Single cell (sc) RNAseq reveals transcriptional states of transgenic MAGE-A1 specific TCR-T CD4+ and CD8+ cells in NSG tumor model

We have studied the efficacy of MAGE-A1 specific transgenic T cells against A375F NSG model by flow cytometry and IHC analysis. We then employed a scRNAseq approach to characterize the transcriptional states of transgenic CD4+ and CD8+ TCR-T cells that infiltrated into A375F tumor by extracting these cells from tumors. This scRNAseq study allow us to compare the transcriptional differences between our groups ( T-TCR-MA1-CD8ab, T-TCR-MA1-CD8/28) Overall design: CD4+ and CD8+ T-TCR-MA1-CD8ab, T-TCR-MA1-CD8/28 were infused into in vivo murine models with A375F tumor cell line as described in methods. Four of the following group were compared: CD4+ T-TCR-MA1-CD8ab only , CD8+ T-TCR-MA1-CD8ab only, CD4+ and CD8+ T-TCR-MA1-CD8ab combo, and CD4+ and CD8+ T-TCR-MA1-CD8/28 combo. Tumors (n=6) were digested and CD4+ and CD8+ T cells were sorted from single cell suspension. CD4+ only and CD8+ only T-TCR-MA1-CD8ab (pooled as one sample) (NSG_CD8_4), CD4+ and CD8+ T-TCR-MA1-CD8ab combo(NSG_WT), and CD4+ and CD8+ T-TCR-MA1-CD8/28 combo(NSG_G) were then continued for scRNA seq library prep.