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ABSTRACT The mechanism of B cell–antigen encounter in lymphoid tissues is incompletely understood. It is also unclear how immune complexes are transported to follicular dendritic cells.
Here, using real-time two-photon microscopy we noted rapid delivery of immune complexes through the lymph to macrophages in the lymph node subcapsular sinus. B cells captured immune
complexes by a complement receptor–dependent mechanism from macrophage processes that penetrated the follicle and transported the complexes to follicular dendritic cells. Furthermore,
cognate B cells captured antigen-containing immune complexes from macrophage processes and migrated to the T zone. Our findings identify macrophages lining the subcapsular sinus as an
important site of B cell encounter with immune complexes and show that intrafollicular B cell migration facilitates the transport of immune complexes as well as encounters with cognate
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support SIMILAR CONTENT BEING VIEWED BY OTHERS DYNAMIC ENCOUNTERS WITH RED BLOOD CELLS TRIGGER SPLENIC MARGINAL ZONE B CELL RETENTION AND FUNCTION Article Open access 04 December 2023
SPATIAL MAPPING OF INNATE LYMPHOID CELLS IN HUMAN LYMPHOID TISSUES AND LYMPHOMA AT SINGLE-CELL RESOLUTION Article Open access 15 May 2025 LONG-TERM RETENTION OF ANTIGENS IN GERMINAL CENTERS
IS CONTROLLED BY THE SPATIAL ORGANIZATION OF THE FOLLICULAR DENDRITIC CELL NETWORK Article 13 July 2023 REFERENCES * Nossal, G.J., Abbot, A., Mitchell, J. & Lummus, Z. Antigens in
immunity. XV. Ultrastructural features of antigen capture in primary and secondary lymphoid follicles. _J. Exp. Med._ 127, 277–290 (1968). Article CAS Google Scholar * Tew, J.G., Phipps,
R.P. & Mandel, T.E. The maintenance and regulation of the humoral immune response: persisting antigen and the role of follicular antigen-binding dendritic cells as accessory cells.
_Immunol. Rev._ 53, 175–201 (1980). Article CAS Google Scholar * Szakal, A.K., Kosco, M.H. & Tew, J.G. Microanatomy of lymphoid tissue during humoral immune responses: structure
function relationships. _Annu. Rev. Immunol._ 7, 91–109 (1989). Article CAS Google Scholar * Nossal, G.J., Ada, G.L., Austin, C.M. & Pye, J. Antigens in immunity. 8. Localization of
125-I-labelled antigens in the secondary response. _Immunology_ 9, 349–357 (1965). CAS PubMed PubMed Central Google Scholar * Fossum, S. The architecture of rat lymph nodes. IV.
Distribution of ferritin and colloidal carbon in the draining lymph nodes after foot-pad injection. _Scand. J. Immunol._ 12, 433–441 (1980). Article CAS Google Scholar * Carroll, M.C. The
role of complement and complement receptors in induction and regulation of immunity. _Annu. Rev. Immunol._ 16, 545–568 (1998). Article CAS Google Scholar * Qin, D. et al. Fcγ receptor
IIB on follicular dendritic cells regulates the B cell recall response. _J. Immunol._ 164, 6268–6275 (2000). Article CAS Google Scholar * Heyman, B. Regulation of antibody responses via
antibodies, complement, and Fc receptors. _Annu. Rev. Immunol._ 18, 709–737 (2000). Article CAS Google Scholar * Szakal, A.K., Holmes, K.L. & Tew, J.G. Transport of immune complexes
from the subcapsular sinus to lymph node follicles on the surface of nonphagocytic cells, including cells with dendritic morphology. _J. Immunol._ 131, 1714–1727 (1983). CAS PubMed Google
Scholar * Tew, J.G., Mandel, T.E., Phipps, R.P. & Szakal, A.K. Tissue localization and retention of antigen in relation to the immune response. _Am. J. Anat._ 170, 407–420 (1984).
Article CAS Google Scholar * Gretz, J.E., Norbury, C.C., Anderson, A.O., Proudfoot, A.E. & Shaw, S. Lymph-borne chemokines and other low molecular weight molecules reach high
endothelial venules via specialized conduits while a functional barrier limits access to the lymphocyte microenvironments in lymph node cortex. _J. Exp. Med._ 192, 1425–1440 (2000). Article
CAS Google Scholar * 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 * Martinez-Pomares, L. et al. Fc chimeric protein containing the cysteine-rich domain of the murine mannose receptor binds to
macrophages from splenic marginal zone and lymph node subcapsular sinus and to germinal centers. _J. Exp. Med._ 184, 1927–1937 (1996). Article CAS Google Scholar * Mueller, C.G. et al.
