HBsAg阴性慢性乙型肝炎患者停用核苷酸类似物治疗后的血清和

StephenW

Serum and intrahepatic HBV markers and HBV-specific CD8 T cell responses after nucleos(t)ide analog therapy discontinuation in HBeAg-negative chronic hepatitis B patients

EASL 2020 Aug 27-29 virtual

Mireia García-López1, Sabela Lens1, Laura J Pallett2, Barbara Testoni3, Zoe Mariño1, Sergio Rodriguez-Tajes1, Concepció Bartres1, Ester García-Pras1, Thais Leonel1, Elena Perpiñan1, Juanjo Lozano4, Francisco Rodríguez-Frías5, George Koutsoudakis1, Fabien Zoulim3, Mala Maini2, Xavier Forns1, Sofía Pérez-del-Pulgar1
1Liver Unit, Hospital Clínic, University of Barcelona, IDIBAPS, CIBERehd, Barcelona, Spain, 2Division of Infection and Immunity, Institute of Immunity and Transplantation, University College London, London, United Kingdom, 3Cancer Research Center of Lyon (CRCL), University of Lyon, UMR_S1052, UCBL, INSERM, U1052, Lyon, France, 4Bioinformatics Platform, CIBERehd, Barcelona, Spain, 5Liver Pathology Unit, Department of Biochemistry and Microbiology, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, CIBERehd, Barcelona, Spain


Background and aims: Previous studies have reported that up to 20% of patients with chronic hepatitis B (CHB) may achieve functional cure (HBsAg loss) after nucleos(t)ide analog (NA) treatment withdrawal. CD8 T cells play an important role in the immune control of HBV infection. The objective of this study was to analyze HBV-specific CD8 responses in parallel with peripheral and intrahepatic virological markers after NUC discontinuation in patients with HBeAg-negative (HBeAg-) CHB.
Method: Twenty-seven HBeAg- CHB patients with complete viral suppression (>3 years) and without cirrhosis were prospectively studied. A liver biopsy was taken at the time of treatment withdrawal (baseline). PBMC and serum samples were collected at baseline and various time-points during follow-up. Intrahepatic HBV-DNA (iHBV-DNA), covalently closed circular DNA (cccDNA) and serum HBV-DNA, HBsAg, core-related antigen (HBcrAg) and pregenomic RNA (pgRNA) levels were determined. HBV-specific T cell responses (IFNy, TNF and CD107a) were analyzed by multiparametric flow cytometry after in vitro expansion in the presence of overlapping peptides (OLP) spanning core, envelope and polymerase.

Results: After a median follow-up of 34 months (IQR 26-37), 22 (81%) patients remain off-therapy, with 8 (30% of the total cohort) losing HBsAg; whilst 5 (19%) required NA reintroduction due to relapse. Although all patients were iHBV-DNA and cccDNA positive at baseline, only 41% and 48% had detectable serum pgRNA and HBcrAg, respectively.

see slides below for actual data presented

Baseline HBsAg levels correlated significantly with iHBV-DNA (r = 0.7, p < 0.0001) and both markers were lower in patients who lost HBsAg (p < 0.001). Baseline intrahepatic (iHBV-DNA, cccDNA) or serum (HBsAg, HBcrAg or pgRNA) viral markers did not show any association with peripheral CD8 T cell responses. Importantly, degranulating CD8 T cells (CD107a+) or those co-producing IFNy and TNF in response to stimulation with core OLP were significantly higher (p = 0.05 and p = 0.039, respectively) at baseline in patients remaining off-therapy compared to those requiring NA reintroduction. Interestingly, the enhanced frequency of CD8 T cells co-producing IFNy and TNF persisted up to 1 year of follow-up (p = 0.009). Notably, CD8 T cell responses to polymerase or envelope failed to associate with outcome as robustly as those against core.

Conclusion: NA discontinuation is feasible in a high proportion of HBeAg- patients, particularly in those with low HBsAg levels. Higher frequencies of CD8 T cells with cytotoxic and non-cytolytic anti-HBV (core) reactivity are detectable at baseline in those patients who maintain viral control after therapy withdrawal. These data support HBsAg levels and HBV-specific CD8 T cell frequencies as correlates of HBV control off-therapy, requiring validation in larger studies.


