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Keith E. Mostov - One of the best experts on this subject based on the ideXlab platform.
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the Polymeric Immunoglobulin Receptor
Seminars in Cell Biology, 2013Co-Authors: Charlotte S. Kaetzel, Keith E. MostovAbstract:IgA is the primary Immunoglobulin found in a wide range of epithelial and mucosal secretions, including milk, saliva, intestinal secretions, bile, respiratory secretions, and tears [1]. Over 30 years ago it was observed that IgA isolated from such secretions contains an extra polypeptide of about 70 kDa, known as secretory component (SC) [2]. SC is synthesized by the epithelial cell and associates with dimeric IgA (dIgA) as it is transported across the epithelial cell. It was subsequently discovered that SC is a proteolytic fragment of an integral membrane protein, known as the Polymeric Immunoglobulin Receptor (pIgR) [3]. The basic model for the transepithelial transport of dIgA by the pIgR is shown in Figure 5.1. The pIgR is synthesized as an integral membrane protein in the rough endoplasmic reticulum and then travels to the Golgi apparatus. In the last station of the Golgi, known as the trans-Golgi network (TGN), the pIgR is sorted into vesicles that deliver it to the basolateral surface of the epithelial cell. At that surface the pIgR can bind to dIgA that is produced by plasma cells in the lamina propria underlying the epithelium. The pIgR and bound dIgA are then endocytosed and delivered to endosomes. The Receptor and ligand move through a series of endocytotic and transcytotic vesicles and are ultimately delivered to the apical plasma membrane. There the extracellular, ligand binding portion of the pIgR is cleaved and released together with the dIgA into external secretions. This cleaved fragment is the SC.
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Chapter 12 – Immunoglobulin Transport and the Polymeric Immunoglobulin Receptor
Mucosal Immunology, 2005Co-Authors: Charlotte S. Kaetzel, Keith E. MostovAbstract:The Polymeric Immunoglobulin Receptor plays a unique role in the mucosal immune system, acting both as an epithelial transporter and as an integral component of secretory Immunoglobulins. Major advances in the past several years with use of the approaches of immunology, protein chemistry, cell biology, and molecular biology have increased the understanding of the structure, function, and regulation of Polymeric Immunoglobulin Receptor (pIgR). Production of mice genetically deficient in pIgR expression has allowed a more defined characterization of its biological functions. Molecular characterization of the gene encoding pIgR in humans and several animal species has provided important insights into the mechanisms by which its transcription is regulated by the immune and endocrine systems. Further characterization of molecular mechanisms of pIgR regulation, at the level of both synthesis and transcytosis, would increase the understanding of the integration of this important molecule with other effectors of the mucosal immune system.
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chapter 12 Immunoglobulin transport and the Polymeric Immunoglobulin Receptor
Mucosal Immunology (Third Edition), 2005Co-Authors: Charlotte S. Kaetzel, Keith E. MostovAbstract:The Polymeric Immunoglobulin Receptor plays a unique role in the mucosal immune system, acting both as an epithelial transporter and as an integral component of secretory Immunoglobulins. Major advances in the past several years with use of the approaches of immunology, protein chemistry, cell biology, and molecular biology have increased the understanding of the structure, function, and regulation of Polymeric Immunoglobulin Receptor (pIgR). Production of mice genetically deficient in pIgR expression has allowed a more defined characterization of its biological functions. Molecular characterization of the gene encoding pIgR in humans and several animal species has provided important insights into the mechanisms by which its transcription is regulated by the immune and endocrine systems. Further characterization of molecular mechanisms of pIgR regulation, at the level of both synthesis and transcytosis, would increase the understanding of the integration of this important molecule with other effectors of the mucosal immune system.
