Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines
Author(s)Lee Szeto, Gregory; Alejandro, Brian; Park, Clara; Frew, Kirubel; Brefo, Mavis S.; Mao, Shirley; Heimann, Megan; Van Egeren, Debra S.; Sharei, Armon Reza; Worku, Hermoon A.; Langer, Robert S; Irvine, Darrell J; Jensen, Klavs F; Szeto, Gregory; ... Show more Show less
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B-cells are promising candidate autologous antigen-presenting cells (APCs) to prime antigen-specific T-cells both in vitro and in vivo. However to date, a significant barrier to utilizing B-cells as APCs is their low capacity for non-specific antigen uptake compared to “professional” APCs such as dendritic cells. Here we utilize a microfluidic device that employs many parallel channels to pass single cells through narrow constrictions in high throughput. This microscale “cell squeezing” process creates transient pores in the plasma membrane, enabling intracellular delivery of whole proteins from the surrounding medium into B-cells via mechano-poration. We demonstrate that both resting and activated B-cells process and present antigens delivered via mechano-poration exclusively to antigen-specific CD8[superscript +]T-cells, and not CD4[superscript +]T-cells. Squeezed B-cells primed and expanded large numbers of effector CD8[superscript +]T-cells in vitro that produced effector cytokines critical to cytolytic function, including granzyme B and interferon-γ. Finally, antigen-loaded B-cells were also able to prime antigen-specific CD8[superscript +]T-cells in vivo when adoptively transferred into mice. Altogether, these data demonstrate crucial proof-of-concept for mechano-poration as an enabling technology for B-cell antigen loading, priming of antigen-specific CD8[superscript +]T-cells, and decoupling of antigen uptake from B-cell activation.
DepartmentMassachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemical Engineering; Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science; Massachusetts Institute of Technology. Department of Materials Science and Engineering; Ragon Institute of MGH, MIT and Harvard; Koch Institute for Integrative Cancer Research at MIT
Nature Publishing Group
Lee Szeto, Gregory, Debra Van Egeren, Hermoon Worku, Armon Sharei, Brian Alejandro, Clara Park, Kirubel Frew, et al. “Microfluidic Squeezing for Intracellular Antigen Loading in Polyclonal B-Cells as Cellular Vaccines.” Sci. Rep. 5 (May 22, 2015): 10276. © 2015 Macmillan Publishers Limited
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