细针抽吸全面采样肝内免疫

StephenW

Fine needle aspirates comprehensively sample intrahepatic immunity

    Upkar S Gill1, Laura J Pallett2, Niclas Thomas2, Alice R Burton2, Amit A Patel3, Simon Yona3, Patrick T F Kennedy1, Mala K Maini2

Author affiliations

    Barts Liver Centre, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK
    Division of Infection and Immunity, Institute of Immunity and Transplantation, University College London, London, UK
    Division of Medicine, University College London, London, UK

    Correspondence to Professor Mala K Maini, Division of Infection and Immunity, Institute of Immunity and Transplantation, UCL, London, UK; [email protected]

Abstract

Objective In order to refine new therapeutic strategies in the pipeline for HBV cure, evaluation of virological and immunological changes compartmentalised at the site of infection will be required. We therefore investigated if liver fine needle aspirates (FNAs) could comprehensively sample the local immune landscape in parallel with viable hepatocytes.

Design Matched blood, liver biopsy and FNAs from 28 patients with HBV and 15 without viral infection were analysed using 16-colour multiparameter flow cytometry.

Results The proportion of CD4 T, CD8 T, Mucosal Associated Invariant T cell (MAIT), Natural Killer (NK) and B cells identified by FNA correlated with that in liver biopsies from the same donors. Populations of Programmed Death-1 (PD-1)hiCD39hi tissue-resident memory CD8 T cells (CD69+CD103+) and liver-resident NK cells (CXCR6+T-betloEomeshi), were identified by both FNA and liver biopsy, and not seen in the blood. Crucially, HBV-specific T cells could be identified by FNAs at similar frequencies to biopsies and enriched compared with blood. FNAs could simultaneously identify populations of myeloid cells and live hepatocytes expressing albumin, Scavenger Receptor class B type 1 (SR-B1), Programmed Death-Ligand 1 (PD-L1), whereas hepatocytes were poorly viable after the processing required for liver biopsies.

Conclusion We demonstrate for the first time that FNAs identify a range of intrahepatic immune cells including locally resident sentinel HBV-specific T cells and NK cells, together with PD-L1-expressing hepatocytes. In addition, we provide a scoring tool to estimate the extent to which an individual FNA has reliably sampled intrahepatic populations rather than contaminating blood. The broad profiling achieved by this less invasive, rapid technique makes it suitable for longitudinal monitoring of the liver to optimise new therapies for HBV.

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See: https://creativecommons.org/licenses/by/4.0/.

http://dx.doi.org/10.1136/gutjnl-2018-317071

StephenW

细针抽吸全面采样肝内免疫

    Upkar S Gill1，Laura J Pallett2，Niclas Thomas2，Alice R Burton2，Amit A Patel3，Simon Yona3，Patrick T F Kennedy1，Mala K Maini2

作者隶属关系

    Barts肝脏中心，Blizard研究所，Barts和伦敦医学和牙科学院，伦敦大学玛丽皇后学院，英国伦敦
    英国伦敦大学学院免疫与移植研究所感染与免疫科
    英国伦敦大学学院医学系

    英国伦敦伦敦大学学院免疫与移植研究所感染与免疫科Mala K Maini教授的通讯; [email protected]

抽象

目的为了改进HBV治愈方法的新治疗策略，需要对感染部位区分的病毒学和免疫学变化进行评估。因此，我们研究了肝细针抽吸物（FNAs）是否可以与活肝细胞平行地全面地采样局部免疫景观。

设计使用16色多参数流式细胞仪分析来自28名HBV患者和15名无病毒感染患者的血液，肝脏活组织检查和FNA。

结果通过FNA鉴定的CD4T，CD8T，粘膜相关不变T细胞（MAIT），自然杀伤（NK）和B细胞的比例与来自相同供体的肝活组织检查中的比例相关。程序性死亡-1（PD-1）hiCD39hi组织驻留记忆CD8 T细胞（CD69 + CD103 +）和肝脏驻留NK细胞（CXCR6 + T-betloEomeshi）的群体通过FNA和肝脏活组织检查鉴定，未见在血液中。至关重要的是，HBV特异性T细胞可以通过FNA以与活组织检查相似的频率进行鉴定，并且与血液相比可以富集。 FNA可以同时鉴定骨髓细胞群和表达白蛋白的活肝细胞，Scavenger Receptor B类1型（SR-B1），程序性死亡 - 配体1（PD-L1），而肝细胞在肝脏活组织检查所需的处理后很差。

结论我们首次证明了FNA识别一系列肝内免疫细胞，包括局部驻留的哨兵HBV特异性T细胞和NK细胞，以及表达PD-L1的肝细胞。此外，我们提供了一种评分工具，用于估计个体FNA对肝内人群进行可靠取样而不是污染血液的程度。通过这种侵入性较小的快速技术实现的广泛分析使其适用于肝脏的纵向监测，以优化HBV的新疗法。

这是根据知识共享署名4.0 Unported（CC BY 4.0）许可分发的开放获取文章，允许其他人出于任何目的复制，再分发，重新混合，转换和构建此作品，前提是原始作品被正确引用，给出了许可证的链接，并指出是否进行了更改。请参阅：https：//creativecommons.org/licenses/by/4.0/。

http://dx.doi.org/10.1136/gutjnl-2018-317071

StephenW

https://gut.bmj.com/content/gutj ... 018-317071.full.pdf