythrocyte antigen; Hemolytic transfusion reaction.

Figure 1

Dog erythrocyte antigen (DEA) 1,DEA 4 and DEA 7 typing results. (A) Dog erythrocyte antigen (DEA) 1 immunochromatographic typing strip results were graded from positive (strong, moderate, and weak) to negative (no band). (B) Typing results of 2 DEA 4+ donors showing many small agglutinates by the paper card method. No DEA 4− dogs were found. (C) DEA 7 typing results of 3 donors by the gel column method. Blood samples agglutinating at the top of (4+) and/or within the gel (2+; 3+) were typed as DEA 7+, whereas when all RBCs resided at the bottom, the sample was considered DEA 7−. For all samples, a negative control column with phosphate‐buffered saline (PBS) was included.

Figure 2

Example of Dog erythrocyte antigen (DEA) 1 and Cross‐match test results for recipient R13 and its donor D35 pre‐ and post‐transfusion by the gel column method. For each series of cross‐match tests, a negative control (CT‐; plasma from a nonalloimmunized dog), a positive control (CT+; DEA 4 antisera), and a DEA 1 blood typing (DEA 1) were performed on the RBCs from recipient R13 and its donor D35. Both recipient R13 and its donor D35 are DEA 1+. (A) Donor D35 RBCs: major cross‐match with recipient R13 plasma pre‐ (0) and post‐transfusion at days 19 and 70 (19 and 70). (B) Recipient R13 RBCs: autocontrol pre‐ (0) and post‐transfusion at days 19 and 70 (19 and 70).

Figure 3

Example of different cross‐match test results with the direct antiglobulin‐enhanced immunochromatographic strip and gel column methods. (A) Cross‐match strip method: Any band intensity at “XM” is considered incompatible (graded 1+ to 4+). (B) Cross‐match gel column method: For each series of cross‐matches, a negative control (CT‐) and a positive control (CT+) were performed with the donor RBCs. In the absence of agglutination, all RBCs resided at the bottom of the gel, which was scored as “compatible” (0), whereas agglutination on the top of or within the gel was considered “incompatible” (graded 1+ to 4+).

Figure 4

Comparison of pre‐ and post‐transfusion immunochromatographic and gel column cross‐match test results for 26 recipient dogs. Cross‐match strip and gel grading is shown linearly. Each bullet (◆) represents results from both methods compared for each sample with a linear regression (—). The bullets were overlapping for several dogs. The number of dogs allocated to each bullet is written next to each bullet.