Lee M G, Abina M A, Haddada H, Perricaudet M
Laboratoire de Génétique de Virus Oncogènes, URA 1301 CNRS, Institut Gustave Roussy PR2, Villejuif, France.
Gene Ther. 1995 Jun;2(4):256-62.
The immune response against cells infected by gene therapy vectors may be a major hindrance for gene therapy, destroying infected cells thus limiting the length of exogene expression and quickly eliminating infected cells on repeat administration. Adenoviruses and many other pathogens have evolved strategies for escape from immune surveillance, including the gp19k gene, found in the adenovirus E3 region, known to down-regulate major histocompatibility complex class 1 expression on the cell surface, and thus reduce lysis of the infected cells by cytotoxic T cells. We have constructed an adenoviral vector expressing the genes for beta-galactosidase and gp19k both under the control of constitutive promoters, and compared the capacity of lymphocytes from DBA/2 mice previously injected with the virus or with Ad-beta gal, a virus expressing beta-galactosidase but not gp19k, to lyse target cells infected with various viruses. Lymphocytes raised against Ad-beta gal fail to lyse target cells infected with Ad-beta gal-gp19k significantly, whereas Ad-beta gal infected target cells and a beta-galactosidase expressing cell line are strongly lysed. The administration of Ad-beta gal-gp19k fails to stimulate the proliferation of anti-vector lymphocytes, and thus these lymphocytes show poor cytotoxic activity against Ad-beta gal or Ad-beta gal-gp19k infected cells.
针对基因治疗载体感染细胞的免疫反应可能是基因治疗的主要障碍，它会破坏感染细胞，从而限制外源基因表达的时长，并在重复给药时迅速清除感染细胞。腺病毒和许多其他病原体已经进化出逃避免疫监视的策略，包括在腺病毒E3区域发现的gp19k基因，已知该基因可下调细胞表面主要组织相容性复合体I类的表达，从而减少细胞毒性T细胞对感染细胞的裂解。我们构建了一种腺病毒载体，在组成型启动子的控制下表达β-半乳糖苷酶和gp19k基因，并比较了先前注射该病毒或Ad-βgal（一种表达β-半乳糖苷酶但不表达gp19k的病毒）的DBA/2小鼠淋巴细胞裂解感染各种病毒的靶细胞的能力。针对Ad-βgal产生的淋巴细胞不能有效裂解被Ad-βgal-gp19k感染的靶细胞，而被Ad-βgal感染的靶细胞和一个表达β-半乳糖苷酶的细胞系则被强烈裂解。给予Ad-βgal-gp19k不能刺激抗载体淋巴细胞的增殖，因此这些淋巴细胞对被Ad-βgal或Ad-βgal-gp19k感染的细胞显示出较差的细胞毒性活性。
