Toh Myew-Ling, Hong Saw-See, van de Loo Fons, Franqueville Laure, Lindholm Leif, van den Berg Wim, Boulanger Pierre, Miossec Pierre
Department of Immunology and Rheumatology, Mixed Unit Civil Hospital of Lyon-BioMérieux, Edouard Herriot Hospital, Lyon, France.
J Immunol. 2005 Dec 1;175(11):7687-98. doi: 10.4049/jimmunol.175.11.7687.
Rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) do not express the coxsackie-adenovirus (Ad) receptor and are poorly permissive to Ad serotype 5 (Ad5). Genetically modified, coxsackie-Ad receptor-independent Ad5 vectors were studied for gene delivery in human RA FLS and synovium explants and murine collagen-induced arthritis. Short-fiber Ad5 vectors with seven fiber shaft repeats Ad5GFP-R7-knob, Ad5GFP-R7-arginine-glycine-aspartic acid (RGD) (RGD-liganded), and Ad5GFPDeltaknob (knob-deleted) were compared with Ad5GFP-FiWT, a conventional wild-type (WT) Ad5 vector. Gene transfer by Ad5GFP-R7-knob and Ad5GFP-R7-RGD was 40- to 50-fold and 25-fold higher, respectively, than Ad5GFP-FiWT in FLS. Ad5GFPDeltaknob was more efficacious than its knob-bearing version Ad5GFP-R7-knob in FLS transduction. Virus attachment and entry required RGD- and LDV-binding integrins including alpha(v), alpha(v)beta3, a(v)beta5, and beta1. Ad5GFP-R7-knob infection of FLS was partially neutralized by synovial fluid (SF), but remained 30- to 40-fold higher than Ad5GFP-FiWT in the presence of SF. Ad5GFPDeltaknob was partially neutralized by SF at low virus input, but escaped viral neutralization by SF at higher virus input. Gene transfer to human synovium ex vivo explants and murine collagen-induced arthritis in vivo was also more efficient with short fiber-modified vectors (with and without the knob domain) than Ad5GFPFiWT. Gene transfer by short fiber-modified vectors was enhanced by inflammatory cytokines in vitro and in the presence of inflammation in murine synovium in vivo. Our data indicated that the highly efficient gene delivery RA was mediated by RGD- and non-RGD-binding integrins and enhanced by inflammation. Short fiber modifications with knob ablation may be a strategy to enhance gene delivery, reducing vector dose and vector-induced inflammation and toxicity.
类风湿关节炎（RA）的成纤维样滑膜细胞（FLS）不表达柯萨奇腺病毒（Ad）受体，对Ad5血清型病毒的易感性较低。研究了基因改造的、不依赖柯萨奇Ad受体的Ad5载体在人RA FLS、滑膜外植体及小鼠胶原诱导性关节炎中的基因递送情况。将具有7个纤维柄重复序列的短纤维Ad5载体Ad5GFP-R7- knob、Ad5GFP-R7-精氨酸-甘氨酸-天冬氨酸（RGD）（RGD配体化）和Ad5GFPΔknob（缺失柄）与传统野生型（WT）Ad5载体Ad5GFP-FiWT进行比较。在FLS中，Ad5GFP-R7- knob和Ad5GFP-R7-RGD的基因转移效率分别比Ad5GFP-FiWT高40至50倍和25倍。在FLS转导中，Ad5GFPΔknob比其带有柄的版本Ad5GFP-R7- knob更有效。病毒附着和进入需要RGD和LDV结合整合素，包括α(v)、α(v)β3、α(v)β5和β1。滑膜液（SF）可部分中和FLS对Ad5GFP-R7- knob的感染，但在有SF存在的情况下，其感染效率仍比Ad5GFP-FiWT高30至40倍。在低病毒输入量时，Ad5GFPΔknob可被SF部分中和，但在高病毒输入量时可逃避SF的病毒中和作用。与Ad5GFPFiWT相比，短纤维修饰载体（有或无柄结构域）在体外人滑膜外植体和体内小鼠胶原诱导性关节炎中的基因转移效率也更高。在体外及小鼠滑膜存在炎症的体内环境中，炎性细胞因子可增强短纤维修饰载体的基因转移。我们的数据表明，RA中高效的基因递送由RGD和非RGD结合整合素介导，并受炎症增强。缺失柄的短纤维修饰可能是一种增强基因递送的策略，可降低载体剂量以及载体诱导的炎症和毒性。
