摘要
目的应用壳聚糖(CS)包载含CpG序列的寡脱氧核苷酸(ODN),制备CS CpG-ODN基因载体,并研究其免疫活性。方法采用复凝法制备CS Cp-ODN基因载体。24只小鼠随机分为4组,每组6只,包括对照组、CS组、CpG-ODN组和CS CpG-ODN组,均通过后大腿肌内注射进行免疫接种。用MTT法检测淋巴细胞增殖情况,ELISA方法检测血清中IgG、IL-2和IL-12水平,流式细胞仪检测外周血CD4+和CD8+T淋巴细胞亚群。结果免疫接种CS CpG-ODN能提高小鼠淋巴细胞增殖水平;血清中IL-2和IL-12的含量明显升高,能获得高水平的保护性IgG;同时外周血T细胞亚群分析显示,CS CpG-ODN能更有效上调CD4+T细胞百分比以及CD4+/CD8+比值。结论CS CpG-ODN基因载体能有效提高CpG-ODN免疫促进作用。
Objective To prepare chitosan-CpG-ODN gene carriers with the use of chitosan （CS） as the material to carry oligodeoxyribonucleotide （ODN） containing CpG sequences and to study their immunological competence in mice. Methods The CS CpG-ODN carriers were prepared with the method of complex coacervation. 24 male Balb/c mice were randomly divided into four equal groups ： （1）control group, （2）CS group, （3）CpG-ODN group and （4）CS CpG-ODN group. Mice of the group （1） to （4） were given each an I. M. injection of 200 μL of normal saline, 31.25 μg of CS, 15 μg of CpG-ODN and 15μg of CpG-ODN ＋ 31.25 μg of CS, respectively, as an initial immunization. A booster immunization was carried out 2 weeks later with the same method. 2 weeks thereafter, blood samples were taken from behind the orbit for examination. Lymphocyte proliferation was assayed with the MTT test. Serum IgG, IL-2 and IL-12 contents were determined with ELISA and peripheral blood T lymphocyte subsets CD^＋ 4 and CD^＋ 8 were detected with flow cytometry. Results Enhanced lymphocyte proliferation, striking increased serum contents of IL-2 and IL-12 , high levels of protective serum IgG as well as very effective up-regulation of the percentage of CD^＋ 4 T cells and CD^＋ 4 / CD^＋ 8 ratio in the peripheral blood lymphocytes were demonstrated in the blood of mice immunized with CS CpG-ODN. Conclusion CS CpG-ODN gene carriers were shown to effectively improve the immunoenhancing effect of CpG-ODN.
出处
《医药导报》
CAS
2007年第4期356-359,共4页
Herald of Medicine
基金
湖北省自然科学基金资助项目(基金编号:2006ABA134)
