TLR3 Is Essential for the Induction of Protective Immunity against Punta Toro Virus Infection by the Double-Stranded RNA (dsRNA), Poly(I:C12U), but not Poly(I:C): Differential Recognition of Synthetic dsRNA Molecules1

In the wake of RNA virus infections, dsRNA intermediates are often generated. These viral pathogen-associated molecular patterns can be sensed by a growing number of host cell cytosolic proteins and TLR3, which contribute to the induction of antiviral defenses. Recent evidence indicates that melanoma differentiation-associated gene-5 is the prominent host component mediating IFN production after exposure to the dsRNA analog, poly(I:C). We have previously reported that Punta Toro virus (PTV) infection in mice is exquisitely sensitive to treatment with poly(I:C12U), a dsRNA analog that has a superior safety profile while maintaining the beneficial activity of the parental poly(I:C) in the induction of innate immune responses. The precise host factor(s) mediating protective immunity following its administration remain to be elucidated. To assess the role of TLR3 in this process, mice lacking the receptor were used to investigate the induction of protective immunity, type I IFNs, and IL-6 following treatment. Unlike wild-type mice, those lacking TLR3 were not protected against PTV infection following poly(I:C12U) therapy and failed to produce IFN-α, IFN-β, and IL-6. In contrast, poly(I:C) treatment significantly protected TLR3−/− mice from lethal challenge despite some deficiencies in cytokine induction. There was no indication that the lack of protection was due to the fact that TLR3-deficient mice had a reduced capacity to fight infection because they were not found to be more susceptible to PTV. We conclude that TLR3 is essential to the induction of antiviral activity elicited by poly(I:C12U), which does not appear to be recognized by the cytosolic sensor of poly(I:C), melanoma differentiation-associated gene-5.

[1]  Gunther Hartmann,et al.  5'-Triphosphate RNA Is the Ligand for RIG-I , 2006, Science.

[2]  A. Pichlmair,et al.  RIG-I-Mediated Antiviral Responses to Single-Stranded RNA Bearing 5'-Phosphates , 2006, Science.

[3]  R. Flavell,et al.  TLR3 Deletion Limits Mortality and Disease Severity due to Phlebovirus Infection1 , 2006, The Journal of Immunology.

[4]  D. Smee,et al.  Recombinant Eimeria Protozoan Protein Elicits Resistance to Acute Phlebovirus Infection in Mice but Not Hamsters , 2006, Antimicrobial Agents and Chemotherapy.

[5]  Richard A Flavell,et al.  Essential role of mda-5 in type I IFN responses to polyriboinosinic:polyribocytidylic acid and encephalomyocarditis picornavirus. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Christine A. Biron,et al.  Type 1 Interferons and the Virus-Host Relationship: A Lesson in Détente , 2006, Science.

[7]  K. Ishii,et al.  Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses , 2006, Nature.

[8]  D. Smee,et al.  Protective immunity against acute phleboviral infection elicited through immunostimulatory cationic liposome-DNA complexes. , 2006, Antiviral research.

[9]  D. Davies,et al.  The dsRNA binding site of human Toll‐like receptor 3 , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Akira,et al.  Pathogen Recognition and Innate Immunity , 2006, Cell.

[11]  I. Wilson,et al.  Crystal Structure of Human Toll-Like Receptor 3 (TLR3) Ectodomain , 2005, Science.

[12]  N. Dimitrov,et al.  Protein from intestinal Eimeria protozoan stimulates IL‐12 release from dendritic cells, exhibits antitumor properties in vivo and is correlated with low intestinal tumorigenicity , 2005, International journal of cancer.

[13]  M. Colonna,et al.  Dendritic cells respond to influenza virus through TLR7‐ and PKR‐independent pathways , 2005, European journal of immunology.

[14]  C. Peters,et al.  Pathogenesis of a phleboviral infection (Punta Toro virus) in golden Syrian hamsters , 2005, Archives of Virology.

[15]  S. Goodbourn,et al.  The V proteins of paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-beta promoter. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Shizuo Akira,et al.  The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses , 2004, Nature Immunology.

[17]  D. Smee,et al.  Effect of Interferon-Alpha and Interferon-Inducers on West Nile Virus in Mouse and Hamster Animal Models , 2004, Antiviral chemistry & chemotherapy.

[18]  P. Carmeliet,et al.  The Interferon Inducer Ampligen [Poly(I)-Poly(C12U)] Markedly Protects Mice against Coxsackie B3 Virus-Induced Myocarditis , 2004, Antimicrobial Agents and Chemotherapy.

[19]  Richard A Flavell,et al.  Upregulation of costimulatory molecules induced by lipopolysaccharide and double-stranded RNA occurs by Trif-dependent and Trif-independent pathways , 2003, Nature Immunology.

[20]  A. Yamamoto,et al.  Subcellular Localization of Toll-Like Receptor 3 in Human Dendritic Cells , 2003, The Journal of Immunology.

