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.
出处
《中华肝脏病杂志》
CAS
CSCD
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
