Expression Of MiR-155 In HIV/AIDS Patients And Role In Abnormal Immune Activation

BackgroundNonspecific and extensive immune abnormal activation is an important symbol of human immunodeficiency virus(HIV-1)infected patients progressing to acquired immunodeficiency syndrome(AIDS).Persistent antigenic stimulation and abnormal immune activation during HIV-1 infection can also lead to immune exhaustion,a phenomenon in which effector T cells become dysfunctional and lose effector functionality and proliferative capacity.So the level of abnormal immune activation and immune exhaustion are important symbols of AIDS progression.Long-term effective HAART can inhibit HIV replication and abnormal immune activation and gradually achieve the immune reconstruction,reduce morbidity and mortality,however the virus still persist in the body of infection,the reason is that the latent reservoir(LR)has been established in the early stage of HIV infection.The latent reservoir is mainly composed of resting memory CD4+T cells.Resting memory CD4+ Tcells are largely nonpermissive for viral gene expression and the viral genome persists in a DNA form as an integrated provirus that is not actively transcribed.For this reason,the host immune system can not recognize and eliminate the latent reservoir.However,once HIV-1 infected patients stop HAART the latent virus will continue to activate replication,that is virus rebound,a large mumber of virus replication will lead to abnormal immune activation.The existence of abnormal immune activation and latent reservoir is an important factor of persistent infection with the HIV virus.MiR-155 is an important factor in regulation of the immune response.It can control the immune responses and lymphocyte differentiation level.Its expression is increased after the activation in various immune cell,especially in effect/effect memory CD8+T cells.However,the expression level of MiR-155 is low in central memory effect CD8+T cell.It is found that MiR-155 can regulate the initial regulation of T cells and the activation of CD4+T cells in HIV-1 infection,then affecting the progression of HIV-1 infection.MiR-155 also plays a role in HIV-1 latent reservoir cells.The last studies have found that TRIM32 is the target of miR-155.TRIM32 can make the transient activation of latent reservoir cells back to the latent state.MiR-155 can also interrupt the HIV-1 replication by working on a series of HIV-1 replication process depending on host factors(these host factors involved in the nuclear transport process integration complex).After the treatment o f miR-155,preintegration complexes(late RT products)were increased.However,the current expression of miR-155 in peripheral blood of HIV-1 infected patients and the relationship between miR-155 and the HIV/AIDS immune activation or HIV-1 DNA are not clear.objectiveThis study intends to find out the expression level of miR-155 in peripheral blood and the relationship between miR-155 and immune activation or HIV-1 DNA in HIV/AIDS patients..We also study the effect of miR-155 in the latent cell and HIV infection(H9/HIV ?B)cell.Method1.Subjects:HIV-infected patients(including three subgroups,64 Virologic responders?57 Virologic non-responders?40 HAART-naive)and 35 normal were recruited in the study.Plasma,peripheral blood lymphocytes(PBMC),CD4 T cells and CD8 T cells were collected in all the subjects.2.Detection:Expression levels of miR-155 in plasma,PBMC,CD4 T cells and CD8 T cells were measured by Real-time RT-PCR.We also detect the symbols of activation of immune cells(CD4/CD8 CD38)and immune exhaustion(PD1)by flow cytometry.Furthermore,we analysis the correlation between expression level of miR-155 and immune activation or immune level in CD4+,CD8+T cell.3.We detect the blood HTV-1 DNA in HIV-1 infection by HIV-1DNA quantification kit.We further analysis the correlation between expression level of miR-155 and HIV-1 DNA.4.In vitro studies:Effects of miR-155 on reactivation of HIV provirus and HIV replication were evaluated by overexpression and interference of miR-155 in two cell lines,J-Lat(a cell line with latent HIV infection)and H9 cell line(with stable infection of HIV IIIB).(the level of core antigen P24 is generally considered to be the indirect marker of viral replication,we by detecting P24 to evaluate miR-155 expression regulation of latent cell activation and the effect of HIV replication)5.Statistical analysis:All data were analyzed using SPSS 17.0 statistical software.Test results are used mean ? standard deviation(x±s)said.One-way ANOVA was used to compare the multiple sample groups.Spearman correlation analysis was used to analyze the correlation between the two groups.P<0.05 for the difference was statistically significant.Result1.The expression of miR-155 in PBMC,CD4 + T and CD8 + T cells of HIV-1 infected patients was significantly higher than that of normal controls(P<0.01).The expression of miR-155 was significantly lower in Virologic responders than Virologic non-responders(P<0.05),and close to the normal level.The expression of miR-155 in cell is different from the expression in plasma.2.HTV-1 infection can induce CD4+?CD8+T cell immune activation and immune exhaustion,effective HAART therapy can improve the immune activation and immune exhaustion level.Further correlation analysis found that the expression of miR-155 was positively correlated with the T cell immune activation(CD38)?immune exhaustion(PD-1)in HIV-1 infection,3.In Virologic responders,HIV-1DNA was significantly lower than Virologic non-responders and HAART-naive(P<0.01),HIV-1DNA and CD4+T cell miR-155 expression level was positively correlated with CD4+T immune activation level.4.Overexpression of miR-155 could decrease the expression of P24 antigen after latent cell activation and had no effect on the expression of HIV-1P24 antigen in HIV-1 infected cells(H9/HIV ?B).conclusion1.The expression level of miR-155 was increased in HIV-1 infected patients and positively correlated with immune activation CD38 and PD-1 of T cells in HIV-1 infected patients.MiR-155 plays an important role in the regulation of HIV-1 immune activation and immune exhaustion.2.MiR-155 is positively correlated with HIV-1 DNA,and in vitro studies have demonstrated that overexpression of miR-155 has an inhibitory effect on latent cell activation...