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Biochem Biophys Res Commun. 2015 Jul 31;463(3):216-21. doi: 10.1016/j.bbrc.2015.05.026. Epub 2015 May 19.

Ectopic expression of anti-HIV-1 shRNAs protects CD8(+) T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation.

Author information

1
Division of Hematology-Oncology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA. Electronic address: masa3k@ucla.edu.
2
Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
3
Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
4
Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA; Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA; UCLA AIDS Institute, Los Angeles, CA, USA; AIDS Healthcare Foundation, Los Angeles, CA, USA.
5
Division of Hematology-Oncology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA; Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA; UCLA AIDS Institute, Los Angeles, CA, USA.

Abstract

Chimeric antigen receptors (CARs) are artificially engineered receptors that confer a desired specificity to immune effector T cells. As an HIV-1-specific CAR, CD4ζ CAR has been extensively tested in vitro as well as in clinical trials. T cells modified with this CAR mediated highly potent anti-HIV-1 activities in vitro and were well-tolerated in vivo, but exerted limited effects on viral load and reservoir size due to poor survival and/or functionality of the transduced cells in patients. We hypothesize that ectopic expression of CD4ζ on CD8(+) T cells renders them susceptible to HIV-1 infection, resulting in poor survival of those cells. To test this possibility, highly purified CD8(+) T cells were genetically modified with a CD4ζ-encoding lentiviral vector and infected with HIV-1. CD8(+) T cells were vulnerable to HIV-1 infection upon expression of CD4ζ as evidenced by elevated levels of p24(Gag) in cells and culture supernatants. Concurrently, the number of CD4ζ-modified CD8(+) T cells was reduced relative to control cells upon HIV-1 infection. To protect these cells from HIV-1 infection, we co-expressed two anti-HIV-1 shRNAs previously developed by our group together with CD4ζ. This combination vector was able to suppress HIV-1 infection without impairing HIV-1-dependent effector activities of CD4ζ. In addition, the number of CD4ζ-modified CD8(+) T cells maintained similar levels to that of the control even under HIV-1 infection. These results suggest that protecting CD4ζ-modified CD8(+) T cells from HIV-1 infection is required for prolonged HIV-1-specific immune surveillance.

KEYWORDS:

CD4ζ; Chimeric antigen receptor (CAR); HIV-1; Immunotherapy; shRNA

PMID:
25998390
PMCID:
PMC4686265
DOI:
10.1016/j.bbrc.2015.05.026
[Indexed for MEDLINE]
Free PMC Article

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