瓜氨酸化人乳头状瘤病毒47-E2345-362肽抗体在类风湿关节炎发病中的意义

目的 检测类风湿关节炎(RA)患者血清中与聚丝蛋白原306-324多肽高度同源的人乳头状瘤病毒E2345-362肽及瓜氨酸化E2345-362肽的抗体水平,并探讨其在RA发病中的意义.方法 选取RA患者119例、其他风湿病患者223例、健康对照101名.应用NCBI数据库BLAST方法检索出与聚丝蛋白原306-324肽高度同源的人乳头状瘤病毒(HPV)47型E2蛋白的345-362多肽序列(E2345-362),并以瓜氨酸替换此多肽序列中第348位上的精氨酸.合成并纯化E2345-362肽及瓜氨酸化E2345-362肽.分别以E2345-362肽及瓜氨酸化E2345-362肽为抗原,用酶联免疫吸附法(ELISA)检测患者血清中相关抗体的水平.并研究其与RA患者发病年龄、病程、疾病活动指数(DAS28)评分、抗环瓜氨酸(CCP)抗体、抗肽酰精氨酸脱亚胺酶(PADI)4抗体、C反应蛋白(CRP)、关节x线分期等指标的关系.结果 ①RA患者组血清中抗瓜氨酸化E2345-362蛳肽抗体的阳性率(41.2%)及滴度(Au值为105.7)明显高于其他对照组(P均<0.01);RA患者、其他对照组血清中抗E2345-362.肽抗体的阳性率差异无统计学意义.②与CCP抗原预孵育后的血清中抗瓜氨酸化E2345-362肽抗体水平明显下降(P<0.05).③抗瓜氨酸化E2345-362肽抗体阳性组与阴性组抗CCP抗体的阳性率差异有统计学意义(P<0.01);两组间的抗PAD14抗体阳性率及滴度差异有统计学意义(P<0.01);相关性分析显示抗瓜氨酸化E2345-362肽抗体滴度与抗PADl4抗体滴度具有相关性(r=0.485,P<0.01).④抗瓜氨酸化E2345-362肽抗体阳性患者的DAS28评分、ESR及X线分期均高于阴性患者(P均<0.05);抗瓜氨酸化E2345-362肽抗体阳性组的抗角蛋白抗体(AKA)阳性率、抗核周因子(APF)阳性率及RF均高于阴性组(P均<0.05).结论 RA患者血清中可以检测到抗瓜氨酸化HPV47-E2345-362肽的抗体,该抗体与抗CCP抗体存在部分交叉反应,其滴度与抗PAD14抗体滴度呈正相关,且与疾病活动及病情严重程度相关,提示HPV47-E2345-362肽可能与RA的发病相关,RA患者体内PAD14的增多可能使E2345-362肽瓜氨酸化,继而机体产生相应的抗体,参与RA的发病.

Significance of anti-citrullinated human papilloma virus-47 E2345-362 peptide antibodies in the pathogenesis of rheumatoid arthritis

Objective To detect antibodies against human papilloma virus-47 E2m345-362 peptide whichis homologous to profilaggri306324 peptide and anti-citrullinated human papilloma virus-47 E2345-362 peptide antibodies in rheumatoid arthritis(RA)and to investigate its role in the pathogenesis of RA.Methods Serum samples were obtained from 119 patients with RA, other rheumatic diseases and healthy individuals.We searched the homologus sequence of profilaggrin306-324peptide by using NCBI (the National Center for Bioteehnology Information)BLAST (basic local alignment search tool),and synthesized human papilloma virus-47 E2345-362 peptide which was highly homologous to profilaggrin306-324 peptide and the citrullinated Human papilloma virus-47 E2345-362 peptide.The presence of antibodies against E2345-362 peptide and citrullinated E2345-362 peptide was examined by enzyme-linked immunosorbent assay(ELISA).The associations between these antibodies and the clinical features of RA were evaluated.Results ①(41.2%)and titer (AU was 105.7)of anti citrullinated E2345-362 peptide antibodies in RA were significantly higher than those of patients with other rheumatic diseases and healthy individuals.However,the prevalence of anti-E2345-362 peptide antibodies in RA patients was similar to that of patients with other rheumatic diseases and healthy individuals(P>0.05).②The samples that were pre-incubated with cyclic citrullinated peptide (CCP) had lower titer of anti-citrulllinated E2345-362 peptide antibodies.③The titers of anti-CCP antibodies and anti-PAD14 antibodies in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negtive patients.It showed significant correlations between anti-citrulllinated E2345-362 peptide antibodies and anti-PAD14 antibodies(r=0.485,P<0.01).④ DAS28 score,ESR,X-ray stages,AKA in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negative patients.Conclusion The presence of anti-citrullinated E2345-362 peptide antibodies in RA indicates that HPV-47 E2 may act as an auto-antigen which may play an important role in the pathogenesis of RA.The increasing of PAD14 may make it easy for HPV-47 E2 to be citrullinated and may induce the subsequent auto-immune reactions.