中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
2期
85-88
,共4页
施敏骅%邢玉斐%张增利%黄建安%陈永井
施敏驊%邢玉斐%張增利%黃建安%陳永井
시민화%형옥비%장증리%황건안%진영정
肺肿瘤%程序性死亡配体1%T淋巴细胞%细胞增殖%细胞免疫%免疫逃逸
肺腫瘤%程序性死亡配體1%T淋巴細胞%細胞增殖%細胞免疫%免疫逃逸
폐종류%정서성사망배체1%T림파세포%세포증식%세포면역%면역도일
Lung neoplasms%PD-L1%T-lymphocytes%Cell proliferation%Cellular immunity%Immune escape
目的 观察可溶性程序性死亡配体1(sPD-L1)在不同肺癌细胞中的表达特性,并探讨其对人T淋巴细胞生物学功能的影响.方法 采用流式细胞术检测肺癌细胞表面膜型PD-L1(mPD-L1)和人外周血T淋巴细胞表面程序性死亡受体1(PD-1)的表达水平,采用酶联免疫吸附试验(ELISA)检测肺癌细胞分泌的sPD-L1的表达,采用CCK-8法检测mPD-L1和sPD-L1对T细胞增殖的抑制作用.结果 PD-1在人外周血静止T淋巴细胞中的表达水平为(16.08±2.28)％,而在活化T淋巴细胞中的表达水平为(78.06±7.21)％.mPD-L1在H1299、HO8910、SPAC-1、H460和H446细胞中的表达水平分别为(78.34±10.25)％、(68.17±11.56)％、(45.32±7.98)％、(47.52 ±9.62)％和(40.95±8.56)％,而在A549细胞中的表达水平为(16.02±6.28)％.sPD-L1在H1299、HO8910、SPAC-1、H460和H446细胞中的表达水平分别是(0.17±0.01) ng/ml、(0.30±0.03) ng/ml、(0.59±0.03) ng/ml、(0.34±0.02)ng/ml和(0.57±0.03) ng/ml,而在A549细胞中不表达.CCK-8体外实验结果显示,活化T淋巴细胞+ H1299细胞+mIgG组和活化T淋巴细胞+H1299细胞+α-PD-L1组的A值分别为0.92±0.09和1.28±0.13,差异有统计学意义(P＜0.05).活化T淋巴细胞+H1299细胞上清+ α-PD-L1组和活化T淋巴细胞+H1299细胞上清+mIgG组的A值分别为0.92±0.03和0.68±0.03,差异有统计学意义(P＜0.05).结论 肺癌细胞表达的mPD-L1和sPD-L1均可有效抑制T淋巴细胞增殖,发挥细胞免疫的负性调节作用,这可能导致肿瘤抗原特异性T淋巴细胞失活和肿瘤细胞免疫逃逸.
目的 觀察可溶性程序性死亡配體1(sPD-L1)在不同肺癌細胞中的錶達特性,併探討其對人T淋巴細胞生物學功能的影響.方法 採用流式細胞術檢測肺癌細胞錶麵膜型PD-L1(mPD-L1)和人外週血T淋巴細胞錶麵程序性死亡受體1(PD-1)的錶達水平,採用酶聯免疫吸附試驗(ELISA)檢測肺癌細胞分泌的sPD-L1的錶達,採用CCK-8法檢測mPD-L1和sPD-L1對T細胞增殖的抑製作用.結果 PD-1在人外週血靜止T淋巴細胞中的錶達水平為(16.08±2.28)％,而在活化T淋巴細胞中的錶達水平為(78.06±7.21)％.mPD-L1在H1299、HO8910、SPAC-1、H460和H446細胞中的錶達水平分彆為(78.34±10.25)％、(68.17±11.56)％、(45.32±7.98)％、(47.52 ±9.62)％和(40.95±8.56)％,而在A549細胞中的錶達水平為(16.02±6.28)％.sPD-L1在H1299、HO8910、SPAC-1、H460和H446細胞中的錶達水平分彆是(0.17±0.01) ng/ml、(0.30±0.03) ng/ml、(0.59±0.03) ng/ml、(0.34±0.02)ng/ml和(0.57±0.03) ng/ml,而在A549細胞中不錶達.CCK-8體外實驗結果顯示,活化T淋巴細胞+ H1299細胞+mIgG組和活化T淋巴細胞+H1299細胞+α-PD-L1組的A值分彆為0.92±0.09和1.28±0.13,差異有統計學意義(P＜0.05).活化T淋巴細胞+H1299細胞上清+ α-PD-L1組和活化T淋巴細胞+H1299細胞上清+mIgG組的A值分彆為0.92±0.03和0.68±0.03,差異有統計學意義(P＜0.05).結論 肺癌細胞錶達的mPD-L1和sPD-L1均可有效抑製T淋巴細胞增殖,髮揮細胞免疫的負性調節作用,這可能導緻腫瘤抗原特異性T淋巴細胞失活和腫瘤細胞免疫逃逸.
