Chikungunya viruses that escape monoclonal antibody therapy are clinically attenuated, stable, and not purified in mosquitoes

Journal of Virology

Volumn 88, Issue 15, 2014, Pages 8213-8226

  Pal, Pankaj ^a   Fox, Julie M ^a   Hawman, David W ^b   Huang, Yan Jang S ^c   Messaoudi, Ilhem ^d,e,f,g   Kreklywich, Craig ^d   Denton, Michael ^d   Legasse, Alfred W ^f   Smith, Patricia P ^d   Johnson, Syd ^h   Axthelm, Michael K ^f   Vanlandingham, Dana L ^c   Streblow, Daniel N ^d,e,f   Higgs, Stephen ^c   Morrison, Thomas E ^b   Diamond, Michael S ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b UNIVERSITY OF COLORADO SCHOOL OF MEDICINE   (United States)

^c KANSAS STATE UNIVERSITY   (United States)

^d OREGON HEALTH AND SCIENCE UNIVERSITY   (United States)

^e OREGON HEALTH AND SCIENCE UNIVERSITY   (United States)

^f OREGON HEALTH AND SCIENCE UNIVERSITY   (United States)

^g UNIVERSITY OF CALIFORNIA   (United States)

^h Analytical Sciences   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

MONOCLONAL ANTIBODY; MONOCLONAL ANTIBODY CHK 152; MONOCLONAL ANTIBODY CHK 166; UNCLASSIFIED DRUG;

AEDES ALBOPICTUS; ANIMAL EXPERIMENT; ANIMAL MODEL; ANTIVIRAL RESISTANCE; ARTICLE; CHIKUNGUNYA; CHIKUNGUNYA ALPHAVIRUS; CONTROLLED STUDY; DRUG EFFICACY; DRUG RESPONSE; MOUSE; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; SURVIVAL TIME; VIRUS LOAD; VIRUS MUTATION; VIRUS NEUTRALIZATION; VIRUS REPLICATION; VIRUS RESISTANCE; VIRUS STRAIN; VIRUS SURVIVAL; VIRUS TITRATION; VIRUS TRANSMISSION;

AEDES; ALPHAVIRUS INFECTIONS; ANIMALS; ANTIBODIES, MONOCLONAL; ANTIBODIES, VIRAL; CHIKUNGUNYA VIRUS; DISEASE MODELS, ANIMAL; FEMALE; HOMEODOMAIN PROTEINS; IMMUNOTHERAPY; MACACA MULATTA; MALE; MICE; MICE, INBRED C57BL; MICE, KNOCKOUT; SURVIVAL ANALYSIS; VIRULENCE; VIRUS REPLICATION;

EID: 84903890722     PISSN: 0022538X     EISSN: 10985514     Source Type: Journal    
DOI: 10.1128/JVI.01032-14     Document Type: Article

References (66)

