Gahéry-Ségard H, Molinier-Frenkel V, Le Boulaire C, Saulnier P, Opolon P, Lengagne R, Gautier E, Le Cesne A, Zitvogel L, Venet A, Schatz C, Courtney M, Le Chevalier T, Tursz T, Guillet J G, Farace F
Laboratoire d'Immunologie des Pathologies Infectieuses et Tumorales, INSERM Unité 445, Université R. Descartes, Hôpital Cochin, Paris, France.
J Clin Invest. 1997 Nov 1;100(9):2218-26. doi: 10.1172/JCI119759.
Animal studies indicate that the use of replication-deficient adenovirus for human gene therapy is limited by host antivector immune responses that result in transient recombinant protein expression and blocking of gene transfer when rechallenged. Therefore, we have examined immune responses to an adenoviral vector and to the beta-galactosidase protein in four patients with lung cancer given a single intratumor injection of 10(9) plaque-forming units of recombinant adenovirus. The beta-galactosidase protein was expressed in day-8 tumor biopsies from all patients at variable levels. Recombinant virus DNA was detected by PCR in day-30 and day-60 tumor biopsies from all patients except patient 1. A high level of neutralizing antiadenovirus antibodies was detected in patient 1 before Ad-beta-gal injection whereas it was low (patient 3) or undetectable in the other two patients. All patients developed potent CD4 type 1 helper T cell (Th1) responses to adenoviral particles which increased gradually over time after injection. Antiadenovirus cytotoxic T lymphocyte responses were consistently boosted in the two patients examined (patients 3 and 4). Sustained production of anti-beta-galactosidase IgG was observed in all patients except patient 1. Consistent with anti-beta-gal antibody production, all patients except patient 1 developed intense, dose-dependent Th1 responses to soluble beta-galactosidase which increased over time. Strong beta-galactosidase-specific cytotoxic T lymphocyte responses were detected in patients 2, 3, and 4. Our results clearly show that despite the intensity of antiadenovirus responses, transgene protein expression was sufficient to induce strong and prolonged immunity in three patients. Recombinant adenovirus injected directly into the tumor is a highly efficient vector for immunizing patients against the transgene protein.
动物研究表明，用于人类基因治疗的复制缺陷型腺病毒受到宿主抗载体免疫反应的限制，这种反应会导致重组蛋白短暂表达，并在再次给药时阻断基因转移。因此，我们研究了4例肺癌患者在瘤内单次注射10⁹ 噬斑形成单位重组腺病毒后，对腺病毒载体和β-半乳糖苷酶蛋白的免疫反应。所有患者第8天的肿瘤活检组织中均检测到不同水平的β-半乳糖苷酶蛋白表达。除患者1外，所有患者第30天和第60天的肿瘤活检组织经聚合酶链反应（PCR）均检测到重组病毒DNA。患者1在注射Ad-β-gal前检测到高水平的中和抗腺病毒抗体，而其他两名患者抗体水平较低（患者3）或未检测到。所有患者对腺病毒颗粒均产生了有效的1型辅助性T细胞（Th1）反应，注射后随时间逐渐增强。在接受检测的两名患者（患者3和患者4）中，抗腺病毒细胞毒性T淋巴细胞反应持续增强。除患者1外，所有患者均观察到抗β-半乳糖苷酶IgG的持续产生。与抗β-半乳糖苷酶抗体产生一致，除患者1外，所有患者对可溶性β-半乳糖苷酶均产生了强烈的、剂量依赖性的Th1反应，且随时间增强。在患者2、3和4中检测到强烈的β-半乳糖苷酶特异性细胞毒性T淋巴细胞反应。我们的结果清楚地表明，尽管抗腺病毒反应强烈，但转基因蛋白表达足以在三名患者中诱导强烈且持久的免疫反应。直接注射到肿瘤内的重组腺病毒是一种高效的载体，可使患者针对转基因蛋白产生免疫。
