Scaffolds that mimic antigen-presenting cells enable ex vivo expansion of primary T cells

  • Nature Biotechnology volume 36, pages 160169 (2018)
  • doi:10.1038/nbt.4047
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Therapeutic ex vivo T-cell expansion is limited by low rates and T-cell products of limited functionality. Here we describe a system that mimics natural antigen-presenting cells (APCs) and consists of a fluid lipid bilayer supported by mesoporous silica micro-rods. The lipid bilayer presents membrane-bound cues for T-cell receptor stimulation and costimulation, while the micro-rods enable sustained release of soluble paracrine cues. Using anti-CD3, anti-CD28, and interleukin-2, we show that the APC-mimetic scaffolds (APC-ms) promote two- to tenfold greater polyclonal expansion of primary mouse and human T cells compared with commercial expansion beads (Dynabeads). The efficiency of expansion depends on the density of stimulatory cues and the amount of material in the starting culture. Following a single stimulation, APC-ms enables antigen-specific expansion of rare cytotoxic T-cell subpopulations at a greater magnitude than autologous monocyte-derived dendritic cells after 2 weeks. APC-ms support over fivefold greater expansion of restimulated CD19 CAR-T cells than Dynabeads, with similar efficacy in a xenograft lymphoma model.

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This work was supported by the National Institutes of Health (NIH) (1R01EB015498 U01 CA214369) and the Wyss Institute for Biologically Inspired Engineering at Harvard University. D.K.Y.Z. was supported by the Canadian Institutes of Health Research (CIHR-DFSA). S.T.K. was supported by an HHMI ISRF. This work was performed in part at the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Coordinated Infrastructure Network (NNCI), which is supported by the National Science Foundation under NSF award no. 1541959. CNS is part of Harvard University. We thank the National Institutes of Health (NIH) Tetramer Core Facility for the SIINFEKL/H-2K(b) biotinylated monomer, Alexa Fluor 647-labeled SIINFEKL/H-2K(b) tetramer, CLGGLLTMV/HLA-A*0201 biotinylated monomer, Alexa Fluor 647-labeled CLGGLLTMV/HLA-A*02:01 tetramer, GLCTLVAML/HLA-A*02:01 biotinylated monomer, and Alexa Fluor 647-labeled GLCTLVAML/HLA-A*0201 tetramer, N. Shastri for the B3Z cell line, and G. Freeman for the T2 cell line. We thank Unum Therapeutics for the luciferized Raji cell line and the 19BBz CAR-T cells. We also thank C. Stamoulis from Boston Children′s Hospital and the Harvard Catalyst for her help with statistical analysis; Harvard Catalyst is supported, in part, by the NIH (UL1 TR001102). Lastly, we thank R. Bates, M. Pezone, B. Schultes, L. Edwards, G. Motz, T. Barnitz, T. Hickman, K. McGinness, J. Ritz, M. Maus, T. Snyder, and W.A. Li for valuable scientific discussions.

Author information


  1. John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.

    • Alexander S Cheung
    • , David K Y Zhang
    • , Sandeep T Koshy
    •  & David J Mooney
  2. The Wyss Institute for Biologically Inspired Engineering Harvard University, Cambridge, Massachusetts, USA.

    • Alexander S Cheung
    • , David K Y Zhang
    • , Sandeep T Koshy
    •  & David J Mooney
  3. Harvard-MIT Division of Health Sciences and Technology, Cambridge, Massachusetts, USA.

    • Sandeep T Koshy


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A.S.C. and D.J.M. conceived and designed the experiments. A.S.C., D.K.Y.Z., and S.T.K. performed the experiments. A.S.C., D.K.Y.Z., and D.J.M. analyzed the data. A.S.C., D.K.Y.Z., and D.J.M. wrote the manuscript. All authors discussed the results and commented on the manuscript. The principal investigator is D.J.M.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David J Mooney.

Integrated supplementary information

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–9

  2. 2.

    Life Sciences Reporting Summary

  3. 3.

    Supplementary Table 1

    Enrichment of EBV-specific CD8+ 1 T cells following a single stimulation with APC-ms presenting either CLG or GLC.


  1. 1.

    Formation of APC-ms in culture.

    Individual rods randomlysettled and stacked to form a 3D scaffold in culture.

  2. 2.

    Infiltration of APC-ms by primary T cells.

    The large interparticle spaces formed by 3D scaffold were infiltrated by mouse T cells.