Improving regulatory T cell production through mechanosensing
Lingting Shi
Department of Biomedical Engineering, Columbia University, New York, New York, USA
Search for more papers by this authorJee Yoon Lim
Department of Biological Sciences, Columbia University, New York, New York, USA
Search for more papers by this authorCorresponding Author
Lance C. Kam
Department of Biomedical Engineering, Columbia University, New York, New York, USA
Correspondence
Lance C. Kam, 351 Engineering Terrace, Mail Code 8904, New York, NY 10027, USA.
Email: [email protected]
Search for more papers by this authorLingting Shi
Department of Biomedical Engineering, Columbia University, New York, New York, USA
Search for more papers by this authorJee Yoon Lim
Department of Biological Sciences, Columbia University, New York, New York, USA
Search for more papers by this authorCorresponding Author
Lance C. Kam
Department of Biomedical Engineering, Columbia University, New York, New York, USA
Correspondence
Lance C. Kam, 351 Engineering Terrace, Mail Code 8904, New York, NY 10027, USA.
Email: [email protected]
Search for more papers by this authorAbstract
Induced Tregs (iTregs) have great promise in adoptive immunotherapy for treatment of autoimmune diseases. This report investigates the impacts of substrate stiffness on human Treg induction, providing a powerful yet simple approach to improving production of these cells. Conventional CD4+ human T cells were activated on materials of different elastic modulus and cultured under suppressive conditions. Enhanced Treg induction was observed on softer materials as early as 3 days following activation and persisted for multiple weeks. Substrate stiffness also affected epigenetic modification of Treg specific genes and Treg suppressive capacity. Tregs induced on substrates of an optimal stiffness balance quantity and suppressive quality.
CONFLICT OF INTEREST STATEMENT
The authors declare no conflict of interest.
Open Research
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Supporting Information
| Filename | Description |
|---|---|
| jbma37702-sup-0001-Figures.docxWord 2007 document , 7.6 MB | Figure S1. Contour plots of SSC-A and FSC-A for PDMS substrate to assess T cell activation. Gates were set on the cell population outside of the main population of unstimulated sample. Figure S2. Stiffness dependent Treg induction. (A) Median Fluorescence Intensity (MFI) of FOXP3+ cells, activated on PDMS of the indicated stiffness, on D3 and D6. (B & C) Comparison between 870 kPa PDMS substrate and rigid controls based on (B) Percent (C) Total count of FOXP3+ cells. Data are mean ± SEM., n = 5; * p < .05, ** p < .01, *** p < .001, **** p < .0001, two-way ANOVA with Tukey multiple comparisons. Figure S3. Substrate stiffness modulates T cell suppressive function. (A) Representative CFSE data comparing 2600 kPa with controls. (B) Percent suppression with 1:1, 1:2, and 1:4 ratios of Treg and Tconvs comparing 2600 kPa to plate-bound control and Dynabeads. Data are mean, whiskers are min and max, n = 3 individual experiments; One-way ANOVA; * p < .05, ** p < .01. Figure S4. Differential methylation analysis on Treg epigenetic profiles. (A) Volcano plot of differentially methylation analysis comparing 2,600kPa and 2600 kPa PDMS treated with Y-27632. (B) Volcano plot of differentially methylation analysis comparing Tconvs to plate-bound control. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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