T淋巴细胞亚群在胶质瘤患者外周血及癌组织中水平及意义

Levels and significances of T lymphocyte subsets in the peripheral blood and cancer tissues of glioma patients

目的 观察胶质瘤患者外周血及癌组织T淋巴细胞亚群水平变化,探讨其在胶质瘤中的作用。方法 2022年12月—2023年8月新疆医科大学第二附属医院诊治胶质瘤患者12例为胶质瘤组(根据WHO分级分为低级别组2例和高级别组10例),均行手术切除治疗,术前采集外周血标本,术中收集癌组织及癌旁正常组织标本。健康体检者11例为对照组,体检日采集外周血标本。采用流式细胞术检测外周血和癌组织、癌旁组织CD3^(+)、CD4^(+)、CD8^(+)T淋巴细胞百分比,计算CD4^(+)/CD8^(+);采用免疫组织化学法检测癌组织Ki-67表达情况。比较胶质瘤组与对照组外周血,胶质瘤患者癌组织与癌旁正常组织、外周血与癌组织,低级别组与高级别组外周血、癌组织T淋巴细胞亚群水平。采用Pearson相关法分析胶质瘤患者癌组织T淋巴细胞亚群与Ki-67表达的相关性。结果 胶质瘤组外周血CD4^(+)T淋巴细胞百分比[(41.41±9.16)%]低于对照组[(50.74±0.69)%](t=3.407,P=0.007),CD3^(+)、CD8^(+)T淋巴细胞百分比及CD4^(+)/CD8^(+)与对照组比较差异均无统计学意义(P>0.05)。胶质瘤患者癌组织CD4^(+)、CD8^(+)T淋巴细胞百分比[(58.04±13.76)%、(16.47±9.56)%]均低于癌旁正常组织[(77.43±24.82)%、(25.48±9.01)%](t=5.516,P=0.004;t=2.157,P=0.007),CD3^(+)T淋巴细胞百分比、CD4^(+)/CD8^(+)与癌旁正常组织比较差异均无统计学意义(P>0.05)。高级别组外周血CD4^(+)T淋巴百分比[(43.95±11.53)%]低于低级别组[(49.60±43.95)%](t=2.422,P=0.009),CD8^(+)T淋巴细胞百分比[(29.63±12.48)%]高于低级别组[(22.30±3.59)%](t=2.645,P=0.016),CD3^(+)T淋巴细胞百分比、CD4^(+)/CD8^(+)与低级别组比较差异均无统计学意义(P>0.05);高级别组癌组织CD4^(+)、CD8^(+)T淋巴细胞百分比[(53.80±11.45)%、(13.90±2.81)%]均低于低级别组[(72.90±3.54)%、(28.30±5.52)%](t=2.576,P=0.003;t=3.579,P=0.008),CD4^(+)/CD8^(+)(3.87±0.04)高于低级别组(2.64±0.64)(t=0.645,P=0.030),CD3^(+)T淋巴细胞百分比与低级别组比较差异无统计学意义(P>0.05)。胶质瘤患者癌组织CD3^(+)、CD8^(+)T淋巴细胞百分比均低于外周血(P<0.05),CD4^(+)T淋巴细胞百分比、CD4^(+)/CD8^(+)均高于外周血(P<0.05)。胶质瘤患者癌组织CD3^(+)T淋巴细胞百分比与Ki-67表达呈负相关(r=-0.669,P=0.024),CD4^(+)T淋巴细胞百分比与Ki-67表达呈正相关(r=0.681,P=0.009),CD8^(+)T淋巴细胞百分比、CD4^(+)/CD8^(+)与Ki-67表达均无相关性(r=-0.497,P=0.120;r=0.162,P=0.634)。结论 胶质瘤患者外周血及癌组织T淋巴细胞亚群水平失调,以CD4^(+)T淋巴细胞百分比降低为主;CD4^(+)T淋巴细胞多分布于癌组织,CD8^(+)T淋巴细胞多分布于外周血;癌组织CD4^(+)T淋巴细胞百分比低的胶质瘤患者肿瘤细胞增殖较快。

