Engineered expression of the TLR5 ligand flagellin enhances paramyxovirus activation of human dendritic cell function.
The paramyxovirus simian virus 5 (SV5) is a poor activator of human dendritic cell (DC) maturation pathways in vitro, and infected DC do not upregulate cell surface costimulatory proteins or secretion of immunomodulatory cytokines. We evaluated the hypothesis that activation of SV5-infected DC would be enhanced by engineering SV5 to express a Toll-like-receptor (TLR) ligand. To test this hypothesis, a novel virus was engineered such that the gene encoding an intracellular form of the TLR5 ligand flagellin was expressed from the genome of wild-type (WT) SV5 (SV5-flagellin). Cells infected in vitro with the flagellin-expressing virus released low levels of biologically active flagellin, which was capable of stimulating TLR5 signaling. Infection of human peripheral blood mononuclear cell-derived immature DC with SV5-flagellin resulted in enhanced levels of interleukin-6 (IL-6) and IL-12 compared to infection with DC with the parental virus, WT SV5. In contrast to cytokine induction, the flagellin-expressing virus did not appreciably increase DC surface expression of the costimulatory molecule CD80 or CD86 above the level seen with WT SV5 alone. In mixed-culture assays, DC infected with the flagellin-expressing virus were more effective at activating gamma interferon secretion from both CD8(+) and CD4(+) allogeneic T cells than DC infected with WT SV5. Our results with SV5-directed intracellular expression of flagellin may be applicable to other vectors or pathogenic viruses where overcoming impairment of DC activation could contribute to the development of safer and more effective vaccines.
Arimilli
Subhashini
S
Department of Microbiology and Immunology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157-1064, USA.
Johnson
John B
JB
Clark
Kimberly M
KM
Graff
Aaron H
AH
Alexander-Miller
Martha A
MA
Mizel
Steven B
SB
Parks
Griffith D
GD
D000276:Adjuvants, Immunologic / Q000235:genetics / Q000494:pharmacology*; D018122:B7-1 Antigen / Q000096:biosynthesis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology*; D005408:Flagellin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D018664:Interleukin-12 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D045402:Parainfluenza Virus 5 / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D011995:Recombination, Genetic; D051198:Toll-Like Receptor 5 / Q000276:immunology* / Q000378:metabolism
D000276:Adjuvants, Immunologic; D018122:B7-1 Antigen; D015850:Interleukin-6; D011994:Recombinant Proteins; C495346:TLR5 protein, human; D051198:Toll-Like Receptor 5; D005408:Flagellin; D018664:Interleukin-12
Vaccine. 1996 Jan;14(1):85-92
8821654
J Exp Med. 1997 Sep 15;186(6):801-12
9294135
Vaccine. 2008 Jan 10;26(2):201-14
18063235
J Gen Virol. 2002 May;83(Pt 5):1167-1172
11961272
Virology. 2008 Jul 5;376(2):416-28
18456301
J Immunol. 2004 Mar 1;172(5):2861-9
14978087
J Virol. 2005 May;79(9):5507-15
15827165
J Virol. 2006 Apr;80(7):3128-34
16537581
Virology. 2007 Aug 15;365(1):144-56
17459446
J Virol. 2008 Jan;82(2):828-39
17977969
J Virol. 2006 Jul;80(13):6295-304
16775317
J Immunol. 1999 Dec 15;163(12):6762-8
10586075
J Immunol. 2003 May 15;170(10):5165-75
12734364
J Virol. 2007 Oct;81(20):11116-27
17686837
Virology. 2007 May 25;362(1):139-50
17254623
J Virol. 2002 Oct;76(20):10109-21
12239285
J Virol. 1999 Dec;73(12):9928-33
10559305
Infect Immun. 2000 Oct;68(10):5525-9
10992449
Blood. 2002 Apr 15;99(8):2913-21
11929782
Mech Ageing Dev. 2008 May;129(5):271-81
18367233
Immunity. 2001 May;14(5):495-8
11371351
J Virol. 2006 Nov;80(21):10293-306
17041210
Nat Immunol. 2000 Nov;1(5):398-401
11062499
Nat Immunol. 2005 Feb;6(2):163-70
15654341
Nature. 2001 Oct 18;413(6857):732-8
11607032
Nature. 2001 Apr 26;410(6832):1099-103
11323673
Nat Immunol. 2004 Oct;5(10):987-95
15454922
J Virol. 2004 Aug;78(15):7867-73
15254159
Infect Immun. 2004 Nov;72(11):6676-9
15501801
J Virol. 2006 Apr;80(7):3416-27
16537609
Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5598-603
15034168
Virology. 1964 Jun;23:224-33
14192298
J Virol. 2005 Jan;79(1):401-9
15596833
Virology. 2002 Nov 10;303(1):15-32
12482655
Nat Immunol. 2006 Jun;7(6):549-51
16715062
J Virol. 2003 Apr;77(8):4588-96
12663765
J Gen Virol. 1999 Apr;80 ( Pt 4):823-833
10211949
Annu Rev Immunol. 2000;18:767-811
10837075
Nat Med. 2007 May;13(5):552-9
17479101
J Biol Chem. 2005 Mar 11;280(10):9482-8
15632166
J Virol. 2003 Jun;77(12):7124-30
12768033
J Infect Dis. 2003 Dec 1;188(11):1630-8
14639532
J Virol. 2002 Sep;76(17):8729-36
12163593
Immunol Res. 2005;33(1):83-101
16120974
J Exp Med. 1999 Apr 19;189(8):1315-28
10209048