Mannose receptor ligand-positive cells express the metalloprotease decysin in the B cell follicle. _J. Immunol._ 167, 5052–5060 (2001). Article CAS Google Scholar * 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 * 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 CAS Google Scholar * Banerji, S. et al. LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan. _J. Cell Biol._ 144,
789–801 (1999). Article CAS Google Scholar * Hadjantonakis, A.K., Macmaster, S. & Nagy, A. Embryonic stem cells and mice expressing different GFP variants for multiple non-invasive
reporter usage within a single animal. _BMC Biotechnol._ 2, 11 (2002). Article Google Scholar * Allen, C.D., Okada, T., Tang, H.L. & Cyster, J.G. Imaging of germinal center selection
events during affinity maturation. _Science_ 315, 528–531 (2007). Article CAS Google Scholar * Molina, H. et al. Markedly impaired humoral immune response in mice deficient in complement
receptors 1 and 2. _Proc. Natl. Acad. Sci. USA_ 93, 3357–3361 (1996). Article CAS Google Scholar * Cascalho, M., Ma, A., Lee, S., Masat, L. & Wabl, M. A quasi-monoclonal mouse.
_Science_ 272, 1649–1652 (1996). Article CAS Google Scholar * Miller, J.J. & Nossal, G.J.V. Antigens in Immunity VI. The phagocytic reticulum of lymph node follicles. _J. Exp. Med._
120, 1075–1095 (1964). Article Google Scholar * Mitchell, J. & Abbot, A. Ultrastructure of the antigen-retaining reticulum of lymph node follicles as shown by high-resolution
autoradiography. _Nature_ 208, 500–502 (1965). Article CAS Google Scholar * Bergtold, A., Desai, D.D., Gavhane, A. & Clynes, R. Cell surface recycling of internalized antigen permits
dendritic cell priming of B cells. _Immunity_ 23, 503–514 (2005). Article CAS Google Scholar * Veerman, A.J. & van Rooijen, N. Lymphocyte capping and lymphocyte migration as
associated events in the _in vivo_ antigen trapping process. An electron-microscopic autoradiographic study in the spleen of mice. _Cell Tissue Res._ 161, 211–217 (1975). Article CAS
Google Scholar * Gray, D., Kumararatne, D.S., Lortan, J., Khan, M. & MacLennan, I.C. Relation of intra-splenic migration of marginal zone B cells to antigen localization on follicular
dendritic cells. _Immunology_ 52, 659–669 (1984). CAS PubMed PubMed Central Google Scholar * Groeneveld, P.H., Erich, T. & Kraal, G. The differential effects of bacterial
lipopolysaccharide (LPS) on splenic non-lymphoid cells demonstrated by monoclonal antibodies. _Immunology_ 58, 285–290 (1986). CAS PubMed PubMed Central Google Scholar * Ferguson, A.R.,
Youd, M.E. & Corley, R.B. Marginal zone B cells transport and deposit IgM-containing immune complexes onto follicular dendritic cells. _Int. Immunol._ 16, 1411–1422 (2004). Article CAS
Google Scholar * Brown, J.C., De Jesus, D.G., Holborow, E.J. & Harris, G. Lymphocyte-mediated transport of aggregated human gamma-globulin into germinal centre areas of normal mouse
spleen. _Nature_ 228, 367–369 (1970). Article CAS Google Scholar * Heinen, E. et al. Transfer of immune complexes from lymphocytes to follicular dendritic cells. _Eur. J. Immunol._ 16,
167–172 (1986). Article CAS Google Scholar * Whipple, E.C., Shanahan, R.S., Ditto, A.H., Taylor, R.P. & Lindorfer, M.A. Analyses of the _in vivo_ trafficking of stoichiometric doses
of an anti-complement receptor 1/2 monoclonal antibody infused intravenously in mice. _J. Immunol._ 173, 2297–2306 (2004). Article CAS Google Scholar * Schwickert, T.A. et al. _In vivo_
imaging of germinal centres reveals a dynamic open structure. _Nature_ 446, 83–87 (2007). Article CAS Google Scholar * Boes, M. et al. Enhanced B-1 cell development, but impaired IgG
antibody responses in mice deficient in secreted IgM. _J. Immunol._ 160, 4776–4787 (1998). CAS PubMed Google Scholar * Ehrenstein, M.R., O'Keefe, T.L., Davies, S.L. & Neuberger,
M.S. Targeted gene disruption reveals a role for natural secretory IgM in the maturation of the primary immune response. _Proc. Natl. Acad. Sci. USA_ 95, 10089–10093 (1998). Article CAS
Google Scholar * Kraal, G. Cells in the marginal zone of the spleen. _Int. Rev. Cytol._ 132, 31–73 (1992). Article CAS Google Scholar * Mombaerts, P. et al. RAG-1-deficient mice have no
