Smith J S, Keller J R, Lohrey N C, McCauslin C S, Ortiz M, Cowan K, Spence S E
Laboratories of Molecular Immunoregulation, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702, USA.
Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):8855-60. doi: 10.1073/pnas.96.16.8855.
The inability of adenovirus to infect primitive hematopoietic cells presents an obstacle to the use of adenovirus vectors for gene transfer to these cell types. Therefore, expanding the tropism of adenovirus vectors to unique cell surface antigens would be an important development for gene therapy protocols. In this study, we sought to redirect infection of adenovirus vectors to primitive human hematopoietic cells that universally express the c-Kit receptor on their cell surface. To accomplish this, a vector was constructed by covalently linking biotin molecules to recombinant adenovirus, followed by addition of the biotinylated ligand for the c-Kit receptor, stem cell factor (SCF), through an avidin bridge. Gene transfer was directed specifically to c-Kit-positive hematopoietic cell lines, resulting in up to a 2,440-fold increase in luciferase expression with frequencies equivalent to recombinant virus infection of permissive cells. Substitution of biotinylated antibodies directed against c-Kit, CD34 (binds L-selectin), and CD44 (hyaluronate receptor) receptors for biotinylated SCF resulted in 50-, 8-, and 260-fold increases in reporter gene expression, respectively, demonstrating that infection also could be redirected through antibody-antigen interactions and through antigens other than growth factor receptors. The versatility of this vector was demonstrated further by infection of primary T cells with vectors targeted with antibodies to CD44 (resting and activated T cells) and biotinylated IL-2 (activated T cells only). Taken together, directly biotinylated adenovirus vectors represent a versatile and efficient method for redirection of virus infection to specific cells.
腺病毒无法感染原始造血细胞，这为将腺病毒载体用于向这些细胞类型进行基因转移设置了障碍。因此，扩大腺病毒载体对独特细胞表面抗原的嗜性将是基因治疗方案的一项重要进展。在本研究中，我们试图将腺病毒载体的感染重定向至原始人类造血细胞，这些细胞在其细胞表面普遍表达c-Kit受体。为实现这一点，构建了一种载体，通过将生物素分子共价连接到重组腺病毒上，随后通过抗生物素蛋白桥添加c-Kit受体的生物素化配体——干细胞因子（SCF）。基因转移被特异性地导向c-Kit阳性造血细胞系，导致荧光素酶表达增加高达2440倍，其频率与重组病毒感染允许细胞相当。用针对c-Kit、CD34（结合L-选择素）和CD44（透明质酸受体）受体的生物素化抗体替代生物素化SCF，分别导致报告基因表达增加50倍、8倍和260倍，表明感染也可通过抗体-抗原相互作用以及通过生长因子受体以外的抗原进行重定向。通过用针对CD44的抗体（静息和活化T细胞）和生物素化IL-2（仅活化T细胞）靶向的载体感染原代T细胞，进一步证明了该载体的多功能性。综上所述，直接生物素化的腺病毒载体代表了一种将病毒感染重定向至特定细胞的通用且高效的方法。
