Abstract
A characteristic mucosal immune response involves the production of antigen-specific secretory immunoglobulin A (SIgA) antibodies. In order to study transcytosis by mimicking the SIgA secretion and to screen for SIgA secretion-promoting substances, we developed a model system of a transfected Madin-Darby canine kidney (MDCK) cell line that expresses the human polymeric immunoglobulin receptor (pIgR). We thus isolated the human dIgA (dimeric IgA)/pIgA (polymeric IgA) complex as the binding ligands. In the present study, a recombinant vector encoding the human pIgR gene was constructed and infected into MDCK cells. Following reverse transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunofluorescence staining, we confirmed that pIgR was expressed in the transfectant MDCK-pIgR cells and was located at the basolateral side of the cell surface. We also confirmed the coexistence of the dIgA/pIgA complex in the IgA myeloma serum. The covalent dIgA/pIgA complex was then isolated from the serum of an IgA multiple myeloma patient using an ÄKTA purifier operation system with a HiPrep 16/60 Sephacryl S-300 HR column, in order to utilize the complex as transcytosis ligands for human pIgR. Finally, we confirmed the uptake of the isolated human dIgA/pIgA complex into MDCK-pIgR cells. We demonstrated that the human dIgA/pIgA complex was transcytosed into the apical side of the monolayer cells. Therefore, our MDCK-human pIgR cell transcytosis model is an operational system and can be used for screening functional food components that promote dIgA/pIgR transcytosis as well as SIgA secretion.
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Acknowledgments
We thank Dr. Wayne I. Lencer (Boston Children’s Hospital and Harvard Medical School, MA, USA) for providing the plasmid pSK-pIgR encoding human pIgR and the plasmid pEGFP-N1. This work was supported by the National Marine Public Welfare Scientific Research Project of China (201105029), the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT: IRT1188), and the Chinese Scholarship Council of the Ministry of Education, People’s Republic of China.
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There is no conflict of interest.
Compliance with ethical standards
The human subject was conducted in compliance with applicable laws in China and the ethical standards that the international community recognizes and expects from leading multinational corporations. All aspects of the experiment were conducted according to guidelines provided by the ethical committee both at the Ocean University of China (Qingdao, China) and the Affiliated Hospital of Medical College of Qingdao University (Qingdao, China).
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Zuo, T., Feng, X., Zhang, N. et al. Establishment of a functional secretory IgA transcytosis model system in vitro for functional food screening. Appl Microbiol Biotechnol 99, 5535–5545 (2015). https://doi.org/10.1007/s00253-015-6501-9
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DOI: https://doi.org/10.1007/s00253-015-6501-9