Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and present them to antiviral B cells

Nature volume 450, pages 110–114 (2007)Cite this article

Abstract

Lymph nodes prevent the systemic dissemination of pathogens such as viruses that infect peripheral tissues after penetrating the body’s surface barriers. They are also the staging ground of adaptive immune responses to pathogen-derived antigens1,2. It is unclear how virus particles are cleared from afferent lymph and presented to cognate B cells to induce antibody responses. Here we identify a population of CD11b+CD169+MHCII+ macrophages on the floor of the subcapsular sinus (SCS) and in the medulla of lymph nodes that capture viral particles within minutes after subcutaneous injection. Macrophages in the SCS translocated surface-bound viral particles across the SCS floor and presented them to migrating B cells in the underlying follicles. Selective depletion of these macrophages compromised local viral retention, exacerbated viraemia of the host, and impaired local B-cell activation. These findings indicate that CD169+ macrophages have a dual physiological function. They act as innate ‘flypaper’ by preventing the systemic spread of lymph-borne pathogens and as critical gatekeepers at the lymph–tissue interface that facilitate the recognition of particulate antigens by B cells and initiate humoral immune responses.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Capture of lymph-borne VSV by SCS macrophages.
Figure 2: Macrophage-mediated transfer of lymph-borne VSV across the SCS floor alters virus-specific B cell behaviour.
Figure 3: SCS macrophages are required for early activation of VSV-specific B cells in lymph nodes.

References

  1. von Andrian, U. H. & Mempel, T. R. Homing and cellular traffic in lymph nodes. Nature Rev. Immunol. 3, 867–878 (2003)

    Article  CAS  Google Scholar 

  2. Karrer, U. et al. On the key role of secondary lymphoid organs in antiviral immune responses studied in alymphoplastic (aly/aly) and spleenless (Hox11-/-) mutant mice. J. Exp. Med. 185, 2157–2170 (1997)

    Article  CAS  Google Scholar 

  3. Mead, D. G., Ramberg, F. B. & Mare, C. J. Laboratory vector competence of black flies (Diptera: Simuliidae) for the Indiana serotype of vesicular stomatitis virus. Ann. NY Acad. Sci. 916, 437–443 (2000)

    Article  ADS  CAS  Google Scholar 

  4. Bachmann, M. F., Hengartner, H. & Zinkernagel, R. M. T helper cell-independent neutralizing B cell response against vesicular stomatitis virus: role of antigen patterns in B cell induction? Eur. J. Immunol. 25, 3445–3451 (1995)

    Article  CAS  Google Scholar 

  5. Mempel, T. R., Henrickson, S. E. & von Andrian, U. H. T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Nature 427, 154–159 (2004)

    Article  ADS  CAS  Google Scholar 

  6. Clark, S. L. The reticulum of lymph nodes in mice studied with the electron microscope. Am. J. Anat. 110, 217–258 (1962)

    Article  Google Scholar 

  7. Farr, A. G., Cho, Y. & De Bruyn, P. P. The structure of the sinus wall of the lymph node relative to its endocytic properties and transmural cell passage. Am. J. Anat. 157, 265–284 (1980)

    Article  CAS  Google Scholar 

  8. Ochsenbein, A. F. et al. Protective T cell-independent antiviral antibody responses are dependent on complement. J. Exp. Med. 190, 1165–1174 (1999)

    Article  CAS  Google Scholar 

  9. Ochsenbein, A. F. et al. Control of early viral and bacterial distribution and disease by natural antibodies. Science 286, 2156–2159 (1999)

    Article  CAS  Google Scholar 

  10. Taylor, P. R. et al. Macrophage receptors and immune recognition. Annu. Rev. Immunol. 23, 901–944 (2005)

    Article  CAS  Google Scholar 

  11. Delemarre, F. G., Kors, N., Kraal, G. & van Rooijen, N. Repopulation of macrophages in popliteal lymph nodes of mice after liposome-mediated depletion. J. Leukoc. Biol. 47, 251–257 (1990)

    Article  CAS  Google Scholar 

  12. Ludewig, B. et al. Induction of optimal anti-viral neutralizing B cell responses by dendritic cells requires transport and release of virus particles in secondary lymphoid organs. Eur. J. Immunol. 30, 185–196 (2000)

    Article  CAS  Google Scholar 

  13. Qi, H., Egen, J. G., Huang, A. Y. & Germain, R. N. Extrafollicular activation of lymph node B cells by antigen-bearing dendritic cells. Science 312, 1672–1676 (2006)

