Shen S.-S.YA-WUN YANG2021-01-062021-01-0620150264410Xhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-84939572330&doi=10.1016%2fj.vaccine.2015.04.051&partnerID=40&md5=0dcfb08bfd1775be9728c8a60eb9d4b3https://scholars.lib.ntu.edu.tw/handle/123456789/539285This 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. ? 2015 Elsevier Ltd.Antigen delivery; CTL; Pluronic L121; Vaccine adjuvant[SDGs]SDG3adjuvant; antigen; bovine serum albumin; fluorescein isothiocyanate; L121 adjuvant; lipocortin 5; propidium iodide; unclassified drug; antigen; caspase; immunological adjuvant; ovalbumin; poloxamer; protein; animal cell; animal experiment; antigen presenting cell; Article; cell compartmentalization; confocal microscopy; controlled study; cytotoxic T lymphocyte; dendritic cell; endoplasmic reticulum; flow cytometry; Golgi complex; immunization; intracellular transport; lymph node; lysosome; macrophage; mouse; nonhuman; priority journal; target cell; thymoma cell line; animal; C57BL mouse; immunology; metabolism; transport at the cellular level; ultrastructure; Adjuvants, Immunologic; Animals; Antigen-Presenting Cells; Antigens; Biological Transport; Caspases; Dendritic Cells; Flow Cytometry; Immunization; Lymph Nodes; Macrophages; Mice, Inbred C57BL; Microscopy, Confocal; Ovalbumin; Poloxamer; Proteins; T-Lymphocytes, Cytotoxicjournal article10.1016/j.vaccine.2015.04.051259176782-s2.0-84939572330