High efficiency gene transfer to normal and malignant hematopoietic precursor cells using a chimeric adenovirus

Patricia Yotnda, Haroaki Onishi, Helen Heslop, Malcolm Brenner, Dmitri Shayakhmetov, Andre Lieber, Alan Davis

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Almost all studies of Adenoviral vector gene transfer have made use of the Adenovinis type 5 serotype. Unfortunately, Ad5 has generally been ineffective at transducirg hemopoieitic progenitor cells (HPC). Chimeric Adenovinis Type 5 vectors that have bei n engineered to substitute the shorter-shafted fiber protein from Adenovinis type 35 ci.n transduce cells apparently lacking CAR or alpha(v) integrins required for Ad5 binding. We find that these vectors have the ability to rapidly transduce even the most phenotypical y primitive subset of HPC when they are used at low viral concentration even in the absem ;e of growth factors. An AdSGFP and Ad5/35GFP vector was added to CD34+ and o CD34- lineage- human marrow progenitor cells. Transduction used a 6 hr co-incubation 3f the cells with the virus (1000 vp) in the absence of growth factors. Twenty-four hours after infection, cells were analyzed by flow cytometry for eGFP expression. Only 5-20% of CD34+ and CD34-lineage - cells expressed eGFP after Ad5 exposure. In contrast, with the Ad35 pseudotyped vector, 30-70% of the CD34+ and 50-70%CD34-lineage-cells were positive for eGFP expression. The eGFP expression was detectable as soon as 6hr post-infection, when 24hr was necessary to reach discernible expression for Ad5 infectpd cells. Because of these improved results, we also analyzed the ability of the chimeric virLis to infect the Hoechst negative Side Profile population of CD34- marrow cells, which appear to be amongst the very earliest hematopoietic progenitor cells (Goodell MA et al Nat Med. 1997 Dec;3(12): 1337-45). Between 51% and 80% of SP bone marrow ce Is expressed eGFP 24-hr post-infection. The transduced CD34+ and CD34- lin - populatio is retained their ability to form colonies in short and long term culture systems, with no significant loss of viability. Moreover, a high level of expression was also obtained with the chimeric vector but not with Ad5 in unstimulated malignant blasts from patients wi th CD34+ and CD34- AML and in the CDS positive B cells of patients with B-CLL. T le ability of chimeric Ad5/35F to deliver transgenes to normal and malignant hematopoiei ic stem cells with high efficiency and low toxicity in the absence of growth factors providîs an improved means of studying the consequences of transient gene expression in thepe cells.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART I
StatePublished - Dec 1 2000

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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