title: Non-covalent pomegranate (Punica granatum) hydrolyzable tannin-protein complexes modulate antigen uptake, processing and presentation by a T-cell hybridoma line co-cultured with murine peritoneal macrophages.
abstract: In this work we characterize the interaction of pomegranate hydrolyzable tannins (HT) with hen egg-white lysozyme (HEL) and determine the effects of non-covalent tannin-protein complexes on macrophage endocytosis, processing and presentation of antigen. We isolated HT from pomegranate and complex to HEL, the resulting non-covalent tannin-protein complex was characterized by gel electrophoresis and MALDI-TOF MS. Finally, cell culture studies and confocal microscopy imaging were conducted on the non-covalent pomegranate HT-HEL protein complexes to evaluate its effect on macrophage antigen uptake, processing and presentation to T-cell hybridomas. Our results indicate that non-covalent pomegranate HT-HEL protein complexes modulate uptake, processing and antigen presentation by mouse peritoneal macrophages. After 4 h of pre-incubation, only trace amounts of IL-2 were detected in the co-cultures treated with HEL alone, whereas a non-covalent pomegranate HT-HEL complex had already reached maximum IL-2 expression. Pomegranate HT may increase rate of endocytose of HEL and subsequent expression of IL-2 by the T-cell hybridomas.
authors.affiliation: a National Center for Biotechnological Innovations (CENIBiot), CeNAT-CONARE, Edificio Franklin Chang Diaz , San Jose , Costa Rica ;
authors.forename: Sergio
authors.initials: S
authors.lastname: Madrigal-Carballo
authors.affiliation: b Department of Animal Sciences, Reed Research Group , University of Wisconsin-Madison , Madison , WI , USA ;
authors.forename: Linda
authors.initials: L
authors.lastname: Haas
authors.affiliation: c Mass Spectrometry Facility, Department of Chemistry , University of Wisconsin-Madison , Madison , WI , USA.
authors.forename: Martha
authors.initials: M
authors.lastname: Vestling
authors.affiliation: b Department of Animal Sciences, Reed Research Group , University of Wisconsin-Madison , Madison , WI , USA ;
authors.forename: Christian G
authors.initials: CG
authors.lastname: Krueger
authors.affiliation: b Department of Animal Sciences, Reed Research Group , University of Wisconsin-Madison , Madison , WI , USA ;
authors.forename: Jess D
authors.initials: JD
authors.lastname: Reed
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D018920:Coculture Techniques; D019587:Dietary Supplements / Q000032:analysis; D004527:Egg Proteins / Q000737:chemistry / Q000276:immunology; D055951:Functional Food / Q000032:analysis; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D047348:Hydrolyzable Tannins / Q000737:chemistry* / Q000276:immunology*; D029561:Lythraceae / Q000737:chemistry* / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology; D051379:Mice; D046912:Multiprotein Complexes / Q000737:chemistry / Q000276:immunology; D009113:Muramidase / Q000737:chemistry* / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D004527:Egg Proteins; D047348:Hydrolyzable Tannins; D046912:Multiprotein Complexes; C115642:punicalagin; D009113:Muramidase
keywords: Tannins; polyphenols; pomegranate juice; proteins