AML不同疾病阶段患者T细胞耗竭及相关细胞因子表达水平实验研究

Experimental Study on T Cell Exhaustion and Related Cytokine Expression Levels in Patients with AML at Different Disease Stages

目的探讨急性髓系白血病(acute myeloid leukemia,AML)不同疾病阶段患者T细胞耗竭(T cell exhaustion,Tex)及相关细胞因子的表达水平。方法选择2017年2月~2020年6月接受治疗的AML患者120例,同期选择60例体检健康者为对照组(HC组)。根据AML不同疾病阶段患者可分为初诊(initial diagnosis,ND)45例、完全缓解(complete remission,CR)35例、未缓解(ono-remission,NR)24例和复发(recurrence,HR)16例,采用流式细胞仪检测各组外周血和骨髓不同免疫检查点Tex(PD1+TIM-3+T,PD1+T和TIM-3+T),IL-2和IFN-γ细胞因子的表达水平。结果AML不同疾病阶段患者外周血和骨髓中CD4+PD1+TIM-3+T和CD8+PD1+TIM-3+T细胞水平均高于HC组(F=9.149,6.068,3.438,4.807,均P<0.001),CR患者外周血和骨髓中CD4+PD1+TIM-3+T细胞(t=6.320,8.841,37.420,均P<0.001)和CD8+PD1+TIM-3+T细胞(t=5.417,6.096,12.610,均P<0.001)水平均明显低于ND,NR和HR患者,差异均有统计学意义。AML患者外周血和骨髓中CD8+PD1+TIM-3+T细胞数明显高于CD4+PD1+TIM-3+T细胞数(t=8.227,7.289,均P<0.001),且ND患者骨髓CD4+PD1+TIM-3+T和CD8+PD1+TIM-3+T细胞水平明显高于外周血(t=6.297,7.527,均P<0.001),差异均有统计学意义。ND患者外周血和骨髓中CD4+PD1+T和CD8+PD1+T细胞水平明显高于HC患者(F=13.810,8.029,7.541.10.540,均P<0.001),CR患者外周血和骨髓中CD4+PD1+T细胞(t=4.576,3.164,5.625,均P<0.001)和CD8+PD1+T细胞(t=5.293,3.091,5.091,均P<0.001)明显低于ND和HR患者,差异均有统计学意义。AML患者外周血和骨髓中CD4+TIM-3+T和CD8+TIM-3+T细胞水平明显高于HC组(F=24.230,9.101,16.010,12.540,均P<0.001),且CR患者中CD4+TIM-3+T和CD8+TIM-3+T细胞水平明显低于HR组(t=3.986,4.621,均P<0.001),差异均有统计学意义;HR患者外周血和骨髓中PD1+T细胞和TIM-3+T细胞所产生的IL-2和IFN-γ细胞因子明显低于HC组和ND患者(F=6.218,5.925,5.841,5.017,均P<0.001)。结论AML不同疾病阶段患者的外周血和骨髓中Tex动态变化与其临床特点、缓解复发及预后密切相关。

Objective To investigate the expression levels of T cell exhaustion(Tex)and related cytokines in patients with acute myeloid leukemia(AML)at different disease states.Methods 120 patients with AML who received treatment from February 2017 to June 2020 were selected,and 60 healthy subjects were selected as control group(HC group)during the same period.AML patients at different disease states were divided into initial diagnosis(ND)45 cases,complete remission(CR)35 cases,non-remission(NR)24 cases,and recurrence(HR)16 cases,respectively,the expression levels of Tex expression levels(PD1+TIM-3+T,PD1+T and TIM-3+T),IL-2 and IFN-γcytokines were detected by flow cytometry at different immune checkpoints in each group.Results CD4+PD1+TIM-3+T and CD8+PD1+TIM-3+T cells levels in peripheral blood and bone marrow of patients with different disease stages of AML were higher than those in HC group(F=9.149,6.068,3.438,4.807,all P<0.001).CD4+PD1+TIM-3+T(t=6.320,8.841,37.420,all P<0.001)and CD8+PD1+TIM-3+T(t=5.417,6.096,12.610,all P<0.001)cells levels in peripheral blood and bone marrow of patients with CR were significantly lower than those of patients with ND,NR and HR,the differences were statistically significant,respectively.CD4+PD1+TIM-3+T cells in peripheral blood and bone marrow were significantly higher than CD4+PD1+TIM-3+T cells(t=8.227,7.289,all P<0.001),and CD4+PD1+TIM-3+T and CD8+PD1+TIM-3+T cells levels in bone marrow of newly diagnosed AML patients were significantly higher than those in peripheral blood(t=6.297,7.527,all P<0.001),the differences were statistically significant,respectively.CD4+PD1+T and CD8+PD1+T cells in peripheral blood and bone marrow of ND patients were significantly higher than those in HC patients(F=13.810,8.029,7.541,10.540,all P<0.001),CD4+PD1+T cells in peripheral blood and bone marrow of patients with CR(t=4.576,3.164,5.625,all P<0.001)and CD8+PD1+T cells(t=5.293,3.091,5.091,all P<0.001)were significantly lower than those in ND and HR group,the differences were statistically significant,respectively.The levels of CD4+TIM-3+T and CD8+TIM-3+T cells in peripheral blood and bone marrow of AML patients were significantly higher than those of HC patients(F=24.230,9.101,16.010,12.540,all P<0.001),and CD4+TIM-3+T and CD8+TIM-3+T cells in CR patients were significantly lower than those in HR group(t=3.986,4.621,all P<0.001),the differences were statistically significant,respectively.The IL-2 and IFN-γcytokines secretion of PD1+T and TIM-3+T cells levels in peripheral blood and bone marrow of HR patients were significantly lower than those of HC group and ND patients(F=6.218,5.925,5.841,5.017,all P<0.001).Conclusion Tex dynamic changes in peripheral blood and bone marrow at different disease states were closely related to clinical characteristics,remission of recurrence and prognosis of AML patients.