白细胞介素-7促进原发性肾病综合征患儿外周血CD14^+单核细胞活性  

Interleukin-7 promotes the activity peripheral CD14^+monocytes in children with idiopathic nephrotic syndrome

作　　者：刘君 冯仕品 LIU Jun;FENG Shipin(Department of Nephrology,Chengdu Women’s and Children’s Central Hospital,School of Medicine,University of Electronic Science and Technology of China,Chengdu 610017,Sichuan,China)

机构地区：[1]电子科技大学医学院附属妇女儿童医院,成都市妇女儿童中心医院儿童肾脏内科,四川成都610017

出　　处：《临床儿科杂志》2020年第11期861-866,共6页Journal of Clinical Pediatrics

基　　金：四川省卫生厅科研课题(No.120513)。

摘　　要：目的观察白细胞介素-7(IL-7)对儿童原发性肾病综合征(INS)患者CD14^+单核细胞功能的调控作用。方法入组33例INS患儿(INS组)和15例健康儿童(对照组)。分选外周血CD14^+单核细胞和CD4^+T细胞,采用酶联免疫吸附试验(ELISA)检测血清IL-7水平,采用流式细胞术和实时定量PCR法检测CD14^+单核细胞中CD127的平均荧光强度(MFI)和CD127 mRNA相对表达量。使用重组人IL-7刺激INS组CD14^+单核细胞,ELISA法检测培养上清中细胞因子的表达。建立INS组CD14^+单核细胞和CD4^+T细胞间接接触、直接接触共培养系统,流式细胞术检测分泌干扰素-γ(IFN-γ)的Th1细胞和分泌IL-17的Th17细胞比例。结果INS组血清IL-7水平较对照组升高,差异有统计学意义(P<0.001)。INS组纯化的CD14^+单核细胞经IL-7刺激后分泌IL-1β、IL-6、IL-8、单核细胞趋化蛋白-1和肿瘤坏死因子-α的水平高于无IL-7刺激组,差异有统计学意义(P<0.05)。INS组的直接接触培养系统中,无IL-7刺激和经IL-7刺激的CD14^+单核细胞均可诱导CD4^+T细胞中Th1和Th17细胞比例升高,且经IL-7刺激的CD14^+单核细胞诱导Th1和Th17细胞比例高于无IL-7刺激的CD14^+单核细胞,差异有统计学意义(P<0.05)。抗IL-6中和抗体抑制IL-7对INS组CD14^+单核细胞诱导的Th1比例升高。结论IL-7促进INS患儿CD14^+单核细胞分泌炎症细胞因子,IL-7刺激的CD14^+单核细胞可能通过直接接触和分泌IL-6诱导INS患儿CD4^+T细胞活化。Objective To investigate the regulatory function of interleukin-7(IL-7)to CD 14^+monocytes activity in children with idiopathic nephrotic syndrome(INS).Methods Thirty-three INS children and fifteen normal controls(NCs)were enrolled.Peripheral CD 14^+monocytes and CD 4^+T cells were isolated.Serum IL-7 level was measured by ELISA.Mean fluorescence intensity(MFI)and mRNA relative level of CD 127 in CD 14^+monocytes was assessed by flow cytometry and real-time PCR,respectively.CD14^+monocytes from INS children were stimulated with recombinant human IL-7.Cytokine expression in cultured supernatants was measured by ELISA.The indirect and direct contact co-culture system was set up between CD 14^+monocytes and CD 4^+T cells.The percentage of interferon-γ-secreting Th 1 cells and IL-17-secerting Th 17 cells was measured by flow cytometry.Student t test or paired t test was used for comparison between the two groups.Results Serum IL-7 was elevated in INS children compared with NCs(P<0.0001).There were no significant differences of either CD 127 MFI or CD127 mRNA relative level in CD 14^+monocytes between NCs and INS children(P>0.05).IL-1β,IL-6,IL-8,monocyte chemotactic protein-1,and tumor necrosis factor-αexpression was elevated in IL-7 stimulated CD 14^+monocytes from INS children compared with unstimulated cells(P<0.05).In direct contact co-culture system,CD 14^+monocytes from INS children raised Th1 and Th17 percentages in CD4^+T cells from INS children.Moreover,IL-7 stimulated CD14^+monocytes induced higher Th1 and Th17 cells in comparison with unstimulated CD14^+monocytes(P<0.05).In direct contact co-culture system,anti-IL-6 neutralizing antibody suppressed the elevation of Th1 cells,which induced by IL-7 stimulated CD14^+monocytes from INS children.Conclusion IL-7 promoted proinflammatory cytokines production by CD 14^+monocytes from INS children.IL-7 stimulated CD14^+monocytes also induced CD4^+T cells activation via direct contact and IL-6 secretion in INS children.

关 键 词：细胞因子 单核细胞 肾病综合征 免疫调控 

分 类 号：R726.9[医药卫生—儿科]