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J Virol, February 1998, p. 1424-1430, Vol. 72, No. 2
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Entry of Amphotropic and 10A1 Pseudotyped Murine Retroviruses Is Restricted in Hematopoietic Stem Cell Lines

Dorothee von Laer,^1,* Silke Thomsen,¹ Birgit Vogt,¹ Martina Donath,² Joachim Kruppa,² Alan Rein,³ Wolfram Ostertag,¹ and Carol Stocking¹

Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg, 20251 Hamburg,¹ and Institut für Physiologische Chemie, Universität Hamburg, 20146 Hamburg,² Germany, and Laboratory of Molecular Virology and Carcinogenesis, Advanced Bioscience Laboratories, Basic Research Program, National Cancer Institute Frederick Cancer Research and Development Center, Frederick, Maryland 21702-1201³

Received 17 July 1997/Accepted 30 October 1997

Although transduction with amphotropic murine leukemia virus (MLV) vectors has been optimized successfully for hematopoietic differentiated progenitors, gene transfer to early hematopoietic cells (stem cells) is still highly restricted. A similar restriction to gene transfer was observed in the mouse stem cell line FDC-Pmix compared with transfer in the more mature myeloid precursor cell line FDC-P1 and the human erythroleukemia cell line K562. Gene transfer was not improved when the vector was pseudotyped with gp70^SU of the 10A1 strain of MLV, which uses the receptor of the gibbon ape leukemia virus (Pit1), in addition to the amphotropic receptor (Pit2). Although 10A1 and amphotropic gp70^SU bound to FDC-P1, K562, and fibroblasts, no binding to FDC-Pmix cells was detected. This indicates that FDC-Pmix cells lack functional Pit2 and Pit1 receptors. Pseudotyping with the vesicular stomatitis virus G protein improved transduction efficiency in FDC-Pmix stem cells by 2 orders of magnitude, to fibroblast levels, confirming a block to retroviral infection at the receptor level.

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^* Corresponding author. Mailing address: Heinrich-Pette-Institut, Martinistr. 52, D-20251 Hamburg, Germany. Phone: 49-40-48051274. Fax: 49-40-48051187. E-mail: laer{at}hpi.uni-hamburg.de.

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