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PMID: 8676458 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Transduction of human CD34+ hematopoietic progenitor cells by a retroviral vector expressing an RRE decoy inhibits human immunodeficiency virus type 1 replication in myelomonocytic cells produced in long-term culture.

Journal of virology ·Vol. 70 ·No. 7 ·1996-07-00 ·Pages 4352-60

Bahner I, Kearns K, Hao QL, Smogorzewska EM, Kohn DB

Abstract

Genetic modification of hematopoietic stem cells with a synthetic "anti-human immunodeficiency virus type 1 (HIV-1) gene" which inhibits replication of HIV-1 may allow production of mature lymphoid and monocytic cells resistant to HIV-1 growth after autologous transplantation. Because productive HIV-1 replication requires binding of the Rev protein to the Rev-responsive element (RRE) within the viral transcripts for the HIV-1 structural proteins, anti-HIV-1 gene products which interfere with Rev-RRE interactions may inhibit HIV-1 replication. One such strategy involves overexpression of the RRE sequences in transcripts derived from retroviral vectors to act as decoys to sequester Rev protein and prevent its binding to the RRE element in HIV-1 transcripts. We developed an in vitro model to test the efficacy of this gene therapy approach in primary human hematopoietic cells. Human CD34+ hematopoietic progenitor cells from normal bone marrow or umbilical cord blood were transduced with retroviral vectors carrying RRE decoy sequences as part of a long terminal repeat-directed transcript expressing the neo gene (L-RRE-neo) or with a control vector expressing only the neo gene (LN). The transduced progenitors were allowed to differentiate into mature myelomonocytic cells which were able to support vigorous growth of the monocytotropic isolate of HIV-1, JR-FL. HIV-1 replication was measured in unselected cell populations and following G418 selection to obtain uniformly transduced cell populations. Inhibition of HIV-1 replication in the unselected cell cultures was between 50.2 and 76.7% and was highly effective (99.4 to 99.9%) in the G418-selected cultures. Progenitors transduced by either the L-RRE-neo vector or the control LN vector were identical with respect to hematopoietic growth and differentiation. These findings demonstrate the ability of an RRE decoy strategy to inhibit HIV-1 replication in primary human myelomonocytic cells after transduction of CD34+ progenitor cells, without adverse effects on hematopoietic cell function.

MeSH Terms
3T3 Cells Animals Antigens, CD34 Base Sequence Cell Differentiation Cell Division Cells, Cultured Gene Products, rev/antagonists & inhibitors,metabolism Gene Transfer Techniques Genetic Therapy/methods Genetic Vectors HIV Core Protein p24/biosynthesis HIV-1/metabolism,physiology Hematopoietic Stem Cells/immunology,virology Humans Mice Molecular Sequence Data RNA, Viral T-Lymphocytes/cytology,virology Virus Replication/genetics rev Gene Products, Human Immunodeficiency Virus
Chemicals
Antigens, CD34 Gene Products, rev HIV Core Protein p24 RNA, Viral rev Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bahner I
Department of Pediatrics, University of Southern California School of Medicine, California 90027, USA.
Kearns K
Hao Q L
Smogorzewska E M
Kohn D B
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.70.7.4352-4360.1996
Published
1996-07-00
Pages
4352-60
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC190368
Subset
IM
Grants
NIAID NIH HHS · AI 1U19AI36606 · United States
NIAID NIH HHS · AI RFA-89-AI-19 A125959 · United States
External Links
PubMed source DOI: 10.1128/JVI.70.7.4352-4360.1996 PMC Full Text View journal details
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