Cellular HIV-1DNA And γ-cytokine Responses To48Week Highly Active Antiretroviral Therapy In Chinese HIV Infected Individuals

The introduction of Highly Active Antiretroviral Therapy (HAART) has dramatically changed the course of the HIV infection. HIV/AIDS has gone from untreatable disease to a curable disease. However, despite the effectiveness of HAART, the virus has developed different mechanisms such as the drug resistance and the genetic mutations to escape to the most efficient strategies. The resting memory CD4+T have been shown to integrate the HIV-1DNA and enter in quiescence for more than44month however the infected monocytes were shown to be resistant to apoptosis. These mechanisms help the virus to escape from antiretroviral drugs and persist for longer time in infected patients. Researches on HIV-1DNA have been conducted for years abroad but still no data are available in this field in Chinese HIV infected individuals. PARTIDetection of the cellular HIV-1DNA and its relationship with peripheral blood HIV-1RNA and CD4+Tcell count in Chinese HIV-AIDS patients undergoing48weeks’HAARTOBJECTIVE:The aim of this study was to verify that T lymphocytes and monocytes were the virus reservoir cells, and to observe the dynamic changes of these two viral reservoir cells and its relationship with main immunological parameters (peripheral CD4+cell count) and the virological parameters (plasma HIV-1RNA level) in Chinese HIV-infected patients receiving highly-active antiretroviral treatment (HAART) for48weeks. Design:To measure the blood HIV-1RNA virus load, CD4+T cell count, HIV-1DNA level in latently infected monocytes and T lymphocytes, and the relationship among them in chronic HIV-infected patients before,24, and48weeks after the initiation of HAART. DESIGN:Measuring the HIV-1DNA in chronic HIV infected patients before treatment,6and12months after treatment using the following combination of antiretroviral HAART.METHODS:35chronic HIV-1-infected adults (aged from20to 62, mean39.5±10years old) initiated HAART between June2009and June2010were enrolled in this study, with24male and11female. The patients didn’t occur with opportunistic infection or other diseases which might influence the immune response (such as T cell count (CD3+), monocytes (CD14+), or cytokines) during the study period. After obtaining the ethical approval,20milliliter peripheral whole blood was obtained from each patient at baseline (0week),24, and48weeks. The amount of T cell subgroup was detected with flow cytometry, T lymphocytes and monocytes were separated from peripheral blood mononuclear cells (PBMC) using bead sorting method, and DNA was extracted with human DNA kit. Real-time fluorescent quantitative PCR was used to detect the serum HIV-1RNA, HIV-1DNA in CD3+and CD14+cells at baseline (0week),24, and48weeks. SPSSvl8software was used to analyze the data collected.RESULTS:35chronic HIV-infected adult patients who had not received antiviral treatment were included in the study. The average HIV-1RNA level in peripheral blood was4.18±1.1(Log10copies/ml),≤1.69(Log10copies/ml), and≤1.69(Log10copies/ml) at0week,24, and48weeks after the initiation of HAART, respectively; average CD4+cells were204.42±97/μl;336.65±101/μl, and394.37±141/μl, respectively. Statistical analysis revealed that HIV-1RNA and CD4+cell count were negatively correlated. HIV-1DNA in T lymphocyte was3.62±0.65(Log10copies/106cells),2.91±0.49(Log10copies/106cells), and2.37±0.39(Log10copies/106cells) at0week,24, and48weeks after the initiation of HAART, respectively; while in mononuclear cells, HIV-1DNA level was2.78±0.71(Log10copies/106cells),2.015±0.27(Log10copies/106cells), and1.99±0.206(Log10copies/106cells). Statistical analysis showed that HIV-1RNA level was positively correlated with HIV-1DNA level in peripheral blood T lymphocytes and HIV-1DNA level in monocytes; while the number of CD4+cells and the number of CD8+cells were not significantly correlated. Detailed data analysis showed that HIV-1DNA level in monocytes and T lymphocytes declined significantly slowly than the decline rate of peripheral blood HIV-1RNA, and the HIV-1DNA level in T lymphocytes decreased more slowly than the decline rate of HIV-1DNA in monocytes. The peripheral blood CD4+cells and HIV-1DNA load in T lymphocyte was negatively correlated either before or after the initiation of HAART; however, CD4+cell count and HIV-1DNA level in monocytes were not significantly correlated.CONCLUSIONS:1.48weeks after initiating anti-retroviral therapy (HAART), peripheral blood HIV-1RNA level in HIV-1-infected patients was significantly decreased, and the number of CD4+T cells significantly increased, indicating a significant negative correlation.2. Blood T lymphocytes and monocytes in Chinese patients infected with HIV-1may serve as HIV-1reservoir cells. During48weeks of HAART, with the decreasing of HIV-1RNA level in peripheral blood, HIV-1DNA in T lymphocytes and HIV-1RNA in monocytes gradually reduced; HIV-1DNA in T lymphocytes and plasma HIV-1RNA level were positively correlated, while HIV-1DNA in monocytes and plasma HIV-1RNA load were not significantly correlated.3. This study firstly observed peripheral blood HIV-1RNA level, HIV-1DNA level in monocytes and T lymphocytes, and the percentage of HIV-1DNA which could still be detected were decreased at the same pace in Chinese HIV-1-infected patients during HAART treatment. However, the decline rate of HIV-1DNA level in monocytes and T lymphocytes was significantly slower than that of peripheral blood HIV-1RNA, and HIV-1DNA level in T lymphocytes decreased more slowly than HIV-1DNA in monocytes. These results suggested that these two cells are HIV pro-DNA reservoirs, and T lymphocytes played a more important role.4. This study firstly observed that peripheral blood CD4+cell count was negatively correlated with HIV-1DNA load in T lymphocytes either before or during HAART in Chinese HIV patients; However, CD4+cell count and HIV-1DNA level in monocytes were not obviously correlated before or after the initiation of HAART.PART ⅡDynamics of gamma cytokines during48week antiretroviral therapy and its significanceOBJECTIVES:The objective of this study was to investigate the course of certain common gamma cytokines (IL-2, IL-7and IL-21) and their role on the control of the viral infection in a short term antiviral therapy.MATERIAL AND METHODS:A total of35adults with chronic HIV infection, responding to combined antiretroviral therapy (cART) guideline criteria were enrolled for a one year follow-up study. After signing an informed consent,20ml blood were collected from each patient at base line, week0, week24and week48. One ml serum collected from each patient was kept at-80℃until use. Serum concentration of IL-2, IL-7and IL-21were determined using ELISA kit from "ebioscience and Dia clone biotech Beijing". CD4+and CD8+cell count were quantified using flux cytometry, serum HIV-1RNA quantified using real time PCR.RESULTS:CD4+and CD8+T counts showed a significant increase after HAART initiation, however the plasma viral HIV-1RNA load showed a significant decrease and reached the undetectable levels within six months after HAART initiation. The HIV-1RNA correlated negatively with CD4cell count but did not show any correlation with the CD8+T cell count. IL-2and IL-21showed a significant increase and correlated positively with CD4+T cell count after HAART initiation; however the IL-7showed a significant decrease after HAART initiation but did not show any correlation with CD4cell count. We noted no correlation between IL-2,11-21and CD4+T count from6month after HAART.CONCLUSION:In this study we found a close relationship between the increase of the CD4+T cell count and the IL-2and IL-21levels, however the IL-7level did not show obvious relationship with CD4+T count, suggesting a key role of these cytokines in the immune reconstitution and viral control.