    Article  ADS  CAS  Google Scholar 

  14. Roost, H. P., Haag, A., Burkhart, C., Zinkernagel, R. M. & Hengartner, H. Mapping of the dominant neutralizing antigenic site of a virus using infected cells. J. Immunol. Methods 189, 233–242 (1996)

    Article  CAS  Google Scholar 

  15. Hangartner, L. et al. Antiviral immune responses in gene-targeted mice expressing the immunoglobulin heavy chain of virus-neutralizing antibodies. Proc. Natl Acad. Sci. USA 100, 12883–12888 (2003)

    Article  ADS  CAS  Google Scholar 

  16. Rossbacher, J. & Shlomchik, M. J. The B cell receptor itself can activate complement to provide the complement receptor 1/2 ligand required to enhance B cell immune responses in vivo . J. Exp. Med. 198, 591–602 (2003)

    Article  CAS  Google Scholar 

  17. Okada, T. et al. Antigen-engaged B cells undergo chemotaxis toward the T zone and form motile conjugates with helper T cells. PLoS Biol. 3, e150 (2005)

    Article  Google Scholar 

  18. Dang, L. H. & Rock, K. L. Stimulation of B lymphocytes through surface Ig receptors induces LFA-1 and ICAM-1-dependent adhesion. J. Immunol. 146, 3273–3279 (1991)

    CAS  PubMed  Google Scholar 

  19. Vascotto, F. et al. Antigen presentation by B lymphocytes: how receptor signaling directs membrane trafficking. Curr. Opin. Immunol. 19, 93–98 (2007)

    Article  CAS  Google Scholar 

  20. Pape, K. A., Catron, D. M., Itano, A. A. & Jenkins, M. K. The humoral immune response is initiated in lymph nodes by B cells that acquire soluble antigen directly in the follicles. Immunity 26, 491–502 (2007)

    Article  CAS  Google Scholar 

  21. Barchet, W. et al. Virus-induced interferon α production by a dendritic cell subset in the absence of feedback signaling in vivo . J. Exp. Med. 195, 507–516 (2002)

    Article  CAS  Google Scholar 

  22. Shiow, L. R. et al. CD69 acts downstream of interferon-α/β to inhibit S1P1 and lymphocyte egress from lymphoid organs. Nature 440, 540–544 (2006)

    Article  ADS  CAS  Google Scholar 

  23. Reif, K. et al. Balanced responsiveness to chemoattractants from adjacent zones determines B-cell position. Nature 416, 94–99 (2002)

    Article  ADS  Google Scholar 

  24. Wessels, M. R. et al. Studies of group B streptococcal infection in mice deficient in complement component C3 or C4 demonstrate an essential role for complement in both innate and acquired immunity. Proc. Natl Acad. Sci. USA 92, 11490–11494 (1995)

    Article  ADS  CAS  Google Scholar 

  25. Boes, M. et al. T-cell engagement of dendritic cells rapidly rearranges MHC class II transport. Nature 418, 983–988 (2002)

    Article  ADS  CAS  Google Scholar 

  26. Wright, D. E. et al. Cyclophosphamide/granulocyte colony-stimulating factor causes selective mobilization of bone marrow hematopoietic stem cells into the blood after M phase of the cell cycle. Blood 97, 2278–2285 (2001)

    Article  CAS  Google Scholar 

  27. Casola, S. et al. B cell receptor signal strength determines B cell fate. Nature Immunol. 5, 317–327 (2004)

    Article  CAS  Google Scholar 

  28. Whelan, S. P., Ball, L. A., Barr, J. N. & Wertz, G. T. Efficient recovery of infectious vesicular stomatitis virus entirely from cDNA clones. Proc. Natl Acad. Sci. USA 92, 8388–8392 (1995)

    Article  ADS  CAS  Google Scholar 

  29. Leopold, P. L. et al. Fluorescent virions: dynamic tracking of the pathway of adenoviral gene transfer vectors in living cells. Hum. Gene Ther. 9, 367–378 (1998)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G. Cheng, M. Flynn and D. Baumjohann for technical support; R. M. Zinkernagel and H. Hengartner for providing VI10YEN mice; A. Wagers for providing Act(EGFP) mice; M. Ericsson and E. Benecchi for expert support in electron microscopy studies; S. Behnke for immunohistochemistry; and D. Cureton for help and advice with VSV preparations. This work was supported by grants from the NIH-NIAID (to U.H.v.A.), a Pilot and Feasibility Grant from the Harvard Skin Disease Research Center (to T.J. and U.H.v.A.), a stipend from the Swiss National Science Foundation (to T.J.) and a NIH T32 Training Grant in Hematology (to E.A.M.).

