Identification And Clinical Significance Of Tumor Infiltrating Lymphocytes Cell Subsets In Laryngeal Carcinoma

Objective(s):Tumor Infiltrating Lymphocytes(TIL)therapy mainly relies on lymphocytes within or near tumors that have the ability to recognize and kill tumor cells,which are isolated,amplified in vitro and transfused into patients to expand the immune response and enhance the killing effect of tumors.Adoptive Cell Therapy(ACT)has become a hot research topic.This article through in vitro in patients with laryngeal cancer TIL analysis and testing its cells,provide new ideas for laryngeal cancer immunotherapy.Methods:TIL was cultured in vitro from laryngeal cancer tissues,and was stimulated to amplify TIL by adding CD3 antibody and irradiated peripheral blood mononuclear cells.We set TIL group,TIL+CD3 group,TIL+PBMC group,TIL+CD3+PBMC(later referred to as TIL++group)as the control group.Flow cytometry was used to detect the expression of each cell subgroup in different culture time and different feeders.In this study,Graph Pad Prism 8.0.2 was used for data processing and analysis.P<0.05,which indicates that the data difference have statistical significance.Results:1.TIL compared to in vitro culture day 14 more days 0,CD3~+T cells,CD3~+CD~+T cells,Treg cells,Tcm cells,Teff cells and CD137 expression level is on the rise,and the difference is statistically significant(P<0.01,P<0.05,P<0.01,P<0.01,P<0.001,P<0.01,respectively),Tnaive cells and Tem cells expression level is on the decline,and the difference is statistically significant(P<0.05,P<0.05,respectively),TIL compared to the seventh day of in vitro cultured from day 0CD3~+CD4~+T cells expression level is on the decline,Teff cells and PD-1 expression level is on the rise,and the difference is statistically significant(P<0.05,P<0.01,P<0.05,respectively);TIL in vitro culture of 14 days more than 7 days,CD3~+T cells and CD137 expression level is on the rise,and the difference is statistically significant(P<0.05,P<0.01,respectively).2.TIL in vitro cultured T++group compared with TIL group,the expression of Treg cells,Tnaive cells and CD137 level is on the rise,and the difference is statistically significant(P<0.01,P<0.05,P<0.01,respectively),PD-1 expression level is on the decline,and the difference is statistically significant(P<0.05);TIL in vitro cultured T++group compared with TIL+CD3 group,and Tcm cells,Tnaive cells and CD137 expression level is on the rise,and the difference is statistically significant(P<0.01,P<0.05,P<0.01,respectively);TIL the in vitro cultured TIL+PBMC groups compared with TIL group,expression level of Treg cells and CD137 is on the rise,and the difference is statistically significant(P<0.05,P<0.05,respectively).3.7 days TIL cells on the expression of PD-1 is higher than that of 0 days,14days,namely the PD-1 a downward trend after rising first,statistically significant differences(P<0.0001,P<0.001);14 days TIL cells on the expression of CD137 is higher than that of 0,7 days,statistically significant differences(P<0.01,P<0.01).Conclusion(s):1.On the 14th day of in vitro culture,the growth level of cell subsets of TIL showed an increasing trend,compared with 0,7 days for optimal incubation time.2.The growth level of cell subsets in TIL cultured in vitro with CD3 antibody and irradiated PBMC group showed an increased trend.Compared with TIL group,TIL plus CD3 antibody group and TIL plus irradiated PBMC group,it was the optimal culture scheme.3.TIL cells have distinct phenotypic characteristics in vitro culture.These CD3~+CD4~+T,CD3~+CD8~+T,Treg and Tem cell subgroup percentage is higher.