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Title: Ectopic expression of anti-HIV-1 shRNAs protects CD8{sup +} T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation

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{sup +} T cells renders them susceptible to HIV-1 infection, resulting in poor survival of those cells. To test this possibility, highly purified CD8{sup +} T cells were genetically modified with a CD4ζ-encoding lentiviral vector and infected with HIV-1. CD8{sup +} T cells were vulnerable to HIV-1 infection upon expression of CD4ζ as evidenced by elevated levels of p24{sup Gag} in cells and culture supernatants. Concurrently, the number of CD4ζ-modified CD8{sup +} 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.more » In addition, the number of CD4ζ-modified CD8{sup +} T cells maintained similar levels to that of the control even under HIV-1 infection. These results suggest that protecting CD4ζ-modified CD8{sup +} T cells from HIV-1 infection is required for prolonged HIV-1-specific immune surveillance. - Highlights: • Ectopic expression of CD4ζ CAR in CD8{sup +} T cells renders them susceptible to HIV-1 infection. • Co-expression of two anti-HIV-1 shRNAs protects CD4ζ CAR-modified CD8{sup +} T cells from HIV-1 infection. • Protecting CD4ζ CAR-modified CD8{sup +} T cells from HIV-1 infection suppresses its cytopathic effect.« less

Authors:
 [1];  [2];  [3]; ; ; ;  [2];  [2];  [1]
  1. Division of Hematology-Oncology, David Geffen School of Medicine at UCLA, Los Angeles, CA (United States)
  2. Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA (United States)
  3. Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA (United States)
Publication Date:
OSTI Identifier:
22462116
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 463; Journal Issue: 3; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AIDS VIRUS; ANTIGENS; CELL PROLIFERATION; IMMUNOTHERAPY; IN VITRO; IN VIVO; INSPECTION; MEDICAL SURVEILLANCE; MONITORING; PATIENTS; RECEPTORS; RNA; SPECIFICITY; VECTORS

Citation Formats

Kamata, Masakazu, Kim, Patrick Y., Ng, Hwee L., Ringpis, Gene-Errol E., Kranz, Emiko, Chan, Joshua, O'Connor, Sean, Yang, Otto O., Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA, UCLA AIDS Institute, Los Angeles, CA, AIDS Healthcare Foundation, Los Angeles, CA, Chen, Irvin S.Y., Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, and UCLA AIDS Institute, Los Angeles, CA. Ectopic expression of anti-HIV-1 shRNAs protects CD8{sup +} T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation. United States: N. p., 2015. Web. doi:10.1016/J.BBRC.2015.05.026.
Kamata, Masakazu, Kim, Patrick Y., Ng, Hwee L., Ringpis, Gene-Errol E., Kranz, Emiko, Chan, Joshua, O'Connor, Sean, Yang, Otto O., Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA, UCLA AIDS Institute, Los Angeles, CA, AIDS Healthcare Foundation, Los Angeles, CA, Chen, Irvin S.Y., Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, & UCLA AIDS Institute, Los Angeles, CA. Ectopic expression of anti-HIV-1 shRNAs protects CD8{sup +} T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation. United States. doi:10.1016/J.BBRC.2015.05.026.
Kamata, Masakazu, Kim, Patrick Y., Ng, Hwee L., Ringpis, Gene-Errol E., Kranz, Emiko, Chan, Joshua, O'Connor, Sean, Yang, Otto O., Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA, UCLA AIDS Institute, Los Angeles, CA, AIDS Healthcare Foundation, Los Angeles, CA, Chen, Irvin S.Y., Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA, and UCLA AIDS Institute, Los Angeles, CA. Fri . "Ectopic expression of anti-HIV-1 shRNAs protects CD8{sup +} T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation". United States. doi:10.1016/J.BBRC.2015.05.026.
@article{osti_22462116,
title = {Ectopic expression of anti-HIV-1 shRNAs protects CD8{sup +} T cells modified with CD4ζ CAR from HIV-1 infection and alleviates impairment of cell proliferation},
author = {Kamata, Masakazu and Kim, Patrick Y. and Ng, Hwee L. and Ringpis, Gene-Errol E. and Kranz, Emiko and Chan, Joshua and O'Connor, Sean and Yang, Otto O. and Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, CA and UCLA AIDS Institute, Los Angeles, CA and AIDS Healthcare Foundation, Los Angeles, CA and Chen, Irvin S.Y. and Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA and UCLA AIDS Institute, Los Angeles, CA},
abstractNote = {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{sup +} T cells renders them susceptible to HIV-1 infection, resulting in poor survival of those cells. To test this possibility, highly purified CD8{sup +} T cells were genetically modified with a CD4ζ-encoding lentiviral vector and infected with HIV-1. CD8{sup +} T cells were vulnerable to HIV-1 infection upon expression of CD4ζ as evidenced by elevated levels of p24{sup Gag} in cells and culture supernatants. Concurrently, the number of CD4ζ-modified CD8{sup +} 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{sup +} T cells maintained similar levels to that of the control even under HIV-1 infection. These results suggest that protecting CD4ζ-modified CD8{sup +} T cells from HIV-1 infection is required for prolonged HIV-1-specific immune surveillance. - Highlights: • Ectopic expression of CD4ζ CAR in CD8{sup +} T cells renders them susceptible to HIV-1 infection. • Co-expression of two anti-HIV-1 shRNAs protects CD4ζ CAR-modified CD8{sup +} T cells from HIV-1 infection. • Protecting CD4ζ CAR-modified CD8{sup +} T cells from HIV-1 infection suppresses its cytopathic effect.},
doi = {10.1016/J.BBRC.2015.05.026},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 3,
volume = 463,
place = {United States},
year = {2015},
month = {7}
}