Anti-human immunodeficiency virus type 1 gene therapy in human central nervous system-based cells: an initial approach against a potential viral reservoir.
Studies have demonstrated that human immunodeficiency virus type 1 (HIV-1) infection of central nervous system (CNS)-based cells in vivo results in a series of devastating clinical conditions collectively termed acquired immune deficiency syndrome (AIDS) dementia complex (ADC). Gene therapy for these neurovirological disorders necessitates utilization of a vector system that can mediate in vivo delivery and long-term expression of an antiretroviral transgene in nondividing/postmitotic CNS cellular elements. The present studies focus on the transfer of an anti-HIV-1 gene to primary isolated CNS microvascular endothelial cells (MVECs) and neuronal-based cells, for its effects in protecting these cells from HIV-1 infection. By using an HIV-1-based vector system, it was possible to efficiently transduce and maintain expression of a marker transgene, beta-galactosidase (beta-Gal), in human CNS MVECs, human fetal astrocytes, plus immature and mature (differentiated) NT2 cells. Significant transduction of the marker gene, beta-Gal, in CNS-based cells prompted the utilization of this system with an anti-HIV-1 gene therapeutic construct, RevM10, a trans-dominant negative mutant Rev protein. Initially, it was not possible to generate any HIV-1 vector particles with the RevM10 gene in the transducing construct, because of inhibitory effects on the HIV-1 vector by this gene product. However, the vector could be partially rescued by adding an additional construct that supplied wild-type rev, in trans, during a multiple construct transfection in the packaging 293T cells. Thus, it was possible to significantly improve the titer of RevM10-expressing viral particles generated from these cells. Moreover, this RevM10 vector transduced the neuronal precursor cell line NT2, retinoic acid-differentiated human neurons (hNT) from the precursor cells, and primary isolated human brain MVECs with high efficiency. RevM10 generated from the HIV-1-based vector system potently inhibited replication of diverse HIV-1 strains in human CNS MVECs and neuronal cells. The data generated from these studies represent an initial approach for future development of anti-HIV-1 gene therapy in the CNS.
Mukhtar
M
M
Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Duke
H
H
BouHamdan
M
M
Pomerantz
R J
RJ
D015526:AIDS Dementia Complex / Q000628:therapy; D001921:Brain / Q000378:metabolism*; D002454:Cell Differentiation; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D015757:Genes, rev / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008833:Microcirculation / Q000378:metabolism*; D008957:Models, Genetic; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D016366:Open Reading Frames; D013997:Time Factors; D014212:Tretinoin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D019065:Virus Assembly / Q000235:genetics
D014212:Tretinoin