Electroporation of a nanoparticle-associated DNA vaccine induces higher inflammation and immunity compared to its delivery with microneedle patches in pigs
Electroporation of a nanoparticle-associated DNA vaccine induces higher inflammation and immunity compared to its delivery with microneedle patches in pigs
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DOI：
10.1016/j.jconrel.2019.06.041
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发表时间：
2019-08-28
影响因子：
10.8
通讯作者：
Schwartz-Cornil, Isabelle
中科院分区：
文献类型：
--
作者：
Bernelin-Cottet, Cindy;Urien, Celine;Schwartz-Cornil, Isabelle
DNA 疫苗接种是一项有吸引力的技术，因为其成熟的制造工艺、安全性、快速对抗大流行病原体的适应性以及环境温度下的稳定性；然而，DNA 的最佳递送方法仍有待确定。由于猪是人类的相关模型，我们比较评估了使用可溶性微针贴片、针皮内接种（ID）、表面电穿孔（EP）以及与或不与阳离子聚乳酸共乙醇酸纳米颗粒（NP）结合的DNA将疫苗DNA递送到猪体内的效率。我们使用荧光素酶编码质粒（pLuc）作为报告基因，并使用编码猪繁殖与呼吸综合征病毒（PRRSV）（一种具有临床意义的猪动脉病毒）抗原的疫苗质粒。贴片成功诱导皮肤中荧光素酶的表达，尽管水平低于 EP。 EP 诱导粒细胞、MHC2(pos)CD172A(pos) 骨髓细胞和 1 型常规树突细胞的皮肤募集，与局部产生 IL-1β、IL-8 和 IL-17 相关；这些局部反应对于 ID 来说更加有限，并且对于补丁来说是无法检测到的。 EP中添加NP尤其促进了MHC2(pos)CD172A(pos)CD163(int)和CD163(neg)骨髓亚群的募集。值得注意的是，我们通过 EP 传递的 DNA + NP 获得了针对一组 PRRSV 抗原的最强和最广泛的 IFN γ T 细胞反应，而贴片和 ID 则无效。独立于 NP 的 EP 给药的抗 PRRSV IgG 反应最高，ID 给药的反应轻微，而贴剂给药时的抗 PRRSV IgG 反应无法检测到。这些结果与之前在带有贴片的小鼠中诱导的免疫原性和功效形成对比。这项研究的结论是，通过电穿孔和贴片可以在猪的皮肤上成功注射 DNA 疫苗，但只有前者诱导局部炎症、体液和细胞免疫，在使用 NP 时效力最高。这一发现表明了在与猪等人类相关的临床前模型中评估小鼠模型之外的 DNA 疫苗的递送和免疫原性的重要性，并揭示了与 DNA 结合的 EP 与 NP 诱导强免疫原性。
DNA vaccination is an attractive technology, based on its well-established manufacturing process, safety profile, adaptability to rapidly combat pandemic pathogens, and stability at ambient temperature; however an optimal delivery method of DNA remains to be determined. As pigs are a relevant model for humans, we comparatively evaluated the efficiency of vaccine DNA delivery in vivo to pigs using dissolvable microneedle patches, intradermal inoculation with needle (ID), surface electroporation (EP), with DNA associated or not to cationic poly-lactic-co-glycolic acid nanoparticles (NPs). We used a luciferase encoding plasmid (pLuc) as a reporter and vaccine plasmids encoding antigens from the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), a clinically-significant swine arterivirus. Patches were successful at inducing luciferase expression in skin although at lower level than EP. EP induced the cutaneaous recruitment of granulocytes, of MHC2(pos)CD172A(pos) myeloid cells and type 1 conventional dendritic cells, in association with local production of IL-1 beta, IL-8 and IL-17; these local responses were more limited with ID and undetectable with patches. The addition of NP to EP especially promoted the recruitment of the MHC2(pos)CD172A(pos) CD163(int) and CD163(neg) myeloid subsets. Notably we obtained the strongest and broadest IFN gamma T-cell response against a panel of PRRSV antigens with DNA + NPs delivered by EP, whereas patches and ID were ineffective. The anti-PRRSV IgG responses were the highest with EP administration independently of NPs, mild with ID, and undetectable with patches. These results contrast with the immunogenicity and efficacy previously induced in mice with patches. This study concludes that successful DNA vaccine administration in skin can be achieved in pigs with electroporation and patches, but only the former induces local inflammation, humoral and cellular immunity, with the highest potency when NPs were used. This finding shows the importance of evaluating the delivery and immunogenicity of DNA vaccines beyond the mouse model in a preclinical model relevant to human such as pig and reveals that EP with DNA combined to NP induces strong immunogenicity.