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Expression and Transport Functionality of FcRn within Rat Alveolar Epithelium: A Study in Primary Cell Culture and in the Isolated Perfused Lung

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Purpose

The neonatal constant region fragment receptor (FcRn) binds and transports IgG. FcRn expression in the upper tracheobronchial airways of the lung is recognized. In this study, we sought to characterize the functional expression of FcRn within alveolar regions of lung tissue.

Methods

FcRn immunohistochemistry was performed on intact rat lung. FcRn expression [Western blot, reverse transcription–polymerase chain reaction (RT-PCR), and immunofluorescence microscopy] and IgG transport functionality were assessed in an in vitro rat alveolar epithelial primary cell culture model. An isolated perfused rat lung model was used to examine IgG transport across pulmonary epithelium from airspace to perfusate.

Results

FcRn is expressed in intact alveolar epithelium, substantiated by expression and functionality in an in vitro alveolar epithelial model within which IgG transport was temperature sensitive, concentration dependent, and inhibited by excess unlabeled IgG and, to a disproportionate level, by anti-FcRn antibody. Saturable IgG transport across pulmonary epithelium was evident in an isolated perfused rat lung, inhibitable by competing IgG, and displayed a relatively low maximal net IgG absorptive rate of approximately 80 ng/h.

Conclusion

Pulmonary epithelium expresses functional FcRn providing an absorption pathway potentially important for highly potent Fcγ-fusion proteins but unlikely to be of quantitative significance for the systemic delivery of inhaled therapeutic monoclonal IgGs.

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Abbreviations

AE:

alveolar epithelial

BSA:

bovine serum albumin

cDNA:

complementary deoxyribonucleic acid

DMEM:

Dulbecco's modified Eagle medium

EDTA:

ethylenediaminetetraacetic acid

ELISA:

enzyme-linked immunosorbent assay

FBS:

fetal bovine serum

FcRn:

neonatal Fc receptor

FITC:

fluorescein isothiocyanate

HRP:

horseradish peroxide

IgG:

immunoglobulin G

IPRL:

isolated perfused rat lung

MDI:

metered dose inhaler

MHC:

major histocompatibility complex

MWCO:

molecular weight cutoff

NIH:

National Institute of Health

RT:

reverse transcriptase

RT-PCR:

reverse transcription–polymerase chain reaction

SDS-PAGE:

sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TBST:

Tris-buffered saline with Tween 20

tRNA:

total RNA

UV:

ultraviolet

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Acknowledgments

The authors are grateful to Pamela J. Björkman, Ph.D., Division of Biology, California Institute of Technology, Pasadena, CA, USA, for her gift of truncated rFcRn. This research began during MS's 2-month research leave at Cardiff University (CU) and was supported in part by the Medical College of Virginia Foundation, Richmond, VA, USA, and by MG's research funds at Cardiff University, UK. MS and MG acknowledge the encouragement of Peter Byron, Ph.D., School of Pharmacy Virginia Commonwealth University, throughout this work.

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  1. Yadollah Omidi & Jaleh Barar

    Present address: Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

Authors and Affiliations

  1. Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, Virginia, 23298-0533, USA

    Masahiro Sakagami

  2. Pharmaceutical Cell Biology, Welsh School of Pharmacy, Cardiff University, Cardiff, CF10 3XF, UK

    Yadollah Omidi, Lee Campbell, Lana E. Kandalaft, Christopher J. Morris, Jaleh Barar & Mark Gumbleton

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  1. Masahiro Sakagami
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  2. Yadollah Omidi
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Correspondence to Mark Gumbleton.

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Sakagami, M., Omidi, Y., Campbell, L. et al. Expression and Transport Functionality of FcRn within Rat Alveolar Epithelium: A Study in Primary Cell Culture and in the Isolated Perfused Lung. Pharm Res 23, 270–279 (2006). https://doi.org/10.1007/s11095-005-9226-0

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