mature B and T lymphocytes. _Cell_ 68, 869–877 (1992). Article CAS Google Scholar * Phan, T.G. et al. B cell receptor-independent stimuli trigger immunoglobulin (Ig) class switch
recombination and production of IgG autoantibodies by anergic self-reactive B cells. _J. Exp. Med._ 197, 845–860 (2003). Article CAS Google Scholar * 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 Download references ACKNOWLEDGEMENTS We thank
M. Krummel for help with the two-photon microscope; F. Schaufele (Diabetes Endocrinology Research Center Imaging Core) for help with the confocal microscope; C. Allen for advice; T. Gerdes
and M. Wabl (University of California, San Francisco) for QM mice, J. Atkinson and X. Wu (University of Washington) for _Cr2__−/−_ mice; T. Kinoshita (Osaka University) and S. Boackle
(University of Colorado Health Sciences Center) for the 8C12 hybridoma; G. Kraal (Vrije University Medical Center) for the MOMA-1 hybridoma; M. Cooper (University of Alabama at Birmingham)
for monoclonal antibody BP-3; and G. Cinamon and L. Shiow for comments on the manuscript. Supported by the National Health and Medical Research Council and American Australian Association
(T.G.P.), the Cancer Research Institute (I.G.), the Howard Hughes Medical Institute (J.G.C.), the National Institutes of Health (AI45073 and AI40098) and a Sandler New Technology Award.
AUTHOR INFORMATION Author notes * Takaharu Okada Present address: Present address: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto
University, Katsura Campus, Nishikyo-ku, Kyoto, Japan., AUTHORS AND AFFILIATIONS * Howard Hughes Medical Institute and Department of Microbiology and Immunology, University of California,
San Francisco, 94143, California, USA Tri Giang Phan, Irina Grigorova, Takaharu Okada & Jason G Cyster Authors * Tri Giang Phan View author publications You can also search for this
author inPubMed Google Scholar * Irina Grigorova View author publications You can also search for this author inPubMed Google Scholar * Takaharu Okada View author publications You can also
search for this author inPubMed Google Scholar * Jason G Cyster View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS T.G.P. and J.G.C. designed
and conceptualized the research; T.G.P. did the experiments; I.G. and T.O. assisted with the two-photon microscopy; and T.G.P., I.G. and J.G.C. analyzed the data and prepared the manuscript.
CORRESPONDING AUTHOR Correspondence to Jason G Cyster. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY
TEXT AND FIGURES Supplementary Figure 1 (PDF 1687 kb) SUPPLEMENTARY MOVIE 1 Intravital imaging of IC capture in the SCS. Time-lapse image sequence of 10 μm z-projection shows capture of
PE-ICs by cells lining the subcapsular sinus (annotated SCS). The SCS is visible between the collagen fibers (blue; second harmonic) of the capsule and the BP-3+ follicular stromal network
(green). PE-ICs were generated in vivo by subcutaneous injection of PE into a mouse that had previously received anti-PE IgG. Within minutes large PE-ICs (red) drain to the lymph node and
are visible flowing and accumulating on cells lining the SCS. The autofluorescent macrophage (labeled Mφ in the first few frames) protrudes into the lumen and is seen to rapidly accumulate
PE-ICs on its surface. PE-ICs can also be seen accumulating along a process of the macrophage (arrowhead). B cells (cyan) are not associated with any PE signal at this early time. Elapsed
time is shown as hh:mm:ss from the time of PE injection. Data are representative of three independent experiments. (MPG 3379 kb) SUPPLEMENTARY MOVIE 2 Real-time imaging of B cell migration
and interaction with SCS macrophages. Time-lapse image sequence of 20 μm z-projection from an inguinal lymph node explant 3 h after PE injection. By now the subcapsular space is coated with
PE-ICs (red) captured on SCS macrophages. B cells (cyan) continuously migrate to the SCS and interact with macrophages some of which are autofluorescent in the green channel. Capsule and SCS
