Single Cell RNA-seq on Human PBMC

To assess the impact of IgG1 and IgG4 variants of a TIGIT-Fc-LIGHT fusion protein on human PBMC expression, each fusion protein or an untreated control were incubated with human PBMC for 2 days in AIMV lymphoproliferative media. Both IgG1 and IgG4 variants of TIGIT-Fc-LIGHT induced broad immune cell activation on T, NK, and myeloid cells. Human PBMC were purchased from StemCell Technologies and cultured either alone or with 150nM of human TIGIT-Fc(IgG1)-LIGHT or TIGIT-Fc(IgG4)-LIGHT fusion proteins for 2 days in AIMV media. After 2 days, floating and adherent cells were isolated and processed using the 10x Genomics Chromium Handler. Single cell libraries were generated using 10x Genomics Chromium Next GEM Single Cell 3’ v3.1: Dual Index reagents. Libraries were sequenced on an Illumina NovaSeq 6000.