电脉冲和布吡卡因增强A型肉毒毒素DNA核酸疫苗的免疫效果

目的：考查DNA疫苗注射免疫后电脉冲和布吡卡因佐剂化DNA疫苗递送方式对A型肉毒毒素DNA核酸疫苗免疫效果的影响。方法：A型肉毒毒素DNA复制子疫苗和传统DNA疫苗肌肉注射免疫小鼠后电脉冲和布吡卡因佐剂化DNA后再肌肉注射免疫小鼠；检测免疫小鼠的抗体和细胞水平，并分析抗体亚类。结果：电脉冲和布吡卡因这二种递送方式均增强DNA复制子疫苗和传统DNA疫苗的体液免疫和细胞免疫效果；电脉冲提高DNA疫苗免疫效果更为明显，并且电脉冲和布吡卡因组合这种递送方式增强DNA疫苗体液免疫和细胞免疫水平最高；与传统DNA疫苗相比，A型肉毒毒素DNA复制子疫苗在这些递送方式下均诱导产生了更好的特异性体液免疫和细胞免疫应答，并且这些递送方式没有改变DNA疫苗的Th1/Th2免疫应答特性，即DNA复制子疫苗诱导产生Th1/Th2混合免疫应答但偏向于Th2途经，而传统DNA疫苗则完全偏向于Th2途经。结论：电脉冲和布吡卡因增强DNA复制子疫苗和传统DNA疫苗的免疫效果，是提高DNA疫苗免疫原性的良好策略。

Enhanced effects of BoNT/A DNA vaccines by electric pulses and bupivacaine

Objective: To determine if suitable electric pulses-mediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of botulinum neurotoxin serotype A (BoNT/A) DNA vaccines in mouse model. Methods: Vaccination of mice i.m. with plasmid DNA replicon vaccine pSCARSHc and conventional plasmid DNA vaccine pcDNASHc following electric pulses and with DNA and bupivacaine complexes. AHc-specific group antibody ELISA titers and lymphocyte proliferative responses of mice were detected and IgG1 and IgG2a isotype profiles were assayed. Results: Immune effects of DNA vaccines were enhanced following electric pulses and bupivacaine delivery. Effects of DNA vaccines following electric pulses were better than that of DNA vaccines formulated with bupivacaine, and the combined delivery technology of electric pulses and bupivacaine induced the highest level of specific antibodies and lymphocyte proliferative responses. Plasmid DNA replicon vaccine pSCARSHc induced relatively higher AHc-specific antibodies and lymphocyte proliferative responses in immunized Balb/c mice than conventional plasmid DNA vaccine pcDNASHc in these DNA delivery technologies. And vaccine pSCARSHc induced Th2/Th1-type immune responses with a general bias to Th2-type, and vaccine pcDNASHc induced Th2-type immune responses. Conclusion: Suitable electric pulsesmediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of BoNT/A DNA vaccines in mouse model. Therefore, the methods described here potentially provide suitable strategies in developing an efficacious vaccine against botulinum neurotoxin serotype A.