The CD28-transmembrane domain mediates chimeric antigen receptor heterodimerization with CD28
Yannick D. Muller,
Duy P. Nguyen,
Roxxana Beltran Valencia,
Frederic Van Gool,
James A Wells,
Jeffrey A. Bluestone,
Posted 19 Sep 2020
bioRxiv DOI: 10.1101/2020.09.18.296913
Posted 19 Sep 2020
Anti-CD19 chimeric antigen receptor (CD19-CAR)-engineered T cells are approved therapeutics for malignancies. The impact of the hinge (HD) and transmembrane (TMD) domains between the extracellular antigen-targeting and the intracellular signaling modalities of CARs has not been systemically studied. Here, a series of CD19-CARs differing only by their HD (CD8/CD28/IgG4) and TMD (CD8/CD28) was generated. CARs containing a CD28-TMD, but not a CD8-TMD, formed heterodimers with the endogenous CD28 in human T cells, as shown by co-immunoprecipitation and CAR-dependent proliferation to anti-CD28 stimulation. This dimerization depended on polar amino-acids in the CD28-TMD. CD28-CAR heterodimerization was more efficient in CARs containing a CD8-HD or CD28-HD as compared to an IgG4-HD. CD28-CAR heterodimers did not respond to CD80 and CD86 stimulation but led to a significant reduction of CD28 cell-surface expression. These data unveil a new property of the CD28-TMD and suggest that TMDs can modulate CAR T-cell activities by engaging endogenous partners. ![Figure]</img> ### Competing Interest Statement Conflict of interest. A provisional patent has been submitted. JAB and QT are co-founders of Sonoma Biotherapeutics. AM and TR are co-founders of Arsenal Biosciences. AM is also a co-founder of Spotlight Therapeutics.. JAB and AM have served as advisors to Juno Therapeutics. AM was a member of the scientific advisory board at PACT Pharma and was an advisor to Trizell. QT, JAB, and AM have received sponsored research support from Juno Therapeutics. AM has received research support from Epinomics, Sanofi, GlaxoSmithKline, and gifts from Gilead and Anthem. JAW is co-Founder of Soteria Biotherapeutics developing small molecule switchable biologics, on the SAB of Spotlight, and recipient of sponsored research from Bristol Myers Squibb. JE is an advisor for Mnemo Therapeutics and Cytovia and received research support from Cytovia. The authors declare no other relevant conflict of interest. * CAR : Chimeric Antigen Receptor HD : Hinge domain ICD : Intracellular signaling domain scFv : Single chain variable fragment TMD : Transmembrane domain, : pending:yes
- Downloaded 1,208 times
- Download rankings, all-time:
- Site-wide: 24,674
- In bioengineering: 443
- Year to date:
- Site-wide: 44,135
- Since beginning of last month:
- Site-wide: 26,260
Downloads over time
Distribution of downloads per paper, site-wide
- 27 Nov 2020: The website and API now include results pulled from medRxiv as well as bioRxiv.
- 18 Dec 2019: We're pleased to announce PanLingua, a new tool that enables you to search for machine-translated bioRxiv preprints using more than 100 different languages.
- 21 May 2019: PLOS Biology has published a community page about Rxivist.org and its design.
- 10 May 2019: The paper analyzing the Rxivist dataset has been published at eLife.
- 1 Mar 2019: We now have summary statistics about bioRxiv downloads and submissions.
- 8 Feb 2019: Data from Altmetric is now available on the Rxivist details page for every preprint. Look for the "donut" under the download metrics.
- 30 Jan 2019: preLights has featured the Rxivist preprint and written about our findings.
- 22 Jan 2019: Nature just published an article about Rxivist and our data.
- 13 Jan 2019: The Rxivist preprint is live!