Analysis Of Differential Expressedgene Of Immune Regulation In HIV Infected Adults Andfollow-Up Study Of Liver Function In Patients With Long-Term Haart

With the application of high-efficiency antiretroviral therapy(HAART),the life expectancy of HIV-infected people has been improved,and morbidity and mortality have decreased significantly.As a result,chronic inflammation and abnormal immune activation caused by long-term HIV infection and its non-AIDS-related complications and accumulated toxic side effects of long-term HAARThave attracted more attention.Therefore,we conducted the following research:In the first part,in order to explore genes related to chronic immune abnormalities caused by HIV infection,we selected 65 immune regulation-related genes and performed m RNA analysis in peripheral blood mononuclear cell(PBMC)samples from36 HIV-infected adults and healthy controls.We screened out 13 differentially expressed genes.Then we selected the three genes with the most significant expression differences:A20,S1PR1,and USP18.We continued to examine the PBMCs of 48HAART patients with different HIV viral loads,and compared the results with viral loads and CD4~+T cells count.Correlation analysis was performed and the correlation was found.In the second part,in order to study the role of A20 in HIV infection,we screened H9,an HIV target cell line that highly expresses A20,and knocked out the A20 gene of H9 using CRISPR/Cas9.It was found that after knockout of A20,the expression of p-P65 protein increased indicates that NF-?B activation level increased,and virus replication increased after cells infected with HIV.In the third part,we statistically analyzed 15 years of liver function related follow-up data from 765 HAART patients,and found that 84.9%of patients before treatment were at a normal level of alanine aminotransferase(ALT),with an average CD4~+T cell of 182/?l.Viral loads were>10,000copies/ml;ALT was normal in 90.1%of patients after treatment,CD4~+T cells were 382/?l,and the average viral load was75.8 copies/ml.After long-term HAART treatment,the patient's liver function did not appear to be worse,and CD4~+T lymphocytes<200/?l were risk factors related to severe liver dysfunction after HAART treatment(HR:2.457;P=0.042),and women were protective factors(HR:0.081;P=0.014).In summary,this study can draw the following conclusions:(1)Compared with healthy people,the expression levels of 13 immuno regulation associated genes(A20,CYLD,DDX24,MARCH5,MKRN2,PTP1B,RNF125, S1PR1,SOCS1,IFI35,RBCK1,TTLL12,USP18)in PBMC of HIV-infected (untreated)people have changed significantly.(2)In HAART patients,CD4~+T cell counts were positively correlated with A20 and S1PR1 expression levels;viral load was positively correlated with USP18 expression levels and negatively correlated with A20 and S1PR1.(3)The virus replication was enhanced when H9 cells were infected with HIV after knocking out the A20 gene,which may be caused by increased activation of NF-?B.(4)Under the premise of effective follow-up management and regular medication to achieve successful virologicalsuppression,liver function in patients with long-term HAART has no significant changes compared with before treatment.