Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines
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 de...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
| Published: |
Nature Publishing Group
2015
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| Online Access: | http://hdl.handle.net/1721.1/97090 https://orcid.org/0000-0003-1833-9822 https://orcid.org/0000-0001-7604-1333 https://orcid.org/0000-0001-7192-580X https://orcid.org/0000-0003-4255-0492 |
| _version_ | 1811074903649026048 |
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| author | 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 |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering 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 |
| author_sort | Lee Szeto, Gregory |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T09:57:06Z |
| format | Article |
| id | mit-1721.1/97090 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:57:06Z |
| publishDate | 2015 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/970902022-09-26T14:45:33Z Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines 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 Massachusetts 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 Szeto, Gregory Lee Brefo, Mavis S. Irvine, Darrell J. Van Egeren, Debra S. Park, Clara Sharei, Armon Reza Heimann, Megan Langer, Robert Worku, Hermoon A. Alejandro, Brian Mao, Shirley Jensen, Klavs F. 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. Kathy and Curt Marble Cancer Research Fund (Frontier Research Programme Grant) National Cancer Institute (U.S.) (Cancer Center Support (Core) Grant P30-CA14051) National Institutes of Health (U.S.) (Ruth L. Kirschstein National Research Service Award 1F32CA180586) 2015-05-28T16:57:23Z 2015-05-28T16:57:23Z 2015-05 2015-02 Article http://purl.org/eprint/type/JournalArticle 2045-2322 http://hdl.handle.net/1721.1/97090 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 https://orcid.org/0000-0003-1833-9822 https://orcid.org/0000-0001-7604-1333 https://orcid.org/0000-0001-7192-580X https://orcid.org/0000-0003-4255-0492 en_US http://dx.doi.org/10.1038/srep10276 Scientific Reports Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group Nature |
| spellingShingle | 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 Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title | Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title_full | Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title_fullStr | Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title_full_unstemmed | Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title_short | Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines |
| title_sort | microfluidic squeezing for intracellular antigen loading in polyclonal b cells as cellular vaccines |
| url | http://hdl.handle.net/1721.1/97090 https://orcid.org/0000-0003-1833-9822 https://orcid.org/0000-0001-7604-1333 https://orcid.org/0000-0001-7192-580X https://orcid.org/0000-0003-4255-0492 |
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