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The mammalian retromer regulates transcytosis of the Polymeric Immunoglobulin Receptor
Nature cell biology, 2004Co-Authors: Marcel Verges, Frédéric Luton, Carmen Gruber, Frank Tiemann, Lorri G. Reinders, Lan Huang, Alma L. Burlingame, Carol Renfrew Haft, Keith E. MostovAbstract:Epithelial cells have separate apical and basolateral plasma membrane domains with distinct compositions. After delivery to one surface, proteins can be endocytosed and then recycled, degraded or transcytosed to the opposite surface. Proper sorting into the transcytotic pathway is essential for maintaining polarity, as most proteins are endocytosed many times during their lifespan. The Polymeric Immunoglobulin Receptor (pIgR) transcytoses Polymeric IgA (pIgA) from the basolateral to the apical surface of epithelial cells and hepatocytes. However, the molecular machinery that controls polarized sorting of pIgR-pIgA and other Receptors is only partially understood. The retromer is a multimeric protein complex, originally described in yeast, which mediates intracellular sorting of Vps10p, a Receptor that transports vacuolar enzymes. The yeast retromer contains two sub-complexes. One includes the Vps5p and Vps17p subunits, which provide mechanical force for vesicle budding. The other is the Vps35p-Vps29p-Vps26p subcomplex, which provides cargo specificity. The mammalian retromer binds to the mannose 6-phosphate Receptor, which sorts lysosomal enzymes from the trans-Golgi network to the lysosomal pathway. Here, we show a function for the mammalian Vps35-Vps29-Vps26 retromer subcomplex in promoting pIgR-pIgA transcytosis.
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Direct Interaction between Rab3b and the Polymeric Immunoglobulin Receptor Controls Ligand-Stimulated Transcytosis in Epithelial Cells
Developmental cell, 2002Co-Authors: Sven C.d. Van Ijzendoorn, Michael J. Tuvim, Thomas Weimbs, Burton F. Dickey, Keith E. MostovAbstract:We have examined the role of rab3b in epithelial cells. In MDCK cells, rab3b localizes to vesicular structures containing the Polymeric Immunoglobulin Receptor (pIgR) and located subjacent to the apical surface. We found that GTP-bound rab3b directly interacts with the cytoplasmic domain of pIgR. Binding of dIgA to pIgR causes a dissociation of the interaction with rab3b, a process that requires dIgA-mediated signaling, Arg657 in the cytoplasmic domain of pIgR, and possibly GTP hydrolysis by rab3b. Binding of dIgA to pIgR at the basolateral surface stimulates subsequent transcytosis to the apical surface. Overexpression of GTP-locked rab3b inhibits dIgA-stimulated transcytosis. Together, our data demonstrate that a rab protein can bind directly to a specific cargo protein and thereby control its trafficking.
Vera Jankowski - One of the best experts on this subject based on the ideXlab platform.
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P0724ASSOCIATIONS OF URINARY Polymeric Immunoglobulin Receptor PEPTIDES IN THE CONTEXT OF CARDIO-RENAL SYNDROM
Nephrology Dialysis Transplantation, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:Abstract Background and Aims The Polymeric Immunoglobulin Receptor (pIgR) which transports Immunoglobulins from the basolateral to the apical surface of epithelial cells was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to show the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to get information on how the SC will be released. Method We investigated urinary peptides of 2707 individuals available in the Human Urine Proteome Database using capillary electrophoresis coupled to mass spectrometry. Results The mean abundance of 23 different pIgR peptides correlates negatively with the estimated glomerular filtration rate (eGFR, r=-0.314, p<0.0001). Furthermore, pIgR peptides are significantly increased in coronary artery disease after adjustment for eGFR. We further predicted the proteases involved in urinary peptide generation using the Proteasix tool. Peptide cleavage site analysis suggests that several, and not one, proteases are involved in the generation of the SC. Conclusion In this large cohort, we could demonstrate that pIgR is associated with the cardio-renal syndrome and provide more detailed insights on how pIgR can be potentially cleaved to release the SC.
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Associations of urinary Polymeric Immunoglobulin Receptor peptides in the context of cardio-renal syndrome
Scientific Reports, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:The Polymeric Immunoglobulin Receptor (pIgR) transports Immunoglobulins from the basolateral to the apical surface of epithelial cells. PIgR was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface of epithelial cells where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to evaluate the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to obtain information on how the SC is released. We investigated urinary peptides of 2964 individuals available in the Human Urine Proteome Database generated using capillary electrophoresis coupled to mass spectrometry. The mean amplitude of 23 different pIgR peptides correlated negatively with the estimated glomerular filtration rate (eGFR, rho = -0.309, p
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Associations of urinary Polymeric Immunoglobulin Receptor peptides in the context of cardio-renal syndrome.
