Fine needle aspirates comprehensively sample intrahepatic immunity
Fine needle aspirates comprehensively sample intrahepatic immunity
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DOI：
10.1136/gutjnl-2018-317071
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发表时间：
2019-08-01
期刊：
影响因子：
24.5
通讯作者：
Maini, Mala K.
中科院分区：
文献类型：
--
作者：
Gill, Upkar S.;Pallett, Laura J.;Maini, Mala K.
目的为了改进治疗乙肝病毒的新策略，需要对感染部位的病毒学和免疫学变化进行评估。采用16色多参数流式细胞术对28例乙肝患者和15例无病毒感染患者的血液、肝活检和细针抽吸物(FNAs)进行分析。结果FNA检测的CD4T、CD8T、粘膜相关不变T细胞(MAIT)、自然杀伤细胞(NK)和B细胞的比例与同一供者肝活检组织中CD4T、CD8T、MAIT、NK和B细胞的比例相关。FNA和肝活检均可鉴定出程序性死亡-1(PD-1)(Hi)、CD39(Hi)、组织驻留记忆CD8 T细胞(CD69(+)CD103(+))和肝脏驻留NK细胞(CXCR6(+)T-bet(Lo)Eomeshi)。至关重要的是，病毒特异性T细胞可以被FNAs以与活组织检查相似的频率识别，并与血液相比有所丰富。FNAs能同时识别表达白蛋白、清道夫受体B型(SR-B1)、程序性死亡配体1(PD-L1)的髓系细胞和肝细胞，而肝细胞经肝活检后活性较差。结论首次证实FNAs可识别多种肝内免疫细胞，包括局部驻留的前哨病毒特异性T细胞和NK细胞，以及表达PD-L1的肝细胞。此外，我们提供了一个评分工具来估计单个FNA在多大程度上可靠地采样了肝内人群，而不是污染了血液。这种侵入性较小、快速的技术所实现的广泛图谱使其适合于对肝脏进行纵向监测，以优化治疗乙肝病毒的新疗法。
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)(hi)CD39(hi) tissue-resident memory CD8 T cells (CD69(+)CD103(+)) and liver-resident NK cells (CXCR6(+) T-bet(lo)Eomeshi), 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.