| Chronic active Epstein-Barr virus infection?CAEBV?is a special condition of chronic EBV infection that refers to persistent or fulminant infectious mononucleosis like symptoms,abnormal EBV-specific antibody titers and markedly elevated levels of EBV DNA in the peripheral blood after infection with Epstein-Barr virus.The lesions involve multiple functional organs.It usually occurs in children and adolescents with complex clinical manifestations.Patients are prone to die under various complications,which lead to high mortality and poor prognosis.The diagnosis of the disease is still difficult due to unknown pathogenesis and lack of specific symptoms.What's worse,no therapy for this disease has proven to be broadly effective.Here we analyzed and summarized the general condition and features of adult patients with CAEBV so as to improve the awareness of CAEBV.Then we detected the serum protein expression?Th1/Th2 cytokines?,EBV-positive lymphocyte proliferation properties?TCR gene rearrangement?and identified the differential genes by genome-wide association studies in adult patients with CAEBV.The findings suggest that there is an unbalanced cytokine profile and some differential expressed single nucleotide polymorphisms,which may provide reference and help for early diagnosis of the disease and related etiology research.Part one:Detection of Th1 and Th2 Cytokines in Patients with CAEBVObjective:To evaluate the changes and clinical significance of Th1/Th2 cytokines in patients with CAEBV via detecting the serum levels of TNF-?,IFN-?,IL-4 and IL-10.Methods:Serum samples were collected from 6 patients with CAEBV and 16 healthy volunteers who visited the Fuzhou General Hospital.Enzyme-linked immunosorbent assay?ELISA?was used to detect the levels of T helper?Th?1?IFN-?,TNF-??and Th2?IL-4,IL-10?type cytokines.Statistical analysis was performed using the Mann-Whitney U test for non-parametric data.Results:Serum levels of IFN-?,TNF-?and IL-10 were significantly higher in patients with CAEBV than in healthy volunteers?IFN-?:median 129.9 pg/ml,range31.87-16520,versus median 0 pg/ml,range 0-27.08,p<0.001;TNF-?:median 79.98pg/ml,range 44.67-1080,versus median 7.865 pg/ml,range 6.463-11.44,p<0.001;IL-10:median 48.41 pg/ml,range 15.47-496.4,versus median 1.565 pg/ml,range1.320-2.410,p<0.001?.There is no difference of the serum levels of IL-4 between patients with CAEBV and healthy volunteers?IL-4:median 2.385pg/ml,range2.028-3.605,versus median 1.565 pg/ml,range 1.320-2.935,p=0.302?.Conclusion:The expression of TNF-?,IFN-?and IL-10 were increased in patients with CAEBV,suggesting that there is an“unbalanced cytokine profile.”Part two:Detection of T Cell Receptor Beta and T Cell Receptor Gamma Gene Rearrangement in Patients with CAEBVObjective:To assess the diagnostic value of T cell receptor gene rearrangement in patients with CAEBV by detecting the TCR beta and TCR gamma rearrangement.Methods:The genomic DNA was extracted from the peripheral blood of 6 adult patients with CAEBV who visited Fuzhou General Hospital.Then the T cell receptor beta and T cell receptor gamma rearrangement were detected by using Biomed-2multiplex polymerase chain reaction?PCR?and capillary electrophoresis gene scan.Results:All the 6 DNA samples were qualified.Multiplex PCR demonstrated monoclonal TCR beta gene rearrangement in 1 of 6?16.7%?and 3 of 6?50%?in patients with CAEBV.Conclusion:Monoclonicity of TCR?and TCR?is an important feature of CAEBV,but the results of monoclonal rearrangement need to be combined with the patient's clinical symptom and laboratory examination.Part three:Genomic SNP Association Analysis in Adult Patients with Chronic EBV InfectionObjective:To screen and analyze the single nucleotide polymorphisms?SNPs?and differentially expressed genes in patients with chronic EBV infection.Methods:The 120 DNA samples of 46 patients with chronic EBV infection and 74healthy volunteers who visited the Fuzhou General Hospital were scanned by Illumina SNP chips to detect the whole genome genotyping,then we filtrate variants that doesn't make sense by PLINK software.Genome-wide association study?GWAS?was used to screen out the SNPs and differentially expressed genes.The SNP loci with significant differences in the two groups were analyzed by GO enrichment and KEGG enrichment.Results:A total of 14185 SNPs were found to be of significant difference between cases and control groups?P<0.05?.The major differentially expressed SNPs that are significantly associated with chronic EBV infection(P<10-5)are shown as follows:rs80019788,rs10142901,rs1172402,rs3116565,rs5942250 and rs77220782.Gene enrichment analysis of the differentially expressed SNPs indicated that these genes were mainly located in cell junction and cell membrane,which were significantly correlated with the process of cell adhesion and calcium ion transmembrane transport.The molecular function mainly lay in actin binding,calmodulin binding,calcium ion binding and calcium channel activity,mainly related to focal adhesion,calcium signaling pathway,inflammatory mediator regulation of TRP channels,PI3K-Akt signaling pathway and MAPK signaling pathway.Conclusion:There are some differential SNPs associated with the development of chronic EBV infection.Enrichment analysis showed that the differential genes are mainly located in cell junction and cell membrane.Calcium signaling pathway and MAPK signaling pathway may be involved in the chronicity of EBV infection. |
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