Dual CD4-based CAR T cells with distinct costimulatory domains mitigate HIV pathogenesis in vivo

Nat Med. 2020 Nov;26(11):1776-1787. doi: 10.1038/s41591-020-1039-5. Epub 2020 Aug 31.


An effective strategy to cure HIV will likely require a potent and sustained antiviral T cell response. Here we explored the utility of chimeric antigen receptor (CAR) T cells, expressing the CD4 ectodomain to confer specificity for the HIV envelope, to mitigate HIV-induced pathogenesis in bone marrow, liver, thymus (BLT) humanized mice. CAR T cells expressing the 4-1BB/CD3-ζ endodomain were insufficient to prevent viral rebound and CD4+ T cell loss after the discontinuation of antiretroviral therapy. Through iterative improvements to the CAR T cell product, we developed Dual-CAR T cells that simultaneously expressed both 4-1BB/CD3-ζ and CD28/CD3-ζ endodomains. Dual-CAR T cells exhibited expansion kinetics that exceeded 4-1BB-, CD28- and third-generation costimulated CAR T cells, elicited effector functions equivalent to CD28-costimulated CAR T cells and prevented HIV-induced CD4+ T cell loss despite persistent viremia. Moreover, when Dual-CAR T cells were protected from HIV infection through expression of the C34-CXCR4 fusion inhibitor, these cells significantly reduced acute-phase viremia, as well as accelerated HIV suppression in the presence of antiretroviral therapy and reduced tissue viral burden. Collectively, these studies demonstrate the enhanced therapeutic potency of a novel Dual-CAR T cell product with the potential to effectively treat HIV infection.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Humanized / immunology
  • Antibodies, Monoclonal, Humanized / pharmacology
  • Bone Marrow / immunology
  • Bone Marrow / virology
  • CD3 Complex / antagonists & inhibitors
  • CD4 Antigens / administration & dosage
  • CD4 Antigens / immunology*
  • Gene Expression Regulation / immunology
  • HIV Envelope Protein gp41 / antagonists & inhibitors
  • HIV Envelope Protein gp41 / immunology
  • HIV Infections / immunology
  • HIV Infections / pathology
  • HIV Infections / therapy*
  • HIV Infections / virology
  • HIV-1 / immunology
  • HIV-1 / pathogenicity
  • Humans
  • Immunotherapy, Adoptive*
  • Liver / immunology
  • Liver / virology
  • Mice
  • Peptide Fragments / antagonists & inhibitors
  • Peptide Fragments / immunology
  • Protein Domains / immunology
  • Receptors, CXCR4 / antagonists & inhibitors
  • Receptors, CXCR4 / immunology
  • Receptors, Chimeric Antigen / administration & dosage
  • Receptors, Chimeric Antigen / immunology*
  • T-Lymphocytes / immunology
  • Thymus Gland / immunology
  • Thymus Gland / virology
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / antagonists & inhibitors


  • Antibodies, Monoclonal, Humanized
  • CD3 Complex
  • CD3 antigen, zeta chain
  • CD4 Antigens
  • CXCR4 protein, mouse
  • HIV Envelope Protein gp41
  • Peptide Fragments
  • Receptors, CXCR4
  • Receptors, Chimeric Antigen
  • Tumor Necrosis Factor Receptor Superfamily, Member 9
  • peptide C34