Cell death induced by vaccine adjuvants containing surfactants
Journal
Vaccine
Journal Volume
22
Journal Issue
44512
Pages
1524-1536
Date Issued
2004
Author(s)
YA-WUN YANG
Wu C.-A.
Morrow W.J.W.
Abstract
Many vaccine adjuvants contain surface-active agents, but the immunological roles played by these components have been essentially ignored. The objective of this study was to examine possible apoptotic and necrotic effects of the surface-active agents, Pluronic L121 and Tween 80, which are components of L121-adjuvant (a formulation we synthesized with the aim of representing several commercially produced adjuvants), on EL4 lymphoma cells. Cell viability and cytolytic effects were analyzed using the MTT and LDH release assays, and the distribution of cells in different stages of the cell cycle after treatment with these agents was analyzed by propidium iodide (PI) staining and flow cytometry. L121-adjuvant was shown to induce cell cycle arrest and inhibit cell proliferation. Treatment of EL4 cells with surface-active agents resulted in a concentration-dependent increase in the apoptotic/necrotic cell populations. Fluorescence microscopy using Hoechst 33342 staining demonstrated chromosome condensation and DNA fragmentation in cells treated with surfactants or adjuvant. The apoptotic and necrotic effects of vaccine adjuvant containing surface-active agents were confirmed by Annexin V/propidium iodide staining and flow cytometric analysis. Pretreatment of EL4 cells with zVAD-fmk, a broad range caspase inhibitor, partially prevented apoptosis induced by Pluronic L121, but did not prevent the cell death induced by Tween 80 or L121-adjuvant. These findings suggested that Tween 80 and L121-adjuvant induced apoptosis in EL4 cells via a "non-classical" caspase-independent pathway. Results presented in this study suggest mechanisms of elicitation of CD8+, class I-restricted CTL response by soluble antigens mediated by the vaccine adjuvant containing surface-active agents. ? 2003 Elsevier Ltd. All rights reserved.
Subjects
3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide; Antigen-presenting cell; APC; CTL; Cytotoxic T-lymphocyte; DCs; Deionized water; Dendritic cells; DI; L121; Lactate dehydrogenase; LDH; Major histocompatibility complex; MHC; MTT; PI; Pluronic L121
SDGs
Other Subjects
3 (4,5 dimethyl 2 thiazolyl) 2,5 diphenyltetrazolium bromide; benzyloxycarbonylvalylalanylaspartyl fluoromethyl ketone; caspase inhibitor; CD8 antigen; DNA fragment; hoe 33342; immunological adjuvant; lactate dehydrogenase; lipocortin 5; pluronic l121; polysorbate 80; propidium iodide; surfactant; unclassified drug; vaccine; animal cell; apoptosis; article; cancer adjuvant therapy; cell death; cell subpopulation; cell viability; chromosome condensation; concentration response; controlled study; cytolysis; drug formulation; enzyme assay; flow cytometry; fluorescence microscopy; immunopharmacology; lymphoma cell; mitosis inhibition; mouse; nonhuman; priority journal; staining; Adjuvants, Immunologic; Animals; Apoptosis; Caspases; Cell Cycle; Cell Death; Cell Line; Coloring Agents; Diploidy; Flow Cytometry; Fluorescent Dyes; L-Lactate Dehydrogenase; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence; Necrosis; Propidium; Surface-Active Agents; T-Lymphocytes, Cytotoxic; Tetrazolium Salts; Thiazoles
Type
journal article