[21]  H. Guzmán,et al.  Induction of severe disease in hamsters by two sandfly fever group viruses, Punta toro and Gabek Forest (Phlebovirus, Bunyaviridae), similar to that caused by Rift Valley fever virus. , 2003, The American journal of tropical medicine and hygiene.

[22]  S. Akira,et al.  Role of Adaptor TRIF in the MyD88-Independent Toll-Like Receptor Signaling Pathway , 2003, Science.

[23]  E. De Clercq,et al.  Interferons, Interferon Inducers, and Interferon-Ribavirin in Treatment of Flavivirus-Induced Encephalitis in Mice , 2003, Antimicrobial Agents and Chemotherapy.

[24]  D. Smee,et al.  Viruses of the Bunya- and Togaviridae families: potential as bioterrorism agents and means of control. , 2003, Antiviral research.

[25]  R. Flavell,et al.  Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3 , 2001, Nature.

[26]  D. Liggitt,et al.  Lipid-DNA complexes induce potent activation of innate immune responses and antitumor activity when administered intravenously. , 1999, Journal of immunology.

[27]  A. Aguzzi,et al.  Deficient signaling in mice devoid of double‐stranded RNA‐dependent protein kinase. , 1995, The EMBO journal.

[28]  D. Smee,et al.  Antiviral and immunomodulating inhibitors of experimentally-induced Punta Toro virus infections. , 1994, Antiviral research.

[29]  S. Ida,et al.  In vivo antiviral effects of mismatched double‐stranded RNA on duck hepatitis B virus , 1994, Journal of medical virology.

[30]  D. Smee,et al.  Potential Role of Immunomodulators for Treatment of Phlebovirus Infections of Animals a , 1992, Annals of the New York Academy of Sciences.

[31]  M. Kende,et al.  Effect of human, recombinant interleukin 2 on Punta Toro virus infections in C57BL/6 mice. , 1991, Antiviral research.

[32]  D. Smee,et al.  Prophylactic and therapeutic activities of 7-thia-8-oxoguanosine against Punta Toro virus infections in mice , 1991, Antiviral Research.

[33]  D. Gillespie,et al.  Detection of Poly(I): Poly(C12U), Mismatched Double‐stranded RNA, by Rapid Solution Hybridization: Blood Values after Intravenous Infusion , 1990, The Journal of pharmacy and pharmacology.

[34]  R. Sidwell,et al.  Effects of ribamidine, a 3-carboxamidine derivative of ribavirin, on experimentally induced Phlebovirus infections. , 1988, Antiviral research.

[35]  R. Sidwell,et al.  In vitro and in vivo Phlebovirus inhibition by ribavirin , 1988, Antimicrobial Agents and Chemotherapy.

[36]  J. Smith,et al.  Punta Toro virus infection of C57BL/6J mice: a model for phlebovirus-induced disease. , 1987, Microbial pathogenesis.

[37]  D. Montefiori,et al.  CLINICAL, IMMUNOLOGICAL, AND VIROLOGICAL EFFECTS OF AMPLIGEN, A MISMATCHED DOUBLE-STRANDED RNA, IN PATIENTS WITH AIDS OR AIDS-RELATED COMPLEX , 1987, The Lancet.

[38]  D. Gillespie,et al.  Molecular hybridization with RNA probes in concentrated solutions of guanidine thiocyanate. , 1987, Analytical biochemistry.

[39]  D. Montefiori,et al.  Antiviral activity of mismatched double-stranded RNA against human immunodeficiency virus in vitro. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[40]  J. J. Greene,et al.  Augmentation of human natural killer cell activity by polyinosinic acid-polycytidylic acid and its nontoxic mismatched analogues. , 1980, Journal of immunology.

[41]  Levy Hb.  Induction of interferon in vivo by polynucleotides. , 1977 .

[42]  H. Levy.  Induction of interferon in vivo by polynucleotides. , 1977, Texas reports on biology and medicine.

[43]  W. Carter,et al.  An integrated and comparative study of the antiviral effects and other biological properties of the polyinosinic-polycytidylic acid duplex and its mismatched analogues. III. Chronic effects and immunological features. , 1976, Molecular pharmacology.

[44]  J. Alderfer,et al.  An integrated and comparative study of the antiviral effects and other biological properties of the polyinosinic acid-polycytidylic acid and its mismatched analogues. , 1976, Molecular pharmacology.

[45]  P. Pitha,et al.  Structural requirements of the rI n -rC n complex for induction of human interferon. , 1972, Journal of molecular biology.

[46]  M. Hilleman,et al.  Inducers of interferon and host resistance. II. Multistranded synthetic polynucleotide complexes. , 1967, Proceedings of the National Academy of Sciences of the United States of America.

[47]  L. Reed,et al.  A SIMPLE METHOD OF ESTIMATING FIFTY PER CENT ENDPOINTS , 1938 .