Paediatric Gastroenterology-Hepatology, Koç University Hospital.
Liver Transplantation Centre, Koç University.
J Pediatr Gastroenterol Nutr. 2022 Feb 1;74(2):200-207. doi: 10.1097/MPG.0000000000003353.
Liver biopsy is the standard in diagnosing liver diseases. Yet, it provides little space to perform comprehensive immune profiling of the liver. Hence, we explored whether fine needle aspirates (FNAs) could be used to elucidate the hepatic immunity in children.
We enrolled 74 children undergoing diagnostic (n = 17) or protocol biopsy (n = 57) following liver transplantation (LT). Matched blood and FNAs were obtained. Additionally, explant liver tissue was collected from children (n = 14) undergoing LT. Immune cells were isolated from peripheral blood, FNAs and explanted livers. Immune-phenotypical profiling was done by flow cytometry.
Biopsied patients (58% female) were at a median age of 46 months (interquartile range [IQR]: 12-118) and LT patients (71% female) were 48 months (IQR: 21-134, P = 0.78) old. CD69+, a hallmark of tissue-resident immune cells was expressed in 1.3% of CD3+ T cells from blood being higher in FNA (20%) and tissue (49%, P < 0.001). CD4+ T-cell frequencies in tissue (13%) and FNAs (20%) were lower compared to blood (35%, P < 0.001) whereas CD8+ T cells in tissue (33.5%) and FNA (32%) were higher than in blood (25%, P < 0.01). Mucosal associated invariant T cells were enriched in liver tissue (8.8%) and in the FNA (4.4%) compared to blood (1.7%, P < 0.001). Whereas the percentage of total Tregs (CD4+CD25+FOXP3+CD127low/-) decreased, the proportion of activated Tregs (CD4+CD45RA-FOXP3high) increased in FNA and explant. Breg (CD19+CD20+CD24highCD38high) frequencies were similar in all groups.
FNA is a practical method to sample the liver immune system collecting even small cell subsets such as regulatory T/B cells.
肝活检是诊断肝脏疾病的标准方法。然而，它提供的空间很小，无法对肝脏进行全面的免疫分析。因此，我们探讨了细针穿刺（FNA）是否可用于阐明儿童的肝免疫。
我们纳入了 74 名接受诊断性（n=17）或移植后方案性肝活检（n=57）的儿童。采集了匹配的血液和 FNA，此外，还从接受肝移植（LT）的儿童（n=14）中收集了肝移植组织。从外周血、FNA 和移植组织中分离免疫细胞，通过流式细胞术进行免疫表型分析。
活检患者（58%为女性）的中位年龄为 46 个月（IQR：12-118），LT 患者（71%为女性）为 48 个月（IQR：21-134，P=0.78）。CD3+T 细胞中表达组织驻留免疫细胞的标志 CD69+，在血液中的表达率为 1.3%，在 FNA（20%）和组织（49%，P<0.001）中更高。组织（13%）和 FNA（20%）中的 CD4+T 细胞频率低于血液（35%，P<0.001），而组织（33.5%）和 FNA（32%）中的 CD8+T 细胞高于血液（25%，P<0.01）。黏膜相关不变 T 细胞在肝组织（8.8%）和 FNA（4.4%）中比血液（1.7%，P<0.001）更丰富。虽然总 Treg（CD4+CD25+FOXP3+CD127low/-）的百分比下降，但 FNA 和移植组织中激活的 Treg（CD4+CD45RA-FOXP3high）的比例增加。所有组的 Breg（CD19+CD20+CD24highCD38high）频率相似。
FNA 是一种实用的方法，可以采集肝免疫系统，甚至可以采集到调节性 T/B 细胞等小细胞亚群。