Scientific reports, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:The Polymeric Immunoglobulin Receptor (pIgR) transports Immunoglobulins from the basolateral to the apical surface of epithelial cells. PIgR was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface of epithelial cells where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to evaluate the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to obtain information on how the SC is released. We investigated urinary peptides of 2964 individuals available in the Human Urine Proteome Database generated using capillary electrophoresis coupled to mass spectrometry. The mean amplitude of 23 different pIgR peptides correlated negatively with the estimated glomerular filtration rate (eGFR, rho = -0.309, p < 0.0001). Furthermore, pIgR peptides were significantly increased in cardiovascular disease (coronary artery disease and heart failure) after adjustment for eGFR. We further predicted potential proteases involved in urinary peptide generation using the Proteasix algorithm. Peptide cleavage site analysis suggested that several, and not one, proteases are involved in the generation of the SC. In this large cohort, we could demonstrate that pIgR is associated with the cardio-renal syndrome and provided a more detailed insight on how pIgR can be potentially cleaved to release the SC.
Charlotte S. Kaetzel - One of the best experts on this subject based on the ideXlab platform.
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cooperativity among secretory iga the Polymeric Immunoglobulin Receptor and the gut microbiota promotes host microbial mutualism
Immunology Letters, 2014Co-Authors: Charlotte S. KaetzelAbstract:Abstract Secretory IgA (SIgA) antibodies in the intestinal tract form the first line of antigen-specific immune defense, preventing access of pathogens as well as commensal microbes to the body proper. SIgA is transported into external secretions by the Polymeric Immunoglobulin Receptor (pIgR). Evidence is reported here that the gut microbiota regulates production of SIgA and pIgR, which act together to regulate the composition and activity of the microbiota. SIgA in the intestinal mucus layer helps to maintain spatial segregation between the microbiota and the epithelial surface without compromising the metabolic activity of the microbes. Products shed by members of the microbial community promote production of SIgA and pIgR by activating pattern recognition Receptors on host epithelial and immune cells. Maternal SIgA in breast milk provides protection to newborn mammals until the developing intestinal immune system begins to produce its own SIgA. Disruption of the SIgA-pIgR-microbial triad can increase the risk of infectious, allergic and inflammatory diseases of the intestine.
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The Polymeric Immunoglobulin Receptor
eLS, 2013Co-Authors: Charlotte S. KaetzelAbstract:Secretory Immunoglobulin A (SIgA) antibodies represent the first line of antigen-specific immune defence protecting the mucosal surfaces against environmental pathogens and antigens and maintaining homoeostasis with the commensal microbiota. The Polymeric Ig Receptor (pIgR) plays the dual role of transporting locally produced dimeric IgA across mucosal epithelia and serving as the precursor of the secretory component moiety of SIgA. The complex regulation of pIgR expression and transcytosis by host and microbial factors is finely tuned to optimise the role of SIgA in mucosal immunity. Recent discoveries highlight the dynamic cross-talk between pIgR, SIgA and the commensal microbiota that populate our mucosal surfaces. Dysregulation of pIgR expression and/or function can result in profound consequences for the pathogenesis of infectious, inflammatory and neoplastic diseases. Future research into the function and regulation of pIgR and SIgA may offer new insights into the prevention and treatment of diseases that originate at mucosal surfaces. Key Concepts: Secretory IgA (SIgA) antibodies represent the first line of antigen-specific immune defence at mucosal surfaces. The Polymeric Immunoglobulin Receptor (pIgR) transports dimeric IgA across mucosal epithelial