AIM： To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder （Thymelaeaceae）. METHOD： The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking. RESULTS： Wikstroelide M potently inhibited different HIV-1 strains, including HIV-lmn, HIV-1AI7, and HIV-19495, induced a cytopathic effect, with ECs0 values ranging from 3.81 to 15.65 ng.mL-I. Wikstroelide M also had high inhibitory activities against HIV-2noD and HIV-2cBL_20-induced cytopathic effects with ECs0 values of 18.88 and 31.90 ng.mL 1. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with ECs0 values ranging from 15.16 to 35.57 ng.mL-1. Wikstroelide M also potently inhibited HIV-lnm induced cytolysis in MT-4 cells, with an ECs0 value of 9.60 ng.mL ~. The mechanistic assay showed that wikstroelide M targeted HIV-I reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75. CONCLUSION： Wikslroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear Iranslocation through dismpting the interaction between integrase and LEDGF/p75.
Non-human primates such as Chinese rhesus macaques are the favorable models for preclinical study of potential therapeutic drugs,vaccines and mechanisms of human diseases.Little is known about the normal levels of leukocyte subpopulations of Chinese rhesus macaques.To obtain these data,100 blood samples from Chinese rhesus macaques were collected.The normal range of major leukocyte subpopulations,such as T lymphocytes,B lymphocytes,monocytes,myeloid dendritic cells(mDCs)and plasmacytoid dendritic cells(pDCs),were quantitatively analyzed by flow cytometry through BD trucount tubes.The influence of age and sex on the cell counts of leukocyte subpopulations was analyzed.The counts of CD3^(+)T cells,CD3+CD4^(+)T cells,CD3+CD8^(+)T cells and B cells decreased with age,but those of monocytes,mDCs and pDCs had no significant correlation with age.Significant differences existed in the cell counts of most leukocyte subpopulations between the male and female groups except pDCs.Furthermore the values of the females were higher than those of the males.The study provided basic information about the leukocyte subpopulations of Chinese rhesus macaques,and it may be valuable for immunobiological study of Chinese rhesus macaques.
It is currently widely accepted that immune activation in HIV-infected individuals leads to a severe loss of CD4＋ T cells and the progression to AIDS. However, the underlying mechanism of this immune activation remains unclear. Experimental data suggest that the activation of plasmacytoid dendritic cells （pDCs） by plasma viremia may play a critical role in HIV-induced immune activation. In this study, we found that the level of immune activation was higher in the late phase of SIVmac239 infection compared with chronic infection, which suggests that immune activation might be related to disease progression in SIVmac239-infected non-human primate models. Our work also showed that chloroquine could effectively inhibit the activation of pDCs in vitro and in vivo. However, chloroquine treatment of SIVmac239-infected macaques had no significant influence on the Cellular composition of peripheral blood in these animals.
