Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published October 29, 2019 | Published + Supplemental Material
Journal Article Open

4D electron microscopy of T cell activation

Abstract

T cells can be controllably stimulated through antigen-specific or nonspecific protocols. Accompanying functional hallmarks of T cell activation can include cytoskeletal reorganization, cell size increase, and cytokine secretion. Photon-induced near-field electron microscopy (PINEM) is used to image and quantify evanescent electric fields at the surface of T cells as a function of various stimulation conditions. While PINEM signal strength scales with multiple of the biophysical changes associated with T cell functional activation, it mostly strongly correlates with antigen-engagement of the T cell receptors, even under conditions that do not lead to functional T cell activation. PINEM image analysis suggests that a stimulation-induced reorganization of T cell surface structure, especially over length scales of a few hundred nanometers, is the dominant contributor to these PINEM signal changes. These experiments reveal that PINEM can provide a sensitive label-free probe of nanoscale cellular surface structures.

Additional Information

© 2019 National Academy of Sciences. Published under the PNAS license. Edited by Pamela J. Bjorkman, California Institute of Technology, Pasadena, CA, and approved September 22, 2019 (received for review August 15, 2019). PNAS first published October 14, 2019. We thank D. Baltimore for providing MART-1-specific Jurkat cells and K. A. Woodrow for providing the Zetasizer. Funding: This work was supported by the National Cancer Institute of the National Institutes of Health (U01CA217655). The authors thank the Parker Institute for Cancer Immunotherapy (PICI) for support. A.H.C.N. is supported by a Banting Postdoctoral Fellowship from the Government of Canada. M.M.L. acknowledges the Welch Foundation for support. Author contributions: Y.L., B.-K.Y., A.H.C.N., J.K., J.T., A.H.Z., and J.R.H. designed research; Y.L., B.-K.Y., A.H.C.N., and J.K. performed research; Y.L., B.-K.Y., A.H.C.N., S.Y., and A.H.Z. contributed new reagents/analytic tools; Y.L., B.-K.Y., A.H.C.N., S.Y., J.T., M.M.L., and J.R.H. analyzed data; and Y.L., B.-K.Y., A.H.C.N., and J.R.H. wrote the paper. The authors declare no competing interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1914078116/-/DCSupplemental.

Attached Files

Published - 22014.full.pdf

Supplemental Material - pnas.1914078116.sapp.pdf

Files

22014.full.pdf
Files (2.3 MB)
Name Size Download all
md5:d1e0d15d51887730bb35e896442bde8e
1.6 MB Preview Download
md5:203befe3e70f37274c7e1797dea3c027
644.6 kB Preview Download

Additional details

Created:
August 22, 2023
Modified:
October 18, 2023