The Study On The Correlation Of CCR5,CXCR3,CD25 And CD81 With HCV/HIV Co-infection In China

Hepatitis C ( HC) is caused by hepatitis C virus ( HCV) to be a complicated pathologic status. HCV infection has a wide distribution in the world. And 3. 2% Chinese are HCV seropositive in China. The majority of the individuals infected with HCV develop chronic infection, progressive liver injury, fibrosis and cirrhosis, and even hepatocellular carcinoma. HCV infection is a major public health problem now.The mechanism of HCV infection has not been clear. It has been proposed that the immune response results in the liver injury. It is suggested that the HCV injury mechanism is closely correlative with the host immune response. The number of HIV infectors has been increased in China and the majority of infected individuals are injection drug users (IDUs). Furthermore 94% HCV infectors co - infect HIV in IDUs. HIV infection can impair the immune system. There is close correlation between the expressions of CD81 ,CCR5,CXCR3 and CD25 on CD4+ and CD8+ T lymphocytes and the immune responses to chronic viral infection. So far, the study on how the immune response alters when HCV/HIV co -infection is too limited. In this study, to investigate the relationship between the immune response and HCV/HIV co - infection, we analyzed the levels of CD81, CCR5, CXCR3 and CD25 expressed on the surface of CD4+ and CD8 +T lymphocytes from individuals with HCV infection, HIV infection and HCV/HTV co - infection differently.MATERIALS AND METHODS1. Study subjects14 HTV/AIDS patients are diagnosed by AIDS research institution of China Medical University, the Center for disease prevention and control in Iiaoning and Jilin province. Diagnosis of HIV - 1 infection was made on the basis of a positive anti - HIV ELISA ( Vironostika, Organon Tednika, The Netherlands) and confirmed by a positive Western immunoblot ( Genelab Diagnostics, Singapore ). 21 HCV infectors were from the outpatients and inpatients with anti -HCV IgG ( + ) and HCV RNA > 104 copies/ml in the First Affiliated Hospital of China Medical University. And 28 patients infect both HCV and HIV. 30 health controls were blood donors from the Blood Bank Center of Shenyang with normal blood routine test, liver and renal function and without diseases of the immune system and other hepatitis.2. Determination of CD4+ lymphocytes absolute countPipette 20μl of TriTEST CD4FTTC/CD8PE/CD3PerCP reagent and add 50μl anticoagulated whole blood into the bottom of the TruCOUNT Tube using reverse pipetting and incubate for 15 minutes in dark at room temperature (20 -25℃. Add 450μl IX FACS Lysing Solution to the tube, incubate for 15minutes in the dark at room temperature and analyze the samples on the flow cytometer using FACS MULTISET software to acquire T lymphocyte counts.3. Cell surface receptors stainingAccording to the manual of cell surface staining kits of BD Company, 100, 000 cells were acquired and expression of cell surface markers on PBMC cells were analyzed using CELLQUEST software by FACSCalibur flow cytometer.4. Data and statistical analysisDetermine the group of lymphocytes by FSC and SSC. Using CD3 +T lymphocytes and CD4 + T lymphocytes or CD8+T lymphocytes as gates, analysis expressions of CD8UCCR5,CXCR3 and CD25. SPSS 10. 0 software was used to make the statistical analysis. Comparisons of means of different groups were done using t — test and comparisons of means within one group were done usingpaired t — test.RESULT1. The absolute count of CD4+ and CD8 + TlymphocytesThe absolute count of CD4+ T lymphocytes in HIV infectors and HCV/HIV co - infectors were lower than it in health controls (P<0.01;P<0. 