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FcRn-mediated antibody transport across epithelial cells revealed by electron tomography

Nature volume 455, pages 542–546 (2008)Cite this article

Abstract

The neonatal Fc receptor (FcRn) transports maternal IgG across epithelial barriers1,2, thereby providing the fetus or newborn with humoral immunity before its immune system is fully functional. In newborn rats, FcRn transfers IgG from milk to blood by apical-to-basolateral transcytosis across intestinal epithelial cells. The pH difference between the apical (pH 6.0–6.5) and basolateral (pH 7.4) sides of intestinal epithelial cells facilitates the efficient unidirectional transport of IgG, because FcRn binds IgG at pH 6.0–6.5 but not at pH 7 or more1,2. As milk passes through the neonatal intestine, maternal IgG is removed by FcRn-expressing cells in the proximal small intestine (duodenum and jejunum); remaining proteins are absorbed and degraded by FcRn-negative cells in the distal small intestine (ileum)3,4,5,6. Here we use electron tomography to make jejunal transcytosis visible directly in space and time, developing new labelling and detection methods to map individual nanogold-labelled Fc within transport vesicles7 and simultaneously to characterize these vesicles by immunolabelling. Combining electron tomography with a non-perturbing endocytic label allowed us to conclusively identify receptor-bound ligands, resolve interconnecting vesicles, determine whether a vesicle was microtubule-associated, and accurately trace FcRn-mediated transport of IgG. Our results present a complex picture in which Fc moves through networks of entangled tubular and irregular vesicles, only some of which are microtubule-associated, as it migrates to the basolateral surface. New features of transcytosis are elucidated, including transport involving multivesicular body inner vesicles/tubules and exocytosis through clathrin-coated pits. Markers for early, late and recycling endosomes each labelled vesicles in different and overlapping morphological classes, revealing spatial complexity in endo-lysosomal trafficking.

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Figure 1: Au-Fc uptake in intestinal cells.
Figure 2: Gold-containing tubular vesicles from jejunal regions 1 and 2.
Figure 3: Gold-containing irregular vesicles in jejunal regions 2 and 3.
Figure 4: Gold-containing MVBs in jejunal cells.
Figure 5: Jejunal LIS and schematic pathways.

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References

  1. Rodewald, R. & Kraehenbuhl, J.-P. Receptor-mediated transport of IgG. J. Cell Biol. 99, S159–S164 (1984)

    Article  Google Scholar 

  2. Simister, N. E. & Mostov, K. E. An Fc receptor structurally related to MHC class I antigens. Nature 337, 184–187 (1989)

    Article  ADS  CAS  Google Scholar 

  3. Rodewald, R. Selective antibody transport in the proximal small intestine of the neonatal rat. J. Cell Biol. 45, 635–640 (1970)

    Article  CAS  Google Scholar 

  4. Jones, E. A. & Waldmann, T. A. The mechanism of intestinal uptake and transcellular transport of IgG in the neonatal rat. J. Clin. Invest. 51, 2916–2927 (1972)

    Article  CAS  Google Scholar 

  5. Rodewald, R. Intestinal transport of antibodies in the newborn rat. J. Cell Biol. 58, 189–211 (1973)

    Article  CAS  Google Scholar 

  6. Rodewald, R. Distribution of immunoglobulin G receptors in the small intestine of the young rat. J. Cell Biol. 85, 18–32 (1980)

    Article  CAS  Google Scholar 

  7. He, W. et al. A freeze substitution fixation-based gold enlarging technique for EM studies of endocytosed Nanogold-labeled molecules. J. Struct. Biol. 160, 103–111 (2007)

    Article  CAS  Google Scholar 

  8. Benlounes, N. et al. Intestinal transport and processing of immunoglobulin G in the neonatal and adult rat. Biol. Neonate 67, 254–263 (1995)

    Article  CAS  Google Scholar 

  9. McIntosh, J. R., Nicastro, D. & Mastronarde, D. N. New views of cells in 3D: an introduction to electron tomography. Trends Cell Biol. 15, 43–51 (2005)

    Article  CAS  Google Scholar 

  10. Morphew, M., He, W., Bjorkman, P. J. & McIntosh, J. R. Silver enhancement of nanogold particles during freeze substitution fixation for electron microscopy. J. Microsc. 230, 263–267 (2008)

