qAnti-HBc与慢性乙型肝炎儿童抗病毒疗效的相关性分析及可能免疫机制探索

Correlation of qAnti-HBc with antiviral efficacy in children with chronic hepatitis B and exploration of its possible immune mechanism

目的﹑比较行抗病毒治疗的HBeAg阳性慢性乙型肝炎(CHB)患儿无应答组和应答组间基线乙型肝炎核心抗体定量(qAnti-HBc)水平的差异,以探讨不同qAnti-HBc水平患儿外周血CD_(8)^(+)记忆T细胞亚群比例及其功能活性。方法︰回顾性检测2018年6月至2020年12月于重庆医科大学附属儿童医院感染科就诊的85例HBeAg阳性CHB患儿基线qAnti-HBc水平。分析其中37例抗病毒治疗患儿基线qAnti-HBc水平与HBeAg血清学应答的关系。流式细胞检测59例患儿基线外周血CD_(8)^(+)记忆T细胞亚群比例及干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α分泌水平,分析qAnti-HBc水平与CD_(8)^(+)记忆T细胞亚群比例及其功能活性间的关系。计数资料比较采用Pearson's Chi-square检验,两组或多组间计量资料比较采用Mann-Whitney U检验或Kruskal-Wallis检验,连续性变量间的相关性采用Spearman秩相关分析。结果﹐在37例接受恩替卡韦(ETV,21/37)或聚乙二醇干扰素)(Peg-IFN,16/37)治疗的患儿中,有18例发生HBeAg血清学转换(ETV组10/21,Peg-IFN组8/16),应答组患儿基线qAnti-HBc水平[4.71(4.64~4.81)log_(10)IU/ml]显著高于无应答组[4.54(4.45~4.64)log_(10)IU/ml,Z=-3.316,P=0.001]。高qAnti-HBc组CD8^(+)Tem,CD38^(+)CD8^(+)Tem、CD38^(+)CD8^(+)Temra细胞比例及CD8^(+)T细胞分泌的IFN-γ、TNF-α水平显著高于低qAnti-HBc组(P<0.05);ALT>1×正常值上限(ULN)组CD8^(+)Tem、CD38^(+)CD8^(+)Tem,CD38^(+)CD8^(+)Temra细胞比例显著高于ALT≤1×ULN组(P<0.05),但两组CD8T细胞分泌的IFN-γ、TNF-α水平差异无统计学意义(P>0.05)。Spearman相关分析显示,qAnti-HBc与CD8^(+)Tem、CD38^(+)CD8^(+)Tem、CD38^(+)CD8^(+)Temra细胞比例及CD8^(+)T细胞分泌的IFN-γ水平呈正相关(P<0.05),ALT仅与CD38^(+)CD8^(+)Tem、CD38^(+)CD8^(+)Temra细胞比例呈正相关(P<0.05)。结论﹑较高的基线qAnti-HBc水平与CHB儿童患者抗病毒治疗HBeAg血清学应答相关﹔qAnti-HBc较高的CHB患儿外周血效应CD8T细胞表现出更强的表型及功能活化特点,此研究可能对CHB儿童抗病毒治疗疗效相关的潜在免疫机制作出一定阐释。

Objective To compare the baseline difference in the quantitative hepatitis B core antibody levels(qAnti-HBc)between non-response and response group in children with HBcAg-positive chronic hepatitis B(CHB)who received antiviral therapy,and further explore the proportion and flinctional activity of CDS+memory T lymphocyte subsets with different qAnti-HBC levels in peripheral blood of children.Methods The baseline anti-FI Be quantification(qAnti-HBc)levels of 85 children with HBeAg-positive CHB who visited the Department of Infectious Diseases,Children's Hospital of Chongqing Medical University from June 2018 to December 2020 were detected retrospectively.The relationship between the baseline qAnti-HBc level and HBeAg serological response in 37 children who received antiviral therapy was analyzed.The proportion of CD8 memory T lymphocyte subsets and the secretion levels of interferon(IFN)γ,and tumor necrosis factor(TNF)α in peripheral blood of 59 children at baseline were detected by flow cytometry.The relationship between qAnti-HBc level and the proportion and functional activity of CD8 memory T lymphocyte subsets was analyzed.Pearson's Chi-square test was used to compare the count data.Mann-Whitney U test or Kruskal-Wallis test was used to compare measurement data between two or more groups,and Spearman's rank correlation analysis was used for the correlation between continuous variables.Results Among 37 children who received entecavir(ETV,21/37 cases)or pegylated interferon(Peg-IFN,16/37 cases),18 cases had developed HBeAg seroconversion(10/21 cases in the ETV group,8/16 cases in the Peg-IFN group).The baseline qAnti-HBc level was significantly higher in the response group[4.71(4.64〜4.81)log10IU/ml]than the non-response group children[4.54(4.45〜4.64)log10IU/ml,Z=-3.316,P=0.001].The proportion of CD8 Tem,CD38^(+)TD8 ^(+)Tem,CD38^(+)CD^(+)Temra cells and the levels of IFNγ and TNFα secreted by CD8^(+) T lymphocytes were significantly higher in the high-qAnti-HBc group than the low-qAnti-HBc group(P<0.05).The proportion of CD8^(+) Tem,CD38^(+) CD8^(+) Tem and CD38^(+) CD8 ^(+)Temra cells was significantly higher in ALT>1× upper limit of normal value(ULN)group than ALT≤1×ULN group(P<0.05).However,there were no significant differences in the levels of IFNy and TNFa secreted by CD8^(+)T lymphocytes between the two groups(P>0.05).Spearman's correlation analysis showed that qAnti-HBc was positively correlated with the proportion of CD8^(+) Tem,CD38^(+) CD8^(+) Tem,CD38^(+) CD8^(+) Temra cells and the level of IFNy secreted by CD8^(+) T lymphocytes(/><0.05).Additionally,ALT was only positively correlated with the proportion of CD38^(+) CD8^(+)TEM and CD38^(+)CD8^(+) Temra cells(P<0.05).Conclusion Raised baseline qAnti-HBc level is related to the HBeAg serological response to antiviral therapy in children with CHB.Peripheral blood efifector CD8+T lymphocytes of CHB children with higher qAnti-HBc show stronger phenotype and flinctional activation characteristics,which may shed some light on the underlying immune mechanism related to antiviral therapy efficacy in children with CHB.