乙型肝炎表面抗原与粒细胞巨噬细胞集落刺激因子融合表达质粒诱导小鼠特异性免疫应答  

Specific immune responses in Balb/c mice induced by plasmid coexpressing hepatitis B surface antigen and granulocyte-macrophage colony stimulating factor

作　　者：卿玉玲[1] 凌宁[1] 赵嘉将[1] 彭明利[1] 许红梅[1] 任红[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,400010

出　　处：《中华肝脏病杂志》2003年第8期474-476,共3页Chinese Journal of Hepatology

基　　金：重庆市科委应用基金

摘　　要：目的 观察乙型肝炎表面抗原(HBsAg)与粒细胞巨噬细胞集落刺激因子(GM CSF)融合表达质粒诱导特异性免疫应答的效果。方法 以HBsAg与GM-CSF融合表达质粒DNA免疫BALB/C小鼠,用酶链免疫吸附方法检测质粒诱导BALB/C小鼠产生抗HBs及脾细胞经HBsAg体外刺激后分泌白细胞介素(IL)-2、4及γ干扰素(IFNγ)的水平;并用^(21)Cr释放法检测HHsAg特异性细胞毒性T淋巴细胞(CTL)反应。结果 各组小鼠加强免疫后血清抗体水平(P/N值)A组(PcDNA3.1-S)16.1+20.3,B组(PcDNA3.1 GM-CSF-S)28.3±19.7,F组(重组酵母乙型肝炎疫苗)29.5±21.5,C组(PcDNA3.1S-GM-CSF)0.3±0.0,D组(PcDNA3.1—S+PcDNA3.1-GM-CSF)4.5+5.9,E组(PcDNA3.1—GM-CSF)0.9±1.2,G组(PcDNA3.1)及H组(PBS)为阴性(F-4.176,P<0.01);IL-2分泌水平A组(26.6±1.9)、B组(56.0±2.2)、C组(4.3±1.2)、D组(33.5±1.7)、F组(3.5+1.4)、F组(7.5±2.0)、G组(1.5±0.7),H组未检测到IL-2分泌(F=31.188,P<0.01);IFN-γ分泌水平A组(64.0±7.5)、B组(139.0±17.0)、C组(11.0±5.0)、D组(53.0±9.5)、E组(9.0±3.0)、F组(13.0±2.0)、G组(8.0±2.0,P(0.05),H组未检测到IFN—γ泌(F=31.796,P<0.01);HBsAg特异性CTL活性,效靶比为30:1及10:1时各组音有差异有显著性(F值分别为26.891。Objective To investigate the improvement of specific immune responses induced by plasmid coexpressing hepatitis B surface antigen (HBsAg) and granulocyte-macrophage colony stimulating factor (GM-CSF). Methods All Balb/c (H-2d) mice were immunized with pGM-CSF/S, pS/GM-CSF, pS or control plasmids. 4 weeks later, anti-HBs titer and the levels of IL-2, IL-4 and IFN- y in the supernatant of splenocytes were detected using enzyme-linked immunosorbent assay (ELJSA), and HBsAg-specific cytotoxic T lymphocytes (CTL) activity was measured with a 51Cr release assay, using P815/S transfectants as target cells. Results The anti-HBs antibody titers in the serum, the levels of IL-2 and IFN- y , and the CTL aclivity in pcDNA3.1-GM-CSF-S immuned mice were higher than those in PcDNA3.1-S immunized mice (F = 4.176, P <0.01; F= 31.188, P < 0.01; F = 31.796, P < 0.01; F < 26.891, P < 0.01). Conclusion It will improve the specific immune responses induced by HBsAg DNA vaccine after it is binded to the gene of GM-CSF.

关 键 词：乙型肝炎表面抗原 粒细胞 巨噬细胞集落刺激因子 融合表达质粒 诱导 小鼠 免疫应答 

分 类 号：R392[医药卫生—免疫学]