Study On Chemokine Alterations Of HIV Infected Individuals And The Effects And Mechanisms On Function Of Killer Cells And Infection Of HIV

Objective:Plasma chemokine levels significantly altered after HIV infection,but the effects of chemokine,the largest subfamily of cytokine,on variety of events releted to HIV pathology,such as HIV disease progression,immune reconstitution,dysfunction of immune cells and process of HIV infection are still unknown.The current study aim at exploring the alteration of the spectra of chemokine prior to and after HIV infection and defining the key chemokine and further clarify its roles and mechanisms on HIV pathology,which may provide novel insights and methods for HIV functional cure and target therapy.Methods:In the first part,we chose 54 participants to explore the alteration of chemokine panorama prior to and after HIV infection.Bio-Plex Human Chemokine Panel 40-Plex kits were used to evulate the plasma chemokine levels.We then analyzed the correlations between chemokine levels and index related to HIV disease progression and immune reconstitution.Through summarizing the obtained results,the key chemokine was determined and was studied in the next step.In the second part,we chose untreated HIV⁺subjects?HIV⁺?,ART treated HIV⁺subjects?HIV⁺on ART?,and normal controls?NC?,to determine the effect of high levels of IP-10 on killer cell,NK cell,receptor expression and function.The cell culture and flow cytometry were used in comparing the function of chemokine-treated NK cells and receptor expression on NK cell subfamily.In the third part,we separated resting memory CD4⁺T cells mainly from NC subjects,the flow cytometry,cell culture,in vitro viral infection,P24 detection,digital droplets PCR and confocal microscope were used in evaluating the effects and mechanisms of chemokines in process of HIV infection of resting memory CD4⁺T cells.Results:1.By detecting spectra of chemokine prior to and after HIV infection,we found16 chemokine levels significantly altered,and some of them were associated with HIV disease progression and immune reconstitution,and IP-10 was the most important chemokine.2.High levels of IP-10 could decrease the secretion of IFN-?of NK cells from HIV⁺and NC subjects.CXCR3 expression on total NK,CD56^-CD16⁺NK and CD56^dim CD16⁺NK cells were significantly elevated and IP-10 played roles mainly through combined with CXCR3 receptor on the NK cells.3.High levels of IP-10 could promote HIV infection of the resting memory CD4⁺T cells,which was mainly through promoting HIV entry and integration,but not promoting the activation and proliferation of resting memory CD4⁺T cells.After blocking of IP-10 and CXCR3,the effect of IP-10on HIV infection was inhibited,which indicated that it was IP-10 promoted HIV infection of resting memory CD4⁺T cells.In exploreing the further mechanism,we found IP-10 effect on actin and cofilin in resting memory CD4⁺T cells,moreover,IP-10may mainly use LIMK-cofilin signaling pathways to affect the polarization state of the actin and further impact on HIV infection.LIMK inhibitor might counteract the effect of the promoted infection by IP-10.Conclusion:1.16 chemokine levels were significantly altered during primary HIV infection,among them,IP-10 was significantly increased during primary HIV infection,and correlated with PHI VL,HIV disease progression and immune reconstitution.2.IP-10 could inhibited NK cell function mainly through inhibited the function of CXCR3⁺NK cells.3.IP-10 through influence the activity of actin to promote HIV infection of resting memory CD4⁺T cells,which further promoted the establishment of HIV reservoir.