Tonsillar HLACs provide a good source of cells for panel validation because T cell antigens are often differentially expressed in B cells, and tonsils provide an abundant source of both T and B cells (
Cavrois et al., 2017). Shown are data representative of one of 10 independent donors. (
A) As demonstrated schematically in the upper left, the top population of each plot corresponds to T cells (CD3+) while the bottom population corresponds to B cells (CD3-). Each antibody in the panel was validated by standard two-dimensional plots showing expression of CD3 (y-axis) versus each indicated antibody (x-axis). The observed expression patterns are consistent with known expression patterns on T cells and B cells. Results are gated on live, singlet cells. (
B) Some antigens were further validated by demonstrating differential expression in naïve (Tn) vs. memory (Tm) CD4+ T cells. As demonstrated schematically on the left, in the first row the top population of each plot corresponds to Tm cells while the bottom population corresponds to Tn cells, while in the second row the top population of each plot corresponds to Tn cells while the bottom population corresponds to Tm cells. Results s are gated on live, singlet, CD3+CD8- cells.