用蛋白质转导的树突状细胞进行疫苗接种可引发对丙型肝炎病毒抗原的持续反应。

Vaccination with protein-transduced dendritic cells elicits a sustained response to hepatitis C viral antigens.

作者信息

Kuzushita Noriyoshi, Gregory Stephen H, Monti Nola A, Carlson Rolf, Gehring Stephan, Wands Jack R

机构信息

The Liver Research Center, Department of Medicine, Rhode Island Hospital and Brown Medical School, Providence, 02903, USA.

出版信息

Gastroenterology. 2006 Feb;130(2):453-64. doi: 10.1053/j.gastro.2005.10.048.

DOI:10.1053/j.gastro.2005.10.048

PMID:16472599

Abstract

BACKGROUND & AIMS: Professional antigen-presenting dendritic cells are capable of eliciting a vigorous antiviral response in naive T cells. The administration of antigen-loaded dendritic cells offers a potential approach to induce high-level immunity against hepatitis C virus.

METHODS

The dendritic cell population in mice was expanded in vivo by hydrodynamic delivery of naked DNA that encoded the secreted form of human fms-like tyrosine kinase 3 ligand. The CD11c-enriched dendritic cell population obtained from the spleen was transduced in vitro with recombinant hepatitis C virus core and nonstructural 5 proteins by using macromolecular-based protein delivery. Vaccine efficacy was assessed with a cytotoxic T-lymphocyte assay, cytokine enzyme-linked immunosorbent assays, and intracellular cytokine staining in vitro and by a tumor challenge model in vivo.

RESULTS

Relative to mice inoculated with nontransduced dendritic cells, splenocytes derived from mice immunized with either hepatitis C virus core-transduced or nonstructural 5-transduced dendritic cells showed 3- to 5-fold greater antigen-specific cytotoxic T lymphocyte activity. The CD4(+) T cells obtained from mice immunized with nonstructural 5-transduced dendritic cells produced interferon gamma, but not interleukin 4, when stimulated with nonstructural 5. In contrast, T cells derived from mice immunized with hepatitis C virus core-transduced dendritic cells produced neither interferon gamma nor interleukin 4 when stimulated with core protein. Mice vaccinated with nonstructural 5-transduced dendritic cells, but not a nonstructural 5-expressing plasmid, showed a sustained antiviral response to nonstructural 5 as evidenced by reduced growth of nonstructural 5-expressing tumor cells inoculated 10 weeks after vaccination.

CONCLUSIONS

These findings suggest that vaccination with protein-transduced dendritic cells may constitute an important antiviral strategy for hepatitis C virus.

摘要

背景与目的

专业的抗原呈递树突状细胞能够在初始T细胞中引发强烈的抗病毒反应。给予负载抗原的树突状细胞为诱导针对丙型肝炎病毒的高水平免疫提供了一种潜在方法。

方法

通过水动力递送编码人fms样酪氨酸激酶3配体分泌形式的裸DNA，在体内扩增小鼠的树突状细胞群体。使用基于大分子的蛋白质递送方法，将重组丙型肝炎病毒核心蛋白和非结构5蛋白在体外转导从脾脏获得的富含CD11c的树突状细胞群体。通过细胞毒性T淋巴细胞测定、细胞因子酶联免疫吸附测定、体外细胞内细胞因子染色以及体内肿瘤攻击模型评估疫苗效力。

结果

相对于接种未转导树突状细胞的小鼠，用丙型肝炎病毒核心转导或非结构5转导的树突状细胞免疫的小鼠来源的脾细胞显示出高3至5倍的抗原特异性细胞毒性T淋巴细胞活性。用非结构5转导的树突状细胞免疫的小鼠获得的CD4(+)T细胞在用非结构5刺激时产生干扰素γ，但不产生白细胞介素4。相比之下，用丙型肝炎病毒核心转导的树突状细胞免疫的小鼠来源的T细胞在用核心蛋白刺激时既不产生干扰素γ也不产生白细胞介素4。用非结构5转导的树突状细胞而不是表达非结构5的质粒接种的小鼠，对非结构5显示出持续的抗病毒反应，这通过接种疫苗10周后接种表达非结构5的肿瘤细胞的生长减少来证明。