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.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
von Andrian, U. H. & Mempel, T. R. Homing and cellular traffic in lymph nodes. Nature Rev. Immunol. 3, 867–878 (2003)
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)
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)
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)
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)
Clark, S. L. The reticulum of lymph nodes in mice studied with the electron microscope. Am. J. Anat. 110, 217–258 (1962)
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)
Ochsenbein, A. F. et al. Protective T cell-independent antiviral antibody responses are dependent on complement. J. Exp. Med. 190, 1165–1174 (1999)
Ochsenbein, A. F. et al. Control of early viral and bacterial distribution and disease by natural antibodies. Science 286, 2156–2159 (1999)
Taylor, P. R. et al. Macrophage receptors and immune recognition. Annu. Rev. Immunol. 23, 901–944 (2005)
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)
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)
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)
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)
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)
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)
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)
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)
Vascotto, F. et al. Antigen presentation by B lymphocytes: how receptor signaling directs membrane trafficking. Curr. Opin. Immunol. 19, 93–98 (2007)
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)
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)
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)
Reif, K. et al. Balanced responsiveness to chemoattractants from adjacent zones determines B-cell position. Nature 416, 94–99 (2002)
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)
Boes, M. et al. T-cell engagement of dendritic cells rapidly rearranges MHC class II transport. Nature 418, 983–988 (2002)
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)
Casola, S. et al. B cell receptor signal strength determines B cell fate. Nature Immunol. 5, 317–327 (2004)
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)
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)
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.
The authors declare no competing financial interests.
The file contains Supplementary Figures 1-10 with Legends and Legends to Supplementary Movies 1-8. (PDF 22824 kb)
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)
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)
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)
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)
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)
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)
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)
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)
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
Differential retention of lymph-borne CD8 memory T cell subsets in the subcapsular sinus of resting and inflamed lymph nodes
Cellular & Molecular Immunology (2021)
Nature Reviews Immunology (2021)
Dual-targeting nanoparticle vaccine elicits a therapeutic antibody response against chronic hepatitis B
Nature Nanotechnology (2020)
Nature Nanotechnology (2020)
Nature Reviews Immunology (2020)