Mei Ya-Fang, Segerman Anna, Lindman Kristina, Hörnsten Per, Wahlin Anders, Wadell Göran
Department of Virology, Umeå University, S-901 85 Umeå, Sweden.
Virology. 2004 Oct 25;328(2):198-207. doi: 10.1016/j.virol.2004.07.018.
Gene transfer into human hematopoietic stem cells using Ad5 is inefficient due to lack of the primary receptor CAR and the secondary receptors alphavbeta3 integrin and alphavbeta5 integrin, and due to the high seroprevalence of Ad5 antibodies in most adults, resulting in diminished gene transduction. In the present study, we screened six species (species A-F) of adenovirus, displaying different tropisms for interaction with CD34+ cells, at the level of virus attachment and expression. Virus particles were biotinylated and their binding capacity was determined by FACS analysis using streptavidin-FITC. Ad11p, Ad35, and Ad3 (species B) showed high binding affinity, while Ad7, Ad11a (species B), and Ad37 (species D) displayed intermediate affinity. Virions of Ad4 (species E), Ad5 (species C), Ad31 (species A), and Ad41 (species F) hardly bound to hematopoietic progenitor cells. Using a double-labeling system, we demonstrated that adenoviruses bind to quiescent CD34+ cells. Ad11p virions showed the highest affinity among the adenoviruses detected. We further confirmed that virus fiber-specific receptors were present on the hematopoietic progenitor cell surface, because both recombinant fiber of Ad11p and specific antiserum against rfiber could block virus attachment. The ability of the adenoviruses to infect hematopoietic cells was studied by immunofluorescence staining. The adenoviruses from species B and Ad37 showed higher infectivity than Ad31, Ad5, Ad4, and Ad41. Among the studied species B adenoviruses, Ad11p manifested a superior infectivity. Thus, we have confirmed that these cells have high-affinity receptors for species B:2 human adenovirus, Ad11p, and this virus may be used as candidate vector to target therapeutic genes to hematopoietic stem cells.
由于缺乏主要受体柯萨奇病毒和腺病毒受体（CAR）以及次要受体αvβ3整合素和αvβ5整合素，并且由于大多数成年人中腺病毒5型（Ad5）抗体的高血清阳性率，导致基因转导效率降低，因此利用Ad5将基因导入人类造血干细胞的效率较低。在本研究中，我们在病毒附着和表达水平上筛选了六种腺病毒（A - F型），它们对与CD34 +细胞的相互作用表现出不同的嗜性。病毒颗粒用生物素标记，其结合能力通过使用链霉亲和素 - FITC的流式细胞术分析来确定。Ad11p、Ad35和Ad3（B型）显示出高结合亲和力，而Ad7、Ad11a（B型）和Ad37（D型）表现出中等亲和力。Ad4（E型）、Ad5（C型）、Ad31（A型）和Ad41（F型）的病毒粒子几乎不与造血祖细胞结合。使用双标记系统，我们证明腺病毒与静止的CD34 +细胞结合。在检测到的腺病毒中，Ad11p病毒粒子表现出最高的亲和力。我们进一步证实造血祖细胞表面存在病毒纤维特异性受体，因为Ad11p的重组纤维和针对重组纤维的特异性抗血清都可以阻断病毒附着。通过免疫荧光染色研究了腺病毒感染造血细胞的能力。B型腺病毒和Ad37比Ad31、Ad5、Ad4和Ad41表现出更高的感染性。在所研究的B型腺病毒中，Ad11p表现出卓越的感染性。因此，我们已经证实这些细胞对B型人类腺病毒Ad11p具有高亲和力受体，并且这种病毒可作为将治疗基因靶向造血干细胞的候选载体。