    Article  MathSciNet  CAS  Google Scholar 

  11. Hull, B. E. & Staehelin, L. A. The terminal web. A reevaluation of its structure and function. J. Cell Biol. 81, 67–82 (1979)

    Article  CAS  Google Scholar 

  12. Schwartz, S. L., Cao, C., Pylypenko, O., Rak, A. & Wandinger-Ness, A. Rab GTPases at a glance. J. Cell Sci. 120, 3905–3910 (2007)

    Article  CAS  Google Scholar 

  13. Stoorvogel, W., Oorschot, V. & Geuze, H. J. A novel class of clathrin-coated vesicles budding from endosomes. J. Cell Biol. 132, 21–33 (1996)

    Article  CAS  Google Scholar 

  14. McCarthy, K. M., Yoong, Y. & Simister, N. E. Bidirectional transcytosis of IgG by the rat neonatal Fc receptor expressed in a rat kidney cell line: a system to study protein transport across epithelia. J. Cell Sci. 113, 1277–1285 (2000)

    CAS  PubMed  Google Scholar 

  15. Murk, J. L. A. N. et al. Endosomal compartmentalization in three dimensions: Implications for membrane fusion. Proc. Natl Acad. Sci. USA 100, 13332–13337 (2003)

    Article  ADS  CAS  Google Scholar 

  16. Kleijmeer, M. et al. Reorganization of multivesicular bodies regulates MHC class II antigen presentation by dendritic cells. J. Cell Biol. 155, 53–63 (2001)

    Article  CAS  Google Scholar 

  17. Mari, M. et al. SNX1 defines an early endosomal recycling exit for sortilin and mannose 6-phosphate receptors. Traffic 9, 380–393 (2008)

    Article  CAS  Google Scholar 

  18. Altstiel, L. & Branton, D. Fusion of coated vesicles with lysosomes: measurement with a fluorescence assay. Cell 32, 921–929 (1983)

    Article  CAS  Google Scholar 

  19. Brodsky, F. M., Chen, C. Y., Knuehl, C., Towler, M. C. & Wakeham, D. E. Biological basket weaving: formation and function of clathrin-coated vesicles. Annu. Rev. Cell Dev. Biol. 17, 517–568 (2001)

    Article  CAS  Google Scholar 

  20. Conner, S. D. & Schmid, S. L. Regulated portals of entry into the cell. Nature 422, 37–44 (2003)

    Article  ADS  CAS  Google Scholar 

  21. Bonifacino, J. S. & Glick, B. S. The mechanisms of vesicle budding and fusion. Cell 116, 153–166 (2004)

    Article  CAS  Google Scholar 

  22. Greene, L. E. & Eisenberg, E. Dissociation of clathrin from coated vesicles by the uncoating ATPase. J. Biol. Chem. 265, 6682–6687 (1990)

    CAS  PubMed  Google Scholar 

  23. Fesce, R., Grohovaz, F., Valtorta, F. & Meldolesi, J. Neurotransmitter release: fusion or ‘kiss-and-run’? Trends Cell Biol. 4, 1–4 (1994)

    Article  CAS  Google Scholar 

  24. Ober, R. J., Martinez, C., Lai, X., Zhou, J. & Ward, E. S. Exocytosis of IgG as mediated by the receptor, FcRn: an analysis at the single-molecule level. Proc. Natl Acad. Sci. USA 101, 11076–11081 (2004)

    Article  ADS  CAS  Google Scholar 

  25. Sokac, A. M. & Bement, W. M. Kiss-and-coat and compartment mixing: coupling exocytosis to signal generation and local actin assembly. Mol. Biol. Cell 17, 1495–1502 (2006)

    Article  CAS  Google Scholar 

  26. Granseth, B., Odermatt, B., Royle, S. J. & Lagnado, L. Clathrin-mediated endocytosis is the dominant mechanism of vesicle retrieval at hippocampal synapses. Neuron 51, 773–786 (2006)

    Article  CAS  Google Scholar 

  27. Ghetie, V. & Ward, E. S. Multiple roles for the major histocompatibility complex class I-related FcRn. Annu. Rev. Immunol. 18, 739–766 (2000)

    Article  CAS  Google Scholar 

  28. Mastronarde, D. N. Dual-axis tomography: an approach with alignment methods that preserve resolution. J. Struct. Biol. 120, 343–352 (1997)