cells and serves as the precursor for the secretory component of SIgA. Transcription of the PIGR gene in mucosal epithelial cells is regulated by signalling pathways initiated by host cytokines and microbial factors, involving the cytoplasmic adapter protein, myeloid differentiation primary response protein 88 (MyD88). SIgA mediates immune protection by immune exclusion, intracellular neutralisation of antigens and pathogens and excretion of IgA-containing immune complexes by pIgR-mediated epithelial transcytosis. Cross-talk between pIgR, SIgA and commensal bacteria regulates the composition of the gut microbiota and promotes intestinal homoeostasis. Dysregulated expression or function of pIgR can contribute to the pathology of infectious, inflammatory and neoplastic diseases. Keywords: Polymeric Immunoglobulin Receptor; secretory IgA; mucosal immunity; epithelial transcytosis; microbiota; cytokines; Toll-like Receptors; NF-κB; infectious diseases; inflammatory bowel disease; cancer
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the Polymeric Immunoglobulin Receptor
Seminars in Cell Biology, 2013Co-Authors: Charlotte S. Kaetzel, Keith E. MostovAbstract:IgA is the primary Immunoglobulin found in a wide range of epithelial and mucosal secretions, including milk, saliva, intestinal secretions, bile, respiratory secretions, and tears [1]. Over 30 years ago it was observed that IgA isolated from such secretions contains an extra polypeptide of about 70 kDa, known as secretory component (SC) [2]. SC is synthesized by the epithelial cell and associates with dimeric IgA (dIgA) as it is transported across the epithelial cell. It was subsequently discovered that SC is a proteolytic fragment of an integral membrane protein, known as the Polymeric Immunoglobulin Receptor (pIgR) [3]. The basic model for the transepithelial transport of dIgA by the pIgR is shown in Figure 5.1. The pIgR is synthesized as an integral membrane protein in the rough endoplasmic reticulum and then travels to the Golgi apparatus. In the last station of the Golgi, known as the trans-Golgi network (TGN), the pIgR is sorted into vesicles that deliver it to the basolateral surface of the epithelial cell. At that surface the pIgR can bind to dIgA that is produced by plasma cells in the lamina propria underlying the epithelium. The pIgR and bound dIgA are then endocytosed and delivered to endosomes. The Receptor and ligand move through a series of endocytotic and transcytotic vesicles and are ultimately delivered to the apical plasma membrane. There the extracellular, ligand binding portion of the pIgR is cleaved and released together with the dIgA into external secretions. This cleaved fragment is the SC.
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Regulation of the Polymeric Immunoglobulin Receptor in intestinal epithelial cells by Enterobacteriaceae: implications for mucosal homeostasis.
Immunological investigations, 2010Co-Authors: Maria E. C. Bruno, Aubrey L. Frantz, Eric Rogier, Andrew T. Stefka, Stephanie N. Thompson, Charlotte S. KaetzelAbstract:The commensal microbiota of the human colon profoundly impacts host gene expression and mucosal homeostasis. Secretory IgA antibodies, which influence the composition of the intestinal microbiota and provide immunity against pathogens, are transported across intestinal epithelial cells (IEC) by the Polymeric Immunoglobulin Receptor (pIgR). To compare the effects of different colonic bacteria on pIgR expression, the human IEC line HT-29 was stimulated with various species representing the 4 major phyla of colonic bacteria. Only bacteria from the family Enterobacteriaceae (phylum Proteobacteria) induced expression of pIgR and other target genes of bacterial pattern recognition Receptors. HT-29 cells responded to purified ligands for Toll-like Receptor (TLR)4 but not TLR2. Expression of pIgR and transport of IgA were significantly reduced in colons of mice deficient in the TLR adaptor MyD88, consistent with a role for TLR signaling in the regulation of pIgR by colonic bacteria. Induction of pIgR expression i...
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Regulation of Polymeric Immunoglobulin Receptor expression by reovirus.
