Intranasal co-delivery of IL-6 gene enhances the immunogenicity of anti-caries DNA vaccine
Intranasal co-delivery of IL-6 gene enhances the immunogenicity of anti-caries DNA vaccine
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DOI：
10.1038/aps.2013.184
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发表时间：
2014-05
影响因子：
8.2
通讯作者：
Ling-kai Su;F. Yu;Zhao-fei Li;Chang Zeng;Qing-an Xu;M. Fan
中科院分区：
文献类型：
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作者：
Ling-kai Su;F. Yu;Zhao-fei Li;Chang Zeng;Qing-an Xu;M. Fan
目的：探讨共递送IL-6表达质粒pCI-IL-6对编码变形链球菌表面蛋白抗原PAc的抗龋齿DNA疫苗pCIA-P免疫原性的影响。方法：将鼠IL-6基因插入pCI载体，构建质粒pCI-IL-6。使用蛋白质印迹分析评估IL-6的体外表达。 BALB/c 小鼠在第 0 天和第 14 天用 pCIA-P 加 pCI-IL-6 进行鼻内联合免疫。通过 ELISA 评估抗 PAc IgG 和分泌型 IgA (sIgA)。用 PAc 蛋白重新刺激小鼠的脾细胞，并使用 ELISA 测量 IFN-γ 和 IL-4 的产生。用流式细胞术分析脾细胞增殖。同样免疫大鼠，采用Keyes法测定龋齿评分。结果：转染pCI-IL-6的COS-7细胞中IL-6明显表达。在pCI-IL-6联合免疫小鼠中，第4周和第8周时，血清中的特异性IgG抗体和唾液中的sIgA抗体显着高于对照小鼠。此外，pCI-IL-6联合免疫小鼠响应PAc蛋白再刺激而从脾细胞中分泌IFN-γ显着高于对照小鼠，而IL-4的分泌没有显着差异。 pCI-IL-6联合免疫小鼠的脾细胞增殖明显高于pCIA-P和pCI载体免疫小鼠。在大鼠龋病模型中，pCI-IL-6联合免疫大鼠的龋病评分低于对照大鼠。结论：鼻内联合递送IL-6基因显着增强了抗龋齿DNA疫苗的免疫原性。
Aim:To investigate the effects of co-delivering IL-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pCIA-P, which encodes the surface protein antigen PAc of Streptococcus mutans.Methods:Plasmid pCI-IL-6 was constructed by inserting the murine IL-6 gene into the pCI vector. Expression of IL-6 in vitro was assessed using Western blot analysis. BALB/c mice were intranasally co-immunized with pCIA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA (sIgA) were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-γ and IL-4 production was measured using ELISA. Splenocyte proliferation was analyzed with flow cytometry. Rats were similarly immunized, and dental caries scores were determined using the Keyes method.Results:Marked expression of IL-6 was found in COS-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and sIgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-γ from splenocytes in response to re-stimulation with PAc protein was significantly higher in the pCI-IL-6 co-immunized mice than that in the control mice, whereas the secretion of IL-4 had no significant difference. The proliferation of splenocytes from the pCI-IL-6 co-immunized mice was significantly higher than that from the mice immunized with pCIA-P and pCI vector. In the rat caries model, the pCI-IL-6 co-immunization rats displayed lower caries scores than the control rats.Conclusion:Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.