Yang Y, Haecker S E, Su Q, Wilson J M
Institute for Human Gene Therapy, University of Pennsylvania Medical Center, Philadelphia 19104, USA.
Hum Mol Genet. 1996 Nov;5(11):1703-12. doi: 10.1093/hmg/5.11.1703.
Skeletal muscle is an attractive target for somatic gene transfer of both acquired and inherited disorders. Direct injection of adenoviral vectors in the skeletal muscle leads to recombinant gene expression in a large number of muscle fibers. Transgene expression has been transient in most organs and associated with substantial inflammation when experiments are performed in adult immune competent mice. In this report, we utilize a variety of in vivo and in vitro models of T and B cell function to characterize the nature of the immune response to adenoviral vectors injected into murine skeletal muscle. Cellular immunity dependent on CD4+ and CD8+ T cells contributes to the loss of recombinant gene expression and the development of localized inflammation. Antigen specific activation of T cells occurs to both viral proteins and the reporter gene beta-galactosidase. Systemic levels of neutralizing antibody to the capsid proteins of the vector are also generated. Destructive immune responses responsible for loss of transgene expression are largely directed against beta-galactosidase in that transgene expression was stable when beta-galactosidase was eliminated as a neoantigen in mice transgenic for lacZ. A strategy to prevent the cellular and humoral immunity to this therapy was developed based on transiently ablating CD4+ T cell activation at the time of vector delivery. Encouraging results were obtained when vector was administered with one of several immune modulating agents including cyclophosphamide, mAb to CD4+ cells, and mAb to CD40 ligand. These studies indicate that cellular and humoral immune responses are elicited in the context of gene therapy directed to skeletal muscle with adenoviral vectors. Transient ablation of CD4+ T cell activation prevents the effects responses of the CD8+ T and B cells.
骨骼肌是获得性和遗传性疾病体细胞基因转移的一个有吸引力的靶点。将腺病毒载体直接注射到骨骼肌中可导致大量肌纤维中的重组基因表达。在成年免疫健全小鼠中进行实验时，转基因表达在大多数器官中是短暂的，并且与大量炎症相关。在本报告中，我们利用多种T细胞和B细胞功能的体内和体外模型来表征对注射到小鼠骨骼肌中的腺病毒载体免疫反应的性质。依赖CD4+和CD8+ T细胞的细胞免疫导致重组基因表达的丧失和局部炎症的发展。T细胞对病毒蛋白和报告基因β-半乳糖苷酶都发生了抗原特异性激活。还产生了针对载体衣壳蛋白的中和抗体的全身水平。导致转基因表达丧失的破坏性免疫反应主要针对β-半乳糖苷酶，因为当β-半乳糖苷酶作为新抗原在转基因小鼠中被消除时，转基因表达是稳定的。基于在载体递送时短暂消除CD4+ T细胞激活，开发了一种预防针对该疗法的细胞和体液免疫的策略。当载体与几种免疫调节剂之一（包括环磷酰胺、抗CD4+细胞单克隆抗体和抗CD40配体单克隆抗体）一起给药时，获得了令人鼓舞的结果。这些研究表明，在使用腺病毒载体对骨骼肌进行基因治疗的背景下会引发细胞和体液免疫反应。CD4+ T细胞激活的短暂消除可防止CD8+ T细胞和B细胞的效应反应。
