Regulation Of CD8~+T Cell Memory By Sorcin

ObjectiveThis study aimed to dissect whether and how Sorcin regulates memory CD8~+T cell formation.The results may provide new targets for tumor immune therapy.MethodsThe phenotypes of CD8~+T cells in C57BL/6(wild-type)and Sorcin-/-mi ce were detected by flow cytometry,including activation,proliferation,apop tosis,T cell memory.The animal models of subcutaneous transplantation t umor were divided into: Experimental group: Sorcin-/-mice at 6-8 weeks,a nd control group: wild-type mice at 6-8 weeks.The Sorcin-/-mice and WT mice were subcutaneously injected with low concentration of MC-38 cells(mouse colon cancer cells).After 30 days of modeling,the gene knockou t mice and wild-type mice were subcutaneously injected with high concentr ation of MC-38 cells.The tumor size of the two kinds of mice was contin uously observed and record,and the growth curve was drawn.When the vloume exceeded 1500,the tumor was dissected and separated,and the tumor weight was weighed and the curve was drawn.The protein express ion of CD8~+T cells in Sorcin-/-and WT mice was analyzed by Western blot.RNA-seq was used to analyze the differential gene expression between wild type and Sorcin-/-mice.Results1.Western blot results showed that Sorcin expression could be detected in wildtype mice,but not in gene knockout mice.2.The intron of Sorcin gene and the marker gene neomycin were amplified by PCR.2% agarose gel electrophoresis showed that Sorcin gene could be amplified in wild-type mice,but the marker gene neomycin could not be amplified.Sorcin-/-knockout mice could amplify the marker gene neomycin,but the sorcin gene could not be amplified.3.Flow cytometry showed that there was no significant difference in the activation and proliferation of CD8~+T cells between the two groups.4.The apoptosis rate of Sorcin-/-group and control group was not significantly different by flow cytometry.5.Flow cytometry showed that there was no difference in the ratio of CD4 and CD8 between wild-type mice and Sorcin-/-mice,but there was significant difference in the ratio of CD62LhighCD44 highpositive central memory CD8~+T between the two groups.6.In the subcutaneous transplanted tumor model,Sorcin-/-group compared with the control group,after the first injection,the tumor growth was inhibited after the second injection.7.CD8~+T cells from wild-type and Sorcin-/-mice were sorted and activated by CD3 and CD28 antibody packs in vitro for 24 hours.The differentially expressed genes between the experimental group and the control group were found by RNA sequencing.They were enriched into physiological an d pathological processes.ConclusionsSorcin regulates the formation of central memory CD8 T cells.When tumor cells were inoculated again,sorcin knockout mice had better antitumor effect than wild-type mice.The results may provide new targets for tumor immune therapy.