Next Generation Sequencing Facilitates Quantitative Analysis of Tumor Infiltrating CD8+T Cells Transcriptomes From Control and Manganese(II) Treated C57BL/6 Wild Type Mice

Purpose: The goals of this study are to compare the differentially expressed genes of tumor infiltrating CD8+T cells between control and Manganese(II) treated mice. We mainliy fouced on the differentiation of trascription and effector related genes. Methods:C57BL/6 mice were injected subcutaneously in the right flank with B16-F10 and divided into two groups followed by treatment with control (saline) or Manganese(II) respectively. After 14days, mice were euthanized with CO2 and tumor infiltrating CD8+T cells were isolated by FACS. Then RNA of sorted CD8+T cells were generated by deep sequencing using BGISEQ-500. Clean tags were mapped to the mouse genome database (GCF_000001635.25_GRCm38.p5) by HISAT (hierarchical indexing for splicing alignment for transcripts) software. Results: Using an optimized data analysis workflow, we mapped about 60 million sequence reads per sample to the mouse genome (GCF_000001635.25_GRCm38.p5). The significance of the differential expression of genes was defined by the bioinformatics service of Beijing Genomics Institute according to the combination of the absolute value of log2-Ratio = 1 and false discovery rate = 0.001. Conclusions: Manganese(II) enhances the function of tumor infiltrating CD8+T cells. Overall design: mRNA profiles of tumor infiltrating CD8+T cells from control and Manganese (II) treated mice were generated by deep sequencing using BGISEQ-500.