05); and the absolute count of CD8 + T lymphocytes in HIV infectors and HCV/HIV co -infectors were higher than it in health controls ( P <0.001). The absolute count of CD4 + and CD8 + T lymphocytes in HCV infectors were higher than them in health controls, but there were no difference between them( P > 0.05 ).2. The expression of CD81 on CD4+ T lymphocytesThe expression of CD81 on CD4+ T lymphocytes in HIV infectors (14.4% ± 12.4) and HCV/HIV co - infectors (23. 8% ± 10.9) were significantly lower than it in health controls(45.7% ± 12.7) ( P <0.01) ; and it in HIV infectors was significantly lower than it in HCV/HIV co - infectors (P<0.001). It in HCV infectors (51.3% ±15. 3) was higher than them in health controls, but there were no difference between them(P >0.05).3. The expression of CD81 on CD8 + T lymphocytesThe expression of CD81 on CD8 + T lymphocytes in HIV infectors (46.2% ±28. 5) and HCV/HIV co - infectors (44. 9% ± 19. 6) were significantly higher than it in health controls(20. 6% ± 10.4) ( P < 0. 001). It in HCV infectors (29.6% ± 13.1) was higher than it in health controls( P <0.01).4. The expression of CXCR3 on CD4 + T lymphocytesThe expression of CXCR3 on CD4 + T lymphocytes in HIV infectors (6.9% ±3. 8) and HCV/HIV co - infectors (9. 4% ±6. 7) were significantly lower than it in health controls ( 18. 7% ± 6. 1) ( P < 0. 001). It in HCV infectors (21.4% ±9.1) was higher than it in health controls, but there were no difference between them( P >0.05 ).5. The expression of CXCR3 on CD8 + T lymphocytesThe expression of CXCR3 on CD8+ T lymphocytes in HIV infectors (31. 6% ±19.5) and HCV/HIV co -infectors (20.3% ±11.1) were significantlyhigher than it in health controls (11.7% ± 6.3 ) ( P < 0.001). It in HCV infectors (14.1% ±6. 8) was higher than it in health controls, but there were no difference between them( P > 0.05 ).6. The expression of CCR5 on CD4 + T lymphocytesThe expression of CCR5 on CD4 + T lymphocytes in HCV infectors (9.3% ±4. 2) and HCV/HIV co - infectors (9. 6% ± 8. 9) were significantly lower than it in health controls ( 22. 8% ± 9. 7 ) ( P < 0. 001). It in HIV infectors (37.3% ±18.8) was higher than it in health controls( P <0.001).7. The expression of CCR5 on CD8 + T lymphocytesThe expression of CCR5 on CD8 + T lymphocytes in HCV infectors (12.8% ±7.7) and HCV/HIV co - infectors (25. 3% ± 14.6) were significantly lower than it in health controls ( 50. 2% ± 9. 0 ) ( P < 0. 001). It in HIV infectors (72.3% ± 12.4) was higher than them in health controls(P <0.001).8. The expression of CD25 on CD4+ T lymphocytesThe expression of CD81 on CD4+ T lymphocytes in HIV infectors (7. 9% ±6. 1) and HCV/HIV co -infectors (7.2% ±3.4) were significantly lower than it in health controls (18% ±9.1) ( P <0.001). It in HCV infectors (19. 2% ±13.4) was higher than them in health controls, but there were no difference between them( P >0.05).CONCLUSION1. The higher level of CD81 expressed on CD8 + T lymphocytes and the lower level of CD81 expressed on CD4+ T lymphocytes in HCV/ HIV infectors showed HIV infection might influence the expression of CD81 on CD4+ and CD8 +T lymphocytes in Chinese HCV/HIV co - infectors.2. The higher level of CXCR3 expressed on CD8 + T lymphocytes and the lower level of CXCR3 expressed on CD4 + T lymphocytes in HCV/ HIV infectors showed HIV infection influenced the expression of CXCR3 on CD4+ and CD8 +T lymphocytes in Chinese HCV/HIV co - infectors.3. The lower level of CCR5 expressed on CD4 + and CD8 + T lymphocytes in HCV/ HIV infectors showed HCV infection influenced the expression of CCR5...