The online version of this article (doi:10.1186/s13045-017-0437-8) contains supplementary material, which is available to authorized users.
Multiple iterations of chimeric antigen receptors (CARs) have been developed, mainly focusing on intracellular signaling modules. However, the effect of non-signaling extracellular modules on the expansion and therapeutic efficacy of CARs remains largely undefined.
We generated two versions of CAR vectors, with or without a hinge domain, targeting CD19, mesothelin, PSCA, MUC1, and HER2, respectively. Then, we systematically compared the effect of the hinge domains on the growth kinetics, cytokine production, and cytotoxicity of CAR T cells in vitro and in vivo.
During in vitro culture period, the percentages and absolute numbers of T cells expressing the CARs containing a hinge domain continuously increased, mainly through the promotion of CD4+ CAR T cell expansion, regardless of the single-chain variable fragment (scFv). In vitro migration assay showed that the hinges enhanced CAR T cells migratory capacity. The T cells expressing anti-CD19 CARs with or without a hinge had similar antitumor capacities in vivo, whereas the T cells expressing anti-mesothelin CARs containing a hinge domain showed enhanced antitumor activities.
Hence, our results demonstrate that a hinge contributes to CAR T cell expansion and is capable of increasing the antitumor efficacy of some specific CAR T cells. Our results suggest potential novel strategies in CAR vector design.
Additional file 1: Figure S1. Hinge incorporation can promote the expansion of CAR T cells. Flow cytometric analysis of the percentage of 19.28z, 19-H.28z T cells, Meso.28z, Meso-H.28z T cells, PSCA.28z, PSCA-H.28z T cells, and GFP control T cells from day 6 to 15 during the in vitro culture period. The data are representative of independent experiments verified with cells from over three individual healthy human donors. (JPG 2876 kb)13045_2017_437_MOESM1_ESM.jpg
Additional file 2: Figure S2. Hinge incorporation promotes CD4+ anti-CD19 CAR T cell expansion. Flow cytometric analysis of the percentage of CD4+ and CD8+ GFP T, 19.28z T, and 19-H.28z T during the in vitro culture period. The data are representative of independent experiments verified with cells from over three individual healthy human donors. (JPG 2320 kb)13045_2017_437_MOESM2_ESM.jpg
Additional file 3: Figure S3. Percentages of CD4+ and CD8+ CAR T cells without a hinge domain both tended to be stable throughout the in vitro culture period. Flow cytometric analysis of the percentage of CD4+ and CD8+ (A) 19.28z T, (B) Meso.28z T, (C) PSCA.28z T, (D) HER2.28z T, and (E) GFP control T cells during the in vitro culture period. The data are representative of independent experiments verified with cells from over three individual healthy human donors. (JPG 924 kb)13045_2017_437_MOESM3_ESM.jpg
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- Incorporation of a hinge domain improves the expansion of chimeric antigen receptor T cells
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