Author Contributions T.J. and U.H.v.A. designed the study; T.J., E.A.M., M.I., S.M. and P.A.L. performed experiments; T.J., E.A.M., M.I. and S.M. collected and analysed data; M.B., K.F., N.C.D.P., D.M.S., N.v.R. and S.P.W. provided reagents and mice; T.J., E.A.M., M.I. and U.H.v.A. wrote the manuscript; S.M., K.F., S.E.H., T.M. and S.P.W. gave technical support and conceptual advice.

Author information

Authors and Affiliations

  1. Immune Disease Institute and Department of Pathology, Harvard Medical School,

    Tobias Junt, E. Ashley Moseman, Matteo Iannacone, Steffen Massberg, Sarah E. Henrickson, Thorsten R. Mempel & Ulrich H. von Andrian

  2. Department of Dermatology, Brigham and Women's Hospital, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA,

    Marianne Boes

  3. Immunopathogenesis of Liver Infections Unit, San Raffaele Scientific Institute, Via Olgettina 58, 20132 Milan, Italy ,

    Matteo Iannacone

  4. Institute of Experimental Immunology, Zurich University Hospital, Schmelzbergstrasse 12, CH-8091 Zurich, Switzerland ,

    Philipp A. Lang

  5. Novartis Institute of Tropical Diseases, 10 Biopolis Road, 138670 Singapore ,

    Katja Fink

  6. Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, Washington 98195, USA,

    Dmitry M. Shayakhmetov & Nelson C. Di Paolo

  7. Department of Molecular Cell Biology, Vrije Universiteit, VUMC, Faculty of Medicine, Van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands,

    Nico van Rooijen

  8. Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, Massachusetts 02115, USA,

    Sean P. Whelan

Authors
  1. Tobias Junt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Ashley Moseman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Matteo Iannacone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Steffen Massberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Philipp A. Lang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marianne Boes
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Katja Fink
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sarah E. Henrickson
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dmitry M. Shayakhmetov
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Nelson C. Di Paolo
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Nico van Rooijen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Thorsten R. Mempel
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Sean P. Whelan
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Ulrich H. von Andrian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ulrich H. von Andrian.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-10 with Legends and Legends to Supplementary Movies 1-8. (PDF 22824 kb)

Supplementary Movie 1

This file contains Supplementary Movie 1 which shows how, following footpad injection, VSV particles rapidly accumulate in a patchy pattern on the floor of the SCS of the draining popliteal LN. 20 µg AlexaFluor-488 labeled UV-irradiated VSV (green) were injected in 20 µl PBS into the hind footpad of an anesthetized mouse while the SCS and superficial parenchyma of the draining popliteal LN was recorded by MP-IVM. Z-stacks of 11 optical sections (z-increment 4 µm) were acquired every 15 seconds over 30 minutes at a pixel density of 256x256. This movie shows a maximum intensity projection, which displays at each pixel only the brightest value encountered along an axial viewing ray in each color channel. The second harmonic signals from collagen fibers in the LN capsule and the adventitia of the adjacent saphenous vein are shown in blue. Fluorescent VSV accumulated rapidly on the floor of the SCS, but did not enter the LN parenchyma. The counter in the upper left corner shows minutes and seconds after VSV injection. Scale bar corresponds to 100 µm. (MOV 2970 kb)

Supplementary Movie 2

This file contains Supplementary Movie 2 which shows entry of VSV particles into the SCS of a popliteal LN in a C57BL/6→Act(EGFP) chimeric mouse. AlexaFluor-568 labeled UV-irradiated VSV particles (red) were injected into a hind footpad of an anesthetized mouse and recorded by MP-IVM as described in the legend to Suppl. Movie 1. The field of view chosen at a lateral aspect of the draining popliteal LN, with the parenchyma on the left, the SCS running vertically across the center and extranodal connective tissue on the right. Non-hematopoietic EGFP+ cells in C57BL/6→Act(EGFP) chimeras, mostly sinus-lining lymphatic endothelial cells, are shown in green. Second harmonic signals from collagen fibers in the LN capsule and extranodal tissue are visible in blue. The counter in the upper left corner shows minutes and seconds after VSV injection. The scale bar corresponds to 50 µm. (MOV 3826 kb)

