Human and Non-Human Primate Intestinal FcRn Expression and Immunoglobulin G Transcytosis

PURPOSE: To evaluate transcytosis of immunoglobulin G (IgG) by the neonatal Fc receptor (FcRn) in adult primate intestine to determine whether this is a means for oral delivery of monoclonal antibodies (mAbs). METHODS: Relative regional expression of FcRn and localization in human intestinal mucosa by RT-PCR, ELISA & immunohistochemistry. Transcytosis of full-length mAbs (sandwich ELISA-based detection) across human intestinal segments mounted in Ussing-type chambers, human intestinal (caco-2) cell monolayers grown in transwells, and serum levels after regional intestinal delivery in isoflurane-anesthetized cynomolgus monkeys. RESULTS: In human intestine, there was an increasing proximal-distal gradient of mucosal FcRn mRNA and protein expression. In cynomolgus, serum mAb levels were greater after ileum-proximal colon infusion than after administration to stomach or proximal small intestine (1–5 mg/kg). Serum levels of wild-type mAb dosed into ileum/proximal colon (2 mg/kg) were 124 ± 104 ng/ml (n = 3) compared to 48 ± 48 ng/ml (n = 2) after a non-FcRn binding variant. In vitro, mAb transcytosis in polarized caco-2 cell monolayers and was not enhanced by increased apical cell surface IgG binding to FcRn. An unexpected finding in primate small intestine, was intense FcRn expression in enteroendocrine cells (chromagranin A, GLP-1 and GLP-2 containing). CONCLUSIONS: In adult primates, FcRn is expressed more highly in distal intestinal epithelial cells. However, mAb delivery to that region results in low serum levels, in part because apical surface FcRn binding does not influence mAb transcytosis. High FcRn expression in enteroendocrine cells could provide a novel means to target mAbs for metabolic diseases after systemic administration

University of Mississippi Jamie L. Whitten Dinner in Washington, D.C., The

The Jamie L. Whitten Dinner in Washington D.C. to honor U.S. Representative Jamie L. Whitten and to celebrate the creation of the Jamie L. Whitten Chair of Law and Government at the University of Mississippi. Introduction by Chancellor R. Gerald Turner. Tributes by W. Frederick Smith (Chairman of Federal Express Corporation); Ray Mabus (Governor of Mississippi); Mary Ann Mobley (National Chairperson of the Ole Miss Annual Fund). Miss Mississippi Toni Seawright sings Mississippi, I Love My Home. William F. Winter (former governor of Mississippi) discusses the University of Mississippi Law School. Will Hickman (Board of Trustees, Mississippi Institutions of Higher Learning) outlines Whitten\u27s education and career. Tributes by U.S. Representative Bob Michel and Speaker of the House Jim Wright. University of Mississippi documentary saluting the career of Jamie L. Whitten, including clips with U.S. Representative Thomas S. Foley; Whitten; U.S. Representative Edward P. Boland; U.S. Representative William H. Natcher; U.S. Representative Silvio Conte. Remarks of Jamie Whitten. Howard McMillan (President of the Ole Miss Alumni Association) presents Centennial Alumni medallion to Jamie Whitten

Jamie Whitten Gala Event for the Jamie Whitten Chair of Law and Government at the University of Mississippi (Part 1)

The Jamie Whitten Gala in Washington D.C. to honor U.S. Representative Jamie L. Whitten and to celebrate the creation of the Jamie L. Whitten Chair of Law and Government at the University of Mississippi (Part 1 of 2). Introduction by Chancellor R. Gerald Turner. Tributes by W. Frederick Smith (Chairman of Federal Express Corporation); Ray Mabus (Governor of Mississippi); Mary Ann Mobley (National Chairperson of the Ole Miss Annual Fund). Mississippi Toni Seawright sings Mississippi, I Love My Home. William F. Winter (former governor of Mississippi) discusses the University of Mississippi Law School. Will Hickman (Board of Trustees, Mississippi Institutions of Higher Learning) outlines Whitten\u27s education and career. Tribute by U.S. Representative Bob Michel

Raw footage of University of Mississippi Jamie Whitten Dinner in Washington, D.C.

