Harvey B G, Hackett N R, El-Sawy T, Rosengart T K, Hirschowitz E A, Lieberman M D, Lesser M L, Crystal R G
Division of Pulmonary and Critical Care Medicine, Weill Medical College of Cornell University-New York Presbyterian Hospital, New York, USA.
J Virol. 1999 Aug;73(8):6729-42. doi: 10.1128/JVI.73.8.6729-6742.1999.
Administration of adenovirus (Ad) vectors to immunologically naive experimental animals almost invariably results in the induction of systemic anti-Ad neutralizing antibodies. To determine if the human systemic humoral host responses to Ad vectors follow a similar pattern, we evaluated the systemic (serum) anti-Ad serotype 5 (Ad5) neutralizing antibodies in humans after administration of first generation (E1(-) E3(-)) Ad5-based gene transfer vectors to different hosts. AdGVCFTR.10 (carrying the normal human cystic fibrosis [CF] transmembrane regulator cDNA) was sprayed (8 x 10(7) to 2 x 10(10) particle units [PU]) repetitively (every 3 months or every 2 weeks) to the airway epithelium of 15 individuals with CF. AdGVCD.10 (carrying the Escherichia coli cytosine deaminase gene) was administered (8 x 10(8) to 8 x 10(9) PU; once a week, twice) directly to liver metastasis of five individuals with colon cancer and by the intradermal route (8 x 10(7) to 8 x 10(9) PU, single administration) to six healthy individuals. AdGVVEGF121.10 (carrying the human vascular endothelial growth factor 121 cDNA) was administered (4 x 10(8) to 4 x 10(9.5) PU, single administration) directly to the myocardium of 11 individuals with ischemic heart disease. Ad vector administration to the airways of individuals with CF evoked no or minimal serum neutralizing antibodies, even with repetitive administration. In contrast, intratumor administration of an Ad vector to individuals with metastatic colon cancer resulted in a robust antibody response, with anti-Ad neutralizing antibody titers of 10(2) to >10(4). Healthy individuals responded to single intradermal Ad vector variably, from induction of no neutralizing anti-Ad antibodies to titers of 5 x 10(3). Likewise, individuals with ischemic heart disease had a variable response to single intramyocardial vector administration, ranging from minimal neutralizing antibody levels to titers of 10(4). Evaluation of the data from all trials showed no correlation between the peak serum neutralizing anti-Ad response and the dose of Ad vector administered (P > 0.1, all comparisons). In contrast, there was a striking correlation between the peak anti-Ad5 neutralizing antibody levels evoked by vector administration and the level of preexisting anti-Ad5 antibodies (P = 0.0001). Thus, unlike the case for experimental animals, administration of Ad vectors to humans does not invariably evoke a systemic anti-Ad neutralizing antibody response. In humans, the extent of the response is dictated by preexisting antibody titers and modified by route of administration but is not dose dependent. Since the extent of anti-Ad neutralizing antibodies will likely modify the efficacy of administration of Ad vectors, these observations are of fundamental importance in designing human gene therapy trials and in interpreting the efficacy of Ad vector-mediated gene transfer.
给免疫未成熟的实验动物施用腺病毒(Ad)载体几乎总是会诱导全身性抗Ad中和抗体的产生。为了确定人类对Ad载体的全身性体液宿主反应是否遵循类似模式,我们评估了向不同宿主施用第一代(E1(-)E3(-))基于Ad5的基因转移载体后,人类血清中抗Ad血清型5(Ad5)的中和抗体。AdGVCFTR.10(携带正常人囊性纤维化[CF]跨膜调节因子cDNA)以8×10⁷至2×10¹⁰颗粒单位(PU)重复(每3个月或每2周一次)喷入15名CF患者的气道上皮。AdGVCD.10(携带大肠杆菌胞嘧啶脱氨酶基因)以8×10⁸至8×10⁹PU(每周一次,共两次)直接施用于5名结肠癌肝转移患者,并通过皮内途径(8×10⁷至8×10⁹PU,单次给药)施用于6名健康个体。AdGVVEGF121.10(携带人血管内皮生长因子121 cDNA)以4×10⁸至4×10⁹.⁵PU(单次给药)直接施用于11名缺血性心脏病患者的心肌。向CF患者的气道施用Ad载体即使重复给药也不会引起或仅引起极少的血清中和抗体。相比之下,向转移性结肠癌患者肿瘤内施用Ad载体导致强烈的抗体反应,抗Ad中和抗体滴度为10²至>10⁴。健康个体对单次皮内Ad载体的反应各不相同,从未诱导出中和性抗Ad抗体到滴度为5×10³。同样,缺血性心脏病患者对单次心肌内载体给药的反应也各不相同,从中和抗体水平极低到滴度为10⁴。对所有试验数据的评估表明,血清中抗Ad中和反应峰值与所施用Ad载体的剂量之间没有相关性(P>0.1,所有比较)。相反,载体给药引起的抗Ad5中和抗体水平峰值与预先存在的抗Ad5抗体水平之间存在显著相关性(P = 0.0001)。因此,与实验动物的情况不同,向人类施用Ad载体并非总是会引起全身性抗Ad中和抗体反应。在人类中,反应程度由预先存在的抗体滴度决定,并因给药途径而改变,但与剂量无关。由于抗Ad中和抗体的程度可能会改变Ad载体给药的疗效,这些观察结果对于设计人类基因治疗试验和解释Ad载体介导的基因转移疗效具有至关重要的意义。