Back to Search
Start Over
Extracellular domain, hinge, and transmembrane determinants affecting surface CD4 expression of a novel anti-HIV chimeric antigen receptor (CAR) construct.
- Source :
-
PloS one [PLoS One] 2024 Aug 12; Vol. 19 (8), pp. e0293990. Date of Electronic Publication: 2024 Aug 12 (Print Publication: 2024). - Publication Year :
- 2024
-
Abstract
- Chimeric antigen receptor (CAR)-T cells have demonstrated clinical potential, but current receptors still need improvements to be successful against chronic HIV infection. In this study, we address some requirements of CAR motifs for strong surface expression of a novel anti-HIV CAR by evaluating important elements in the extracellular, hinge, and transmembrane (TM) domains. When combining a truncated CD4 extracellular domain and CD8α hinge/TM, the novel CAR did not express extracellularly but was detectable intracellularly. By shortening the CD8α hinge, CD4-CAR surface expression was partially recovered and addition of the LYC motif at the end of the CD8α TM fully recovered both intracellular and extracellular CAR expression. Mutation of LYC to TTA or TTC showed severe abrogation of CAR expression by flow cytometry and confocal microscopy. Additionally, we determined that CD4-CAR surface expression could be maximized by the removal of FQKAS motif at the junction of the extracellular domain and the hinge region. CD4-CAR surface expression also resulted in cytotoxic CAR T cell killing of HIV Env+ target cells. In this study, we identified elements that are crucial for optimal CAR surface expression, highlighting the need for structural analysis studies to establish fundamental guidelines of CAR designs.<br />Competing Interests: The authors report no conflict of interest that may arise from this research.<br /> (Copyright: © 2024 Zenere et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Subjects :
- Humans
HIV Infections immunology
Protein Domains
HEK293 Cells
CD8 Antigens immunology
CD8 Antigens metabolism
Cell Membrane metabolism
Amino Acid Motifs
HIV-1 immunology
Receptors, Chimeric Antigen immunology
Receptors, Chimeric Antigen metabolism
Receptors, Chimeric Antigen genetics
CD4 Antigens metabolism
CD4 Antigens immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 19
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 39133676
- Full Text :
- https://doi.org/10.1371/journal.pone.0293990