Journal of General Virology, 2005Co-Authors: Kasturi Pal, Charlotte S. Kaetzel, Kathleen M. Brundage, Cynthia Cunningham, Christopher F. CuffAbstract:Polymeric Immunoglobulin Receptor (pIgR) transcytoses dimeric IgA and IgA-coated immune complexes from the lamina propria across epithelia and into secretions. The effect of reovirus infection on regulation of pIgR expression in the human intestinal epithelial cell line HT-29 was characterized in this report. Both replication-competent and UV-inactivated reovirus at m.o.i. equivalents of 1–100 p.f.u. per cell upregulated pIgR mRNA by 24 h post-infection and intracellular pIgR protein was increased at 48 h following exposure to UV-inactivated virus. Binding of virus to HT-29 cells was required, as pre-incubating virus with specific antiserum, but not non-immune serum, inhibited reovirus-mediated pIgR upregulation. Endosomal acidification leading to uncoating of virus is a required step for pIgR upregulation, as ammonium chloride or bafilomycin A1 pre-treatment inhibited virus-induced pIgR upregulation. Inhibition experiments using the calpain inhibitor N-acetyl-leucyl-leucyl-norleucinal suggested that calpains are involved in reovirus-mediated pIgR upregulation. Upregulation of pIgR following virus infection appears to be an innate immune response against invading pathogens that could help the host clear infection effectively. Signalling induced by microbes and their products may serve to augment pIgR-mediated transcytosis of IgA, linking the innate and acquired immune responses to viruses.
Pamela B. Davis - One of the best experts on this subject based on the ideXlab platform.
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Antibodies to the Polymeric Immunoglobulin Receptor with different binding and trafficking patterns.
American journal of respiratory cell and molecular biology, 2005Co-Authors: Sanhita Gupta, Michael Heacock, Aura Perez, Pamela B. DavisAbstract:The Polymeric Immunoglobulin Receptor (pIgR) has been proposed as a therapeutic target, but its potential depends on the efficiency of uptake and trafficking of the Receptor ligand. Mouse monoclonal antibodies (Mabs) directed against pIgR, selected for strong binding to secretory component (SC) and secretory IgA (sIgA), were tested in a transcytosis assay in 16HBEo− cells (human bronchial epithelial cell line) transfected with human pIgR. Intracellular trafficking was followed by confocal microscopy. Mabs fell into two classes. For two Mabs, transcytosis from basolateral to apical surface is rapid, unidirectional, and little Mab is retained in the cell. For three Mabs, basolateral to apical transcytosis occurs to a significantly lesser extent, reverse transcytosis is permitted, and some of the Mab is retained in the perinuclear region even after 24 h. When tested for their ability to recognize and immunoprecipitate pIgR with systematic truncations and deletions of the five Immunoglobulin (Ig)-like domains, all Mabs bound to the fifth Ig-like domain, but three of them also bound to the C-terminal region of pIgR near the plasma membrane. Different binding sites probably account for the different trafficking of these Mabs and may predict differential therapeutic utility.
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transport of bifunctional proteins across respiratory epithelial cells via the Polymeric Immunoglobulin Receptor
American Journal of Respiratory and Critical Care Medicine, 2000Co-Authors: Thomas W. Ferkol, Elizabeth Eckman, Shadi Swaidani, Catherine L Silski, Pamela B. DavisAbstract:Neutrophil elastase (NE) contributes to progression of the lung disease characteristic of cystic fibrosis (CF). We developed a strategy that permits the delivery of alpha(1)-antitrypsin (alpha(1)-AT) to inaccessible CF airways by targeting the respiratory epithelium via the Polymeric Immunoglobulin Receptor (pIgR). A fusion protein consisting of a single-chain Fv directed against human secretory component (SC) and linked to human alpha(1)-AT was effectively transported in a basolateral-to-apical direction across in vitro model systems of polarized respiratory epithelium consisting of 16HBEo cells transfected with human pIgR complementary DNA, which overexpress the Receptor, and human respiratory epithelial cells grown in primary culture at an air-liquid interface. When applied to the basolateral surface, the anti-SC Fv/alpha(1)-AT fusion protein penetrated the respiratory epithelia, with transcytosis of the fusion protein being related to the amount of SC detected at the apical surface. Significantly less fusion protein crossed the cells in the opposite direction. In addition, because the antihuman SC Fv/alpha(1)-AT fusion protein was transported vectorially and deposited into the small volume of apical surface fluid, the antiprotease component of this protein was concentrated atop the epithelium. Thus, in cell models, this system is capable of concentrating the antiprotease of the fusion protein, in the thin film of epithelial surface fluid to a level expected to be therapeutic in the airways of many patients with CF.
