Department of Vaccine Engineering, Institute of Basic Medical Sciences, Beijing 100850, China.
Mol Biotechnol. 2013 May;54(1):58-67. doi: 10.1007/s12033-012-9544-5.
In order to overcome the instability of CpG ODN in vivo, sequence diversity, and individual differences, eleven CpG ODN fragments were meticulously selected and linked to form a Multi-CpG, which were repeatedly inserted into the cloning vector pUC19 for constructing the recombinant plasmid pUCpGs10 containing ten of Multi-CpG. Using the multi-genotype HCV E1 and multi-epitope complex HCV-T as immunogens, and plasmid pUCpGs10 as the immune adjuvant, Balb/c mice were immunized through nasal and subcutaneous immunization. Strong-specific humoral and cellular immune response were induced, which can obviously inhibit the growth of homograft expressing HCV antigen. The immune adjuvant effect of pUCpGs10 closely matched that of Freund's complete adjuvant. The plasmid pUCpGs10 can significantly improve IgA content in serum and different mucosal extract and systematical T-cell response via intranasal immunization. In conclusions, the newly constructed immunostimulatory plasmid pUCpGs10 is able to effectively activate the humoral and cellular immune activity, and possesses activation on mucosal immune response.
为了克服 CpG ODN 在体内的不稳定性、序列多样性和个体差异，精心选择了 11 个 CpG ODN 片段并连接起来形成多 CpG，将其反复插入克隆载体 pUC19 中构建含有十个多 CpG 的重组质粒 pUCpGs10。以多基因型 HCV E1 和多表位复合物 HCV-T 作为免疫原，质粒 pUCpGs10 作为免疫佐剂，通过鼻内和皮下免疫接种 Balb/c 小鼠。诱导了强烈的特异性体液和细胞免疫反应，明显抑制了表达 HCV 抗原的同种移植物的生长。pUCpGs10 的免疫佐剂作用与弗氏完全佐剂密切匹配。通过鼻内免疫接种，质粒 pUCpGs10 可显著提高血清和不同黏膜提取物中的 IgA 含量以及系统性 T 细胞反应。总之，新构建的免疫刺激质粒 pUCpGs10 能够有效激活体液和细胞免疫活性，并具有激活黏膜免疫反应的作用。