1. Ross RW. 1956. The Newala epidemic. III. The virus: isolation, pathogenic properties and relationship to the epidemic. J. Hyg. (Lond.) 54:177-191.
2. Powers AM, Logue CH. 2007. Changing patterns of chikungunya virus: re-emergence of a zoonotic arbovirus. J. Gen. Virol. 88:2363-2377. http://dx.doi.org/10.1099/vir.0.82858-0.
3. Staples JE, Breiman RF, Powers AM. 2009. Chikungunya fever: an epidemiological review of a re-emerging infectious disease. Clin. Infect. Dis. 49:942-948. http://dx.doi.org/10.1086/605496.
4. Kariuki Njenga M, Nderitu L, Ledermann JP, Ndirangu A, Logue CH, Kelly CH, Sang R, Sergon K, Breiman R, Powers AM. 2008. Tracking epidemic Chikungunya virus into the Indian Ocean from East Africa. J. Gen. Virol. 89:2754-2760. http://dx.doi.org/10.1099/vir.0.2008/005413-0.
5. Borgherini G, Poubeau P, Jossaume A, Gouix A, Cotte L, Michault A, Arvin-Berod C, Paganin F. 2008. Persistent arthralgia associated with chikungunya virus: a study of 88 adult patients on Reunion Island. Clin. Infect. Dis. 47:469-475. http://dx.doi.org/10.1086/590003.
6. Angelini R, Finarelli AC, Angelini P, Po C, Petropulacos K, Silvi G, Macini P, Fortuna C, Venturi G, Magurano F, Fiorentini C, Marchi A, Benedetti E, Bucci P, Boros S, Romi R, Majori G, Ciufolini MG, Nicoletti L, Rezza G, Cassone A. 2007. Chikungunya in north-eastern Italy: a summing up of the outbreak. Euro Surveill. 12:E0711220.2. http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=3313.
7. Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli AC, Panning M, Cordioli P, Fortuna C, Boros S, Magurano F, Silvi G, Angelini P, Dottori M, Ciufolini MG, Majori GC, Cassone A. 2007. Infection with Chikungunya virus in Italy: an outbreak in a temperate region. Lancet 370:1840-1846. http://dx.doi.org/10.1016/S0140-6736(07)61779-6.
8. Tomasello D, Schlagenhauf P. 2013. Chikungunya and dengue autochthonous cases in Europe, 2007-2012. Travel Med. Infect. Dis. 11:274-284. http://dx.doi.org/10.1016/j.tmaid.2013.07.006.
9. Solignat M, Gay B, Higgs S, Briant L, Devaux C. 2009. Replication cycle of Chikungunya: a re-emerging arbovirus. Virology 393:183-197. http://dx.doi.org/10.1016/j.virol.2009.07.024.
10. Weaver SC, Osorio JE, Livengood JA, Chen R, Stinchcomb DT. 2012. Chikungunya virus and prospects for a vaccine. Expert Rev. Vaccines 11: 1087-1101. http://dx.doi.org/10.1586/erv.12.84.
11. Tsetsarkin KA, Vanlandingham DL, McGee CE, Higgs S. 2007. A single mutation in chikungunya virus affects vector specificity and epidemic potential. PLoS Pathog. 3:e201. http://dx.doi.org/10.1371/journal.ppat.0030201.
12. Benedict MQ, Levine RS, Hawley WA, Lounibos LP. 2007. Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. Vector Borne Zoonotic Dis. 7:76-85. http://dx.doi.org/10.1089/vbz.2006.0562.
13. Charrel RN, de Lamballerie X, Raoult D. 2007. Chikungunya outbreaks-the globalization of vectorborne diseases. N. Engl. J. Med. 356: 769-771. http://dx.doi.org/10.1056/NEJMp078013.
14. Suhrbier A, Jaffar-Bandjee MC, Gasque P. 2012. Arthritogenic alphaviruses-an overview. Nat. Rev. Rheumatol. 8:420-429. http://dx.doi.org/10.1038/nrrheum.2012.64.
15. Queyriaux B, Armengaud A, Jeannin C, Couturier E, Peloux-Petiot F. 2008. Chikungunya in Europe. Lancet 371:723-724. http://dx.doi.org/10.1016/S0140-6736(08)60336-0.
16. Jaffar-Bandjee MC, Ramful D, Gauzere BA, Hoarau JJ, Krejbich-Trotot P, Robin S, Ribera A, Selambarom J, Gasque P. 2010. Emergence and clinical insights into the pathology of Chikungunya virus infection. Expert Rev. Anti. Infect. Ther. 8:987-996. http://dx.doi.org/10.1586/eri.10.92.
17. Waymouth HE, Zoutman DE, Towheed TE. 2013. Chikungunya-related arthritis: case report and review of the literature. Semin. Arthritis Rheum. 43:273-278. http://dx.doi.org/10.1016/j.semarthrit.2013.03.003.
