Requirements for T lymphocyte migration in explanted lymph nodes.
Although the requirements for T lymphocyte homing to lymph nodes (LNs) are well studied, much less is known about the requirements for T lymphocyte locomotion within LNs. Imaging of murine T lymphocyte migration in explanted LNs using two-photon laser-scanning fluorescence microscopy provides an opportunity to systematically study these requirements. We have developed a closed system for imaging an intact LN with controlled temperature, oxygenation, and perfusion rate. Naive T lymphocyte locomotion in the deep paracortex of the LN required a perfusion rate of >13 microm/s and a partial pressure of O(2) (pO(2)) of >7.4%. Naive T lymphocyte locomotion in the subcapsular region was 38% slower and had higher turning angles and arrest coefficients than naive T lymphocytes in the deep paracortex. T lymphocyte activation decreased the requirement for pO(2), but also decreased the speed of locomotion in the deep paracortex. Although CCR7(-/-) naive T cells displayed a small reduction in locomotion, systemic treatment with pertussis toxin reduced naive T lymphocyte speed by 59%, indicating a contribution of Galpha(i)-mediated signaling, but involvement of other G protein-coupled receptors besides CCR7. Receptor knockouts or pharmacological inhibition in the adenosine, PG/lipoxygenase, lysophosphatidylcholine, and sphingosine-1-phosphate pathways did not individually alter naive T cell migration. These data implicate pO(2), tissue architecture, and G-protein coupled receptor signaling in regulation of naive T lymphocyte migration in explanted LNs.
Huang
Julie H
JH
Program in Molecular Pathogenesis, Kimmel Center for Biology and Medicine, Skirball Institute, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
Cárdenas-Navia
L Isabel
LI
Caldwell
Charles C
CC
Plumb
Troy J
TJ
Radu
Caius G
CG
Rocha
Paulo N
PN
Wilder
Tuere
T
Bromberg
Jonathan S
JS
Cronstein
Bruce N
BN
Sitkovsky
Michail
M
Dewhirst
Mark W
MW
Dustin
Michael L
ML
D000241:Adenosine / Q000378:metabolism; D058917:Adenosine A2 Receptor Antagonists; D000818:Animals; D002465:Cell Movement / Q000235:genetics; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D008198:Lymph Nodes / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D010100:Oxygen / Q000032:analysis / Q000378:metabolism*; D010313:Partial Pressure; D010477:Perfusion; D043705:Receptor, Adenosine A2A / Q000378:metabolism; D054400:Receptors, CCR7; D019707:Receptors, Chemokine / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
D058917:Adenosine A2 Receptor Antagonists; C515371:Ccr7 protein, mouse; D043705:Receptor, Adenosine A2A; D054400:Receptors, CCR7; D019707:Receptors, Chemokine; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D000241:Adenosine; D010100:Oxygen