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The Transport of Bifunctional Proteins across Respiratory Epithelia in Human Tracheal Xenografts by Exploiting the Polymeric Immunoglobulin Receptor
Pediatric Research, 1999Co-Authors: Elizabeth Eckman, Pamela B. Davis, Jerry Chipuk, Sheri Miller, Thomas W. FerkolAbstract:The Transport of Bifunctional Proteins across Respiratory Epithelia in Human Tracheal Xenografts by Exploiting the Polymeric Immunoglobulin Receptor
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Gene transfer into the airway epithelium of animals by targeting the Polymeric Immunoglobulin Receptor.
The Journal of clinical investigation, 1995Co-Authors: Thomas W. Ferkol, Charlotte S. Kaetzel, Elizabeth Eckman, Jose C. Perales, Richard W. Hanson, Pamela B. DavisAbstract:Genes of interest can be targeted specifically to respiratory epithelial cells in intact animals with high efficiency by exploiting the Receptor-mediated endocytosis of the Polymeric Immunoglobulin Receptor. A DNA carrier, consisting of the Fab portion of polyclonal antibodies raised against rat secretory component covalently linked to poly-L-lysine, was used to introduce plasmids containing different reporter genes into airway epithelial cells in vivo. We observed significant levels of luciferase enzyme activity in protein extracts from the liver and lung, achieving maximum values of 13,795 +/- 4,431 and 346,954 +/- 199,120 integrated light units (ILU) per milligram of protein extract, respectively. No luciferase activity was detected in spleen or heart, which do not express the Receptor. Transfections using complexes consisting of an irrelevant plasmid (pCMV lacZ) bound to the bona fide carrier or the expression plasmid (pGEMluc) bound to a carrier based on an irrelevant Fab fragment resulted in background levels of luciferase activity in all tissues examined. Thus, only tissues that contain cells bearing the Polymeric Immunoglobulin Receptor are transfected, and transfection cannot be attributed to the nonspecific uptake of an irrelevant carrier-DNA complex. Specific mRNA from the luciferase gene was also detected in the lungs of transfected animals. To determine which cells in the lungs are transfected by this method, DNA complexes were prepared containing expression plasmids with genes encoding the bacterial beta-galactosidase or the human interleukin 2 Receptor. Expression of these genes was localized to the surface epithelium of the airways and the submucosal glands, and not the bronchioles and alveoli. Receptor-mediated endocytosis can be used to introduce functional genes into the respiratory epithelium of rats, and may be a useful technique for gene therapy targeting the lung.
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Gene transfer into respiratory epithelial cells by targeting the Polymeric Immunoglobulin Receptor.
The Journal of clinical investigation, 1993Co-Authors: Thomas W. Ferkol, Charlotte S. Kaetzel, Pamela B. DavisAbstract:A system for targeting foreign DNA to epithelial cells in vitro has been developed by exploiting Receptor-mediated endocytosis. The Polymeric Immunoglobulin Receptor transports dimeric Immunoglobulin A and Immunoglobulin M through epithelial cells, including those of the respiratory tract, by binding the Immunoglobulins at the basolateral surface and transporting them across the cell. Fab fragments of antibodies directed against the extracellular portion of the Receptor, secretory component, are similarly transported. Anti-human secretory component Fab fragments were covalently linked to a polycation, and complexed to various expression plasmids. When bound to an expression plasmid containing the Escherichia coli lacZ gene ligated to the Rous sarcoma virus promoter, the complexes transfected HT29.74 human colon carcinoma cells induced to express Polymeric Immunoglobulin Receptor, but not those lacking the Receptor. Primary cultures of human tracheal epithelial cells grown on collagen gels, which induce the expression of Polymeric Immunoglobulin Receptor, were also transfected with the complexes. From 5 to 66% of the respiratory epithelial cells had beta-galactosidase activity after treatment, comparable to the percentage of cultured human tracheal epithelial cells that express Polymeric Immunoglobulin Receptor (8-35%). The addition of excess human secretory component (Fab ligand) to the culture medium at the time of transfection blocked the delivery of DNA. The expression plasmid, either alone, complexed to the polycation, or complexed to a carrier based on an irrelevant Fab fragment, was not effective in transfecting either cell type. This DNA carrier system introduces DNA specifically into epithelial cells that contain pIgR in vitro.
