Adams R M, Wang M, Steffen D, Ledley F D
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
J Virol. 1995 Mar;69(3):1887-94. doi: 10.1128/JVI.69.3.1887-1894.1995.
Retrovirus infection is normally limited to cells within a specific host range which express a cognate receptor that is recognized by the product of the env gene. We describe retrovirus infection of cells outside of their normal host range when the infection is performed in the presence of a replication-defective adenovirus (dl312). In the presence of adenovirus, several different ecotropic vectors are shown to infect human cell lines (HeLa and PLC/PRF), and a xenotropic vector is shown to infect murine cells (NIH 3T3). Infectivity is demonstrated by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining, selection with G418 for neomycin resistance, and PCR identification of the provirus in infected cells. Infectivity is quantitatively dependent upon both the concentration of adenovirus (10(6) to 10(8) PFU/ml) and the concentration of retrovirus. Infection requires the simultaneous presence of adenovirus in the retrovirus infection medium and is not stimulated by preincubation and removal of adenovirus from the cells before retrovirus infection. The presence of adenovirus is shown to enhance the uptake of fluorescently labeled retrovirus particles into cells outside of their normal host range, demonstrating that the adenovirus enhances viral entry into cells in the absence of the recognized cognate receptor. This observation suggests new opportunities for developing safe retroviral vectors for gene therapy and new mechanisms for the pathogenesis of retroviral disease.
逆转录病毒感染通常局限于特定宿主范围内表达同源受体的细胞,该同源受体可被env基因产物识别。我们描述了在复制缺陷型腺病毒(dl312)存在的情况下,逆转录病毒对其正常宿主范围外细胞的感染。在腺病毒存在的情况下,几种不同的嗜亲性载体可感染人细胞系(HeLa和PLC/PRF),一种异嗜性载体可感染鼠细胞(NIH 3T3)。通过5-溴-4-氯-3-吲哚-β-D-吡喃半乳糖苷(X-Gal)染色、用G418选择新霉素抗性以及对感染细胞中的前病毒进行PCR鉴定来证明感染性。感染性在数量上取决于腺病毒的浓度(10^6至10^8 PFU/ml)和逆转录病毒的浓度。感染需要在逆转录病毒感染培养基中同时存在腺病毒,并且在逆转录病毒感染前对细胞进行预孵育并去除腺病毒并不能刺激感染。已证明腺病毒的存在可增强荧光标记的逆转录病毒颗粒进入其正常宿主范围外细胞的摄取,这表明腺病毒在没有识别的同源受体的情况下增强了病毒进入细胞的能力。这一观察结果为开发用于基因治疗的安全逆转录病毒载体提供了新的机会,并为逆转录病毒疾病的发病机制提供了新的机制。