γ链细胞因子对慢性乙型肝炎患者CD8^+T细胞上TIM-3表达的调节

Regulatory effect of gamma-chain cytokines on expression of TIM-3 on CD8^+T cells in patients with chronic hepatitis B

目的通过检测T细胞免疫球蛋白黏蛋白分子(TIM-3)在慢性乙型肝炎(CHB)患者外周血单个核细胞(PBMC)CD8+T细胞上的表达,探讨共用γ链细胞因子对CHB患者CD8+T细胞上TIM-3表达的影响。方法选取2014年1-5月在第四军医大学唐都医院传染科就诊的CHB初治患者15例,以及健康体检者8例。抽取两组全血,利用Ficoll密度梯度离心法分离出PBMC。分别给予γ链细胞因子白细胞介素(IL)2、IL-7、IL-15、IL-21和人抗CD3/CD28刺激,未刺激孔作为阴性对照。培养4 d后,用单克隆抗体染色,采用流式细胞仪检测CD8+T细胞上TIM-3的表达情况。计量资料比较采用成组t检验。结果与未刺激组相比,TIM-3在CD8+T细胞上表达如下:人抗CD3/CD28和IL-2、IL-15、IL-7刺激组显著升高,分别为(9.629±9.916)%,P=0.000 1;(3.817±2.694)%,P=0.000 6;(5.772±4.732)%,P=0.005 4;(3.560±2.045)%,P=0.030 2。IL-21刺激组表达虽有所增加,为(2.503±2.117)%,但差异无统计学意义,P=0.934 1。结论人抗CD3/CD28、γ链细胞因子中的IL-2、IL-7和IL-15都能够有效上调TIM-3在CHB患者CD8+T细胞上的表达,提示通过抑制它们不仅可以下调TIM-3的表达,而且可能会增强CHB患者体内CD8+T细胞的杀伤作用。

Objective To measure the expression of T - cell immunoglobulin - and mucin domain - 3 - containing molecule 3 （ TIM - 3 ） on CD8 ^＋ T cells in peripheral blood mononuclear cells （PBMCs） among patients with chronic hepatitis B （ CHB ） and to investigate the effect of common gamma - chain cytokines on the expression of TIM - 3 on CD8^ ＋ T ceils in these patients. Methods Fifteen previously un- treated patients with CHB who visited the Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, from Janu- ary to May, 2014, as well as 8 healthy controls, were included in the study. Blood was collected from these subjects, and PBMCs were isolated from blood by Ficoll density gradient centrifugation. PBMCs were separately stimulated with gamma - chain cytokines, interleukin （IL）2, IL - 7, IL - 15, and IL - 21, and anti - CD3/CD28, and the untreated cells were used as a negative control. After four days of culture, PB- MCs were stained with monoclonal antibody. Flow cytometry was used to measure the expression of TIM - 3 on CD8 ^＋ T cells. Comparison of continuous data was made by independent - samples t test. Results Compared with the untreated group, the anti - CD3/CD28, IL - 2, IL - 15, and IL -7 groups had significantly increased expression of TIM - 3 on CD8^ ＋ T cells （9. 629%±9. 916% , P =0. 000 1 ; 3. 817% ±2. 694%, P = 0.000 6 ; 5. 772% ± 4. 732%, P = 0. 005 4 ; 3. 560% ±2. 045%, P = 0. 030 2）, while the IL - 21 group had nonsig- nifieantly increased expression of TIM-3 on CD8 ^＋ T cells （2. 503%±2. 117%, P = 0. 934 1 ）. Conclusion Anti- CD3/CD28 and gamma - chain cytokines IL - 2, IL - 7, and IL - 15 can effectively upregulate the expression of TIM - 3 on CD8^ ＋ T cells in patients with CHB. It indicates that the inhibition of them can not only reduce the expression of TIM - 3, but also may enhance the killing function of CD8 ＋^ T cells in patients with CHB.