The clinical importance of choosing the right assay for detection of HLA-specific donor-reactive antibodies.

BACKGROUND We found earlier that there is a close clinical correlation between the presence of histocompatibility leukocyte antigens (HLA) class I-specific donor-reactive antibodies in cross-match serum and a significantly higher frequency of early acute rejection episodes and graft loss during the first year after the transplant. METHODS Specificity determinations of donor-reactive antibodies present in the cross-match serum before allogeneic kidney transplants were performed. In the present study, we compared the suitability and efficiency of (a) platelet absorptions, (b) blocking with anti-HLA monoclonal antibodies in the microcytotoxicity assay, and (c) donor-specific HLA antigen-coated magnetic microbeads in flow cytometric assays for the definition of clinically relevant HLA antibodies and their correlation with early acute rejections and early graft loss. RESULTS We found that the microlymphocytoxicity test using donor splenic B lymphocytes often gave positive reactions in the absence of class I or class II antibodies; in other words, a high frequency of false positive reactions was observed. Flow cytometric tests are more sensitive than microlymphocytotoxicity, not only because they are more sensitive, but also because noncomplement-binding antibodies are detected. Platelet absorptions, which detect only reactivity against HLA class I antigens, is insufficient for use in specificity determinations of donor-reactive antibodies. We found that a positive test for HLA antibodies using paramagnetic beads coated with solubilized donor-derived HLA antigens (class I or class II) correlates with early immunological complications after a transplant (P<0.001). CONCLUSION Assays to determine the specificity of donor-reactive antibodies are now available for use during an acute transplant situation. The introduction of such methods is expected to enhance graft survival and to reduce significantly the frequencies of acute rejections episodes after a transplant.

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