肾癌患者手术前后免疫功能检测及其临床意义

Immune function test and its clinical significance in patients with renal cell carcinoma before and after surgery

目的探讨肾癌患者手术治疗前后免疫功能检测及其临床意义。方法选取肾癌(RCC)患者240例,采用TNM临床分期诊断标准进行分组,其中A组:局限型1(Ⅰ期)、B组:局限型2(Ⅱ期)、C组:局部进展型(Ⅲ期)和D组:转移型(Ⅳ期)4组,每各60例。A组、B组、C组均经根治性手术治疗,D组中48例经手术切除,12例经穿刺病检,病理结果均为肾细胞癌。手术患者术前未做免疫及靶向治疗。对照组80名健康成人。所有患者及对照组均晨起空腹抽取外周静脉血,采血时间分为2次,手术治疗前1周,手术治疗后1个月。流式细胞仪检测免疫相关指标。结果 (1)术前T细胞亚群水平:A组肾癌患者与对照组相比,T细胞亚群CD3^+、CD4^+细胞百分率、CD4^+/CD8^+比值、CD8^+细胞百分率差异无统计学意义(P>0.05)。B组、C组和D组肾癌患者与对照组相比T细胞亚群CD3^+、CD4^+细胞百分率、CD4^+/CD8^+比值明显低于对照组,CD8^+细胞百分率明显高于对照组(P<0.05),B组、C组和D组间相比T细胞亚群水平差异无统计学意义(P>0.05)。(2)术前外周血CD4^+T(Th)细胞分泌细胞因子水平:A组肾癌患者与对照组相比,外周血CD4^+T细胞分泌的各细胞因子差异无统计学意义(P>0.05)。B组、C组和D组肾癌患者与对照组相比,外周血Th1细胞中干扰素(IFN)-γ、白细胞介素(IL)-2、肿瘤坏死因子(TNF)-α表达水平均降低,差异有统计学意义(P<0.01)。外周血Th2细胞中IL-4明显升高(P<0.05),IL-6也明显升高(P<0.01),差异均有统计学意义。B组、C组和D组间相比CD4^+T细胞分泌的各细胞因子水平差异无统计学意义(P>0.05)。(3)术前外周血Treg表达水平:A组、B组肾癌患者与对照组相比,外周血Treg差异无统计学意义(P>0.05)。C组、D组与对照组相比,外周血Treg表达明显增高(P<0.01),C组、D组与A、B组间相比Treg表达明显增高(P<0.01)。A组与B组间及D组与C组间相比Treg表达差异无统计学意义(P>0.05)。(4)术后A组与术前对比差异无统计学意义。B组、C组肾癌患者术后与术前对比,T细胞亚群CD3^+、CD4^+细胞百分率、CD4^+/CD8^+比值、CD8^+细胞百分率、外周血CD4^+T(Th)细胞分泌细胞因子水平、Treg变化不大(P>0.05)。结论 (1)肾癌患者细胞免疫功能状态与其TNM分期有关,Ⅱ期、Ⅲ期及Ⅳ期细胞免疫功能明显降低。(2)Ⅱ期、Ⅲ期肾癌患者术前细胞免疫功能明显低下,根治手术后短期内免疫功能改善不明显,可以行免疫治疗。

Objective To investigate the immune function in patients with renal cell carcinoma（RCC） before and after surgery and its clinical significance. Methods Two hundred and forty RCC patients were enrolled and divided into four groups according to TNM clinical staging（n=60 each）： group A（local type 1, stage Ⅰ）, group B（local type 2, stage Ⅱ）, group C（local progressive type, stage Ⅲ） and group D（metastatic type, stage Ⅳ）. Patients in groups A through C were treated with radical operation. In group D, 48 patients underwent surgery while the other 12 under-went biopsy only and were confirmed to have RCC by pathological study. Before surgery, all patients did not receive immunotherapy or targeted therapy. A control group was set including 80 healthy adults. All patients and healthy controls were collected for fasting peripheral blood in the morning on two occasions： one week prior to surgery and at one month after surgery. Flow cytometry was used to detect immune related indicators in the blood samples. Results（1）The level of T lymphocyte subsets before surgery： there were no significant differences in the percentages of CD3^＋,CD4^＋, CD4^＋/CD8^＋ratio and CD8^＋cells between group A and healthy controls（P〈0.05）. Compared with the healthy controls, the percentages of CD3^＋, CD4^＋cells and CD4^＋/CD8^＋of T cell subsets were significantly lower, and the percentage of CD8^＋cells was significantly higher in groups B, C and D（P〈0.05）. There was no significant difference in T cell subsets between groups B, C and D（P〈0.05）.（2）The level of CD4^＋T（Th）-secreted cytokines in peripheral blood before surgery： compared with healthy control group, the level of cytokines secreted by peripheral blood CD4^＋T cells in group A did not show any significant difference（P〈0.05）. Compared with the control group, groups B, C and D showed significantly lower levels of IFN-γ, IL-2 and TNF-α in peripheral Th1 cells（P〈0.01）, and significantly increased levels of IL-4（P〈0.05） and IL-6 in peripheral Th2 cells（P〈0.01）. There were no significant differences in the levels of CD4^＋T-secreted cytokines between groups B, C and D（P〈0.05）.（3）Treg expression levels in peripheral blood before surgery： compared with the healthy control group, the expression level of peripheral Treg in groups A and B did not show any significant difference（P〈0.05）. Compared with healthy control group, the Treg expression level in peripheral blood was significantly increased in groups C and D（P〈0.01）. The expression level of Treg in groups C and D was significantly higher than that in groups A and B（P〈0.01）. There was no significant difference in Treg expression level between groups A and B or between groups C and D（P〈0.05）.（4）After surgery, group A did not show any change in these findings, while groups B and C showed modest but not significantly changes（ P〈0.05） in percentages of CD3^＋, CD4^＋cells, CD4^＋/CD8^＋ratio, CD8^＋cells, level of CD4^＋T（Th）-secreted cytokines and Treg expression（P〈0.05）. Conclusions（1）The status of cellular immune function in RCC patients is related to TNM staging. Patients with stages Ⅱ, Ⅲ or Ⅳ RCC have significantly lowered cellular immune function.（2）The cellular immune function in patients with stages Ⅱ or Ⅲ RCC is significantly lowered before surgery, and does not show a short-term improvement even after radical operations, suggesting the need for subsequent immunotherapy.