Retrospective evaluation of flow cytometry as a platelet crossmatching procedure

Abstract

We performed a retrospective analysis of flow cytometry as a platelet crossmatching procedure. Sera from 17 alloimmunized refractory patients were tested against 32 donor platelets, which had been stored as platelet-rich plasma for up to 36 months. Overall, 14/32 (44%) crossmatches were positive. The mean 1 h posttransfusion corrected count increments (CCls) were 9,195 and 2,269 for a negative and a positive crossmatch, respectively. The predictive value of a positive crossmatch was 86%, whereas the predictive value of a negative crossmatch was 56%. When samples with low background fluorescence or with high panel-reactive antibody (PRA) levels were evaluated separately, the accuracy of the crossmatch improved from 69% to 80%. When compared to the platelet adhesion immunofluorescence test (PAIFT) and the standard and antiglobulin-enhanced lymphocytotoxicity tests for the detection of HLA antibodies, flow cytometry appeared to be more sensitive. We conclude that flow cytometry is a useful technique for platelet crossmatching, particularly for alloimmunized patients for whom HLA compatible platelets may not be readily available. o 1994 Wiley-Liss, Inc. Key terms: Blood platelets, platelet transfusions, platelet antibody testingThe management of platelet recipients who are refractory to random donor platelet concentrates poses a difficult problem. Many, but not all of these patients are alloimmunized to HLA-A and -B antigens, platelet-specific antigens, or both. Standard practice is to provide these patients with single donor platelets that are partially or completely matched with the recipient's HLA-A and -B antigens (15,19,24). Although HLA-compatible (A an B-U match grade) platelets are preferred, the marked polymorphism of the HLA system limits their availability. Accordingly, most recipients must be supported with single donor platelets that are mismatched for one or more of the recipient's HLA antigens (6).Recently, platelet crossmatching has been proposed as an alternative to HLA-based donor selection ( 3,4,7,10, 1 1,23). The advantages of platelet crossmatching are that antibodies directed against platelet-specific as well as HLA antigens can be detected and the need for a large pool of H1.A typed donors is avoided. We have developed a flow cytometric procedure for platelet antibody detection (27). In this study, we performed a retrospective evaluation of this method as a platelet crossmatching technique. MATERIALS AND METHODS SubjectsThe 17 patients in this study were participants in a multicenter investigation comparing the efficacy of pro-C 1994 Wiley-Liss, Inc.spective platelet crossmatching and HLA-based selection, the results of which were reported previously (21). These patients were refractory to random donor platelet concentrates (posttransfusion corrected platelet count increments at 1 h less than 7,500 on two consecutive occasions) and lacked nonimmune factors associated with refractoriness, the majority of these patients were either bone marrow transplant recipients or patients with acute leukemi...

Improved method for fluorescence cytometric immunohematology testing

“…The availability of this automated workstation will allow us to address other important considerations, including capital equipment costs, disposable and reagent costs, sample turnaround time and throughput, and reproducibility of assay performance in other transfusion service and blood center environments. Furthermore, the availability of this workstation will allow us to investigate complete automation of other pretransfusion tests, including quantitation of residual WBCs after filtration, PLT cross‐matching, bacterial detection, and quantitation of fetal–maternal hemorrhage, all of which have been previously performed by traditional flow cytometry 11‐19 …”

Flow cytometry antibody screening using pooled red cells

“…The isotypes of these antibodies among the 32 CSi/ FC-positive cases with PEG could be identified via twocolor FC (n): IgG only (19), and IgM only (5: anti-E 3; anti-M 1; anti-c 1), both IgG and IgM (8).…”

“…Flow cytometry (FC) has been previously utilized for antibody detection, including the detection of red cell autoantibodies (4,5), HLA alloantibodies (6,7), platelet crossmatching (8)(9)(10), and other immmunohematological applications (11,12). Recently, FC was also introduced for the screening of antibodies to red cells for automation by Roback et al (3,13).…”

Flow cytometric platelet cross-matching to predict platelet transfusion in acute leukemia

“…Platelet cross matching by flow cytometry was addressed in many studies [14,15]. However, the predictive role of flow cytometric platelet cross-matching on transfusion response and its affection by the presence of clinical factors, HLA alloimmunization, and transfusion of HLA-matched platelet were not fully evaluated.…”