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/Kb 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.
Keywords: Adjuvant, antigen delivery, tomatine, vaccine.
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