Objective To observe the changes of T lymphocyte subsets in the peripheral blood and cancer tissues of glioma patients,and to explore its role in glioma.Methods Twelve patients with glioma(glioma group)were diagnosed and treated in the Second Affiliated Hospital of Xinjiang Medical University from December,2022 to August,2023,and were divided into low-grade group(n=2)and high-grade group(n=10)according to the WHO classification.All patients received surgery,the peripheral blood samples were collected before surgery,and the cancer tissues and adjacent normal tissues were harvested during surgery.Another healthy volunteers were collected the peripheral blood samples on the day of physical examination.The flow cytometry was used to detect the percentages of CD3^(+),CD4^(+)and CD8^(+)T lymphocytes in the peripheral blood,cancer tissues and adjacent normal tissues,and CD4^(+)/CD8^(+)was calculated.Immunohistochemistry was used to detect the expression of Ki-67 in the cancer tissues.The levels of T lymphocyte subsets were compared in the peripheral blood between glioma group and control group,between cancer tissues and adjacent normal tissues and between peripheral blood and cancer tissues in glioma group,and in the peripheral blood and cancer tissues between low-grade group and high-grade group.Pearson correlation method was used to analyze the correlation between T lymphocyte subsets and Ki-67 expression in glioma patients.Results The percentage of CD4^(+)T lymphocytes in the peripheral blood was lower in glioma group[(41.41±9.16)%]than that in control group[(50.74±0.69)%](t=3.407,P=0.007),and there were no significant differences in the percentages of CD3^(+)and CD8^(+)T lymphocytes and the CD4^(+)/CD8^(+)between two groups(P>0.05).The percentages of CD4^(+)and CD8^(+)T lymphocytes were lower in the cancer tissues[(58.04+13.76)%,(16.47±9.56)%]than those in the adjacent normal tissues[(77.43±24.82)%,(25.48±9.01)%]in glioma group(t=5.516,P=0.004;t=2.157,P=0.007),and there were no significant differences in the percentage of CD3^(+)T lymphocytes and CD4^(+)/CD8^(+)(P>0.05).The percentage of CD4^(+)T lymphocytes in the peripheral blood was lower in high-grade group[(43.95±11.53)%]than that in low-grade group[(49.60±43.95)%](t=2.422,P=0.009),the percentage of CD8^(+)T lymphocytes was higher in high-grade group[(29.63±12.48)%]than that in low-grade group[(22.30±3.59)%](t=2.645,P=0.016),and the percentage of CD3^(+)T lymphocytes and CD4^(+)/CD8^(+)showed no significant differences(P>0.05).The percentages of CD4^(+)and CD8^(+)T lymphocytes were lower in high-grade group[(53.80±11.45)%,(13.90±2.81)%]than those in low-grade group[(72.90±3.54)%,(28.30±5.52)%](t=2.576,P=0.003;t=3.579,P=0.008),CD4^(+)/CD8^(+)was higher in high-grade group(3.87±0.04)than that in low-grade group(2.64±0.64)(t=0.645.P=0.030),and there was no significant difference in the percentage of CD3^(+)T lymphocytes between two groups(P>0.05).The percentages of CD3+and CD8^(+)T lymphocytes were lower in the cancer tissues than those in the peripheral blood in glioma group(P<0.05),and the percentage of CD4^(+)T lymphocytes and CD4^(+)/CD8^(+)were higher in the cancer tissues than those in the peripheral blood(P<0.05).The percentage of CD3^(+)T lymphocytes in glioma group was negatively correlated with the expression of Ki-67(r=-0.669,P=0.024),the percentage of CD4^(+)T lymphocytes was positively correlated with the expression of Ki-67(r=0.681,P=0.009),and the percentage of CD8^(+)T lymphocytes and CD4^(+)/CD8^(+)were not correlated with Ki-67(r=-0.497,P=0.120;r=0.162,P=0.634).Conclusions The levels of T lymphocyte subsets in the peripheral blood and cancer tissues of glioma patients are out of balance,which is mainly demonstrated by the decease of percentage of CD4^(+)T lymphocytes.CD4^(+)T lymphocytes are mostly distributed in the cancer tissues,and CD8^(+)T lymphocytes are mostly distributed in the peripheral blood.The cancer cells proliferate more rapidly in glioma patients with a low percentage of CD4+T lymphocytes.