18. Schilte C, Staikovsky F, Couderc T, Madec Y, Carpentier F, Kassab S, Albert ML, Lecuit M, Michault A. 2013. Chikungunya virus-associated long-term arthralgia: a 36-month prospective longitudinal study. PLoS Negl. Trop. Dis. 7:e2137. http://dx.doi.org/10.1371/journal.pntd.0002137.
19. Powers AM, Brault AC, Tesh RB, Weaver SC. 2000. Re-emergence of Chikungunya and O'Nyong-nyong viruses: evidence for distinct geographical lineages and distant evolutionary relationships. J. Gen. Virol. 81:471-479.
20. Arankalle VA, Shrivastava S, Cherian S, Gunjikar RS, Walimbe AM, Jadhav SM, Sudeep AB, Mishra AC. 2007. Genetic divergence of Chikungunya viruses in India (1963-2006) with special reference to the 2005-2006 explosive epidemic. J. Gen. Virol. 88:1967-1976. http://dx.doi.org/10.1099/vir.0.82714-0.
21. Levis R, Schlesinger S, Huang HV. 1990. Promoter for Sindbis virus RNA-dependent subgenomic RNA transcription. J. Virol. 64:1726-1733.
22. Mukhopadhyay S, Zhang W, Gabler S, Chipman PR, Strauss EG, Strauss JH, Baker TS, Kuhn RJ, Rossmann MG. 2006. Mapping the structure and function of the E1 and E2 glycoproteins in alphaviruses. Structure 14:63-73. http://dx.doi.org/10.1016/j.str.2005.07.025.
23. Voss JE, Vaney MC, Duquerroy S, Vonrhein C, Girard-Blanc C, Crublet E, Thompson A, Bricogne G, Rey FA. 2010. Glycoprotein organization of Chikungunya virus particles revealed by X-ray crystallography. Nature 468:709-712. http://dx.doi.org/10.1038/nature09555.
24. Li L, Jose J, Xiang Y, Kuhn RJ, Rossmann MG. 2010. Structural changes of envelope proteins during alphavirus fusion. Nature 468:705-708. http://dx.doi.org/10.1038/nature09546.
25. Lescar J, Roussel A, Wien MW, Navaza J, Fuller SD, Wengler G, Rey FA. 2001. The Fusion glycoprotein shell of Semliki Forest virus: an icosahedral assembly primed for fusogenic activation at endosomal pH. Cell 105:137-148. http://dx.doi.org/10.1016/S0092-8674(01)00303-8.
26. Roussel A, Lescar J, Vaney MC, Wengler G, Wengler G, Rey FA. 2006. Structure and interactions at the viral surface of the envelope protein E1 of Semliki Forest virus. Structure 14:75-86. http://dx.doi.org/10.1016/j.str.2005.09.014.
27. Ziegler SA, Lu L, da Rosa AP, Xiao SY, Tesh RB. 2008. An animal model for studying the pathogenesis of chikungunya virus infection. Am. J. Trop. Med. Hyg. 79:133-139.
28. Couderc T, Chretien F, Schilte C, Disson O, Brigitte M, Guivel-Benhassine F, Touret Y, Barau G, Cayet N, Schuffenecker I, Despres P, Arenzana-Seisdedos F, Michault A, Albert ML, Lecuit M. 2008. A mouse model for Chikungunya: young age and inefficient type-I interferon signaling are risk factors for severe disease. PLoS Pathog. 4:e29. http://dx.doi.org/10.1371/journal.ppat.0040029.
29. Gardner J, Anraku I, Le TT, Larcher T, Major L, Roques P, Schroder WA, Higgs S, Suhrbier A. 2010. Chikungunya virus arthritis in adult wild-type mice. J. Virol. 84:8021-8032. http://dx.doi.org/10.1128/JVI.02603-09.
30. Morrison TE, Oko L, Montgomery SA, Whitmore AC, Lotstein AR, Gunn BM, Elmore SA, Heise MT. 2011. A mouse model of chikungunya virus-induced musculoskeletal inflammatory disease: evidence of arthritis, tenosynovitis, myositis, and persistence. Am. J. Pathol. 178:32-40. http://dx.doi.org/10.1016/j.ajpath.2010.11.018.
31. Hawman DW, Stoermer KA, Montgomery SA, Pal P, Oko L, Diamond MS, Morrison TE. 2013. Chronic joint disease caused by persistent chikungunya virus infection is controlled by the adaptive immune response. J. Virol. 87:13878-13888. http://dx.doi.org/10.1128/JVI.02666-13.
32. Schilte C, Couderc T, Chretien F, Sourisseau M, Gangneux N, Guivel-Benhassine F, Kraxner A, Tschopp J, Higgs S, Michault A, Arenzana-Seisdedos F, Colonna M, Peduto L, Schwartz O, Lecuit M, Albert ML. 2010. Type I IFN controls chikungunya virus via its action on nonhematopoietic cells. J. Exp. Med. 207:429-442. http://dx.doi.org/10.1084/jem.20090851.
