Abstract
Through the use of intravital imaging of the liver, we demonstrate a collaborative role for platelets with Kupffer cells (KCs) in eradicating blood-borne bacterial infection. Under basal conditions, platelets, via the platelet-adhesion receptor GPIb, formed transient 'touch-and-go' interactions with von Willebrand factor (vWF) constitutively expressed on KCs. Bacteria such as Bacillus cereus and methicillin-resistant Staphylococcus aureus (MRSA) were rapidly caught by KCs and triggered platelets to switch from 'touch-and-go' adhesion to sustained GPIIb-mediated adhesion on the KC surface to encase the bacterium. Infected GPIbα-deficient mice had more endothelial and KC damage than did their wild-type counterparts, which led to more fluid leakage, substantial polycythemia and rapid mortality. Our study identifies a previously unknown surveillance mechanism by which platelets survey macrophages that rapidly converts to a critical host response to blood-borne bacteria.
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Acknowledgements
We thank T. Chavakis (Dresden University of Technology) for Gp1ba−/− mice; K. McNagny (University of British Columbia) for GPIIb-deficient mice; J. Handelsman (Yale University) for making the green fluorescent protein–expressing B. cereus strain, and R.R. Pompano (University of Chicago) and R.F. Ismagilov (California Institute of Technology) for providing that strain; The Live Cell Imaging Facility funded by the Canadian Foundation for Innovation; and P. Colarusso for training and assistance related to microscopy. Supported by Alberta Innovates Health Solutions (C.H.Y.W., C.N.J. and P.K.), the Canadian Institutes of Health Research (P.K.) and the Canada Research Chairs Program (P.K.).
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C.H.Y.W. and C.N.J. designed and did most of the experiments; C.H.Y.W. prepared the manuscript; B.P. did all of the muscle, skin and ear experiments; N.L.C. contributed some liver-imaging experiments; C.N.J. made all of the manuscript revisions and did additional experiments; and P.K. provided overall supervision, helped design all of the experiments and prepared the manuscript.
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Supplementary Figures 1–4 (PDF 353 kb)
Supplementary Video 1
Platelet form "touch-and-go" interactions within liver sinusoids in vivo. Intravital visualization of circulating platelets within the liver microvasculature of a non-infected wild-type mouse using spinning-disk confocal fluorescence microscopy. Platelets were labeled with PE-conjugated anti-CD49b (red). Kupffer cells were labelled with Alexa Fluor 647-conjugated anti-F4/80. Elapsed time is shown at the top right. The time lapse was recorded at maximum speed and exported to video at 5 frames per second (fps). Objective: UPLANSAPO 20x/0.70. Scale bar, 50 μm. The video is representative of 2 independent experiments of minimum of 3 mice per experiment. (MOV 153 kb)
Supplementary Video 2
Platelet interactions and aggregation within the liver of a B. cereus infected mouse. Intravital visualization of circulating platelets, adhering and forming firm aggregates within the liver microvasculature during the first 10 min of GFP-expressing B. cereus infection using spinning-disk confocal fluorescence microscopy. Platelets were labeled with PE-conjugated anti-CD49b (red). Elapsed time is shown at the top right. The time lapse was recorded at maximum speed and exported to video at 20 frames per second (fps). Objective: UPLANSAPO 10x/0.40. Scale bar, 100 μm. The video is representative of 2 independent experiments of minimum of 3 mice per experiment. (MOV 474 kb)
Supplementary Video 3
Significant reduction of captured bacterium in KC-depleted animal. Intravital visualization of intravenous GFP-expressing B. cereus infection within the liver microvasculature of KC-depleted mouse using spinning-disk confocal fluorescence microscopy. Platelets were labeled with PE-conjugated anti-CD49b (red). Elapsed time is shown at the top right. The time lapse was recorded at maximum speed and exported to video at 5 frames per second (fps). Objective: UPLANSAPO 10x/0.40. Scale bar, 100 μm. The video is representative of 2 independent experiments of minimum of 3 mice per experiment. (MOV 1608 kb)
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Wong, C., Jenne, C., Petri, B. et al. Nucleation of platelets with blood-borne pathogens on Kupffer cells precedes other innate immunity and contributes to bacterial clearance. Nat Immunol 14, 785–792 (2013). https://doi.org/10.1038/ni.2631
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DOI: https://doi.org/10.1038/ni.2631
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