TLR3-specific double-stranded RNA oligonucleotide adjuvants induce dendritic cell cross-presentation, CTL responses, and antiviral protection.
Maturation of dendritic cells (DC) to competent APC is essential for the generation of acquired immunity and is a major function of adjuvants. dsRNA, a molecular signature of viral infection, drives DC maturation by activating TLR3, but the size of dsRNA required to activate DC and the expression patterns of TLR3 protein in DC subsets have not been established. In this article, we show that cross-priming CD8α(+) and CD103(+) DC subsets express much greater levels of TLR3 than other DC. In resting DC, TLR3 is located in early endosomes and other intracellular compartments but migrates to LAMP1(+) endosomes on stimulation with a TLR3 ligand. Using homogeneous dsRNA oligonucleotides (ONs) ranging in length from 25 to 540 bp, we observed that a minimum length of ∼90 bp was sufficient to induce CD86, IL-12p40, IFN-β, TNF-α, and IL-6 expression, and to mature DC into APC that cross-presented exogenous Ags to CD8(+) T cells. TLR3 was essential for activation of DC by dsRNA ONs, and the potency of activation increased with dsRNA length and varied between DC subsets. In vivo, dsRNA ONs, in a size-dependent manner, served as adjuvants for the generation of Ag-specific CTL and for inducing protection against lethal challenge with influenza virus when given with influenza nucleoprotein as an immunogen. These results provide the basis for the development of TLR3-specific adjuvants capable of inducing immune responses tailored for viral pathogens.
Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Ivett
I
Jelinek
Joshua N
JN
Leonard
Graeme E
GE
Price
Kevin N
KN
Brown
Anna
A
Meyer-Manlapat
Paul K
PK
Goldsmith
Yan
Y
Wang
David
D
Venzon
Suzanne L
SL
Epstein
David M
DM
Segal
D056704:Adaptive Immunity / Q000235:genetics; D000276:Adjuvants, Immunologic / Q000627:therapeutic use*; D000818:Animals; D002478:Cells, Cultured; D018899:CpG Islands / Q000276:immunology; D045142:Cross-Priming / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D018984:Epitopes, T-Lymphocyte / Q000096:biosynthesis / Q000276:immunology* / Q000627:therapeutic use; D005260:Female; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009838:Oligodeoxyribonucleotides / Q000627:therapeutic use*; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D012330:RNA, Double-Stranded / Q000627:therapeutic use*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology; D051196:Toll-Like Receptor 3 / Q000172:deficiency / Q000276:immunology* / Q000627:therapeutic use
D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D018984:Epitopes, T-Lymphocyte; D007252:Influenza Vaccines; D009838:Oligodeoxyribonucleotides; D012330:RNA, Double-Stranded; C519703:TLR3 protein, mouse; D051196:Toll-Like Receptor 3
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