儿童EB病毒感染非肿瘤性疾病与病毒潜伏期膜蛋白基因多态性的相关性研究

The correlation between the non-neoplastic diseases of children with EB virus infection and the polymorphisms of LMP1 gene during virus latent period

目的分析儿童EB病毒(EBV)感染相关非肿瘤性疾病与病毒潜伏期膜蛋白(LMP1)基因多态性的相关性。方法选择2017年12月至2020年12月在我院确诊的94例EBV感染相关非肿瘤性疾病患儿为观察组,其中传染性单核细胞增多症(IM)40例,EBV相关噬血细胞性淋巴组织细胞增生症(EBV-HLH)30例,慢性活动性EBV感染(CAEBV)24例,随机选取同期携带EBV病毒的无症状儿童94例为对照组。检测所有对象血清免疫球蛋白水平,RT-PCR检测外周血淋巴细胞EBV DNA载量和LMP1 mRNA水平,Westernblot检测LMP1蛋白表达,巢式PCR检测LMP1基因多态性。Pearson相关性分析LMP1基因多态性与病毒载量、LMP1表达和免疫球蛋白之间的关系。结果IM组患儿血清IgA和IgM水平高于对照组,EBV-DNA载量和LMP1表达水平高于对照组(P<0.05);EBV-HLH组IgG、IgA、IgM、C3和C4水平低于IM组和对照组(P<0.05),EBV-DNA载量和LMP1表达水平高于IM组和对照组(P<0.05);CAEBV组患儿IgG、IgA、C3和C4水平低于IM组和对照组,高于EBV-HLH组(P<0.05),IgM和IgE水平高于EBV-HLH组、IM组和对照组(P<0.05),EBV-DNA载量和LMP1表达水平低于EBV-HLH组,高于IM组和对照组(P<0.05);EBV-HLH组患儿LMP1基因XhoⅠ位点(-)和del-LMP1的发生率明显高于IM组和对照组(P<0.05)。Pearson相关性分析显示,EBV感染相关非肿瘤疾病患儿LMP1基因XhoⅠ位点(-)变异和del-LMP1位点变异与EBV-DNA和LMP1表达呈显著正相关关系(P<0.05)。结论EBV感染相关非肿瘤疾病患儿LMP1基因XhoⅠ位点(-)变异和del-LMP1位点变异与疾病进展有关。

Objective To analyze the relationship between the Epstein-Barr virus(EBV)infection associated non-neoplastic diseases and the polymorphisms of latent membrane protein 1(LMP1)gene in children.Methods A total of 94 children confirmed with EBV infection associated non-neoplastic diseases in our hospital from December 2017 to December 2020 were enrolled as an experiment group,including 40 cases with infectious mononucleosis(IM),30 cases with EBV associated hemophagocytic lymphohistiocytosis(EBV-HLH)and 24 cases with chronic active EBV infection(CAEBV).Another 94 asymptomatic children with EBV during the same period were randomly enrolled as a control group.Levels of serum immunoglobulins in all subjects were detected.EBV DNA load and level of LMP1 mRNA in peripheral blood lymphocytes were detected with RT-PCR.The expression of LMP1 proteins was detected with Western blot.The polymorphisms of LMP1 gene were detected with nested PCR.The relationship between the polymorphisms of LMP1 gene and the viral load,LMP1 expression,and immunoglobulins was analyzed with Pearson correlation analysis.Results The levels of serum IgA and IgM,EBV-DNA load and expression level of LMP1 were higher in the IM group than those in the control group(P<0.05).The levels of IgG,IgA,IgM,C3 and C4 were lower while EBV-DNA load and expression level of LMP1 were higher in the EBV-HLH group than those in the IM group and the control group(P<0.05).The levels of IgG,IgA,C3 and C4 in the CAEBV group were lower than those in the IM group and the control group,and higher than those in the EBV-HLH group(P<0.05).The levels of IgM and IgE in the CAEBV group were higher than those in the EBV-HLH group,the IM group and the control group(P<0.05).EBV-DNA load and expression level of LMP1 in the CAEBV group were lower than those in the EBV-HLH group,while higher than those in the IM group and the control group(P<0.05).The incidence of LMP1 gene at Xho I locus(-)and del-LMP1 in the EBV-HLH group was significantly higher than that in the IM group and the control group(P<0.05).Pearson correlation analysis showed that the variations of LMP1 gene at Xho I(-)and del-LMP1 loci were significantly positively correlated with EBV-DNA and LMP1 in children with EBV infection associated non-neoplastic diseases(P<0.05).Conclusion The variations of LMP1 gene at Xho I(-)and del-LMP1 loci are related to disease progression in children with EBV infection associated non-neoplastic diseases.