title: Transport of chitosan microparticles for mucosal vaccine delivery in a human intestinal M-cell model.
abstract: Uptake of particulate antigen carrier systems by specialized M-cells of the gut-associated lymphoid tissue is still a limiting step in inducing efficient immune responses after oral vaccination. Although transport of soluble drugs over the epithelial barrier of the gut is extensively studied in vitro by using the Caco-2 cell model, this was for long time not possible for particles due to the absence of M-cells. By co-culturing Caco-2 cells with cultured human B-lymphocytes (Raji-cells), cells which are morphologically and functionally similar to M-cells can be induced. This human M-cells model makes it possible to study the uptake of microparticles for oral vaccine delivery. In this way, chitosan microparticles, which have demonstrated to target the Peyer's patches efficiently in vivo, could be tested in vitro. The development of this M-cells model facilitates the optimization of the microparticles in order to target them even more efficiently to the M-cells in the gut. In this study, the integrity of the human M-cell model was investigated by determining the transepithelial electrical resistance (TEER), 14C-mannitol transport and morphology using scanning electron microscopy. The uptake of particles was investigated by measuring transport of both fluorescently labeled microspheres (Fluospheres) and chitosan microparticles using flowcytometry. No discontinuities or abnormalities could be found in the co-culture. Scanning electron microscopy showed that morphologically different cells were present in the human M-cell model. Both commercially available Fluospheres (size 0.2 microm) and chitosan microparticles (size 1.7 microm) for oral vaccine delivery were transported at a significantly higher amount by the human M-cell model compared to the transport by the Caco-2 cell monoculture. Since chitosan microparticles were proven to be taken up by Peyer's patches in mice as well, this human M-cell model is able to predict the M-cell uptake of microparticles for oral vaccine delivery. This M-cell model is a new tool, which can be used to scan, develop and optimize microparticles for oral vaccine delivery. Since the M-cell uptake can now be studied in vitro, the targeting of these cells can be studied more efficiently and can now be done in cells from human origin.
authors.affiliation: Leiden/Amsterdam Center for Drug Research, Division of Pharmaceutical Technology, PO Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: I M
authors.initials: IM
authors.lastname: van der Lubben
authors.forename: F A C
authors.initials: FA
authors.lastname: van Opdorp
authors.forename: M R
authors.initials: MR
authors.lastname: Hengeveld
authors.forename: J J M
authors.initials: JJ
authors.lastname: Onderwater
authors.forename: H K
authors.initials: HK
authors.lastname: Koerten
authors.forename: J C
authors.initials: JC
authors.lastname: Verhoef
authors.forename: G
authors.initials: G
authors.lastname: Borchard
authors.forename: H E
authors.initials: HE
authors.lastname: Junginger
mesh_terms: D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D001692:Biological Transport / Q000502:physiology; D018938:Caco-2 Cells; D002686:Chitin / Q000008:administration & dosage* / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D048271:Chitosan; D018920:Coculture Techniques / Q000379:methods; D016503:Drug Delivery Systems / Q000379:methods*; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008221:Lymphoid Tissue / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008863:Microspheres; D014407:Tumor Cells, Cultured; D014612:Vaccines / Q000008:administration & dosage*
chemical_list: D014612:Vaccines; D002686:Chitin; D048271:Chitosan