Abstract
The neonatal Fc receptor, FcRn, mediates endocytic recycling pathway that prevents degradation of IgG and is expressed in most endothelial cells. The blood–brain barrier (BBB), formed by brain endothelial cells sealed with tight junctions, restricts transport of IgG from the blood to the brain. In contrast, it has been suggested that IgG undergoes efflux from the brain parenchyma via reverse transcytosis across the BBB mediated by FcRn. The fast elimination of therapeutic antibodies from the brain via this route may limit their therapeutic potency. In vitro and in vivo methods described in this chapter were developed to facilitate research into mechanisms and dynamics of brain efflux of compounds, including FcRn-mediated reverse transcytosis across the BBB. The in vitro model uses immortalized adult rat brain endothelial cells which express high levels of FcRn. In vivo models use Prospective optical imaging to measure the clearance rate of intracerebrally injected FcRn-transported molecules tagged with near-infrared fluorescent probes.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ballabh P, Braun A, Nedergaard M (2004) The blood-brain barrier: an overview: structure, regulation, and clinical implications. Neurobiol Dis 16:1–13.
Gaillard PJ, Visser CC, de Boer AG (2005) Targeted delivery across the blood-brain barrier. Expert Opin Drug Deliv 2:299–309.
Boado RJ, Hui EK, Lu JZ, Zhou QH, Pardridge WM (2010) Selective targeting of a TNFR decoy receptor pharmaceutical to the primate brain as a receptor-specific IgG fusion protein. J Biotechnol 146:84–91.
Zhang Y, Pardridge WM (2001) Mediated efflux of IgG molecules from brain to blood across the blood-brain barrier. J Neuroimmunol 114:168–172.
Thorne RG, Nicholson C (2006) In vivo diffusion analysis with quantum dots and dextrans predicts the width of brain extracellular space. PNAS 103:5567–5572.
Weller RO, Galea I, Carare RO, Minagar A (2009) Pathophysiology of the lymphatic drainage of the central nervous system: Implications for pathogenesis and therapy of multiple sclerosis. J Pathophysiology (doi: 10.1015/j.pathophysiol.2009.10.007).
Abbot JN (2004) Evidence for bulk flow of brain interstitial fluid: significance for physiology and pathology. Neurochemistry Inter-national 45:545–552
Schlachetzki F, Zhu C, Pardridge WM (2002) Expression of the neonatal Fc receptor (FcRn) at the blood-brain barrier. J Neurochem 81:203–206.
Leach JL, Sedmak DD, Osborne JM, Rahill B, Lairmore MD, Anderson CL (1996) Isolation from human placenta of the IgG transporter, FcRn, and localization to the syncytiotrophoblast: implications for maternal-fetal antibody transport. J Immunol 157:3317–3322.
Roopenian DC, Christianson GJ, Sproule TJ, Brown AC, Akilesh S, Jung N, Petkova S, Avanessian L, Choi EY, Shaffer DJ, Eden PA, Anderson CL (2003) The MHC class I-like IgG receptor controls perinatal IgG transport, IgG homeostasis, and fate of IgG-Fc-coupled drugs. J Immunol 170:3528–3533.
Yoshida M, Claypool SM, Wagner JS, Mizoguchi E, Mizoguchi A, Roopenian DC, Lencer WI, Blumberg RS (2004) Human neonatal Fc receptor mediates transport of IgG into luminal secretions for delivery of antigens to mucosal dendritic cells. Immunity 20:769–783.
Ober RJ, Martinez C, Lai X, Zhou J, Ward ES (2004) Exocytosis of IgG as mediated by the receptor, FcRn: an analysis at the single-molecule level. PNAS 101:11076–11081.
Roopenian DC, Akilesh S (2007) FcRn: the neonatal Fc receptor comes of age. Nat Rev Immunol 7:715–725.
He W, Ladinsky MS, Huey-Tubman KE, Jensen GJ, McIntosh JR, Björkman PJ (2008) FcRn-mediated antibody transport across epithelial cells revealed by electron tomography. Nature 455:542–546.
Garg A, Balthasar JP (2009) Investigation of the influence of FcRn on the distribution of IgG to the brain. AAPS J 11:553–557.
Deane R, Sagare A, Hamm K, Parisi M, LaRue B, Guo H, Wu Z, Holtzman DM, Zlokovic BV (2005) IgG-assisted age-dependent clearance of Alzheimer’s amyloid beta peptide by the blood-brain barrier neonatal Fc receptor. J Neurosci 25:11495–11503.
Garberg P, Ball M, Borg N, Cecchelli R, Fenart L, Hurst RD, Lindmark T, Mabondzo A, Nilsson JE, Raub TJ, Stanimirovic D, Terasaki T, Oberg JO, Osterberg T (2005) In vitro models for the blood-brain barrier. Toxicol In Vitro 19:299–334.
Muruganandam A, Herx LM, Monette R, Durkin JP, Stanimirovic DB (1997) Development of immortalized human cerebromicrovascular endothelial cell line as an in vitro model of the human blood-brain barrier. FASEB J 11:1187–1197.
Cecchelli R, Dehouck B, Descamps L, Fenart L, Buée-Scherrer VV, Duhem C, Lundquist S, Rentfel M, Torpier G, Dehouck MP (1999) In vitro model for evaluating drug transport across the blood-brain barrier. Adv Drug Deliv Rev 36:165–178.
Paxinos G, Watson C (2005) The rat brain in stereotaxic coordinates. 5th Edition, Elsevier Academic Press, Amsterdam Boston.
Abulrob A, Brunette E, Slinn J, Baumann E, Stanimirovic D (2007) In vivo time domain optical imaging of renal ischemia-reperfusion injury: discrimination based on fluorescence lifetime. Mol Imaging 6:304–314.
Gumbleton M, Audus KL (2001) Progress and limitations in the use of in vitro cell cultures to serve as a permeability screen for the blood-brain barrier. J Pharm Sci 90:1681–1698.
Colgan OC, Collins NT, Ferguson G, Murphy RP, Birney YA, Cahill PA, Cummins PM (2008) Influence of basolateral condition on the regulation of brain microvascular endothelial tight junction properties and barrier function. Brain Res 1193:84–92.
Frangioni JV (2003) In vivo near-infrared fluorescence imaging Curr Opin Chem Biol 7:626–34.
Andersen JT, Sandlie I (2009) the versatile MHC class-I-related FcRn protects Igg and albumin from degradation: implication for development of new diagnostics and therapeutics. Drug Metab Pharmacokinet 24:318–332.
Chaudhury C, Mehnaz, S, Robinson JM, Hayton WL, Pearl DK, Roopenian DC, Anderson CL (2003) The major histocompatibility complex-related Fc receptor for IgG (FcRn) binds albumin and prolongs its lifespan. J Exp Med 197:315–322.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Caram-Salas, N. et al. (2011). In Vitro and In Vivo Methods for Assessing FcRn-Mediated Reverse Transcytosis Across the Blood–Brain Barrier. In: Turksen, K. (eds) Permeability Barrier. Methods in Molecular Biology, vol 763. Humana Press. https://doi.org/10.1007/978-1-61779-191-8_26
Download citation
DOI: https://doi.org/10.1007/978-1-61779-191-8_26
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-190-1
Online ISBN: 978-1-61779-191-8
eBook Packages: Springer Protocols