粒细胞-巨噬细胞集落刺激因子(GM-CSF)与生长抑素(SS)融合表达质粒的构建及其对小鼠的免疫效果  

Construction of Fusion-expression Plasmid Harboring Porcine Granulocyte-macrophage Colony-stimulating Factor (GM-CSF ) and Somatostatin (SS ) and Its Immunization on Mice

作　　者：舒邓群[1] 茆达干[2] 曹少先[2] 杨利国[3] 吴志敏[1] 

机构地区：[1]江西农业大学动物科技学院,南昌330045 [2]南京农业大学动物繁育研究所,南京210095 [3]华中农业大学动物科技学院,武汉430070

出　　处：《农业生物技术学报》2008年第1期41-46,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助项目(No.30270959)资助

摘　　要：从猪(Sus scrofa)真核表达质粒pGM-CSF PCR扩增猪粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因的编码序列,亚克隆到生长抑素真核表达质粒pcS/2SS中,构建GM-CSF与生长抑素(SS)基因的融合表达质粒pGM-CSF/SS,酶切鉴定和DNA测序证明重组的融合表达质粒pGM-CSF/SS构建正确。选择30只小鼠(Mus musculus),随机分为3组,分别肌肉注射生理盐水(第1组)、pcS/2SS(第2组)和pGM-CSF/SS(第3组),剂量50μg/只,2周后以相同剂量加强免疫1次,研究pGM-CSF/SS对小鼠的免疫效果。结果表明,GM-CSF可增强小鼠脾脏淋巴细胞的增殖能力,以pGM-CSF/SS组增殖能力最强,分别比对照组和pcS/2SS提高8.4%(P<0.05)。pcS/2SS和pGM-CSF/SS免疫小鼠生长抑素抗体P(阳性)/N(阴性)值均有所提高,且pGM-CSF/SS组P/N值高于pcS/2SS组。生长抑素抗体阳性鼠的比例以pGM-CSF/SS为最高,达60%,pcS/2SS组为30%,对照组没有出现阳性鼠。GM-CSF对生长抑素DNA疫苗有一定的免疫增强作用,可增强小鼠脾脏淋巴细胞的增殖能力,提高生长抑素抗体的P/N值。Porcine granulocyte-macrophage colony-stimulating factor （GM-CSF） gene amplified from porcine （Sus scrofa ） expression plasmid pGM-CSF by PCR was subcloned into somatostatin eukaryotic expression plasmid pcS/2SS to construct fusion-expression plasmid （pGM-CSF/SS） harboring porcine GM-CSF and somatostatin （SS）. Restriction endonucleases digestion and DNA sequencing showed that pGM-CSF/SS was successfully constructed. To study the immunization effect ofpGM-CSF/SS on mice （Mus musculus）, a total of 30 mice at the age of 22 days were divided into 3 groups and were given two intramuscular injections at interval of 2 weeks with saline solution as control （group 1）, pcS/2SS （group 2） and pGM-CSF/SS （group 3）, respectively. The results showed that lymphocyte proliferation response of pGM-CSF/SS was higher than that of the control group and pcS/2SS group, which increased 8.4%, respectively （P 〈0.05）. pGM-CSF/SS significantly enhanced the P （positive）/N （negative） value of SS antibody compared to the control group and immunization of pcS/2SS. Percentage of mice with positive SS antibody immunized with pGM-CSF/SS was higher than that of other groups, which got to 60%. Percentage immunized with pcS/2SS was 30%. The results suggest that genetic adjuvant GM-CSF can enhance immune response of somatostatin DNA vaccine, and increase lymphocyte proliferation responses and P / N value of SS antibody.

关 键 词：粒细胞-巨噬细胞集落刺激因子 生长抑素 融合表达质粒 免疫 

分 类 号：S188[农业科学—农业基础科学]