Skin targeted DNA vaccine delivery using electroporation in rabbits. I: efficacy.
Skin targeted DNA vaccine delivery using electroporation in rabbits. I: efficacy.
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使用电穿孔对兔子进行皮肤靶向 DNA 疫苗递送。
DOI：
10.1016/j.ijpharm.2004.12.014
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Singh,Jagdish
中科院分区：
文献类型：
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作者：
Medi,BabuM;Hoselton,Scott;Marepalli,RaoB;Singh,Jagdish
通过皮肤进行基因免疫是非常可取的，因为皮肤有大量的抗原提呈细胞(APC)，而且很容易获得。本研究的目的是研究电穿孔脉冲幅度、脉冲长度和脉冲个数对兔皮肤注射质粒DNA疫苗后免疫应答的影响。利用编码β-半乳糖苷酶的报告质粒研究了所传递的质粒的表达。用编码乙肝表面抗原(HBs-Ag)的乙肝DNA疫苗研究DNA疫苗的免疫效果。免疫后分别对血清和外周血单个核细胞(PBMC)进行体液免疫和细胞免疫分析。转基因在皮肤中的表达是一过性的，在200和300V的脉冲作用下，转基因在皮肤中的表达在2天后达到高峰。在200和300V的脉冲作用下，β-半乳糖苷酶的表达水平分别是第2天的48倍和129倍。用X-Gal对皮肤进行原位组织化学染色，发现200和300V的电穿孔脉冲可以定位地表达GFP-半乳糖苷酶。与被动注射相比，电穿孔介导的皮肤DNA疫苗注射显著增强了体液免疫和细胞免疫应答(p<；0.05)。本研究证明了通过局部电穿孔增强DNA疫苗对皮肤和免疫反应的递送。因此，电穿孔介导的经皮DNA疫苗递送有望成为DNA疫苗递送的一种潜在选择。
Genetic immunization through skin is highly desirable as skin has plenty of antigen presenting cells (APCs) and is easily accessible. The purpose of this study was to investigate the effects of electroporation pulse amplitude, pulse length and number of pulses on cutaneous plasmid DNA vaccine delivery and immune responses, following intradermal injection in vivo in rabbits. Expression of the delivered plasmid was studied using a reporter plasmid, coding for β-galactosidase. The efficiency of DNA vaccine delivery was investigated using a DNA vaccine against Hepatitis B, coding for Hepatitis B surface antigen (HBsAg). Serum samples and peripheral blood mononuclear cells (PBMC) were analyzed for humoral and cellular immunity, respectively, following immunization. The expression of transgene in the skin was transient and reached its peak in 2 days post-delivery with 200 and 300V pulses. The expression levels with 200 and 300V pulses were 48- and 129-fold higher, respectively, compared with the passive on day 2. In situ histochemical staining of skin with X-gal demonstrated the localized expression of β-galactosidase with electroporation pulses of 200 and 300V. Electroporation mediated cutaneous DNA vaccine delivery significantly enhanced both humoral and cellular immune responses (p<0.05) to Hepatitis B compared to passive delivery. The present study demonstrates the enhanced DNA vaccine delivery to skin and immune responses by topical electroporation. Hence, electroporation mediated cutaneous DNA vaccine delivery could be developed as a potential alternative for DNA vaccine delivery.