Transcriptome of human peripheral blood regulatory T cells with distinct ex vivo expansion profile

The UCSF Treg cell therapy program manufactured over 60 human regulatory T cell (Treg) products under the GMP condition in support of 9 clinical trials between 2011 and 2021. These products were manufactured using two alternative protocols of either two rounds of polyclonal stimulations (PolyTregs) or first allogeneic B cell stimulation followed by polyclonal stimulation (arTregs). A high degree of individual variability in Treg expansion with both protocols was observed. RNA-seq analysis was performed on freshly isolated peripheral blood Tregs to identify transcriptome profile associated with Treg expansions. A total of 19 individuals with sufficient archived pre-expansion PBMCs were selected for RNA-seq analysis. Among these, 6 were classified as high expanders, 6 were mid expanders, and 7 were low expanders. Tregs were purfied from cryopreserved PBMCs using FACS based on the phenotype of CD4+CD25+CD127lo/-. Total RNA was extracted for RNAseq.