The flow cytometric crossmatch. Dual-color analysis of T cell and B cell reactivities.

摘要: Abstract The flow cytometric crossmatch (FCXM) has become an increasingly utilized method to detect low levels of anti-donor antibodies (e.g., anti-HLA) in potential renal allograft recipients. Anti-donor not apparent the standard complement-dependent crossmatch, but detectable by FCXM, are often associated with increased episodes graft rejection and early failure. In this study we examined several parameters FCXM order establish a standardized methodology. First, observed that optimal staining results were obtained when secondary antibody was Fc-specific F(ab'), anti-human IgG. contrast anti-whole immunoglobulin antibody, anti-Fc specific reagent did react surface on B cells reactive cytophilic present CD16+ cells. Next determined dualcolor analysis superior single-color both for evaluation T cell reactivities discrimination from reactivities. Additionally, dual-color revealed density class I histocompatibility antigens is greater than cells, indicating may be more sensitive target detecting anti-class antibodies. Finally, shift mean fluorescence intensity 10 channels 256-channel, 3-decade log scale indicative positive FCXM. data presented these studies provide basis performing sensitivity specificity.