Su Ling-kai, Yu Fei, Li Zhao-fei, Zeng Chang, Xu Qing-an, Fan Ming-wen
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine of Ministry of Education (KLOBM), School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China.
Center of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Acta Pharmacol Sin. 2014 May;35(5):592-8. doi: 10.1038/aps.2013.184.
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.
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.
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.
Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.
研究共同递送表达白细胞介素-6（IL-6）的质粒pCI-IL-6对编码变形链球菌表面蛋白抗原PAc的抗龋DNA疫苗pCIA-P免疫原性的影响。
通过将小鼠IL-6基因插入pCI载体构建质粒pCI-IL-6。采用蛋白质免疫印迹分析评估IL-6在体外的表达。BALB/c小鼠于第0天和第14天经鼻联合免疫pCIA-P和pCI-IL-6。采用酶联免疫吸附测定（ELISA）评估抗PAc IgG和分泌型IgA（sIgA）。用PAc蛋白再次刺激小鼠脾细胞，采用ELISA检测干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）的产生。用流式细胞术分析脾细胞增殖。对大鼠进行类似免疫，采用凯斯（Keyes）法测定龋齿评分。
在转染pCI-IL-6的COS-7细胞中发现有明显的IL-6表达。在pCI-IL-6联合免疫的小鼠中，第4周和第8周时血清中的特异性IgG抗体和唾液中的sIgA抗体显著高于对照小鼠。此外，pCI-IL-6联合免疫的小鼠脾细胞经PAc蛋白再次刺激后IFN-γ的分泌显著高于对照小鼠，而IL-4的分泌无显著差异。pCI-IL-6联合免疫的小鼠脾细胞增殖显著高于用pCIA-P和pCI载体免疫的小鼠。在大鼠龋齿模型中，pCI-IL-6联合免疫的大鼠龋齿评分低于对照大鼠。
经鼻共同递送IL-6基因可显著增强抗龋DNA疫苗的免疫原性。
