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JoVE Journal > Immunology and Infection
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サインイン又は無料トライアルを申し込む。
Summary
Abstract
Introduction
Protocol
Results
Discussion
Disclosures
Acknowledgements
Materials
References
Immunology and Infection
Tリンパ球接着分子の相互作用の研究のための接着性の下でせん断応力の測定
Published: June 29, 2016
doi:
10.3791/54203
Please note that all translations are automatically generated.
Click here for the English version.
Marianne Strazza
1
,
Inbar Azoulay-Alfaguter
1
,
Michael Peled
1
,
Adam Mor
1,2
1
Department of Medicine
,
New York University School of Medicine
2
Department of Pathology
,
New York University School of Medicine
Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
T cell samples (cell line or primary)
ATCC
TIB-152
Peripheral human T cells
CHO-ICAM-1 cells
ATCC
CRL-2093
µ-Slide VI 0.4 ibiTreat
ibidi
80606
500 ml glass bottle
Fisher
FB800500
250 ml glass bottle
Fisher
FB800250
Silicone tubing 0.8 mm
ibidi
10841
Confocal microscope with incubator chamber
Ziess
700
Any wide field fluorescent microscope
Syringe pump
New Era Pump Systems
NE-300
60 ml syringe
BD
309653
CFSE
eBioscience
65-0850
SDF-1α
R&D
350-NS-010/CF
RPMI
Lonza
12-702F/12
PBS
Lonza
17-516F
Microcentrifuge
Eppendorf
5424
D-Glucose
Sigma Aldrich
G8270
PMA
Sigma Aldrich
16561-29-8
Volocity software
Perkin Elmer
Version 6.2.1
ImageJ software
NIH
Version 1.48V
Tissue-culture treated culture dishes
Falcon
353003
Trypsin-EDTA (0.25%) Phenol Red
Gibco
25200114
Heat Inactivated FBS
Denville
FB5001-H
Penicillin/Streptomycin
Fisher
BP295950
DOWNLOAD MATERIALS LIST
References
Alon, R., Ley, K.
Cells on the run: shear-regulated integrin activation in leukocyte rolling and arrest on endothelial cells.
Curr Opin Cell Biol
.
20
(5), 525-532 (2008).
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Getting to the site of inflammation: the leukocyte adhesion cascade updated.
Nat Rev Immunol
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(9), 678-689 (2007).
Dustin, M. L., Bivona, T. G., Philips, M. R.
Membranes as messengers in T cell adhesion signaling.
Nat Immunol
.
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(4), 363-372 (2004).
Mor, A., Dustin, M. L., Philips, M. R.
Small GTPases and LFA-1 reciprocally modulate adhesion and signaling.
Immunol Rev
.
218
, 114-125 (2007).
Rose, D. M., Alon, R., Ginsberg, M. H.
Integrin modulation and signaling in leukocyte adhesion and migration.
Immunol Rev
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218
, 126-134 (2007).
Alon, R., Feigelson, S. W.
Chemokine-triggered leukocyte arrest: force-regulated bi-directional integrin activation in quantal adhesive contacts.
Curr Opin Cell Biol
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(5), 670-676 (2012).
Su, W., et al.
Rap1 and its effector RIAM are required for lymphocyte trafficking.
Blood
.
126
(25), 2695-2703 (2015).
Judokusumo, E., Tabdanov, E., Kumari, S., Dustin, M. L., Kam, L. C.
Mechanosensing in T lymphocyte activation.
Biophys J
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102
(2), L5-L7 (2012).
Zeltzer, E., et al.
Diminished adhesion of CD4+ T cells from dialysis patients to extracellular matrix and its components fibronectin and laminin.
Nephrol Dial Transplant
.
12
(12), 2618-2622 (1997).
Strazza, M., Azoulay-Alfaguter, I., Pedoeem, A., Mor, A.
Static adhesion assay for the study of integrin activation in T lymphocytes.
J Vis Exp
. (88), (2014).
Ibidi GmbH.
Shear stress and shear rates for ibidi μ-Slides – based on numerical calculations.
Application Note 11
. , (2014).
van Gijsel-Bonnello, M., et al.
Pantethine Alters Lipid Composition and Cholesterol Content of Membrane Rafts, With Down-Regulation of CXCL12-Induced T Cell Migration.
J Cell Physiol
.
230
(10), 2415-2425 (2015).
Tozeren, A., et al.
Micromanipulation of adhesion of phorbol 12-myristate-13-acetate-stimulated T lymphocytes to planar membranes containing intercellular adhesion molecule-1.
Biophys J
.
63
(1), 247-258 (1992).
Mortier, A., Van Damme, J., Proost, P.
Overview of the mechanisms regulating chemokine activity and availability.
Immunol Lett
.
145
(1-2), 2-9 (2012).
Rot, A.
In situ binding assay for studying chemokine interactions with endothelial cells.
J Immunol Methods
.
273
(1-2), 63-71 (2003).
Yang, L., et al.
ICAM-1 regulates neutrophil adhesion and transcellular migration of TNF-alpha-activated vascular endothelium under flow.
Blood
.
106
(2), 584-592 (2005).
Long, E. O.
ICAM-1: getting a grip on leukocyte adhesion.
J Immunol
.
186
(9), 5021-5023 (2011).
Tags
Adhesion
Shear Stress
T Lymphocyte
Adhesion Molecule
Integrin
LFA1
CHO-ICAM
T Cells
CFSE
Flow Chamber
Live Imaging
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Cite This Article
Strazza, M., Azoulay-Alfaguter, I., Peled, M., Mor, A. Assay of Adhesion Under Shear Stress for the Study of T Lymphocyte-Adhesion Molecule Interactions.
J. Vis. Exp.
(112), e54203, doi:10.3791/54203 (2016).
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