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Imaging-based Research

Janis K. Burkhardt, Ph.D.

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Associate Professor of Pathology and Laboratory Medicine
Department: Pathology and Laboratory Medicine
Graduate Group Affiliations

Contact information
Department of Pathology and Laboratory Medicine
Children's Hospital of Philadelphia
Perelman School of Medicine University of Pennsylvania
816D Abramson Research Center
3615 Civic Center Blvd.
Philadelphia, PA 19104
Office: (267) 426-5410
Fax: (267) 426-5165
Lab: (267) 426-5523
Education:
A.B. (Biology, Magna Cum Laude)
Washington University, 1983.
Ph.D. (Microbiology and Immunology)
Duke University, 1989.
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Description of Penn IMIG Expertise

Research:





Burkhardt Research PhotoResearch in our lab addresses the cell biological mechanisms through which regulated actin dynamics promote the function of T cells and antigen presenting cells.  Our long-term goal is to understand how dysregulation of these events leads to immunodeficiency, autoimmune disease, or cancer.
When a T cell is stimulated by an antigen presenting cell (APC), an actin-rich membrane domain termed the immunological synapse is formed at the cell-cell interface. This event is coordinated by several proteins including WASp, WAVE2, and HS1. Using spinning disk confocal microscopy, we analyze the effects of these actin-regulatory proteins and their upstream regulators on the spatiotemporal control of signaling events at the immunological synapse. T cell-APC engagement also leads to formation of a protein complex distal to the site of TCR engagement, termed the distal pole complex (DPC). The DPC shares many components with the uropod of migrating T cells; both are organized by members of the ERM (ezrin, radixin, moesin) protein family. We are studying the formation of the DPC and uropod, and asking how the proteins that organize these structures direct cell movement both in vitro and in vivo.  Recently, the lab has initiated studies aimed at understanding actin-dependent aspects of dendritic cell function.  Our goal is to define the role of actin-regulatory proteins in dendritic cell migration and antigen presentation. 


Lab expertise and resources:



  • Widefield and confocal fluorescence microscopy of fixed cells

  • Spinning disk microscopy of living cells

  • DIC analysis of cell dynamics and migration

  • Experience in electron microscopy including cryo-immuno-electron microscopy

  • Experience in video enhanced DIC analysis of organelle motility

  • Analysis of T cell signaling pathways

  • Analysis of cytoskeletal proteins

  • Analysis of protein trafficking in lymphocytes and dendritic cells


IMIG Collaborations:



  • Burkhardt and Argon: analysis of HS1 function in antigen presentation by dendritic cells


  • Burkhardt and Baumgart and Koretzky: use of PALM imaging and patterned surfaces to analyze microcluster dynamics during T cell activation


  • Burkhardt and Freedman: analysis of the role of actin-regulatory proteins in controlling Ca2+ signaling in lymphocytes


  • Burkhardt and Hunter: 2-photon imaging of T cell and dendritic cell movement in lymphoid organs


  • Burkhardt and Koretzky: analysis of spatiotemporal patterning in lymphocytes


  • Burkhardt, Koretzky and Hammer: Analysis of cytoskeletal anchors and adapter interactions in T cell adhesion and motility




Burkhardt Lab Members

Lab members:



Selected Publications

Shaffer MH, Dupree RS, Zhu P, Saotome I, Schmidt RF, McClatchey AI, Freedman BD, Burkhardt JK.: Ezrin and moesin function together to promote T cell activation. J Immunol 182(2): 1021-1032. Jan 2009 PMCID: PMC3491660.

Huang Y, Comiskey EO, Dupree RS, Li S, Koleske AJ, Burkhardt JK.: The c-Abl tyrosine kinase regulates actin remodeling at the immune synapse. Blood 112(1): 111-119. Jul 2008 PMCID: PMC2435682.

Burkhardt, J.K., Carrizosa, E. and Shaffer, M.H.: The actin cytoskeleton in T cell activation. Ann Rev Immunol 26, 233-259, Nov 2008.

Nolz JC, Gomez TS, Zhu P, Li S, Medeiros RB, Shimizu Y, Burkhardt, JK, Freeman BD, Billadeau, DD. : WAVE2 regulates actin cytoskeletal reorganization and CRAC-mediated calcium entry during T cell activation. Current Biology 16(1): 24-34, Jan 2006.

Gomez TS, McCarney SD, Carrizosa E, Labno CM, Comiskey EO, Nolz JC, Zhu P, Freedman BD, Clark MR, Rawlings DJ, Billadeau DD, Burkhardt, JK. : HS1 functions as an essential actin-regulatory adapter protein at the immune synapse. Immunity 24(6): 741-752, Jun 2006.

Gomez TS., Hamann MJ., McCarney S., Savoy DN., Lubking CM., Heldebrant MP., Labno CM., McKean DJ., McNiven MA., Burkhardt JK., Billadeau DD.: Dynamin 2 regulates T cell activation by controlling actin polymerization at the immunological synapse. Nature Immunology 6(3): 261-270, Mar 2005.

Labno CM., Lewis CM., You D., Leung DW., Takesono A., Kamberos N., Seth A., Finkelstein LD., Rosen MK., Schwartzberg PL., Burkhardt JK.: Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP. Current Biology 13(18): 1619-1624, Sept 2003.

Allenspach EJ, Cullinan P, Tong J, Tang Q, Tesciuba AG, Cannon JL, Takahashi SM, Morgan R, Burkhardt JK, Sperling AI.: ERM-dependent movement of CD43 defines a novel protein complex distal to the immunological synapse. Immunity 15(5): 739-750, Nov 2001.

Cannon JL, Labno CM, Bosco G, Seth A, McGavin MH, Siminovitch KA, Rosen MK, Burkhardt JK.: Wasp recruitment to the T cell:APC contact site occurs independently of Cdc42 activation. Immunity 15(2): 249-259, Aug 2001.

Billadeau, D.D. and Burkhardt, J.K.: Regulation of Cytoskeletal Dynamics at the Immune Synapse: New Stars Join the Actin Troupe. Traffic 7(11): 1451-1460. Nov 2006.

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Last updated: 02/26/2014
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