33. Werneke SW, Schilte C, Rohatgi A, Monte KJ, Michault A, Arenzana-Seisdedos F, Vanlandingham DL, Higgs S, Fontanet A, Albert ML, Lenschow DJ. 2011. ISG15 is critical in the control of Chikungunya virus infection independent of UbE1L mediated conjugation. PLoS Pathog. 7:e1002322. http://dx.doi.org/10.1371/journal.ppat.1002322.
34. Gardner CL, Burke CW, Higgs ST, Klimstra WB, Ryman KD. 2012. Interferon-alpha/beta deficiency greatly exacerbates arthritogenic disease in mice infected with wild-type chikungunya virus but not with the cell culture-adapted live-attenuated 181/25 vaccine candidate. Virology 425: 103-112. http://dx.doi.org/10.1016/j.virol.2011.12.020.
35. Rudd PA, Wilson J, Gardner J, Larcher T, Babarit C, Le TT, Anraku I, Kumagai Y, Loo YM, Gale M, Jr, Akira S, Khromykh AA, Suhrbier A. 2012. Interferon response factors 3 and 7 protect against Chikungunya virus hemorrhagic fever and shock. J. Virol. 86:9888-9898. http://dx.doi.org/10.1128/JVI.00956-12.
36. Gardner CL, Hritz J, Sun C, Vanlandingham DL, Song TY, Ghedin E, Higgs S, Klimstra WB, Ryman KD. 2014. Deliberate attenuation of chikungunya virus by adaptation to heparan sulfate-dependent infectivity: a model for rational arboviral vaccine design. PLoS neglected tropical diseases 8:e2719. http://dx.doi.org/10.1371/journal.pntd.0002719.
37. Labadie K, Larcher T, Joubert C, Mannioui A, Delache B, Brochard P, Guigand L, Dubreil L, Lebon P, Verrier B, de Lamballerie X, Suhrbier A, Cherel Y, Le Grand R, Roques P. 2010. Chikungunya disease in nonhuman primates involves long-term viral persistence in macrophages. J. Clin. Invest. 120:894-906. http://dx.doi.org/10.1172/JCI40104.
38. Messaoudi I, Vomaske J, Totonchy T, Kreklywich CN, Haberthur K, Springgay L, Brien JD, Diamond MS, Defilippis VR, Streblow DN. 2013. Chikungunya virus infection results in higher and persistent viral replication in aged rhesus macaques due to defects in anti-viral immunity. PLoS Negl. Trop. Dis. 7:e2343. http://dx.doi.org/10.1371/journal.pntd.0002343.
39. Lum FM, Teo TH, Lee WW, Kam YW, Renia L, Ng LF. 2013. An essential role of antibodies in the control of Chikungunya virus infection. J. Immunol. 190:6295-6302. http://dx.doi.org/10.4049/jimmunol.1300304.
40. Rabinowitz SG, Adler WH. 1973. Host defenses during primary Venezuelan equine encephalomyelitis virus infection in mice. I. Passive transfer of protection with immune serum and immune cells. J. Immunol. 110: 1345-1353.
41. Boere WA, Benaissa-Trouw BJ, Harmsen M, Kraaijeveld CA, Snippe H. 1983. Neutralizing and non-neutralizing monoclonal antibodies to the E2 glycoprotein of Semliki Forest virus can protect mice from lethal encephalitis. J. Gen. Virol. 64:1405-1408. http://dx.doi.org/10.1099/0022-1317-64-6-1405.
42. Mathews JH, Roehrig JT, Trent DW. 1985. Role of complement and the Fc portion of immunoglobulin G in immunity to Venezuelan equine encephalomyelitis virus infection with glycoprotein-specific monoclonal antibodies. J. Virol. 55:594-600.
43. Hunt AR, Frederickson S, Hinkel C, Bowdish KS, Roehrig JT. 2006. A humanized murine monoclonal antibody protects mice either before or after challenge with virulent Venezuelan equine encephalomyelitis virus. J. Gen. Virol. 87:2467-2476. http://dx.doi.org/10.1099/vir.0.81925-0.
44. Johnson AJ, Hunt AR, Roehrig JT. 1991. Synthetic peptides of Venezuelan equine encephalomyelitis virus E2 glycoprotein. III. Identification of a protective peptide derived from the carboxy-terminal extramembranal one-third of the protein. Virology 185:840-842.
45. Phillpotts RJ. 2006. Venezuelan equine encephalitis virus complexspecific monoclonal antibody provides broad protection, in murine models, against airborne challenge with viruses from serogroups I, II and III. Virus Res. 120:107-112. http://dx.doi.org/10.1016/j.virusres.2006.02.003.
