Background Developing an effective vaccine against human immunodeficiency virus type 1 （HIV-1） remains a grand challenge after more than two decades of intensive effort. It is partially due to the lack of suitable a...Background Developing an effective vaccine against human immunodeficiency virus type 1 （HIV-1） remains a grand challenge after more than two decades of intensive effort. It is partially due to the lack of suitable animal models for screening and prioritizing vaccine candidates. In this study, we aim to develop a mice model to test HIV-1 vaccine efficacy. Methods We constructed a recombinant vaccinia expressing firefly luciferase and HIV-1 Gag fusion protein based on Tiantan strain, an attenuated but replication-competent poxvirus （rTTV-lucgag）. By quantifying the luciferase activity as its read out, we defined the biodistribution of Tiantan strain poxvirus in mice inoculated intraperitoneally and attempted to apply this model to evaluate the HIV-1 vaccine efficacy. Results Our data demonstrated that the rTTV-lucgag was able to express high level of luciferase （〈106 relative luciferase units （RLU）/mg protein） and HIV-1 Gag （〉3 folds increase comparing to the control）. After intraperitoneal inoculation, this virus had dominant replication in the ovary, uterus, and cervix of mice and the luciferase activities in those organs are significantly correlated with viral titers （r^2=0.71, P 〈0.01）. Pre-immunization with an HIV gag DNA vaccine reduced the luciferase activity in ovary from （6006±3141） RLU/mg protein in control group to （1538±463） RLU/mg protein in vaccine group （P=0.1969）. Conclusions The luciferase activity in ovary could represent viral replication in vivo; this rTTV-lucgag/mice model may be suitable to assess the protective efficacy of cytotoxic T-cell responses to HIV Gag with less tedious work and high through-put.展开更多
Background Natural killer （NK） cells play critical roles in host immune defense, while the quantities and subset distributions may vary among different races. To address the difference, we compared these variables a...Background Natural killer （NK） cells play critical roles in host immune defense, while the quantities and subset distributions may vary among different races. To address the difference, we compared these variables among Chinese Han, the Caucasians and the Blacks. The study may provide critical background information for both basic research and clinical investigation. Methods Blood samples collected from populations of different races were tested within 12 hours after collection and subsets of NK cells were characterized using flow cytometry. Results The absolute NK count in the Chinese Han was significantly higher than that in the Caucasian. The Han and Caucasian groups showed higher percentages of cytotoxic subset compared to that of the Black group. The percentage of cytokine-producing subset of Chinese Han group was lower than that of Caucasian and Black groups. Black group had a higher percentage of function-unknown NK subset than that of the Han and Caucasian groups. Conclusion Our data indicated that NK cell count and the distribution of different subsets varied among different races, which should be taken into consideration in related investigations.展开更多
基金This research was supported by the National Natural Science Foundation of China （No. 30771915）, the National Grand Program on Key Infectious Disease （No. 2008ZX10001-02 Project 2 and 2008ZX10001-012） from the China Ministry of Health ＆ Ministry of Sciences and Technology, China.
文摘Background Developing an effective vaccine against human immunodeficiency virus type 1 （HIV-1） remains a grand challenge after more than two decades of intensive effort. It is partially due to the lack of suitable animal models for screening and prioritizing vaccine candidates. In this study, we aim to develop a mice model to test HIV-1 vaccine efficacy. Methods We constructed a recombinant vaccinia expressing firefly luciferase and HIV-1 Gag fusion protein based on Tiantan strain, an attenuated but replication-competent poxvirus （rTTV-lucgag）. By quantifying the luciferase activity as its read out, we defined the biodistribution of Tiantan strain poxvirus in mice inoculated intraperitoneally and attempted to apply this model to evaluate the HIV-1 vaccine efficacy. Results Our data demonstrated that the rTTV-lucgag was able to express high level of luciferase （〈106 relative luciferase units （RLU）/mg protein） and HIV-1 Gag （〉3 folds increase comparing to the control）. After intraperitoneal inoculation, this virus had dominant replication in the ovary, uterus, and cervix of mice and the luciferase activities in those organs are significantly correlated with viral titers （r^2=0.71, P 〈0.01）. Pre-immunization with an HIV gag DNA vaccine reduced the luciferase activity in ovary from （6006±3141） RLU/mg protein in control group to （1538±463） RLU/mg protein in vaccine group （P=0.1969）. Conclusions The luciferase activity in ovary could represent viral replication in vivo; this rTTV-lucgag/mice model may be suitable to assess the protective efficacy of cytotoxic T-cell responses to HIV Gag with less tedious work and high through-put.
文摘Background Natural killer （NK） cells play critical roles in host immune defense, while the quantities and subset distributions may vary among different races. To address the difference, we compared these variables among Chinese Han, the Caucasians and the Blacks. The study may provide critical background information for both basic research and clinical investigation. Methods Blood samples collected from populations of different races were tested within 12 hours after collection and subsets of NK cells were characterized using flow cytometry. Results The absolute NK count in the Chinese Han was significantly higher than that in the Caucasian. The Han and Caucasian groups showed higher percentages of cytotoxic subset compared to that of the Black group. The percentage of cytokine-producing subset of Chinese Han group was lower than that of Caucasian and Black groups. Black group had a higher percentage of function-unknown NK subset than that of the Han and Caucasian groups. Conclusion Our data indicated that NK cell count and the distribution of different subsets varied among different races, which should be taken into consideration in related investigations.
