TY - JOUR
T1 - ROCK regulates the intermittent mode of interstitial T cell migration in inflamed lungs
AU - Mrass, Paulus
AU - Oruganti, Sreenivasa Rao
AU - Fricke, G. Matthew
AU - Tafoya, Justyna
AU - Byrum, Janie R.
AU - Yang, Lihua
AU - Hamilton, Samantha L.
AU - Miller, Mark J.
AU - Moses, Melanie E.
AU - Cannon, Judy L.
N1 - Funding Information:
This work was supported by funding from the following: DARPA P-1070-113237 (MEM), NIH 1R01AI097202 (JLC), the Spatiotemporal Modeling Center (P50 GM085273), the Center for Evolution and Theoretical Immunology 5P20GM103452 (J.L.C.), a James S. McDonnell Foundation grant for the study of Complex Systems (M.E.M.), and by R01 AI077600, U01 AI095550 and the Mucosal Immunity Studies Two-Photon Imaging Web Resources and Training Fund MIST U01 (M.J.M.). Thanks to the UNM Cancer Center Fluorescence Microscopy Facility (P30-CA118100) as well as the BRaIN Imaging Center (P30GM103400) for help with two-photon microscopy. We would also like to thank Dr Ichiko Kinjo and Dr Eliseo Castillo for critical reading of the manuscript.
Funding Information:
Mice and reagents. All experiments were performed with C57BL/6 (Jackson Laboratories) and B6.Ubiquitin-GFP mice60 (Jackson Laboratories). Typically, female animals were used between 8–20 weeks of age. Breeding and maintenance of animals used in this research conform to the principles outlined by the Animal Welfare Act of the National Institutes of Health. The protocol for animal work was approved by the IACUC at the University of New Mexico (protocol # 15-200328-HSC). All efforts were made to minimize suffering with use of ketamine and xylazine when appropriate. Euthanasia was performed by isofluorane overdose. Anti-CD3 (145-2C11, BioXCell) and anti-CD28 (PV-1, BioXCell) antibodies were used for T cell priming. For intravascular T cell staining, anti-CD3-PerCPCy5.5 (145-2C11, eBioscience; flow experiments) and anti-CD3-Alexa 647 (17A2, BDPharmingen; confocal imaging) were used. For intranasal staining, an anti-CD45-PE antibody (30-F11; Molecular Probes) was used. In inhibitor experiments, the ROCK inhibitor Y-27632 (Calbiochem) was used at a final concentration of 20 µM. PTX (Sigma) was used at a final concentration of 100 ng ml−1. Anti-CD8-APC-Cy7 (53-6.7, eBioscience) was used for staining single cell suspensions. For western blotting, anti-p-ERK (Thr202/Tyr204, Cell Signaling Technology #9101, used at 1:1000), anti-ERK (137F5, Cell Signaling Technology, used at 1:1000) and anti-actin antibodies (AC-15, Sigma, used at 1:5000) were used.
Publisher Copyright:
© 2017 The Author(s).
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Effector T cell migration through tissues can enable control of infection or mediate inflammatory damage. Nevertheless, the molecular mechanisms that regulate migration of effector T cells within the interstitial space of inflamed lungs are incompletely understood. Here, we show T cell migration in a mouse model of acute lung injury with two-photon imaging of intact lung tissue. Computational analysis indicates that T cells migrate with an intermittent mode, switching between confined and almost straight migration, guided by lung-associated vasculature. Rho-associated protein kinase (ROCK) is required for both high-speed migration and straight motion. By contrast, inhibition of Gαi signaling with pertussis toxin affects speed but not the intermittent migration of lung-infiltrating T cells. Computational modeling shows that an intermittent migration pattern balances both search area and the duration of contacts between T cells and target cells. These data identify that ROCK-dependent intermittent T cell migration regulates tissue-sampling during acute lung injury.
AB - Effector T cell migration through tissues can enable control of infection or mediate inflammatory damage. Nevertheless, the molecular mechanisms that regulate migration of effector T cells within the interstitial space of inflamed lungs are incompletely understood. Here, we show T cell migration in a mouse model of acute lung injury with two-photon imaging of intact lung tissue. Computational analysis indicates that T cells migrate with an intermittent mode, switching between confined and almost straight migration, guided by lung-associated vasculature. Rho-associated protein kinase (ROCK) is required for both high-speed migration and straight motion. By contrast, inhibition of Gαi signaling with pertussis toxin affects speed but not the intermittent migration of lung-infiltrating T cells. Computational modeling shows that an intermittent migration pattern balances both search area and the duration of contacts between T cells and target cells. These data identify that ROCK-dependent intermittent T cell migration regulates tissue-sampling during acute lung injury.
UR - http://www.scopus.com/inward/record.url?scp=85031937096&partnerID=8YFLogxK
U2 - 10.1038/s41467-017-01032-2
DO - 10.1038/s41467-017-01032-2
M3 - Article
C2 - 29044117
AN - SCOPUS:85031937096
VL - 8
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 1010
ER -