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An intravascular immune response to Borrelia burgdorferi involves Kupffer cells and iNKT cells

Nature Immunology volume 11, pages 295–302 (2010)Cite this article

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Abstract

Here we investigate the dynamics of the hepatic intravascular immune response to a pathogen relevant to invariant natural killer T cells (iNKT cells). Immobilized Kupffer cells with highly ramified extended processes into multiple sinusoids could effectively capture blood-borne, disseminating Borrelia burgdorferi, creating a highly efficient surveillance and filtering system. After ingesting B. burgdorferi, Kupffer cells induced chemokine receptor CXCR3–dependent clustering of iNKT cells. Kupffer cells and iNKT cells formed stable contacts via the antigen-presenting molecule CD1d, which led to iNKT cell activation. An absence of iNKT cells caused B. burgdorferi to leave the blood and enter the joints more effectively. B. burgdorferi that escaped Kupffer cells entered the liver parenchyma and survived despite Ito cell responses. Kupffer cell–iNKT cell interactions induced a key intravascular immune response that diminished the dissemination of B. burgdorferi.

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Figure 1: Distribution of iNKT cells and Kupffer cells in the hepatic microvasculature.
Figure 2: Binding capacity of Kupffer cells, iNKT cells and SECs for beads and bacteria.
Figure 3: Ingestion of B. burgdorferi by Kupffer cells and Ito cells.
Figure 4: Antigen presentation by Kupffer cells and Ito cells.
Figure 5: Changes in iNKT cell activity after B. burgdorferi infection.
Figure 6: Inhibition of iNKT cell cluster formation, average crawling velocity and stationary adhesion by pertussis toxin, anti-CXCR3 and anti-CD1d.
Figure 7: Role of Kupffer cells in B. burgdorferi infection.

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Acknowledgements

We thank D.R. Littman (New York University School of Medicine) for Cxcr6gfp/gfp and Cx3cr1gfp/gfp mice; E. Allen-Vercoe (University of Guelph) for GFP-expressing and nonfluorescent E. coli; G. Chaconas and P.-O. Hardy for help with the preparation of B. burgdorferi; and B. Millen and P. Colarusso for training and assistance related to spinning-disk confocal microscopy. Supported by the Canadian Institutes of Health Research (P.K. and T.J.M. and MOP-53086 to G.C.), the Crohn's and Colitis Foundation of Canada (P.K.), the Canadian Association of Gastroenterology (W.-Y.L.), the Canada Research Chairs Program (G.C. and P.K.) and the Alberta Heritage Foundation for Medical Research (G.C., P.K. and T.J.M.).

Author information

Authors and Affiliations

  1. Department of Physiology & Pharmacology, University of Calgary, Alberta, Canada

    Woo-Yong Lee, Connie H Y Wong, Hong Zhou & Paul Kubes

  2. Department of Biochemistry & Molecular Biology, University of Calgary, Alberta, Canada

    Tara J Moriarty & George Chaconas

  3. Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada

    George Chaconas

  4. Department of Medicine, University of Virginia, Charlottesville, Virginia, USA

    Robert M Strieter

  5. Department of Molecular Cell Biology, Free University, Amsterdam, The Netherlands

    Nico van Rooijen

Authors
  1. Woo-Yong Lee
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  4. Hong Zhou
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Contributions

W.-Y.L. and T.J.M. designed and did most of the experiments and prepared the manuscript; C.H.Y.W. and H.Z. did some intravital and cell culture experiments; R.M.S. provided anti-CXCR3 serum and helped design anti-CXCR3 experiments; N.v.R. provided CLLs and intellectual input; G.C. provided supervision for the preparation of fluorescent B. burgdorferi and prepared the manuscript; and P.K. provided overall supervision, helped design all the experiments and prepared the manuscript.

Corresponding author

Correspondence to Paul Kubes.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Table 1 and Supplementary Methods (PDF 1138 kb)

Supplementary Video 1

Distribution and movement of iNKT cells in the hepatic sinusoids of Cxcr6gfp/+ mouse. Low magnification (×4) video shows the general crawling pattern of iNKT cells. Experimental conditions were as described in the Figure 1a legend and in more detailed Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 923 kb)

Supplementary Video 2

iNKT cell movement in the hepatic sinusoids of Cxcr6gfp/+ mouse. Intermediate magnification (×10) video shows the general crawling pattern of iNKT cells. Experimental conditions were as described in the Figure 1b legend and in more detailed Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 939 kb)

Supplementary Video 3

Differential behavior of iNKT cells and Kupffer cells in the hepatic sinusoids of Cxcr6gfp/+ mouse. Intermediate magnification (×10) video shows the procedure of Kupffer cell labelling and behavioral patterns of iNKT cells/Kupffer cells. Experimental conditions were as described in the Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 2258 kb)

Supplementary Video 4

Distribution and behavior of iNKT cells. Intermediate magnification (×10) video shows that iNKT cells are localized widely within sinusoids, but rarely in venules. iNKT cells that attempted to move into post-sinusoidal venule were often swept away (indicated by arrow). Experimental conditions were as described in the Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 2391 kb)

Supplementary Video 5

Filtering function of Kupffer cells in the hepatic sinusoids I. High magnification (×20) video shows the typical binding pattern of foreign molecules between Kupffer cells and beads. Experimental conditions were as described in the Figure 1c, 2a and 2b legends and in more detailed Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 1428 kb)

Supplementary Video 6

Strikingly different behavior for B. burgdorferi attached to Kupffer cells and endothelium. High magnification (×20) video shows that B. burgdorferi bound to Kupffer cells were relatively immobilized (indicated by arrow) when compared to endothelium (indicated by arrowhead; moved back and forth over 10-20 μm). Experimental conditions were as described in the Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 596 kb)

Supplementary Video 7

The cluster formed by iNKT cells 24 h after B. burgdorferi treatment I. Intermediate magnification (×10) video shows a big iNKT cluster at 24 h after B. burgdorferi treatment. Experimental conditions were as described in the Figure 5f legend and in more detailed Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 567 kb)

Supplementary Video 8

The cluster formed by iNKT cells 24 h after B. burgdorferi treatment II. High magnification (×20) video shows a big iNKT cluster formed on Kupffer cells 24 h after B. burgdorferi treatment and few spirochetes in the liver. Experimental conditions were as described in the Figure 5f and 5g legend and in more detailed Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 770 kb)

Supplementary Video 9

Filtering function of Kupffer cells in the hepatic sinusoids II. At 12 h after B. burgdorferi treatment, high magnification (×20) video shows that Kupffer cell-depleted liver could not clear the pathogens. Experimental conditions were as described in the Supplementary Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.3 fps and exported to video at 30 fps. (MOV 690 kb)

Supplementary Video 10

iNKT crawling in anti-CD1d antibody treated mouse liver at 24 h after B. burgdorferi treatment. Intermediate magnification (×10) video shows that anti-CD1d did not reduce the speed with which the iNKT cells crawl in the sinusoids, while it inhibited iNKT cluster formation. A small cluster of iNKT cells is seen in the video, but iNKT cells are still crawling on Kupffer cells. Experimental conditions were as described in the Methods. Elapsed time is shown at the top right. The time lapse was recorded at 0.2 fps and exported to video at 30 fps. (MOV 515 kb)

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Lee, WY., Moriarty, T., Wong, C. et al. An intravascular immune response to Borrelia burgdorferi involves Kupffer cells and iNKT cells. Nat Immunol 11, 295–302 (2010). https://doi.org/10.1038/ni.1855

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