Gao Lei, Bao Lang, Hao Mu, Zhang Le, Qin Zi-fang
Department of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Jul;39(4):535-9.
To investigate the immunogenicity of vaccine strategies about human interleukin 12 associated with combined DNA (Ag85A and ESAT-6) prime-BCG boost.
BALB/c mice were divided into PBS negative control and 4 immunity groups: BCG group, DNA/BCG group, DNA + IL-12/BCG group and DNA/BCG + IL-12 group. All mice received three immunizations at 2-week interval. Specific IgG antibody in serum of mice was determined with indirect ELISA in 4, 6, 8 weeks respectively after final vaccination. The splenic lymphocytes of mice were separated and stimulated with PPD to measure their proliferation by flow cytometry, and to evaluate the production of interferon-gamma (IFN-gamma) in cell suspensions of spleen cells by ELISA. The levels of CD4+ and CD8+ T-cell on surface of spleens lymphocyte were determined by flow cytometry.
PPD could stimulate specific IgG responses in 4 immunity groups, and the average valences of 4 groups are 1:80, 1:120,1:160,1:160; the splenic lymphocyte proliferation reactions and IFN-gamma production were detectable in 4 immunity groups, and the most significant response occurred in 12 weeks. DNA + IL-12/BCG group and DNA/ BCG+IL-12 group induced higher production than BCG group and DNA/BCG group (P < 0.05), and the effects between DNA + IL-12/BCG group and DNA/BCG + IL-12 group had little difference. The numbers of CD4+ and CD8+ T-cell in 4 immunity groups were much higher than PBS group (P < 0.05), and DNA+IL-12/BCG group and DNA/BCG+IL-12 group were detected much more CD4+ and CD8+ T-cell than BCG group and DNA/BCG group (P < 0.05). The level of T-cell between DNA + IL-12/BCG group and DNA/BCG + IL-12 group had little difference.
Interleukin 12 associated with the strategy of priming with the combined DNA vaccines and boosting with attenuated M. bovis vaccine (BCG) could induce much stronger specific cellular immunity compared with simple DNA/BCG or attenuated M. bovis vaccine alone.
研究人白细胞介素12联合DNA（Ag85A和ESAT-6）初免-卡介苗加强的疫苗策略的免疫原性。
将BALB/c小鼠分为PBS阴性对照组和4个免疫组：卡介苗组、DNA/卡介苗组、DNA +白细胞介素12/卡介苗组和DNA/卡介苗 +白细胞介素12组。所有小鼠每隔2周进行3次免疫。在末次免疫后第4、6、8周分别用间接ELISA法测定小鼠血清中的特异性IgG抗体。分离小鼠脾淋巴细胞，用PPD刺激，通过流式细胞术检测其增殖情况，并用ELISA法评估脾细胞悬液中γ干扰素（IFN-γ）的产生。通过流式细胞术测定脾淋巴细胞表面CD4+和CD8+ T细胞的水平。
PPD能刺激4个免疫组产生特异性IgG反应，4组的平均效价分别为1:80、1:120、1:160、1:160；4个免疫组均能检测到脾淋巴细胞增殖反应和IFN-γ产生，且在第12周反应最为显著。DNA +白细胞介素12/卡介苗组和DNA/卡介苗 +白细胞介素12组诱导产生的水平高于卡介苗组和DNA/卡介苗组（P < 0.05），DNA +白细胞介素12/卡介苗组与DNA/卡介苗 +白细胞介素12组之间的效果差异不大。4个免疫组中CD4+和CD八年级下册英语书单词8+ T细胞的数量均高于PBS组（P < 0.05），DNA +白细胞介素12/卡介苗组和DNA/卡介苗 +白细胞介素12组检测到的CD4+和CD8+ T细胞比卡介苗组和DNA/卡介苗组多（P < 0.05）。DNA +白细胞介素12/卡介苗组与DNA/卡介苗 +白细胞介素12组之间的T细胞水平差异不大。
与单纯DNA/卡介苗或减毒牛分枝杆菌疫苗（卡介苗）相比，白细胞介素12联合DNA疫苗初免和减毒牛分枝杆菌疫苗（卡介苗）加强的策略能诱导更强的特异性细胞免疫。
