Harwood Lisa J, Gerber Heidi, Sobrino Francisco, Summerfield Artur, McCullough Kenneth C
Institute of Virology and Immunoprophylaxis, CH-3147 Mittelhäusern, Switzerland.
J Virol. 2008 Jul;82(13):6379-94. doi: 10.1128/JVI.00021-08. Epub 2008 Apr 30.
Dendritic cells (DC), which are essential for inducing and regulating immune defenses and responses, represent the critical target for vaccines against pathogens such as foot-and-mouth disease virus (FMDV). Although it is clear that FMDV enters epithelial cells via integrins, little is known about FMDV interaction with DC. Accordingly, DC internalization of FMDV antigen was analyzed by comparing vaccine virus dominated by heparan sulfate (HS)-binding variants with FMDV lacking HS-binding capacity. The internalization was most efficient with the HS-binding virus, employing diverse endocytic pathways. Moreover, internalization relied primarily on HS binding. Uptake of non-HS-binding virus by DC was considerably less efficient, so much so that it was often difficult to detect virus interacting with the DC. The HS-binding FMDV replicated in DC, albeit transiently, which was demonstrable by its sensitivity to cycloheximide treatment and the short duration of infectious virus production. There was no evidence that the non-HS-binding virus replicated in the DC. These observations on virus replication may be explained by the activities of viral RNA in the DC. When DC were transfected with infectious RNA, only 1% of the translated viral proteins were detected. Nevertheless, the transfected cells, and DC which had internalized live virus, did present antigen to lymphocytes, inducing an FMDV-specific immunoglobulin G response. These results demonstrate that DC internalization of FMDV is most efficient for vaccine virus with HS-binding capacity, but HS binding is not an exclusive requirement. Both non-HS-binding virus and infectious RNA interacting with DC induce specific immune responses, albeit less efficiently than HS-binding virus.
树突状细胞(DC)对于诱导和调节免疫防御及反应至关重要,是针对口蹄疫病毒(FMDV)等病原体疫苗的关键靶点。尽管已知FMDV通过整合素进入上皮细胞,但对于FMDV与DC的相互作用却知之甚少。因此,通过比较以硫酸乙酰肝素(HS)结合变体为主的疫苗病毒与缺乏HS结合能力的FMDV,分析了DC对FMDV抗原的内化作用。HS结合病毒通过多种内吞途径进行内化,效率最高。此外,内化主要依赖于HS结合。DC对非HS结合病毒的摄取效率要低得多,以至于常常难以检测到与DC相互作用的病毒。HS结合的FMDV在DC中短暂复制,这可通过其对放线菌酮处理的敏感性以及感染性病毒产生的短暂持续时间得到证明。没有证据表明非HS结合病毒在DC中复制。关于病毒复制的这些观察结果可能由DC中病毒RNA的活性来解释。当用感染性RNA转染DC时,仅检测到1%的翻译后病毒蛋白。然而,转染的细胞以及内化了活病毒的DC确实将抗原呈递给淋巴细胞,诱导了FMDV特异性免疫球蛋白G反应。这些结果表明,对于具有HS结合能力的疫苗病毒,DC对FMDV的内化效率最高,但HS结合并非唯一必要条件。与DC相互作用的非HS结合病毒和感染性RNA均能诱导特异性免疫反应,尽管效率低于HS结合病毒。