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目的以一种CpG寡聚核苷酸为HIV-1DNA疫苗候选佐剂,研究该CpG佐剂增强DNA疫苗免疫原性,体外促进DC细胞成熟等特点。方法在Balb/c小鼠模型上连续3次联合免疫HIV-1DNA疫苗及CpG佐剂,通过IFN-γ、IL-2ELISPOT及ELISA检测HIV特异性细胞免疫反应及体液免疫应答强度;体外制备小鼠骨髓来源的树突状细胞,通过FACS技术、高通量细胞因子检测等方法评价CpG佐剂刺激活化DC的能力。结果 CpG能够增强HIV-1DNA疫苗诱导的特异性细胞免疫反应水平,降低DNA疫苗使用剂量;CpG体外刺激原代小鼠骨髓来源的树突状细胞(BMDC),能显著上调CD40、CD80、CD86等BMDC表面共刺激分子的表达,活化BMDC并分泌各型细胞因子IL-5、IL-12p70,促炎症因子IL-1α、IL-1β、IL-6、IL-10、MIP-2、KC、MIG、Eotaxin、GM-CSF等以发挥佐剂效应。结论综合体内体外实验数据,证实该型CpG能够充分活化BMDC,显著提高HIV-1DNA疫苗免疫原性,降低疫苗使用剂量,可成为HIV-1DNA疫苗临床试验用候选佐剂。
A CpG oligonucleotide is a candidate adjuvant for HIV-1 DNA vaccine. This CpG adjuvant enhances the immunogenicity of DNA vaccine and promotes the maturation of DC cells in vitro. Methods The HIV-1 DNA vaccine and CpG adjuvant were co-immunized with Balb / c mouse three times in a row. HIV-specific cellular and humoral immune responses were detected by IFN-γ, IL-2 ELISA and ELISA. Bone marrow-derived dendritic cells were evaluated for their ability to stimulate activated DCs by FACS techniques, high-throughput cytokine assays and the like. Results CpG increased the specific cellular immune response induced by HIV-1 DNA vaccine and decreased the dosage of DNA vaccine. CpG stimulated primary mouse bone marrow-derived dendritic cells (BMDC) in vitro to significantly upregulate CD40, CD80, CD86, etc. The expression of costimulatory molecules on the surface of BMDC activated BMDC and secreted various cytokines such as IL-5, IL-12p70, proinflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, MIP-2, KC, MIG , Eotaxin, GM-CSF, etc. to play an adjuvant effect. Conclusion Based on the in vitro and in vivo experimental data, we conclude that this CpG can fully activate BMDC, significantly improve the immunogenicity of HIV-1 DNA vaccine and reduce the dose of vaccine, which may be a candidate adjuvant for clinical trials of HIV-1 DNA vaccine.