Characteristics And Clinical Significance Of EBV Infection In HIV Infected And AIDS-related Lymphoma Patients

Background and objectives:To investigate the characteristics of epidemic strains of Epstein-Barr virus(EBV) infection in HIV-1 infected patients in eastern China and the expression characteristics and clinical significance of EBV-encoded micro RNAs in AIDS-associated lymphoma(ARL)and provided virology-related reference for clinical occurrence of HIV-1-infected patients and EBV-related diseases such as ARL.Research subjects and methods:1?A total of 186 HIV-1-infected patients and 52 patients without HIV in our hospital were enrolled in this study,saliva and blood samples were collected,and viral load in the blood was determined by fluorescence quantitative PCR.EBV typing was performed using saliva according to polymorphisms in the EBNA3C region.EBV LMP-1 carboxy terminus(C-ter)was sequenced,and the sequencing results were compared with the epidemic strains in the world.Besides,the differences between two groups were analyzed.2?Plasma was collected from 35 ARL patients with EBV infection and 20 HIV-infected patients with EBV infection,and RNA was extracted.The EBV microRNA quantification chip was used to screen differentially expressed EBV micro RNAs between the two groups,and expanded samples to validate and performed target gene prediction analysis.Results:First part1?Among HIV-1 infected patients,EBV was mainly type 1,and type 2 had a higher viral load.There were differences in EBV viral load levels among different EBV subtypes(P value<0.0001).Type 2 had higher viral load than type 1(P value=0.001,<0.05)and mixed type 1,2(P value=0.006,<0.05).2?The distribution of EBV viral load was different between HIV and non-HIV infected controls.When the viral load levels were low to medium(10 to 10~2copies/?l)and medium to high(10~2to 10~3copies/?l),there were differences in the distribution of patient cases,with P values<0.0001 and 0.01,respectively.3?The EBV LMP-1 variants of HIV and non-HIV-infected patients were mainly China1 variants,but the distribution of LMP-1 C-ter variant types,30bp deletion(del30)and 33bp tandem repeats(rep33)were different between the two groups,with P values<0.0001.HIV-infected patients had different nucleic acids positions of del30 deletion,while non-HIV-infected patients only had deletions at the same nucleic acid position(168295-168266);HIV-infected patients had a higher incidence of high rep33 copies than non-HIV-infected patients.Second part1?5 ARL patients and 8 HIV-infected patients without lymphoma had a different EBV miRNA sexpression in screening results in ebv-miR-BART2-5p,ebv-miR-BART8-3p,ebv-miR-BART15,ebv-miR-BART19-5p and ebv-miR-BART9-5p.And the relative differential expression multiples were 219.69,92.67,36.56,14.16,-1.99 respectively.2?q RT-PCR expanded samples validated the screening results of 5 EBV miRNAs and ebv-miR-BART6-3p in 35 ARL patients.Compared with 20 HIV patients without lymphoma,the differential expression of 6 EBV miRNAs was consistent with the screening results.The relative expression of ebv-miR-BART2-5p,ebv-miR-BART8-3p,ebv-miR-BART15,ebv-miR-BART19-5p,bv-miR-BART6-3p was up-regulated,miR-BART9-5p was down-regulated.3?The relative expression levels of ebv-miR-BART8-3p,ebv-miR-BART19-5p, and ebv-miR-BART9-5p in plasma were positively correlated with the international prognostic index(IPI)score,the Eastern Cooperative Oncology Group(ECOG)score and the occurrence of lymphoma B symptoms,with P values=0.03,0.01,and 0.04,respectively.Compared with the effect of IPI score on ARL prognosis evaluation,6EBV miRNAs were better as biomarkers for ARL prognosis evaluation.It was predicted that some target genes and corresponding 20 signal pathways such as I?B/NF-?B signal pathway and T cell receptor signal,which may be related to the pathogenesis of ARL.Conclusion:1.In the eastern China,EBV in HIV-infected patients had higher active virus replication;EBV infection was mainly type 1 in HIV-infected patients,and EBV type 2infection has higher EBV virus load;the LMP-1 variants were mainly China1 variants.Compared with non-HIV-infected patients,there were difference in the distribution of LMP-1 C-ter variant types and the incidence of del30 and rep33 hotspot mutations.2.The expression of ebv-miR-BART2-5p,ebv-miR-BART8-3p,ebv-miR-BART15,ebv-miR-BART19-5p,ebv-miR-BART9-5p and ebv-miR-BART6-3p in ARL patients were highly different;and the clinical characteristics of ARL patients(IPI score,ECOG score,and lymphoma B symptoms)were closely related with the expression of ebv-miR-BART8-3p,ebv-miR-BART19-5p and ebv-miR-BART9-5p.And these 6 EBV miRNAs could be used as biomarkers for the prognosis evaluation of ARL.Its pathogenic effect may be through the regulation of ARL tumor signaling pathways.