Zucchelli S, Capone S, Fattori E, Folgori A, Di Marco A, Casimiro D, Simon A J, Laufer R, La Monica N, Cortese R, Nicosia A
Istituto di Ricerche di Biologia Molecolare P. Angeletti, 00040 Pomezia (Rome), Italy.
J Virol. 2000 Dec;74(24):11598-607. doi: 10.1128/jvi.74.24.11598-11607.2000.
We describe an improved genetic immunization strategy for eliciting a full spectrum of anti-hepatitis C virus (HCV) envelope 2 (E2) glycoprotein responses in mammals through electrical gene transfer (EGT) of plasmid DNA into muscle fibers. Intramuscular injection of a plasmid encoding a cross-reactive hypervariable region 1 (HVR1) peptide mimic fused at the N terminus of the E2 ectodomain, followed by electrical stimulation treatment in the form of high-frequency, low-voltage electric pulses, induced more than 10-fold-higher expression levels in the transfected mouse tissue. As a result of this substantial increment of in vivo antigen production, the humoral response induced in mice, rats, and rabbits ranged from 10- to 30-fold higher than that induced by conventional naked DNA immunization. Consequently, immune sera from EGT-treated mice displayed a broader cross-reactivity against HVR1 variants from natural isolates than sera from injected animals that were not subjected to electrical stimulation. Cellular response against E2 epitopes specific for helper and cytotoxic T cells was significantly improved by EGT. The EGT-mediated enhancement of humoral and cellular immunity is antigen independent, since comparable increases in antibody response against ciliary neurotrophic factor or in specific anti-human immunodeficiency virus type 1 gag CD8(+) T cells were obtained in rats and mice. Thus, the method described potentially provides a safe, low-cost treatment that may be scaled up to humans and may hold the key for future development of prophylactic or therapeutic vaccines against HCV and other infectious diseases.
我们描述了一种改进的基因免疫策略，通过将质粒DNA电基因转移（EGT）到肌肉纤维中，在哺乳动物中引发全谱抗丙型肝炎病毒（HCV）包膜2（E2）糖蛋白反应。肌内注射编码与E2胞外域N端融合的交叉反应性高变区1（HVR1）肽模拟物的质粒，随后以高频、低电压电脉冲形式进行电刺激处理，在转染的小鼠组织中诱导出超过10倍的高表达水平。由于体内抗原产生的大幅增加，在小鼠、大鼠和兔子中诱导的体液反应比传统裸DNA免疫诱导的反应高10至30倍。因此，与未接受电刺激的注射动物的血清相比，经EGT处理的小鼠的免疫血清对天然分离株的HVR1变体表现出更广泛的交叉反应性。EGT显著改善了针对辅助性和细胞毒性T细胞特异性的E2表位的细胞反应。EGT介导的体液和细胞免疫增强与抗原无关，因为在大鼠和小鼠中针对睫状神经营养因子的抗体反应或针对人类免疫缺陷病毒1型gag CD8(+) T细胞的特异性反应得到了类似的增加。因此，所描述的方法可能提供一种安全、低成本的治疗方法，可扩大到人类应用，并可能是未来开发针对HCV和其他传染病的预防性或治疗性疫苗的关键。
