Mastrangeli A, Harvey B G, Yao J, Wolff G, Kovesdi I, Crystal R G, Falck-Pedersen E
Division of Pulmonary and Critical Care Medicine, New York Hospital-Cornell University Medical College, New York, NY 10021, USA.
Hum Gene Ther. 1996 Jan;7(1):79-87. doi: 10.1089/hum.1996.7.1-79.
Recombinant, replication-deficient adenovirus (Ad) vectors have been successfully used to transfer and express the normal human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA in vivo in the respiratory epithelium of experimental animals and humans with cystic fibrosis (CF). Since Ad-directed gene expression wanes over time, repeat administration is necessary to achieve an effective treatment for CF. A major hurdle to such a strategy is the possibility that anti-Ad humoral immunity may prevent gene expression in individuals with pre-existing anti-Ad immunity or following repeat administration. One strategy to circumvent such a problem would be alternating the use of Ad vectors belonging to different subgroups. Neutralizing antibodies developed with the administration of one Ad serotype do not cross-react with an Ad belonging to a second serotype in a manner that blocks infection and gene expression. To test this hypothesis, an immunizing dose of wild-type Ad5 (subgroup C), Ad4 (subgroup E), or Ad30 (subgroup D) was administered intratracheally to experimental animals, followed by an intratracheal administration of a replication-deficient subgroup C-derived vector coding for marker genes (chloramphenicol acetyl transferase or beta-galactosidase) or for the normal human CFTR cDNA. As expected, studies with vectors coding for marker genes or for CFTR cDNA demonstrated that airway administration of a vector does not yield efficient gene transfer, if there has been prior recent airway administration of the same Ad subgroup. In contrast, effective expression from the second administration can be achieved with an adenovirus vector belonging to a subgroup different from the first adenovirus administered. These data support the paradigm of alternating Ad vectors derived from different subgroups as strategy to circumvent anti-Ad humoral immunity, thus permitting the use of Ad vectors as a means to treat the respiratory manifestations of CF.
重组的、复制缺陷型腺病毒（Ad）载体已成功用于在实验动物和患有囊性纤维化（CF）的人类的呼吸道上皮细胞中体内转移和表达正常人囊性纤维化跨膜传导调节因子（CFTR）cDNA。由于Ad介导的基因表达会随时间减弱，因此需要重复给药才能实现对CF的有效治疗。这种策略的一个主要障碍是，抗Ad体液免疫可能会阻止已有抗Ad免疫的个体或重复给药后的个体中的基因表达。规避这一问题的一种策略是交替使用属于不同亚组的Ad载体。用一种Ad血清型给药产生的中和抗体不会以阻断感染和基因表达的方式与属于第二种血清型的Ad发生交叉反应。为了验证这一假设，将免疫剂量的野生型Ad5（C亚组）、Ad4（E亚组）或Ad30（D亚组）经气管内给予实验动物，随后经气管内给予编码标记基因（氯霉素乙酰转移酶或β-半乳糖苷酶）或正常人CFTR cDNA的复制缺陷型C亚组衍生载体。正如预期的那样，对编码标记基因或CFTR cDNA的载体的研究表明，如果近期之前已经经气道给予了相同的Ad亚组，则气道给予载体不会产生有效的基因转移。相反，使用属于与第一次给予的腺病毒不同亚组的腺病毒载体可以实现第二次给药后的有效表达。这些数据支持将源自不同亚组的Ad载体交替使用作为规避抗Ad体液免疫的策略这一模式，从而允许将Ad载体用作治疗CF呼吸道表现的手段。