Justyna Siwy - One of the best experts on this subject based on the ideXlab platform.
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P0724ASSOCIATIONS OF URINARY Polymeric Immunoglobulin Receptor PEPTIDES IN THE CONTEXT OF CARDIO-RENAL SYNDROM
Nephrology Dialysis Transplantation, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:Abstract Background and Aims The Polymeric Immunoglobulin Receptor (pIgR) which transports Immunoglobulins from the basolateral to the apical surface of epithelial cells was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to show the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to get information on how the SC will be released. Method We investigated urinary peptides of 2707 individuals available in the Human Urine Proteome Database using capillary electrophoresis coupled to mass spectrometry. Results The mean abundance of 23 different pIgR peptides correlates negatively with the estimated glomerular filtration rate (eGFR, r=-0.314, p<0.0001). Furthermore, pIgR peptides are significantly increased in coronary artery disease after adjustment for eGFR. We further predicted the proteases involved in urinary peptide generation using the Proteasix tool. Peptide cleavage site analysis suggests that several, and not one, proteases are involved in the generation of the SC. Conclusion In this large cohort, we could demonstrate that pIgR is associated with the cardio-renal syndrome and provide more detailed insights on how pIgR can be potentially cleaved to release the SC.
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Associations of urinary Polymeric Immunoglobulin Receptor peptides in the context of cardio-renal syndrome
Scientific Reports, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:The Polymeric Immunoglobulin Receptor (pIgR) transports Immunoglobulins from the basolateral to the apical surface of epithelial cells. PIgR was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface of epithelial cells where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to evaluate the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to obtain information on how the SC is released. We investigated urinary peptides of 2964 individuals available in the Human Urine Proteome Database generated using capillary electrophoresis coupled to mass spectrometry. The mean amplitude of 23 different pIgR peptides correlated negatively with the estimated glomerular filtration rate (eGFR, rho = -0.309, p
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Associations of urinary Polymeric Immunoglobulin Receptor peptides in the context of cardio-renal syndrome.
Scientific reports, 2020Co-Authors: Justyna Siwy, Jochen Metzger, William Mullen, Harald Mischak, Joost Schanstra, Petra Zürbig, Vera JankowskiAbstract:The Polymeric Immunoglobulin Receptor (pIgR) transports Immunoglobulins from the basolateral to the apical surface of epithelial cells. PIgR was recently shown to be associated with kidney dysfunction. The immune defense is initiated at the apical surface of epithelial cells where the N-terminal domain of pIgR, termed secretory component (SC), is proteolytically cleaved and released either unbound (free SC) or bound to Immunoglobulins. The aim of our study was to evaluate the association of pIgR peptides with the cardio-renal syndrome in a large cohort and to obtain information on how the SC is released. We investigated urinary peptides of 2964 individuals available in the Human Urine Proteome Database generated using capillary electrophoresis coupled to mass spectrometry. The mean amplitude of 23 different pIgR peptides correlated negatively with the estimated glomerular filtration rate (eGFR, rho = -0.309, p < 0.0001). Furthermore, pIgR peptides were significantly increased in cardiovascular disease (coronary artery disease and heart failure) after adjustment for eGFR. We further predicted potential proteases involved in urinary peptide generation using the Proteasix algorithm. Peptide cleavage site analysis suggested that several, and not one, proteases are involved in the generation of the SC. In this large cohort, we could demonstrate that pIgR is associated with the cardio-renal syndrome and provided a more detailed insight on how pIgR can be potentially cleaved to release the SC.