干扰素疗效与慢性乙型肝炎患者外周血CD8<sup>+</sup> T细胞程序性死亡受体-1和T细胞免疫球蛋白黏蛋白分子3表达的关系

doi:10.3877/cma.j.issn.1674-1358.2018.01.008

中华实验和临床感染病杂志(电子版) ›› 2018, Vol. 12 ›› Issue (01) : 40 -45. doi: 10.3877/cma.j.issn.1674-1358.2018.01.008

所属专题：文献；

临床论著

干扰素疗效与慢性乙型肝炎患者外周血CD8⁺ T细胞程序性死亡受体-1和T细胞免疫球蛋白黏蛋白分子3表达的关系

王海燕¹, 王银铃¹, 朱莉¹, 陈慧¹, 钱峰¹, 李明¹, 朱伟¹, 张雪华¹, 胥萍¹, 朱传武¹^,^†(

)

^1. 215007　苏州市，苏州市第五人民医院肝病科

收稿日期:2017-03-20 出版日期:2018-02-15
通信作者: 朱传武
基金资助:
苏州市科技局和苏州市卫生局兴卫基金项目(No. KJXW2011031); 苏州市感染性疾病临床医学中心项目(No. SZZX201508); 苏州市临床重点病种诊疗项目(No. LCZX201315); 江苏省卫生计生委面上项目(No. 2017068)

Relationship between interferon-alpha efficacy and the expression of programmed death 1 and T-cell immunoglobulin and mucin domain-containing molecule 3 on peripheral blood CD8⁺ T cells in patients with chronic hepatitis B

Haiyan Wang¹, Yinling Wang¹, Li Zhu¹, Hui Chen¹, Feng Qian¹, Ming Li¹, Wei Zhu¹, Xuehua Zhang¹, Ping Xu¹, Chuanwu Zhu¹^,^†()

^1. Department of Hepatology, The Fifth People’s Hospital of Suzhou, Suzhou 215007, China

Received:2017-03-20 Published:2018-02-15
Corresponding author: Chuanwu Zhu
About author:
Corresponding author: Zhu Chuanwu, Email: zhuchw@126.com

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引用本文：

王海燕, 王银铃, 朱莉, 陈慧, 钱峰, 李明, 朱伟, 张雪华, 胥萍, 朱传武. 干扰素疗效与慢性乙型肝炎患者外周血CD8⁺ T细胞程序性死亡受体-1和T细胞免疫球蛋白黏蛋白分子3表达的关系[J]. 中华实验和临床感染病杂志(电子版), 2018, 12(01): 40-45.

Haiyan Wang, Yinling Wang, Li Zhu, Hui Chen, Feng Qian, Ming Li, Wei Zhu, Xuehua Zhang, Ping Xu, Chuanwu Zhu. Relationship between interferon-alpha efficacy and the expression of programmed death 1 and T-cell immunoglobulin and mucin domain-containing molecule 3 on peripheral blood CD8⁺ T cells in patients with chronic hepatitis B[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2018, 12(01): 40-45.

目的

探讨干扰素抗病毒疗效与慢性乙型肝炎（CHB）患者外周血CD8⁺T细胞程序性死亡受体1（PD-1）和T细胞免疫球蛋白黏蛋白分子3（Tim3）表达的关系。

方法

以72例HBeAg阳性CHB患者作为研究对象，分别采集干扰素治疗前和治疗12个月的抗凝外周静脉血，同期以30例健康人作为研究对照，用流式细胞技术检测CD8⁺T细胞PD-1和Tim3的表达。

结果

干扰素治疗前，CHB患者外周血CD8⁺T细胞PD-1和Tim3表达的中位百分率分别为35.5%和4.8%，均显著高于健康对照组（29.3%和3.6%），差异均具有统计学意义（U= 805.0、P = 0.043，U= 741.5、P= 0.013）。干扰素治疗12个月时，患者CD8⁺T细胞PD-1和Tim3的表达分别为32.7%和4.1%，较治疗前显著下降，差异具有统计学意义（Z= 3.305、P= 0.001，Z= 2.065、P = 0.039）；在获得HBeAg血清学转换者中，CD8⁺T细胞PD-1和Tim3表达的中位百分率均显著低于无转换者，差异均具有统计学意义（U= 209.0、P <0.001，U = 302.0，P <0.001）；在ALT复常与异常者之间，两项指标的中位百分率差异无统计学意义（U= 229.5、P = 0.635，U= 209.5、P= 0.405）；在HBV DNA载量低于检测下限者中，CD8⁺T细胞PD-1表达的中位百分率显著低于病毒仍可检测的患者（U= 371.5、P= 0.011），而CD8⁺T细胞Tim3的表达两者差异无统计学意义（U= 558.0、P= 0.727）。

结论

CHB患者外周血CD8⁺T细胞PD-1和Tim3呈高表达状态，干扰素治疗可以下调其表达，并且其表达降低与干扰素疗效有关。

关键词: 肝炎，乙型，慢性, 干扰素, 外周血CD8⁺T淋巴细胞, 程序性死亡受体-1, T细胞免疫球蛋白黏蛋白分子3

Objective

To investigate the association of interferon-alpha (IFN-α) antiviral efficacy and the expression of programmed death 1 (PD-1) and T-cell immunoglobulin and mucin domain-containing molecule 3 (Tim3) on peripheral blood CD8⁺T cells in patients with chronic hepatitis B (CHB).

Methods

Total of 72 CHB patients with HBeAg positive were collected, and their anti-coagulation peripheral venous blood were collected from each patient at the beginning and the end of 12 months of IFN-α therapy. Thirty cases of healthy individuals were enrolled as controls. PD-1 and Tim3 on CD8⁺T cells were detected by flow cytometry.

