The Regulatory Function Of TIM3 And PD1 In Macrophage During HBV Infection

BackgroundChronic hepatitis B virus infection is a serious public health problem.Hepatitis B virus infection,if not effectively controlled,can be further deteriorated into cirrhosis or even hepatocellular carcinoma(HCC),which a serious threat to human health.Patients with chronic hepatitis B(CHB)in patients with virus specific adaptive immune function is inactivated,accompanied by the inherent immune dysfunction of hepatitis.At present,during hepatitis B virus(HBV)infection,it is not still unclear about the molecular mechanisms of the change of the innate immunity regulatory function.T cell immunoglobulin and mucin-3(TIM3)and programmed death receptor 1(PD1)are two of the newly discovered immune regulatory molecules in recent years,both expressed in adaptive immune cells such as CD4+T cells and CD8+T cells,and are also expressed on innate immune cells such as monocytes,macrophages,dendritic cells,mast cells.Therefore,TIM3 and PD1 have plenty of immune regulatory functions.Aims(1)to study the expression level changes after blocking TIM3 or the expression level of PD1 after blocking PD1 on the surface of macrophages in patients with chronic hepatitis B or the Health control;(2)to determine the role of TIM3 and PD1 in regulating the polarization of chronic hepatitis B virus;In order to further explore the immunological characteristics of chronic HBV infection and provide a theoretical basis for the new treatment.MethodsPeripheral blood samples from 31 patients with chronic hepatitis B(CHB)and 46 healthy controls were collected,and PBMC was isolated,then stimulated with PMA.Respectively block macrophage surface molecules TIM3 with anti-TIM3 antibody and PD1 with anti-PD1 antibody,flow cytometry was used to detect the cell surface molecule TIM3 and after blocking PD1 and PD1 after blocking TIM3;to detect the surface marker CD 163 and CD206 expression level;Sort CD 14+cells using the MACS sorting,then detect the secretion level of culture supernatant TNF-a,IL-12,IL-6,IL-1 β,IL-8.Results(1)The expression of PD1 in macrophages was decreased after TIM3 blockade,and the expression of TIM3 was not significantly changed after PD1 blockade.(2)In macrophages,anti-TIM3 or anti-PD1 blocking group was significantly lower than that of the control group,the expression level of CD 163 and CD206 decreased.(3)In macrophages,blocking TIM3 pathway can increase the levels of TNF-a,IL-6,IL-10,IL-12,and is of no significant effect on the secretion of IL-1β and IL-8.Blocking the PD1 pathway could increase the secreation levels of TNF-a,IL-6,IL-10 and IL-12,and had no significant effect on the secretion of IL-1β and IL-8.These results suggest that TIM3 and PD1 may have synergistic blockade effect.Conclusions(1)It was confirmed that the PD1 pathway was blocked in the peripheral blood of chronic hepatitis B,and there was no significant change in TIM3.(2)Blocking the TIM3 pathway or blocking the PD1 pathway,can promote the polarization of macrophages to M1.(3)The influence of macrophages to M1 type polarization by blocking the TIM3 pathway is greater than blocking the PD1 pathway.