Supplementary Movie 3

This file contains Supplementary Movie 3 which shows sustained and selective retention of fluorescent VSV on the floor of the SCS. This recording was taken 3h after footpad injection of AlexaFluor-568 labeled UV-VSV (red) in the same preparation as shown in Suppl. Movie 2. The movie shows a projection through a z-stack of 15 optical sections (z-increment of 2 µm) at a pixel density of 512x512. The image is rotated ±20º around the y-axis. Note that the red viral deposits do not overlap with the green EGFP signal associated with non-hematopoietic cells. The scale bar corresponds to 50 µm. (MOV 2479 kb)

Supplementary Movie 4

This file contains Supplementary Movie 4 which shows that In the absence of virus VI10YENand polyclonal B cells have equivalent distributions and velocities throughout the popliteal LN. Purified, CMTMR labeled VI10YEN B cells (red) and CMAC labeled polyclonal B cells (blue) were transferred i.v. at 5-6x106 cells each into C57BL/6 recipients, 18 h prior to recording. A B cell follicle in the popliteal LN was continuously recorded by MP-IVM over 119 frames, at a frame-to-frame interval of 15 seconds, with Z stack of 11 sections (z increment of 4 mm) and a pixel density of 256x256. The counter in the upper left corner indicates minutes and seconds. Scale bar, 50 mm. (MOV 3825 kb)

Supplementary Movie 5

This file contains Supplementary Movie 5 which shows VI10YEN and wildtype B cells have equivalent distributions and velocities after introduction of VSV-NJ. Purified, CMTMR labeled VI10YEN B cells (red) and CMAC labeled polyclonal B cells (blue) were transferred i.v. at 5-6x106 cells each into C57BL/6 recipients, 18 h prior to recording. A B cell follicle in the popliteal LN was continuously recorded by MP-IVM over 119 frames, at a frame-to-frame interval of 15 seconds, with Z stack of 11 sections (z increment of 4 mm) and a pixel density of 256x256. White numbers in upper left corner indicate minutes and seconds, scale bar corresponds to 50 mm. The recipient mouse received a footpad injection of 20µg AlexaFluor-488 labeled VSV-NJ (green) 5 minutes prior to the start of the recording. This 3D time-lapse recording was taken from 5-30 min after VSV-NJ injection. The counter in the upper left corner indicates minutes and seconds. Scale bar, 50 µm. (MOV 4021 kb)

Supplementary Movie 6

This file contains Supplementary Movie 6 which shows VI10YEN B cells rapidly accumulate below and within the SCS floor after introduction of VSV-IND. The experimental protocol and image acquisition was as in Suppl. Movie 5, except the recipient mouse received a footpad injection of 20µg AlexaFluor-488 labeled VSV-IND (green) 5 minutes prior to the start of the recording. This 3D time-lapse recording was taken from 5-30 min after VSV-IND injection. The counter in the upper left corner indicates minutes and seconds. Scale bar, 50 µm. (MOV 3832 kb)

Supplementary Movie 7

This file contains Supplementary Movie 7 which shows VSV-IND induces VI10YEN B cell arrest but not accumulation at the SCS in CLL-treated popliteal LNs. The experimental protocol and image acquisition was as in Suppl. Movie 5, but in mice that received a footpad injection of CLL 7 d prior to imaging. The recipient mouse received a footpad injection of 20µg Alexa-488 labeled VSV-IND (green) 5 minutes prior to the start of the recording. This 3D time-lapse recording was taken from 5-30 min after VSV-NJ injection. The counter in the upper right corner indicates minutes and seconds. Scale bar, 50 µm. Note that fluorescent VSV appears in the flowing lymph, but is not retained at the SCS floor. Only the rare VI10YEN cells in the SCS lumen have access to the lymph-borne virus as evidenced by acquisition of green fluorescence. (MOV 3098 kb)

Supplementary Movie 8

This file contains Supplementary Movie 8 which shows VSV-IND is internalized by VI10YEN B cells and colocalizes with MHC-class II in endocytic compartments. 18 h following intravenous transfer of 5-6x106 purified VI10YENxMHCII(EGFP) B cells (green), 20 mg AlexaFluor-568 labeled UV-VSV-IND (red) was injected s.c. into the hind footpad of recipient mice. LNs were removed 30 minutes after injection of virus. 30mm cryosections were analyzed by confocal microscopy. Movie is Z stack of 25 sections (z increment of 0.5 mm) at a pixel density of 512x512. (MOV 979 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Junt, T., Moseman, E., Iannacone, M. et al. Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and present them to antiviral B cells. Nature 450, 110–114 (2007). https://doi.org/10.1038/nature06287

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature06287

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing