Chen P L, Zhang B Y, Fu X Q, Zhang Y Q, Xie H P
Department of Gastroenterology, Zhumadian Central Hospital, Zhumadian 463000, China.
Department of Gastroenterology, Henan Provincial People's Hospital, Zhengzhou 450003, China.
Zhonghua Yi Xue Za Zhi. 2022 Jan 11;102(2):141-146. doi: 10.3760/cma.j.cn112137-20210428-01022.
To investigate the change of innate lymphoid cells (ILC) subsets in peripheral blood and ascites in liver cirrhotic patients complicated with spontaneous bacterial peritonitis (SBP). The data of 62 patients with liver cirrhosis admited to the Zhumadian Central Hospital from November 2019 to November 2020 were analyzed. Among them, 41 cases were complicated with untainted ascites (untainted ascites group), while the other 21 cases were complicated with SBP (SBP group). Meanwhile, 20 cases of controls who received healthy examination in the same period were also enrolled (control group). Peripheral blood mononuclear cell (PBMC) was isolated from peripheral blood of all patients and controls. Mononuclear cell in ascites was isolated from patients with liver cirrhosis. The percentage of ILC1, ILC2, and ILC3 subsets in PBMC and mononuclear cell in ascites were measured by flow cytometry. CD3CD19CD20CD14 cells (lincells) were purified from ascites and were stimulated with lipopolysaccharide (LPS) for 24 h. The transcription factor T-bet, GATA3, and RORγt mRNA relative level in lincells was semi-quantified by real-time PCR. Cytokine level in the supernatants was measured by enzyme linked immunosorbent assay. Differences of ILC subsets in peripheral blood and ascites were compared among groups. There were twenty-nine males and twelve females in untainted ascites group, aged (,) 49(33, 78) years. There were twelve males and nine females in SBP group, aged 50(37, 76) years. There were eleven males and nine females in control group, aged 48(32, 69) years. linCD45CD161CD127 ILC cells could be detected in both peripheral blood and ascites. There was no significant difference in total ILC percentage within PBMC among untainted ascites group, SBP group, and control group (=0.235). There was also no significant difference of total ILC percentage within mononuclear cells in ascites between untainted ascites group and SBP group (=0.232). The differences were not statistically significant of peripheral CD117CRTh2ILC1, CRTh2ILC2, or CD117CRTh2ILC3 within peripheral ILC among untainted ascites group, SBP group, and control group (all >0.05). ILC1 percentage in ascites was up-regulated in SBP group compared with untainted ascites group (35.69%±3.39% vs 26.40%±3.85%, <0.001), while ILC2 in ascites was down-regulated in SBP group (36.83%±7.70% vs 48.35%±9.45%, <0.001). There was no statistical difference in ILC3 percentage in ascites between the two groups (=0.230). T-bet mRNA relative level and IFN-γ production by lin cells from ascites were elevated in response to LPS stimulation in SBP group compared with untainted ascites group (both <0.001). GATA3 mRNA relative level and IL-5/IL-13 secretion by lincells from ascites were reduced in SBP group compared with untainted ascites group (both <0.05). There was no significant difference of RORγt mRNA relative level or IL-17/IL-22 expression between the two groups (both >0.05). Peripheral ILC subsets do not change in liver cirrhosis patients with SBP. ILC1 percentage is up-regulated, and ILC2 percentage is down-regulated in ascites in liver cirrhosis patients with SBP.
探讨肝硬化合并自发性细菌性腹膜炎(SBP)患者外周血及腹水中固有淋巴细胞(ILC)亚群的变化。分析2019年11月至2020年11月入住驻马店市中心医院的62例肝硬化患者的数据。其中,41例合并无菌性腹水(无菌性腹水组),另外21例合并SBP(SBP组)。同时,纳入同期进行健康体检的20例对照者(对照组)。从所有患者及对照者的外周血中分离外周血单个核细胞(PBMC)。从肝硬化患者腹水中分离腹水中单个核细胞。采用流式细胞术检测PBMC及腹水中单个核细胞中ILC1、ILC2和ILC3亚群的百分比。从腹水中纯化CD3CD19CD20CD14细胞(淋系细胞),并用脂多糖(LPS)刺激24小时。通过实时PCR半定量检测淋系细胞中转录因子T-bet、GATA3和RORγt mRNA的相对水平。采用酶联免疫吸附测定法检测上清液中的细胞因子水平。比较各组外周血及腹水中ILC亚群的差异。无菌性腹水组有29例男性和12例女性,年龄(,)49(33,78)岁。SBP组有12例男性和9例女性,年龄50(37,76)岁。对照组有11例男性和9例女性,年龄48(32,69)岁。外周血和腹水中均可检测到linCD45CD161CD127 ILC细胞。无菌性腹水组、SBP组和对照组PBMC中ILC总百分比无显著差异(=0.235)。无菌性腹水组和SBP组腹水中单个核细胞中ILC总百分比也无显著差异(=0.232)。无菌性腹水组、SBP组和对照组外周ILC中,外周CD117CRTh2ILC1、CRTh2ILC2或CD117CRTh2ILC3的差异均无统计学意义(均>0.05)。与无菌性腹水组相比,SBP组腹水中ILC1百分比上调(35.69%±3.39% vs 26.40%±3.