Methodological aspects of anti-human leukocyte antigen antibody analysis in solid organ transplantation

Abstract

Donor human leukocyte antigen (HLA)-specific antibodies (DSA) play an important role in solid organ transplantation. Preexisting IgG isotype DSA are considered a risk factor for antibody mediated rejection, graft failure or graft loss. The post-transplant development of DSA depends on multiple factors including immunogenicity of mismatched antigens, HLA class Ⅱ typing of the recipient, cytokine gene polymorphisms, and cellular immunoregulatory mechanisms. De novo developed antibodies require special attention because not all DSA have equal clinical significance. Therefore, it is important for transplant clinicians and transplant immunologists to accurately characterize DSA. In this review, the contemporary immunological techniques for detection and characterization of anti-HLA antibodies and their pitfalls are described.

Detection of Anti-HLA Antibodies by Flow Cytometer

“…These new epitopes which are not present on cell surface in vivo, may lead to false positive results. Thus, HLA antibodies determined by SAB technique should be confirmed by cellular techniques [19,33].…”

“…β 2 -microglobulin is required for heavy chain and antigenic peptide association. This molecule stabilizes heavy chain connection (Figure 2A) [17][18][19].…”

“…This variable region is formed by two distinct chains. It can bind to peptides that are more than 11 amino acids in length in order to present them to CD4 + T H cells (Figure 2B) [17][18][19].…”

“…The very first experiments on FCXM method for transplants were initiated in 1983 by Goravay. Bray [19] had significant contributions to the development of multi-colored FCXM method [50][51][52].…”

“…At the end of the assay, a fluorescence peak is generated, which indicates a positive result (binding of antibody to HLA antigens on the beads). We will now examine each of these tests [19].…”