caa a22 4500 605505845 CHVBK 20210128100624.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00253-015-6501-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6501-9 Establishment of a functional secretory IgA transcytosis model system in vitro for functional food screening [Elektronische Daten] [Tao Zuo, Xianqi Feng, Na Zhang, Changhu Xue, Qing-Juan Tang] 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. Springer-Verlag Berlin Heidelberg, 2015 SIgA nationallicence pIgR nationallicence MDCK nationallicence dIgA nationallicence pIgA nationallicence Transcytosis nationallicence Zuo Tao College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut Feng Xianqi The Affiliated Hospital of Medical College of Qingdao University, 16 Jiangsu Road, 266003, Qingdao, Shandong Province, People's Republic of China aut Zhang Na College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut Xue Changhu College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut Tang Qing-Juan College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5535-5545 0175-7598 99:13<5535 2015 99 253 https://doi.org/10.1007/s00253-015-6501-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6501-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zuo Tao College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut NATIONALLICENCE 700 1- Feng Xianqi The Affiliated Hospital of Medical College of Qingdao University, 16 Jiangsu Road, 266003, Qingdao, Shandong Province, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Na College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut NATIONALLICENCE 700 1- Xue Changhu College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut NATIONALLICENCE 700 1- Tang Qing-Juan College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, 266003, Qingdao, Shandong Province, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5535-5545 0175-7598 99:13<5535 2015 99 253