Dynamics of antigen delivery and the functional roles of L121-adjuvant.
This study investigates the intracellular transport of protein antigens facilitated by L121-adjuvants and examines the associated cytotoxic T lymphocyte (CTL) effect. EL4 mouse thymoma cells were treated with L121-adjuvant and stained with AnnexinV-propidium iodide (PI) followed by flow cytometric analysis. The intracellular trafficking dynamics of bovine serum albumin (BSA)-FITC in the J774.A.1 macrophages, influenced by the L121-adjuvant, was visualized by confocal microscopy. The antigen-specific cytotoxic T lymphocyte (CTL) effect induced by the L121-adjuvant was determined by the cleavage-specific fluorogenic caspase substrate. The trafficking of BSA-FITC in the J774A.1 cells by confocal microscopy illustrated that the L121-adjuvant facilitated the intracellular transport of proteins to the subcellular compartments, including the lysosome, endoplasmic reticulum (ER), and the cis-Golgi apparatus. The L121-adjuvant also facilitated antigen delivery to the dendritic cells in the lymph nodes. Immunization of mice with the L121-adjuvant resulted in cell-mediated cytotoxic responses in the target cells, as detected by PhiPhiLux, a fluorogenic caspase substrate. Taken together, the L121-adjuvant improved the dynamics of protein delivery to antigen presenting cells, and also induced caspase activation, thereby illustrating the mechanism of antigen-specific CTL effects.
School of Pharmacy, College of Medicine, National Taiwan University, 33 Linsen South Road, Room 425, Taipei City 10050, Taiwan.
Shan-Shan
SS
Shen
School of Pharmacy, College of Medicine, National Taiwan University, 33 Linsen South Road, Room 425, Taipei City 10050, Taiwan. Electronic address: ywyang@ntu.edu.tw.
Ya-Wun
YW
Yang
D000276:Adjuvants, Immunologic; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D000941:Antigens / Q000276:immunology* / Q000378:metabolism*; D001692:Biological Transport; D020169:Caspases / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology; D005434:Flow Cytometry; D007114:Immunization; D008198:Lymph Nodes / Q000276:immunology; D008264:Macrophages / Q000276:immunology / Q000648:ultrastructure; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D010047:Ovalbumin / Q000276:immunology; D020442:Poloxamer / Q000008:administration & dosage* / Q000378:metabolism; D011506:Proteins / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D000276:Adjuvants, Immunologic; D000941:Antigens; D011506:Proteins; D020442:Poloxamer; D010047:Ovalbumin; D020169:Caspases
Antigen delivery; CTL; Pluronic L121; Vaccine adjuvant