Specificity of Human Lymphocytes Primed against Allogeneic Cells in vitro. I Optimalization of Culture Conditions for Detection of HLA—D Specificity

Abstract

The specificity of lymphocytes primed against allogeneic cells in vitro was examined in order to find the optimal conditions for detection in the restimulating response of HLA-D specificity, as defined by homozygous cell typing. Reducing the number of stimulating cells (S) to less than the number of responding cells (R) (ratio R/S = 4:1) in the priming culture, and increasing the number of stimulating cells at restimulation gave better discrimination. Priming with HLA-D heterozygous stimulating cells, where the R and S shared the other HLA-D determinant, rendered the primed cells somewhat more discriminatory than priming with HLA-D homozygous cells. The optimal length of the secondary culture was about 38 h.