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Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines
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Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines
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Journal Article
Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines
2015
Overview
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
+
T-cells and not CD4
+
T-cells. Squeezed B-cells primed and expanded large numbers of effector CD8
+
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
+
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
+
T-cells and decoupling of antigen uptake from B-cell activation.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 13/106
/ 13/21
/ 13/31
/ 64/60
/ 82/62
/ 96/21
/ Animals
/ Antigens
/ CD8-Positive T-Lymphocytes - cytology
/ CD8-Positive T-Lymphocytes - immunology
/ CD8-Positive T-Lymphocytes - metabolism
/ Humanities and Social Sciences
/ Interferon-gamma - metabolism
/ Mice
/ Microfluidic Analytical Techniques - instrumentation
/ Microfluidic Analytical Techniques - methods
/ Rodents
/ Science