목적 관찰가용성정서성사망배체1(sPD-L1)재불동폐암세포중적표체특성,병탐토기대인T림파세포생물학공능적영향.방법 채용류식세포술검측폐암세포표면막형PD-L1(mPD-L1)화인외주혈T림파세포표면정서성사망수체1(PD-1)적표체수평,채용매련면역흡부시험(ELISA)검측폐암세포분비적sPD-L1적표체,채용CCK-8법검측mPD-L1화sPD-L1대T세포증식적억제작용.결과 PD-1재인외주혈정지T림파세포중적표체수평위(16.08±2.28)％,이재활화T림파세포중적표체수평위(78.06±7.21)％.mPD-L1재H1299、HO8910、SPAC-1、H460화H446세포중적표체수평분별위(78.34±10.25)％、(68.17±11.56)％、(45.32±7.98)％、(47.52 ±9.62)％화(40.95±8.56)％,이재A549세포중적표체수평위(16.02±6.28)％.sPD-L1재H1299、HO8910、SPAC-1、H460화H446세포중적표체수평분별시(0.17±0.01) ng/ml、(0.30±0.03) ng/ml、(0.59±0.03) ng/ml、(0.34±0.02)ng/ml화(0.57±0.03) ng/ml,이재A549세포중불표체.CCK-8체외실험결과현시,활화T림파세포+ H1299세포+mIgG조화활화T림파세포+H1299세포+α-PD-L1조적A치분별위0.92±0.09화1.28±0.13,차이유통계학의의(P＜0.05).활화T림파세포+H1299세포상청+ α-PD-L1조화활화T림파세포+H1299세포상청+mIgG조적A치분별위0.92±0.03화0.68±0.03,차이유통계학의의(P＜0.05).결론 폐암세포표체적mPD-L1화sPD-L1균가유효억제T림파세포증식,발휘세포면역적부성조절작용,저가능도치종류항원특이성T림파세포실활화종류세포면역도일.
Objective To explore the expression of soluble programmed death ligand-1 on lung cancer cells and to clarify its biological function through PD-1/PD-L1 pathway in regulating the function of T lymphocytes.Methods Labeled monoclonal antibody and flow cytometry were used to analyze the expression of PD-L1 and its receptor PD-1 on lung cancer cells and human T lymphocytes,respectively.The level of sPD-L1 in the supernatant of lung cancer cells was determined with an ELISA kit.The inhibition of proliferation of T lymphocytes by mPD-L1 and sPD-L1 was studied using CCK-8 incorporation.Results Low or no expression [(16.08 ± 2.28) ％] of PD-1 was found on resting T lymphocytes from human peripheral blood with flow cytometry,but up-regulated expression of PD-1 [(78.06 ±7.21)％] was found on the surface of activated T lymphocytes.Soluble PD-L1 was found in supernatant of some lung cancer cell lines,such as H1299,H08910,SPCA-1,H460,H446 cells,with PD-L1 expressing on their cell surface [(78.34±10.25)％,(68.17 ± 11.56)％,(45.32 ±7.98)％,(47.52 ±9.62)％ and (40.95 ±8.56)％,respectively],but very low expression on A549 cells [(16.02 ± 6.28) ％].The level of mPD-L1 on H1299 cells was highest [(78.34 ± 10.25) ％],compared with H08910 cells (68.17 ± 11.56) ％,SPCA-1 cells (45.32 ± 7.98) ％,H446 cells (40.95 ± 8.56) ％,and H460 cells (47.52 ± 9.62) ％.At the same time,the sPD-L1 level on H1299 cells was low [(0.17 ± 0.01) ng/ml],compared with HO8910 cells (0.30 ± 0.03) ng/ml,SPCA-1cells (0.59 ±0.03) ng/ml,H446 cells (0.34 ±0.02)ng/ml,and H460 cells (0.57 ±0.03)ng/ml,but not expressed on A549 cells.PD-L1 expressing H1299 cells inhibited the proliferation of T lymphocytes in the coculture system.Supernatant of the cultured PD-L1 + lung cancer cells also inhibited T cell proliferation.Antihuman PD-L1 blocking antibody could partly restore the proliferation capacity of T lymphocytes.Conclusions Membrane-bound PD-L1 and soluble PD-L1 released from lung cancer cells can effectively inhibit the proliferation of T lymphocytes in mixed culture system and down-regulate cell-mediated immunity in vitro.This may lead to inactivation of tumor antigen-specific T cells and immune escape of lung cancer cells.
