title: The role of tomatine adjuvant in antigen delivery for cross- presentation.
abstract: The aim of this study was to investigate the use of tomatine adjuvant to deliver soluble antigen for crosspresentation by bone marrow-derived dendritic cells (BMDCs). BMDCs were incubated with tomatine adjuvantovalbumin (OVA) complex and analyzed for antigen uptake by flow cytometry. Adjuvant-induced cell death was examined in situ by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. To elucidate the effect of antigen internalization on tomatine adjuvant-mediated antigen presentation, BMDCs were treated with several endocytosis inhibitors, and antigen presentation was analyzed by B3Z activity assay. Our data indicated that tomatine adjuvant enhanced antigen internalization by antigen presenting cells (APCs) and induced significant cell death and leukocyte infiltration at the injection sites. In vitro tomatine adjuvant treatment of BMDCs activated Ova/K(b) restricted B3Z T cell hybridomas, whereas this activation was impaired by pretreatment with brefeldin A, cytochalasin B, wortmannin, or ZnCl2. Our results demonstrated the role of tomatine adjuvant in antigen delivery to antigen presenting cells (APCs) and suggested the involvement of phagocytosis and PI3K signaling during the delivery of soluble antigens in the context of MHC class I.
authors.forename: Paul Y-J
authors.initials: PY
authors.lastname: Hsu
authors.forename: Ching-An
authors.initials: CA
authors.lastname: Wu
authors.forename: Shan-Shan
authors.initials: SS
authors.lastname: Shen
authors.affiliation: School of Pharmacy, College of Medicine, National Taiwan University, 33 Linsen South Road, Room 425, Taipei City 10050, Taiwan. ywyang@ntu.edu.tw.
authors.forename: Ya-Wun
authors.initials: YW
authors.lastname: Yang
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000494:pharmacology; D000818:Animals; D017951:Antigen Presentation / Q000276:immunology*; D001854:Bone Marrow Cells / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology; D004705:Endocytosis; D005434:Flow Cytometry; D006825:Hybridomas; D020287:In Situ Nick-End Labeling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000276:immunology; D010587:Phagocytosis / Q000276:immunology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D014053:Tomatine / Q000008:administration & dosage* / Q000494:pharmacology
chemical_list: D000276:Adjuvants, Immunologic; D014053:Tomatine; D010047:Ovalbumin; D019869:Phosphatidylinositol 3-Kinases