    Article  CAS  Google Scholar 

  29. Ladinsky, M. S. & Howell, K. E. Electron tomography of immunolabeled cryosections. Methods Cell Biol. 79, 543–548 (2007)

    Article  CAS  Google Scholar 

  30. Huber, A. H., Kelley, R. F., Gastinel, L. N. & Bjorkman, P. J. Crystallization and stoichiometry of binding of a complex between a rat intestinal Fc receptor and Fc. J. Mol. Biol. 230, 1077–1083 (1993)

    Article  CAS  Google Scholar 

  31. Halliday, R. The absorption of antibodies from immune sera by the gut of the young rat. Proc. R. Soc. Lond. B 143, 408–413 (1955)

    Article  ADS  CAS  Google Scholar 

  32. Danscher, G. Localization of gold in biological tissue. A photochemical method for light and electron microscopy. Histochemistry 71, 1–16 (1981)

    Article  CAS  Google Scholar 

  33. He, W., Cowin, P. & Stokes, D. L. Untangling desmosomal knots with electron tomography. Science 302, 109–113 (2003)

    Article  ADS  CAS  Google Scholar 

  34. Mastronarde, D. N. Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152, 36–51 (2005)

    Article  Google Scholar 

  35. Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional data using IMOD. J. Struct. Biol. 116, 71–76 (1996)

    Article  CAS  Google Scholar 

  36. Ladinsky, M. S., Mastronarde, D. N., McIntosh, J. R., Howell, K. E. & Staehelin, L. A. Golgi structure in three dimensions: functional insights from the normal rat kidney cell. J. Cell Biol. 144, 1135–1149 (1999)

    Article  CAS  Google Scholar 

  37. Tokuyasu, K. T. Application of cryoultramicrotomy to immunocytochemistry. J. Microsc. 143, 139–149 (1986)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank D. Mastronarde for advice about setting up SerialEM on the Caltech microscopes and three-dimensional modelling; C. Kivork for preparing Au-Fc and Au-dextran; B. Tivol for assistance with electron microscopes; M. Murphy for help with schematic figures; M. Morphew for contributions to silver enhancement methods; Y. Nie, A. Feuerabendt, A. Kules, C. Luna, K. McKenzie, R. Sander and L. Zinn-Bjorkman for segmenting tomograms; J. Vielmetter for SPR studies of Au-Fc; and F. Brodsky, F. Maxfield, S. Schmid and D. Tesar for discussions. This work was supported by the National Institutes of Health (2 R37 AI041239-06A1 to P.J.B. and RR000592 to J.R.M.), a Max Planck Research Award (P.J.B.), gifts from the Gordon and Betty Moore Foundation and the Agouron Institute to support electron microscopy at Caltech, and National University of Singapore AcRF start-up funds (R-154-000-339-133 to W.H.).

Author Contributions W.H. and P.J.B. conceived the experiments. Electron microscopy data were collected by W.H. in collaboration with and in the microscopy laboratory established by G.J.J. at Caltech. W.H. configured SerialEM on the Caltech electron microscopes, prepared intestinal samples by chemical fixation or HPF/FSF, conceived and developed the gold enhancement procedures, and collected, processed, interpreted and modelled tomograms. W.H. and K.E.H.T conducted kinetic experiments. Immunolabelling and associated imaging was done by M.S.L. in Boulder with enhanced samples provided by W.H. and K.E.H.T. Initial phases of the project using transfected cells were conducted by P.J.B. in Boulder in collaboration with J.R.M. and the Boulder Laboratory for 3D Microscopy of Cells. All authors discussed and interpreted the results and edited the manuscript.

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Authors and Affiliations

  1. Division of Biology 114-96 and,,

    Wanzhong He, Kathryn E. Huey-Tubman, Grant J. Jensen & Pamela J. Björkman

  2. Howard Hughes Medical Institute, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, USA ,

    Kathryn E. Huey-Tubman & Pamela J. Björkman

  3. Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543,

    Wanzhong He

  4. Department of Molecular, Boulder Laboratory for 3D Electron Microscopy of Cells, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80309, USA,

    Mark S. Ladinsky & J. Richard McIntosh

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Correspondence to Pamela J. Björkman.

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He, W., Ladinsky, M., Huey-Tubman, K. et al. FcRn-mediated antibody transport across epithelial cells revealed by electron tomography. Nature 455, 542–546 (2008). https://doi.org/10.1038/nature07255

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