Modified Shenlingbaizhu Decoction inhibits colon cancer by enhancing the interaction between memory-like Tfh and B cells

SummarySingle-cell RNA sequencing (scRNA-seq) was performed on mouse colon tumor tissues collected on Day 10 (three samples per group). After quality control, 114,653 cells were clustered into 35 distinct populations, which were further classified into 10 major subsets (CD8T cells, CD4T cells, myeloid cells, B cells, NK cells, endothelial cells, fibroblasts, proliferative cells, epithelial cells, and tumor cells). Subsequently, CD4T, CD8T, and myeloid subsets were defined in detail. Ultimately, we discovered and proved that CXCR5+CD62L+ Tfh cells, together with B cells, played a significant role in the treatment of colon cancer in mice with Modified Shenlingbaizhu Decoction (MSD).Experimental designWe performed single cell RNA-seq on Model, MSD, anti-CTLA4 (positive control), and M+anti-CTLA4 (combination therapy) group, each group consisted of three samples. After dissection and tissue digestion, the tumor tissues were dissociated into single-cell suspensions. The qualified cell suspension was generated by the droplet microfluidic device of DNBelab C4 to realize the separation of single cells. mRNA molecules were recovered by demulsification after droplet generation, and cDNA was synthesized by reverse transcription of mRNA molecules, and then sequencing was performed.