associated collagen fibers appear blue (second harmonic). Elapsed time is shown as hh:mm:ss from the time of PE injection. Data are representative of three independent experiments. (MPG
6491 kb) SUPPLEMENTARY MOVIE 3 Real-time imaging of B cells capturing ICs in the subcapsular region. Time-lapse image sequence of 20 μm z-projection from an inguinal lymph node explant 3 h
after PE injection focusing on the subcapsular space which is coated with PE-ICs (red) captured on SCS macrophages. B cells (cyan) interact with SCS macrophages and can be seen to emerge
with associated PE signal on their surface (arrowheads). At one point a B cell (asterisk) approaches and captures a large depot of PE-ICs (open circle) presumably from the nearby SCS
macrophage (Mφ) and is depicted in Fig. 3c. Capsule and SCS associated collagen fibers appear blue (second harmonic). Elapsed time is shown as hh:mm:ss from the time of PE injection. Data is
representative of three independent experiments. (MPG 9500 kb) SUPPLEMENTARY MOVIE 4 Real-time imaging of B cells transporting PE-ICs into the follicle. Time-lapse image sequence of 20 μm
z-projection from an inguinal lymph node explant 4 h after PE injection. B cells (cyan) are highly motile and have dendritic extensions. The PE-ICs (red) are largely localized to the uropod
in the trailing edge of the B cells. The track of several B cells carrying PE-ICs are annotated with dots. Yellow dots shows the track of the B cell depicted in Fig. 3d. Elapsed time is
shown as hh:mm:ss from the time of PE injection. Data are representative of three independent experiments. (MPG 6465 kb) SUPPLEMENTARY MOVIE 5 Real-time imaging showing B cell expression of
complement receptors is required to capture PE-ICs. Time-lapse image sequence of 20 μm zprojection from inguinal lymph node explants 3 h after PE injection. CFSE-labeled (green) Cr1-,
Cr2-deficient (left panel) and wild-type B cells (right panel) in the subcapsular region are shown side-by-side for comparison. Capsule and SCS associated collagen fibers appear blue (second
harmonic). A wild-type B cell is highlighted (yellow arrow, right panel) that is in intimate contact with a macrophage for a prolonged period before emerging with PE-ICs on its surface.
Cr1-, Cr2-deficient B cells that encounter macrophages (highlighted with white arrows, left panel) are not seen to emerge with PE-IC. Note the dendritic morphology of migrating B cells even
in the absence of Cr1 and Cr2. ICs on B cells appear yellow due to co-localization of red and green. Elapsed time is shown as hh:mm:ss from the time of PE injection. Data are representative
of two independent experiments. (MPG 9615 kb) SUPPLEMENTARY MOVIE 6 Real-time imaging of B cells capturing and transporting cognate antigen coated with ICs. Time-lapse image sequence of 20
μm z-projection from inguinal lymph node explants showing efficient capture (left panel) and transport (right panel) of NP-PE ICs by cognate CFSE-labeled QM B cells (green) compared to
polyclonal CFP+ B cells (cyan). In the left panel a QM B cell (yellow arrowhead) can be seen scanning along the NP-PE IC coated process of a nearby macrophage (Mφ). Subsequently it rips off
some ICs and migrates away (white arrowhead) with NP-PE ICs capped at its tail. In the right panel several QM B cells can be seen migrating with large ICs capped at their tail. The QM B cell
tracked with yellow dots is depicted in Fig. 5c. Note the morphology of cognate QM B cells in contrast to polyclonal CFP+ B cells. Elapsed time is shown as hh:mm:ss from the time of NP-PE
injection. Data are representative of two independent experiments. (MPG 9654 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Phan, T., Grigorova, I.,
Okada, T. _et al._ Subcapsular encounter and complement-dependent transport of immune complexes by lymph node B cells. _Nat Immunol_ 8, 992–1000 (2007). https://doi.org/10.1038/ni1494
Download citation * Received: 29 May 2007 * Accepted: 25 June 2007 * Published: 29 July 2007 * Issue Date: September 2007 * DOI: https://doi.org/10.1038/ni1494 SHARE THIS ARTICLE Anyone you
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