Solutions to avoid false positives for rituximab in pre-transplant crossmatches

Abstract

Rituximab (anti-CD20) is commonly used as immunotherapy against B cells, in the context of pre-transplant crossmatches, where the presence of rituximab in the tested sera with donor cells can alter their results both by flow cytometry (FCXM) as complement-dependent cytotoxicity (CDCXM) giving rise to false positives. In the present study, we tested the use of an anti-rituximab monoclonal antibody (10C5, Abnova) as a method to avoid false positives in FCXM and CDCXM. We used the serum from ten patients who received therapy with rituximab, and the cells were incubated with sera treated or untreated with the 10C5 clone. In previous studies, attempts have been made to control these false positives through the use of pronase, although in these cases the alteration of Human Leukocyte Antigen (HLA) molecules has been found to be a limitation. As an alternative, we performed an assay to exclude false positives by a pre-incubation with anti-rituximab antibody (10C5) in 1:5 proportion avoiding the misinterpretation of crossmatches, particularly in patients with specific donor antibodies (DSA) without affecting the HLA molecules.