摘要
目的观察盐酸达拉他韦联合阿舒瑞韦治疗后慢性丙型肝炎患者外周血CD4＋和CD8＋T细胞功能的变化。方法收集21例HLA-A2限制性、丙型肝炎病毒（hepatitis C virus, HCV）1b基因型感染的慢性丙型肝炎患者，接受盐酸达拉他韦联合阿舒瑞韦抗病毒治疗24周，分别于基线、4周和24周采集外周血，分选CD4＋T细胞和CD8＋T细胞，应用流式细胞术检测CD4＋T细胞分泌细胞因子的水平。利用直接接触培养系统和间接接触培养系统将CD8＋T细胞与体外培养的HCV（HCVcc）感染的Huh7.5细胞共培养，通过检测培养上清中乳酸脱氢酶和细胞因子水平分析CD8＋T细胞的细胞杀伤功能和非细胞杀伤功能。结果盐酸达拉他韦联合阿舒瑞韦治疗4周病毒学和生化学应答率分别为71.43%（15/21）和77.78%（14/18），治疗24周病毒学和生化学应答率均为100%。治疗4周和24周时CD4＋T细胞分泌IFN-γ、TNF-α和IL-17的水平显著升高，分泌IL-10的水平与基线比较未见显著变化。但上述细胞因子的表达在治疗4周发生病毒学应答和未发生病毒学应答的患者之间无显著差异。治疗4周和24周时CD8＋T细胞与HCVcc感染的Huh7.5细胞直接接触培养系统中细胞死亡的比例显著增加，但对间接接触培养系统中CD8＋T细胞的杀伤功能无明显影响。治疗4周和24周时IFN-γ在直接和间接接触培养系统中的表达均显著升高，TNF-α的表达在治疗后仅在直接接触培养系统中显著升高，但在间接接触培养系统中的表达无明显变化。在治疗4周发生病毒学应答和未发生病毒学应答的患者之间CD8＋T细胞的细胞杀伤和非细胞杀伤活性均无显著差异。结论盐酸达拉他韦联合阿舒瑞韦治疗对慢性丙型肝炎患者具有极高的临床治愈率，抑制HCV病毒复制提高患者CD4＋和CD8＋T细胞的功能。
ObjectiveTo investigate the changes in CD4＋ and CD8＋ T cell functions in patients with chronic hepatitis C in response to daclatasvir plus asunaprevir therapy.MethodsA total of 21 HLA-A2-restricted patients with chronic hepatitis C virus （HCV） genotype 1b infection were enrolled in this study. All patients were treated with daclatasvir plus asunaprevir for 24 weeks. Peripheral blood samples were collected at baseline, 4 and 24 weeks post-therapy. CD4＋ and CD8＋ T cells were sorted and purified. Cytokines secreted by CD4＋ T cells were measured by flow cytometry. CD8＋ T cells were co-cultured with HCV cell culture （HCVcc）-infected Huh7.5 cells in both direct and indirect contact co-culture systems. The cytolytic and non-cytolytic functions of CD8＋ T cells were analyzed by measuring the levels lactate dehydrogenase and cytokines in the supernatants of cell culture.ResultsThe virological and biochemical response rates were 71.43% （15/21） and 77.78% （14/18） at 4 weeks post-therapy, respectively. Both rates reached 100% at 24 weeks post-therapy. Secretion of IFN-γ, TNF-α and IL-17 by CD4＋ T cells was significantly enhanced at 4 and 24 weeks in response to daclatasvir plus asunaprevir therapy. In contrast, IL-10 secretion by CD4＋ T cells did not change notably post-therapy. However, no significant differences in cytokine secretion were found between patients with and without virological response at 4 weeks post-therapy. Daclatasvir plus asunaprevir therapy increased the percentage of dead cells in direct contact co-culture system of CD8＋ T cells and HCVcc-infected Huh7.5 cells at 4 and 24 weeks post-therapy. However, it did not affect the cytotoxity of CD8＋ T cells in indirect contact co-culture system. Moreover, IFN-γ expression in both direct and indirect contact co-culture systems was significantly increased at 4 and 24 weeks post-therapy. There was also a notable increase in the expression of TNF-α in direct contact co-culture system, while no remarkable change in TNF-α expr
作者
李彧
田颖
苏莉
曹宁家
李竹
刘清娥
王璐
张丹
张鸿
Li Yu;Tian Ying;Su Li;Cao Ningjia;Li Zhu;Liu Qing＇e;Wang Lu;Zhang Dan;Zhang Hong(Department of Infectious Diseases, Shaanxi Provincial People＇s Hospital, Xi＇an 710068, China;Department of Gynecology, Shaanxi Provincial Hospital of Traditional Chinese Medicine, Xi＇an 710003, Chin;School of Public Health, Lanzhou University, Lanzhou 730000, China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2018年第5期381-389,共9页
Chinese Journal of Microbiology and Immunology
基金
陕西省自然科学基金（2013JQ4044）