Results

Before IFN-α treatment, the median percentage of PD-1 and Tim3 on peripheral blood CD8⁺T cells was 35.5% and 4.8% in CHB, respectively, and both were higher than the corresponding index in healthy control group (29.3% and 3.6%), with significant differences (U = 805.0,P = 0.043;U = 741.5,P= 0.013). At the end of 12 months of IFN-α therapy, the median percentage of PD-1 and Tim3 on CD8⁺T cells was 32.7% and 4.1% in CHB patients, respectively, and significantly decreased than those before treatment (Z= 3.305,P= 0.001;Z= 2.065,P= 0.039). The median percentage of the two molecule expression was significantly lower in patients who achieved HBeAg seroconversion than those of patients without HBeAg seroconversion, with significant differences (U= 209.0,P < 0.001;U= 302.0,P <0.001). The two median percentages were without significant difference between ALT normalization and abnormalization groups (U = 229.5,P = 0.635;U= 209.5,P = 0.405). The median percentage of PD-1 on CD8⁺T cells was significantly lower in patients with HBV DNA undetectable than those with detectable HBV DNA (U= 371.5,P = 0.011), which was not significantly different between the two groups of patients (U = 558.0,P = 0.727).

Conclusions

A higher expression of PD-1 and Tim3 on peripheral blood CD8⁺T cells was presented in CHB patients, which could be downregulated by IFN-α therapy, and the decreased expression was associated with the efficacy of IFN-α treatment.

Key words: Chronic hepatitis B, Interferon alpha, Peripheral blood CD8⁺T lymphocyte, Programmed death 1, T-cell immunoglobulin and mucin domain-containing molecule 3

表1 CHB患者和对照组的临床基线资料

组别	例数	男性[例（%）]	年龄（岁，±s）	ALT[U/L，中位数（范围）]	HBeAg阳性[例（%）]	HBV DNA对数值[IU/ml，中位数（范围）]
CHB组	72	58（80.6）	28.94 ± 6.964	142.5（83~880）	72（100.0）	7.5（4.8~9.4）
健康对照组	30	22（73.3）	30.07 ± 6.518	25.5（9~40）	—	—
统计量		χ² = 0.653	t = 0.755	U = 0.000	—	—
P值		0.419	0.452	0.000	—	—

表1 CHB患者和对照组的临床基线资料

组别	例数	男性[例（%）]	年龄（岁，±s）	ALT[U/L，中位数（范围）]	HBeAg阳性[例（%）]	HBV DNA对数值[IU/ml，中位数（范围）]
CHB组	72	58（80.6）	28.94 ± 6.964	142.5（83~880）	72（100.0）	7.5（4.8~9.4）
健康对照组	30	22（73.3）	30.07 ± 6.518	25.5（9~40）	—	—
统计量		χ² = 0.653	t = 0.755	U = 0.000	—	—
P值		0.419	0.452	0.000	—	—

表2 CHB患者与对照组外周血CD8⁺ T细胞PD-1和Tim3的表达

组别	例数	CD8⁺PD-1⁺ T细胞[%，中位数（范围）]	CD8⁺Tim3⁺ T细胞[%，中位数（范围）]
健康对照组	30	29.3（17.1~49.6）	3.6（2.2~9.8）
CHB组	72	35.5（12.4~78.0）	4.8（1.5~9.8）
U值		805.000	471.500
P值		0.043	0.013

表2 CHB患者与对照组外周血CD8⁺ T细胞PD-1和Tim3的表达

组别	例数	CD8⁺PD-1⁺ T细胞[%，中位数（范围）]	CD8⁺Tim3⁺ T细胞[%，中位数（范围）]
健康对照组	30	29.3（17.1~49.6）	3.6（2.2~9.8）
CHB组	72	35.5（12.4~78.0）	4.8（1.5~9.8）
U值		805.000	471.500
P值		0.043	0.013

表3 干扰素治疗前后CHB患者外周血CD8⁺ T细胞PD-1和Tim3的表达

组别	例数	CD8⁺PD1⁺ T细胞[%，中位数（范围）]	CD8⁺Tim3⁺ T细胞[%，中位数（范围）]
干扰素治疗前	72	35.5（12.4~78.0）	4.8（1.5~9.8）
干扰素治疗12月	72	32.7（7.6~75.2）	4.1（1.2~8.8）
Z值		3.305	2.065
P值		0.001	0.039

表3 干扰素治疗前后CHB患者外周血CD8⁺ T细胞PD-1和Tim3的表达

组别	例数	CD8⁺PD1⁺ T细胞[%，中位数（范围）]	CD8⁺Tim3⁺ T细胞[%，中位数（范围）]
干扰素治疗前	72	35.5（12.4~78.0）	4.8（1.5~9.8）
干扰素治疗12月	72	32.7（7.6~75.2）	4.1（1.2~8.8）
Z值		3.305	2.065
P值		0.001	0.039

图1 CHB患者干扰素治疗不同应答患者外周血CD8⁺ T细胞PD-1和Tim3的表达

图2 HBeAg血清学转换组与健康对照组外周血CD8⁺ T细胞PD-1和Tim3的表达

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Relationship between interferon-alpha efficacy and the expression of programmed death 1 and T-cell immunoglobulin and mucin domain-containing molecule 3 on peripheral blood CD8⁺ T cells in patients with chronic hepatitis B

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Relationship between interferon-alpha efficacy and the expression of programmed death 1 and T-cell immunoglobulin and mucin domain-containing molecule 3 on peripheral blood CD8+ T cells in patients with chronic hepatitis B

Relationship between interferon-alpha efficacy and the expression of programmed death 1 and T-cell immunoglobulin and mucin domain-containing molecule 3 on peripheral blood CD8⁺ T cells in patients with chronic hepatitis B