Raw, unedited footage of University of Mississippi Jame Whitten Diner in Washington, D.C. used in creation of final product, Whitten VHS 8. Introduction by Chancellor R. Gerald Turner. Tributes by W. Frederick Smith (Chairman of Federal Express Corporation); Ray Mabus (Governor of Mississippi); Mary Ann Mobley (National Chairperson of the Ole Miss Annual Fund). Miss Mississippi Toni Seawright sings Mississippi, I Love My Home. William F. Winter (former governor of Mississippi) discusses the University of Mississippi Law School. Will Hickman (Board of Trustees, Mississippi Institutions of Higher Learning) outlines Whitten\u27s education and career. Tributes by U.S. Representative Bob Michel and Speaker of the House Jim Wright. University of Mississippi documentary saluting the career of Jamie L. Whitten, including clips with U.S. Representative Thomas S. Foley; Whitten; U.S. Representative Edward P. Boland; U.S. Representative William H. Natcher; U.S. Representative Silvio Conte. Remarks of Jamie Whitten. Howard McMillan (President of the Ole Miss Alumni Association) presents Centennial Alumni medallion to Jamie Whitten

Alternative Functional In Vitro Models of Human Intestinal Epithelia

Physiologically relevant sources of absorptive intestinal epithelial cells are crucial for human drug transport studies. Human adenocarcinoma-derived intestinal cell lines, such as Caco-2, offer conveniences of easy culture maintenance and scalability, but do not fully recapitulate in vivo intestinal phenotypes. Additional sources of renewable physiologically relevant human intestinal cells would provide a much needed tool for drug discovery and intestinal physiology. We sought to evaluate and compare two alternative sources of human intestinal cells, commercially available primary human intestinal epithelial cells (hInEpCs) and induced pluripotent stem cell (iPSC)-derived intestinal cells to Caco-2, for use in in vitro transwell monolayer intestinal transport assays. To achieve this for iPSC-derived cells, our previously described 3-dimensional intestinal organogenesis method was adapted to transwell differentiation. Intestinal cells were assessed by marker expression through immunocytochemical and mRNA expression analyses, monolayer integrity through Transepithelial Electrical Resistance (TEER) measurements and molecule permeability, and functionality by taking advantage the well-characterized intestinal transport mechanisms. In most cases, marker expression for primary hInEpCs and iPSC-derived cells appeared to be as good as or better than Caco-2. Furthermore, transwell monolayers exhibited high TEER with low permeability. Primary hInEpCs showed molecule efflux indicative of P-glycoprotein transport. Primary hInEpCs and iPSC-derived cells also showed neonatal Fc receptor-dependent binding of immunoglobulin G variants. Primary hInEpCs and iPSC-derived intestinal cells exhibit expected marker expression and demonstrate basic functional monolayer formation, similar to or better than Caco-2. These cells could offer an alternative source of human intestinal cells for understanding normal intestinal epithelial physiology and drug transport

The contribution of cell surface FcRn in monoclonal antibody serum uptake from the intestine in suckling rat pups

The neonatal Fc Receptor (FcRn) in intestinal epithelium is the primary mechanism for transfer of maternal immunoglobulin G (IgG) from suckled milk to serum; but the factors contributing to the rapid uptake of IgG are poorly understood. These studies help to determine the contribution of cell-surface FcRn in IgG uptake in 2-week old rat pups by varying local pH and binding conditions. Variants of a human wild-type IgG monoclonal antibody (mAb WT) were assessed for binding affinity (KD) to rat (r)FcRn at pH6.0 and subsequent off-rate at pH7.4 (1/s) by Surface Plasmon Resonance. Selected mAbs were administered intra-intestinally in isofluorane-anesthetized 2-week rat pups. Full-length mAb in serum was quantified by immunoassay, (r)FcRn mRNA expression by RT-PCR, and mAb epithelial localization was visualized by immunohistochemistry. After duodenal administration, serum levels of mAb variants correlated with their rFcRn off-rate at pH7.4, but not their affinity at pH6.0. The greatest serum levels of IgG were measured when mAb was administered in the duodenum where rFcRn mRNA expression is greatest, and was increased further by duodenal administration in pH6.0 buffer. More intense human IgG immunostaining was detected in epithelium than the same variant administered at higher pH. These data suggest an increased contribution for cell-surface receptor. We conclude that, in the neonate duodenum, receptor off-rates are as important as affinities for FcRn mediated uptake, and cell surface binding of IgG to rFcRn plays contributes to IgG uptake alongside pinocytosis; both of which responsible for increased IgG uptake