Study On The Relationship Between The Characteristics Of Intestinal Injury And Immune Reconstitution In Patients With Chronic HIV-1 Infection After ART

Objective:By detecting the intestinal tissue of immune responders(IRs)and immune nonresponders(INRs)who have been treated with antiretroviral therapy(ART)for more than 2 years after being infected with HIV-1 virus,the correlation and clinical significance between the changes of intestinal injury and immune reconstitution in IRs and INRs groups were explored.Methods:A total of 18 HIV-1 infected patients who have been treated with ART for more than 2 years were enrolled in this study,including 9 in IRs group,7 in INRs group,and 6 in health controls(HCs).(1)Immunohistochemical and H&E staining techniques were used to explore the intestinal appearance characteristics,intestinal tight junction protein(claudin-1,ZO-1),intestinal injury(I-FABP),intestinal apoptosis(caspase-3),intestinal inflammation(TNF-α)and other relevant molecules to detect.,compare and analyze the differences of these indicators in patients of HCs,IRs and INRs,and analyze their correlations with the main immune indicators;(2)Immunofluorescence technology was used to detect the cells(CD4+T,Treg,Th17)in intestinal tissue and analyze their differences and correlations in HCs,IRs,INRs;(3)RNAscope in situ was used to detect the expression of HIV RNA in HCs,IRs and INRs in intestine.(4)Quantitative real-time polymerase chain reaction(qPCR)was used to detect HIV DNA,cell-associated(CA)HIV RNA and intestinal tissue related molecules(claudin-1,ZO1,TNF-α)in intestine and analyze their differences and correlations in HCs,IRs and INRs;(5)Enzyme-linked immunosorbent assay(ELISA)was used to detect intestinal related molecules(claudin-1,ZO-1,I-FABP,TNF-α)in plasma,and explore their change characteristics and correlation analysis in HCs,IRs and INRs.Results:(1)In H&E staining,compared with IRs and INRs groups,the intestinal glands in HCs group were arranged more orderly and tightly,the intestinal mucosa structure was more complete and continuity,interstitial edema was found in IRs group and fibrosis in INRs group;The number of intestinal glands was counted under the same size microscope,compared with the IRs and INRs groups,the number of intestinal glands in the HCs group was the highest,the number of intestinal glands in the IRs group was higher than that in the INRs group,and the number of intestinal glands in the INRs group was the lowest.(2)Compared with HCs group,the apoptosis of epithelial cells in IRs group and INRs group increased in intestinal caspase-3 immunohistochemical staining.(3)Whether in the intestine or peripheral blood,claudin-1:HCs>IRs>INRs.(4)In intestinal tissue,compared with HCs and IRs groups,the content ofZO-1 in INRs group decreased significantly,and the detection of ZO-1 in peripheral blood was as follows:HCs>IRs>INRs.(5)Through immunohistochemical staining,we found that IFABP in intestinal tissue was higher in INRs group than in HCs group;In the detection of I-FABP in peripheral blood,we found that both HCs and IRs groups were lower than INRs group.(6)Whether in the intestine or peripheral blood,the TNF-α increased in IRs and INRs groups compared with HCs group.(7)We located and found HIV RNA in the intestine of patients in IRs and INRs groups by RNAscope in situ.At the same time,through the quantitative detection of the virus,we found HIV DNA:INRs>IRs,CA HIV RNA:INRs>IRs in both intestinal tissues and PBMCs.(8)Immunofluorescence staining of CD4+T cells in intestinal samples of three groups subjects:HCs>IRs>INRs;Th17 cells:compared with INRs group,Th17 cells in IRs group and HCs groups increased;Treg cells:compared with IRs group,Treg cells in INRs group increased;Th17/Treg ratio:compared with INRs group,Th17/Treg ratio in IRs group and HCs group increased.(9)At the same time,we analyzed the correlation between the above intestinal injury indicators and immune reconstitution indicators,as well as Th17 cells,Treg cells and Th17/Treg ratio,and found that the patients with poor immune reconstitution(lower CD4+T cells,higher virus-related indicators)had more severe intestinal injury(lower claudin-1,lower ZO-1,lower number of intestinal glands,and higher I-FABP in peripheral blood),and Th17/Treg ratio had more significant correlation with intestinal injury and immune reconstitution than Th17 and Treg cells,The patients with poor immune reconstitution(lower CD4+T cells and higher virusrelated indicators)not only suffered severe intestinal injury,but also decreased in Th17/Treg ratio.Conclusions:(1)After long-term antiviral treatment,HIV-1 infected patients with successful or failed immune reconstitution have different degree of intestinal injury,and the intestinal injury of patients with failed immune reconstitution is more serious;(2)The degree of immune reconstitution in HIV-1 infected people after ART treatment is closely related to the degree of intestinal injury;(3)Th17/Treg ratio is negatively correlated with the degree of intestinal injury and positively correlated with the degree of immune reconstitution.Whether it can be used as an indicator for the evaluation and prediction of intestinal injury is worth studying.