抗CCL20单克隆抗体对血清淀粉样蛋白A相关的结节病中促炎性细胞因子的抑制作用  

Anti-CCL20 monoclonal antibody suppresses the overexpression of pro-inflammatory cytokines in sarcoidosis accompanied by serum amyloid A

作　　者：魏佳 马成星 郑颖 那雨琪 丁晶晶 WEI Jia;MA Chengxing;ZHENG Ying;NA Yuqi;DING Jingjing(Department of Respiratory and Critical Care Medicine,Nanjing Drum Tower Hospital,Clinical College of Nanjing University of Chinese Medicine,Nanjing 210008,Jiangsu,China;Department of Respiratory and Critical Care Medicine,Nanjing Drum Tower Hospital,The Affiliated Hospital of Nanjing University Medical School,Nanjing 210008,Jiangsu,China;Department of Respiratory and Critical Care Medicine,Nanjing Drum Tower Hospital,Clinical College of Nanjing Medical University,Nanjing 210008,Jiangsu,China)

机构地区：[1]南京中医药大学鼓楼临床医学院呼吸与危重症医学科,南京210008 [2]南京大学医学院附属鼓楼医院呼吸与危重症医学科,南京210008 [3]南京医科大学鼓楼临床医学院呼吸与危重症医学科,南京210008

出　　处：《医学研究与战创伤救治》2024年第1期1-6,共6页Journal of Medical Research & Combat Trauma Care

基　　金：国家自然科学基金(82170077);南京市医药卫生科研课题(YKK20051)。

摘　　要：目的探讨抗CCL20单克隆抗体是否可抑制高水平血清淀粉样蛋白A(SAA)相关的结节病外周血单个核细胞(PBMC)中促炎性细胞因子的表达。方法提取健康人和结节病患者的PBMC,并进行分组。除空白对照组(正常人PBMC)和结节病PBMC对照组(患者PBMC)外,另将结节病组PBMC中分别加入SAA、SAA+抗CCL20单克隆抗体、SAA+NF-κB抑制剂BAY11-7082或地塞米松,经培养48 h后收集各组细胞,分别作为PBMC+SAA刺激组、PBMC+SAA+NF-κB抑制剂BAY11-7082组、PBMC+SAA+抗CCL20单抗组、PBMC+地塞米松干预组,使用ELISA分别测定各组细胞上清液中细胞因子IL-17A、IL-23、IL-6、CCL20和TGF-β1的表达,使用RT-PCR测定各组细胞中CCR6、IL-17、ROR-γt和Foxp3的mRNA表达水平,使用Western blot检测各组细胞中p-NF-κB和NF-κB蛋白的表达水平。结果PBMC+SAA+抗CCL20单抗组与PBMC+SAA组相比,细胞上清液中的IL-17A、IL-23和IL-6的水平显著降低(P<0.01),而TGF-β1的含量显著升高(P<0.01)。PBMC+SAA+抗CCL20单抗组相较于PBMC+SAA组,细胞中CCR6、IL-17A、ROR-γtmRNA水平显著降低(P<0.01),而Foxp3 mRNA差异无统计学意义(P>0.05)。PBMC+SAA+抗CCL20单抗组中p-NF-κB蛋白表达水平较PBMC+SAA组明显降低(P<0.01)。结论抗CCL20单克隆抗体可抑制高水平SAA相关的结节病PBMC中促炎性细胞因子的表达。Objective This study aims to investigate whether the anti-CCL20 monoclonal antibody inhibits the expression of pro-inflammatory cytokines in peripheral blood mononuclear cells(PBMCs)with high levels of serum amyloid A(SAA)associated with sarcoidosis.Methods PBMCs were extracted from both healthy individuals and patients with sarcoidosis.In addition to the blank control group and the sarcoidosis PBMC controlgroup,the PBMCs from the sarcoidosis group were separately treated with SAA,SAA+anti-CCL20 monoclonal antibody,SAA+NF-κB inhibitor BAY11-7082,or dexamethasone.After cell culture for 48 hours,cells from each group were collected.The expression levels of cytokines(IL-17A,IL-23,IL-6,CCL20,and TGF-β1)in the cell supernatant were determined using ELISA.The mRNA expression levels of CCR6,IL-17,ROR-γt,and Foxp3 in the cells were assessed using RT-PCR.The protein expression levels of p-NF-κB and NF-κB in the cells were examined by Western blot.Results In the PBMC+SAA+anti-CCL20 monoclonal antibody group,the levels of IL-17A,IL-23,and IL-6 in the culture supernatant were significantly decreased compared to that in the PBMC+SAA group(P<0.01),while TGF-β1 content was significantly increased(P<0.01).In comparison to the PBMC+SAA group,the PBMC+SAA+anti-CCL20 monoclonal antibody group exhibited a significant reduction in the mRNA levels of CCR6,IL-17A,and ROR-γt(P<0.01),with no significant difference of Foxp3 mRNA(P>0.05).The expression level of p-NF-κB protein in the PBMC+SAA+anti-CCL20 monoclonal antibody group was significantly lower than that in the PBMC+SAA group(P<0.01).Conclusion Anti-CCL20 monoclonal antibody can effectively inhibit the expression of pro-inflammatory cytokines in sarcoidosis PBMCs with elevated levels of SAA.

关 键 词：结节病 血清淀粉样蛋白A 核因子ΚB 抗CCL20单克隆抗体 

分 类 号：R392[医药卫生—免疫学]