ELISpot: a new tool for detecting human HLA--I antibody

B cell lysate was applied as source of HLA antigen. When antigen concentration was 0.25 mg/ml, the spot's frequency was the highest among the other three concentrations (0.125, 0.5 and 1.0 mg/ml, respectively, P 0.05). When B cell lysate was applied as source of HLA antigen and concentration was 0.25 mg/ml, coated antibody's concentration was 1.8 mg/ml and dilution was 1:125, and B cell incubation time was 24 h, the established ELISpot assays represent a sensitive, specific and reliable tool for detecting human HLA class I antibodies.