The Study Of Tumour Infiltrating Lymphocytes On Malignant Pleural Effusion Of Lung Cancer About TCR Vβ Subfamily Expression And Cytotoxicity In Vitro

Objective:To study the expression characteristics of T cell receptor (TCR) Vβ subfamily abouttumor infiltrating lymphocytes (TIL) on malignant pleural effusion of lung cancer and thechanges of killing TIL with interleukin-2（IL2）in vitro. This study would provide theexperiment basis for clinical immunotherapy.Methods:1. Using attachment method to Isolate TIL and tumor cells of on malignant pleuraleffusion of lung cancer, then the expressions of TCR Vβ subfamilies were detected by flowcytometry (FCM).2. To use IL2to stimulate the TIL on malignant pleural effusion of lung cancer andcounte TIL proliferative index according the routine way in3d,7d,14d,21d,28d,35d,thendetect the expression of CD4,CD8,CD56cells by FCM, tested the cytotoxic activity withMTT method after cultured TIL with tumor cells24hours.Results:1.Comparing with PBMC of the healthy controls,6TCRVβ subfamilies wereidentified in experiment of lung cancer with pleural effusion.The expression of clonal expansive subfamilies TCR including TCRVβ5.2,Vβ5.3,Vβ9,Vβ13.1,Vβ14,Vβ18werehigher than others,It had statistical significance (P<0.05);2.After3days culture with IL2,TIL cells began amplification and peaked value at28days,which multiplying expansion from190±50times.In vitro experiments showed thatthe TIL stimulating with IL2were cytotoxicity significantly greater about70±10%at28d. CD8, CD56cells were signifjcantly higher than other days, which had statisticalsignificance (P<0.05).Conclusions:1. The amound of abnormal and clonal expansion of TIL cells TCRVβ subfamily couldbe found on the lung cancer with malignant pleural effusion, but CD4, CD8TIL cellsexpressed differely.2. TIL cells in vitro by the IL2proliferated obviously and increased acxtivity.It couldprovide the experiment basis for clinical immunotherapy.