title: Nanoemulsion-based mucosal adjuvant induces apoptosis in human epithelial cells.
abstract: Nanoemulsions (NEs) are adjuvants that enhance antigen penetration of the nasal mucosa, increase cellular uptake of antigens by both epithelial and dendritic cells, and promote the migration of antigen-loaded dendritic cells to regional lymph nodes within 24-h of vaccine administration. The objective of this study was to elucidate cell death caused by W805EC NE and identify caspases and genes associated with death pathways. Consistent with this aim, we show that exposure of human epithelial cells (EC), both RPMI 2650 and FaDu, to NE results in the activation of caspases (1, 3/7, 6, 8, and 9) and the expression of genes involved in apoptotic as well as authophagy and necrosis pathways. Interestingly, the NE activates caspase 8 which promotes "immunogenic apoptosis". The rescue assay was employed to investigate the fate of RPMI 2650 cells treated with W805EC NE. After four-hour treatment with as little as 0.03% of NE no cells were rescued at 72h. Remarkably, immediately after four-hour treatment, the cells morphologically resembled untreated cells and most of the cells were alive. Altogether, these results suggest that NE induces death of human ECs through multiple pathways. Epithelial cell death caused by W805EC may have further implications on antigen uptake, processing, and presentation by DC's.
authors.affiliation: Department of Immunology of Infectious Diseases, Laboratory of Virology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland.
authors.forename: Beata U
authors.initials: BU
authors.lastname: Orzechowska
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, United States.
authors.forename: Jolanta F
authors.initials: JF
authors.lastname: Kukowska-Latallo
authors.affiliation: The University of Michigan, College of Literature, Science and the Arts, Ann Arbor, MI, United States.
authors.forename: Alexa D
authors.initials: AD
authors.lastname: Coulter
authors.affiliation: The University of Michigan, College of Literature, Science and the Arts, Ann Arbor, MI, United States.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Szabo
authors.affiliation: Department of Immunology of Infectious Diseases, Laboratory of Virology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland; Wroclaw Research Center EIT+, Wroclaw, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Gamian
authors.affiliation: Department of Immunology of Infectious Diseases, Laboratory of Virology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland; Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, United States; Faculty of Biological Sciences, University of Zielona Gora, Zielona Gora, Poland. Electronic address: myca@umich.edu.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Myc
mesh_terms: D000276:Adjuvants, Immunologic / Q000633:toxicity*; D017209:Apoptosis; D002460:Cell Line; D004655:Emulsions / Q000633:toxicity*; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology*; D006801:Humans; D053758:Nanoparticles / Q000633:toxicity*
chemical_list: D000276:Adjuvants, Immunologic; D004655:Emulsions
keywords: Autophagy; Caspases; FaDu; Necroptosis; Necrosis; Pyroptosis; RPMI 2650
references.citation: Clin Vaccine Immunol. 2008 Feb;15(2):348-58
references.pmid: 18057181
references.citation: Clin Cancer Res. 2010 Jun 15;16(12):3100-4
references.pmid: 20421432
references.citation: Vaccine. 2004 Jun 2;22(17-18):2316-27
references.pmid: 15149791
references.citation: Nat Med. 1999 Nov;5(11):1249-55
references.pmid: 10545990
references.citation: Expert Rev Vaccines. 2007 Oct;6(5):723-39
references.pmid: 17931153
references.citation: PLoS One. 2011;6(7):e21823
references.pmid: 21789184
references.citation: J Exp Med. 2000 Feb 7;191(3):423-34
references.pmid: 10662788
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Vaccine. 2003 Sep 8;21(25-26):3801-14
references.pmid: 12922114
references.citation: Vaccine. 2004 Mar 29;22(11-12):1524-36
references.pmid: 15063578
references.citation: Nature. 2011 Sep 15;477(7364):335-9
references.pmid: 21921917
references.citation: J Autoimmun. 2014 May;50:1-11
references.pmid: 24559657
references.citation: Int J Inflam. 2012;2012:819467
references.pmid: 22229105
references.citation: Cell Death Differ. 2003 Oct;10(10):1156-64
references.pmid: 14502239
references.citation: Lupus. 2009 Nov;18(13):1217-25
references.pmid: 19880572
references.citation: Cell Microbiol. 2004 Nov;6(11):1019-25
references.pmid: 15469431
references.citation: J Autoimmun. 2013 Dec;47:1-16
references.pmid: 24238833
references.citation: Nature. 1998 Mar 5;392(6671):86-9
references.pmid: 9510252
references.citation: Med Microbiol Immunol. 2010 May;199(2):81-92
references.pmid: 19967396
references.citation: PLoS Biol. 2004 Aug;2(8):E241
references.pmid: 15314657
references.citation: Vaccine. 2012 Jan 5;30(2):307-16
references.pmid: 22079079
references.citation: Vaccine. 2010 Aug 31;28 Suppl 3:C25-36
references.pmid: 20713254
references.citation: J Pharm Sci. 2012 Apr;101(4):1391-9
references.pmid: 22213631
references.citation: EMBO J. 2009 Mar 4;28(5):578-90
references.pmid: 19165151
references.citation: Eur J Immunol. 2012 Aug;42(8):2073-86
references.pmid: 22653620
references.citation: Cytometry. 2001 May 1;44(1):65-72
references.pmid: 11309810
references.citation: Vaccine. 2013 Feb 4;31(7):1072-9
references.pmid: 23273511
references.citation: Crit Rev Immunol. 2010;30(2):189-99
references.pmid: 20370629
references.citation: PLoS One. 2008;3(8):e2954
references.pmid: 18698426
references.citation: J Autoimmun. 2011 Feb;36(1):4-8
references.pmid: 20708902
references.citation: AIDS Res Hum Retroviruses. 2008 Feb;24(2):271-81
references.pmid: 18260780
references.citation: Trends Microbiol. 2001 Feb;9(2):64-7
references.pmid: 11173244
references.citation: Clin Dev Immunol. 2013;2013:374769
references.pmid: 24151516
references.citation: Hum Vaccin. 2010 Jul;6(7):585-94
references.pmid: 20421727
references.citation: J Immunol. 2009 Apr 15;182(8):4538-46
references.pmid: 19342627
references.citation: Infect Immun. 2007 Aug;75(8):4020-9
references.pmid: 17502384
references.citation: Vaccine. 2007 Nov 7;25(45):7763-72
references.pmid: 17928111
references.citation: Infect Immun. 2005 Apr;73(4):1907-16
references.pmid: 15784530
references.citation: Rheumatology (Oxford). 1999 Dec;38(12):1177-83
references.pmid: 10587542
references.citation: J Exp Med. 1998 Oct 5;188(7):1359-68
references.pmid: 9763615