EB病毒在系统性红斑狼疮患儿外周血单个核细胞中表达的意义

Expression of EB virus in peripheral blood mononuclear cells in children with systemic lupus erythematosus

目的 通过研究EB病毒（EBV）在系统性红斑狼疮（SLE）患儿外周血单个核细胞（PBMCs）中DNA拷贝数及EBV基因的表达,探讨EBV在儿童SLE发病中的作用.方法 1.分离30例SLE患儿和12例健康对照儿童PBMCs,提取DNA.2.用B95.8 EBV株与提取的PBMCs共同培养12 d,提取培养前后的PBMCsRNA.3.实时荧光定量PCR法检测PBMCs EBV DNA拷贝数.4.用反转录PCR的方法检测EBV基因的表达.结果 1.SLE组PBMCs EBV DNA[（658.6±183.6）拷贝/μg]显著高于健康对照组[（40.1±11.6）拷贝/μg]（P＜0.05）;SLE活动组PBMCs EBV DNA[（785.2±179.2）拷贝/μg]显著高于非活动组[（586.0±193.1）拷贝/μg] （P ＜0.05）.2.EBV PBMCs EBV DNA拷贝数与SLE病情活动程度评分不存在相关性（r=0.03,P＞0.05）.3.用EBV培养PBMCs后,结果发现SLE组和健康对照组潜伏期基因和裂解期基因表达均有不同程度的增加,潜伏期基因潜伏膜蛋白1（LMP1）、LMP2、EBV核抗原1和裂解期基因BCRF1、BLLF1的表达与健康对照组比较差异均有统计学意义（P均＜0.05）.结论 SLE患儿EBV DNA拷贝数显著增加,并且EBV基因异常表达,提示EBV可能参与SLE的发病过程.

Objective To discuss the role of EB virus （EBV）in the pathogenesis of systemic lupus erythematosus（SLE） in children through investigating the copies of EBV DNA and expression of EBV genes in peripheral blood mononuclear cells（PBMCs）.Methods （1）PBMCs were isolated from 30 patients with SLE and 12 healthy normal controls respectively and DNA was extracted from PBMCs.（2） PBMCs were co-cultured with EBV for 12 days and RNA was extracted from PBMCs.（3）Real-time fluorescence quantitative PCR（Real-time PCR） was applied to detect the copies of EBV DNA in PBMCs.（4）Reverse transcription PCR was applied to detect expression of EBV genes.Results （1） Compared with the healthy control group [（40.1 ± 11.6） copies/μg],a significant increase of EBV DNA copies was observed in SLE group[（658.6 ± 183.6） copies/μg] （P 〈0.05）.The EBV DNA copies in the active SLE group [（785.2 ± 179.2） copies/μg] were significantly higher than those in the non-active SLE group [（586.0 ± 193.1） copies/μg] （P 〈 0.05）.（2）There was no correlation between EBV DNA copies and systemic lupus erythematosus disease activity index （r =0.03,P 〉 0.05）.（3） After PBMCs got co-cultured with EBV,expression of latent EBV genes and lytic genes were both increased in the patients and healthy controls.The latent EBV genes including latent membrane protein 1 （LMP1）,LMP2,EBV nuclear antigen 1 and the lytic genes including BCRF1,BLLF1 were all increased significantly in the patients compared with the healthy controls （all P 〈 0.05）.Conclusions There is a significant increase of EBV DNA copies and aberrant expression of EBV genes in SLE patients,which suggests that EBV may contribute to the pathogenesis of SLE.