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StephenW

HBsAg阴性慢性乙型肝炎患者停用核苷酸类似物治疗后的血清和肝内HBV标记及HBV特异性CD8 T细胞反应

EASL 2020 8月27-29日虚拟

MireiaGarcía-López1，Sabela Lens1，Laura J Pallett2，Barbara Testoni3，ZoeMariño1，Sergio Rodriguez-Tajes1，ConcepcióBartres1，EsterGarcía-Pras1，Thais Leonel1，ElenaPerpiñgue1，JuanFoíasz，4， ，Mala Maini2，Xavier Forns1，SofíaPérez-del-Pulgar1
1西班牙巴塞罗那大学IDIBAPS，CIBERehd，西班牙巴塞罗那大学医院肝病科，2英国伦敦大学学院免疫与移植研究所感染与免疫科，3英国里昂癌症研究中心（CRCL）里昂，UMR_S1052，UCBL，INSERM，U1052，里昂，法国，4生物信息平台，CIBERehd，西班牙巴塞罗那，5肝病病理科，医院UniversitéVall d'Hebron，巴塞罗那大学Autònomade巴塞罗那大学，CIBERehd，西班牙巴塞罗那


背景与目的：先前的研究报道，多达20％的慢性乙型肝炎（CHB）患者在停用核苷酸类似物（NA）治疗后可以实现功能性治愈（HBsAg丢失）。 CD8 T细胞在HBV感染的免疫控制中起重要作用。这项研究的目的是分析HBeAg阴性（HBeAg-）CHB患者NUC停药后的HBV特异性CD8反应以及外周和肝内病毒学指标。
方法：前瞻性研究了27例完全抑制病毒（> 3年）且无肝硬化的HBeAg-CHB患者。停药时进行肝活检（基线）。在随访期间在基线和各个时间点收集PBMC和血清样品。测定肝内HBV-DNA（iHBV-DNA），共价闭合环状DNA（cccDNA）和血清HBV-DNA，HBsAg，核心相关抗原（HBcrAg）和前基因组RNA（pgRNA）水平。在存在跨越核心，包膜和聚合酶的重叠肽（OLP）的体外扩增后，通过多参数流式细胞术分析了HBV特异性T细胞反应（IFNγ，TNF和CD107a）。

结果：中位随访34个月（IQR 26-37）后，仍有22（81％）名患者停止治疗，其中8名（占总队列的30％）失去了HBsAg。 5（19％）因复发而需要重新引入NA。尽管所有患者基线时iHBV-DNA和cccDNA均为阳性，但分别只有41％和48％的患者可检测到血清pgRNA和HBcrAg。

有关实际数据，请参见下面的幻灯片

基线HBsAg水平与iHBV-DNA显着相关（r = 0.7，p <0.0001），并且在失去HBsAg的患者中两种指标均较低（p <0.001）。基线肝内（iHBV-DNA，cccDNA）或血清（HBsAg，HBcrAg或pgRNA）病毒标志物未显示与外周CD8 T细胞反应相关。重要的是，在停用治疗的患者中，与需要治疗的患者相比，基线时脱颗粒的CD8 T细胞（CD107a +）或响应核心OLP刺激而共同产生IFNγ和TNF的患者明显高于基线（分别为p = 0.05和p = 0.039）。不适用NA。有趣的是，共同产生IFNγ和TNF的CD8 T细胞的频率增加持续了长达1年的随访（p = 0.009）。值得注意的是，CD8 T细胞对聚合酶或包膜的反应未能像对核心反应一样牢固地与预后相关。

结论：NA停药在高比例的HBeAg患者中是可行的，尤其是在HBsAg水平低的患者中。在停药后仍保持病毒控制的那些患者中，基线时可检测到具有细胞毒性和非细胞溶解性抗HBV（核心）反应性的CD8 T细胞，其频率更高。这些数据支持HBsAg水平和HBV特异性CD8 T细胞频率作为HBV控制非治疗的相关因素，需要在更大的研究中进行验证。


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