Simultaneous detection of antibody binding and cytotoxicity in flow cytometry crossmatch for renal transplantation

Abstract

The simultaneous detection of Ab binding and cytotoxicity was possible by the cytotoxic FCXM with the test efficiencies similar to the conventional counterparts. If this new assay is improved through the further studies to optimize the critical assay variables, this may be used as an alternative to the conventional assays to acquire more information on the characteristics of the recipient's HLA alloantibodies.

Effector Mechanisms of Rejection

“…Antibodies can also bind to the surface molecules of target cells and recruit other cells by interactions via their constant domain (Rebellato et al 2006;Won et al 2006). Recently, Hirohashi et al described a new role for NK cells in chronic allograft rejection in which donor-specific antibodies interact with NK cells in the absence of complement and trigger transplant vasculopathy (Hirohashi et al 2012;Li and Baldwin 2012).…”

Cytotoxicity and antibody binding by flow cytometry: A single assay to simultaneously assess two parameters

“…Other investigators previously explored the possibility of a cFCXM for HLA antibodies (8,9). However, those studies were limited in scope.…”

“…Schönemann et al (8) then developed a cell-based complement dependent flow cytometric assay that correlated with the results of conventional cytotoxicity. Subsequently, Won et al (9) made the first attempt to combine antibody binding and cytotoxicity into a single flow cytometric assay. In both of those studies, analysis was restricted to sera that fixed complement by conventional cytotoxicity.…”

“…In both of those studies, analysis was restricted to sera that fixed complement by conventional cytotoxicity. Furthermore, the assays from both studies (8,9) were not optimized for routine clinical application. In this study, sera selected for analysis were based on two criteria, such as (1) antibody specificity as determined by solid phase methodology and (2) a positive FCXM with cells expressing the corresponding HLA antigen(s).…”

Flow cytometry antibody screening using pooled red cells

“…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).…”