46. Levine B, Hardwick JM, Trapp BD, Crawford TO, Bollinger RC, Griffin DE. 1991. Antibody-mediated clearance of alphavirus infection from neurons. Science 254:856-860. http://dx.doi.org/10.1126/science.1658936.
47. Wust CJ, Nicholas JA, Fredin D, Dodd DC, Brideau RJ, Levely ME, Brown A. 1989. Monoclonal antibodies that cross-react with the E1 glycoprotein of different alphavirus serogroups: characterization including passive protection in vivo. Virus Res. 13:101-112. http://dx.doi.org/10.1016/0168-1702(89)90009-9.
48. Schmaljohn AL, Johnson ED, Dalrymple JM, Cole GA. 1982. Nonneutralizing monoclonal antibodies can prevent lethal alphavirus encephalitis. Nature 297:70-72. http://dx.doi.org/10.1038/297070a0.
49. Fric J, Bertin-Maghit S, Wang CI, Nardin A, Warter L. 2013. Use of human monoclonal antibodies to treat chikungunya virus infection. J. Infect. Dis. 207:319-322. http://dx.doi.org/10.1093/infdis/jis674.
50. Warter L, Lee CY, Thiagarajan R, Grandadam M, Lebecque S, Lin RT, Bertin-Maghit S, Ng LF, Abastado JP, Despres P, Wang CI, Nardin A. 2011. Chikungunya virus envelope-specific human monoclonal antibodies with broad neutralization potency. J. Immunol. 186:3258-3264. http://dx.doi.org/10.4049/jimmunol.1003139.
51. Lee CY, Kam YW, Fric J, Malleret B, Koh EG, Prakash C, Huang W, Lee WW, Lin C, Lin RT, Renia L, Wang CI, Ng LF, Warter L. 2011. Chikungunya virus neutralization antigens and direct cell-to-cell transmission are revealed by human antibody-escape mutants. PLoS Pathog. 7:e1002390. http://dx.doi.org/10.1371/journal.ppat.1002390.
52. Pal P, Dowd KA, Brien JD, Edeling MA, Gorlatov S, Johnson S, Lee I, Akahata W, Nabel GJ, Richter MKS, Smit JM, Fremont DH, Pierson TC, Heise MT, Diamond MS. 2013. Development of a highly protective combination monoclonal antibody therapy against Chikungunya virus. PLoS Pathog. 9:e1003312. http://dx.doi.org/10.1371/journal.ppat.1003312.
53. Sun S, Xiang Y, Wataru A, Holdaway HA, Pal P, Zheng X, Diamond MS, Nabel GJ, Rossmann MG. 2013. Structural analyses at pseudo atomic resolution of Chikungunya virus antibody neutralization mechanisms. eLife 2:e00435. http://dx.doi.org/10.7554/eLife.00435.
54. Selvarajah S, Sexton NR, Kahle KM, Fong RH, Mattia KA, Gardner J, Lu K, Liss NM, Salvador B, Tucker DF, Barnes T, Mabila M, Zhou X, Rossini G, Rucker JB, Sanders DA, Suhrbier A, Sambri V, Michault A, Muench MO, Doranz BJ, Simmons G. 2013. A neutralizing monoclonal antibody targeting the acid-sensitive region in chikungunya virus E2 protects from disease. PLoS Negl. Trop. Dis. 7:e2423. http://dx.doi.org/10.1371/journal.pntd.0002423.
55. Goh LY, Hobson-Peters J, Prow NA, Gardner J, Bielefeldt-Ohmann H, Pyke AT, Suhrbier A, Hall RA. 2013. Neutralizing monoclonal antibodies to the E2 protein of chikungunya virus protects against disease in a mouse model. Clin. Immunol. 149:487-497. http://dx.doi.org/10.1016/j.clim.2013.10.004.
56. Tsetsarkin K, Higgs S, McGee CE, De Lamballerie X, Charrel RN, Vanlandingham DL. 2006. Infectious clones of Chikungunya virus (La Reunion isolate) for vector competence studies. Vector Borne Zoonotic Dis. 6:325-337. http://dx.doi.org/10.1089/vbz.2006.6.325.
57. Schilte C, Buckwalter MR, Laird ME, Diamond MS, Schwartz O, Albert ML. 2012. Cutting edge: independent roles for IRF-3 and IRF-7 in hematopoietic and nonhematopoietic cells during host response to Chikungunya infection. J. Immunol. 188:2967-2971. http://dx.doi.org/10.4049/jimmunol.1103185.
58. Rulli NE, Rolph MS, Srikiatkhachorn A, Anantapreecha S, Guglielmotti A, Mahalingam S. 2011. Protection from arthritis and myositis in a mouse model of acute chikungunya virus disease by bindarit, an inhibitor of monocyte chemotactic protein-1 synthesis. J. Infect. Dis. 204:1026-1030. http://dx.doi.org/10.1093/infdis/jir470.
59. Gorchakov R, Wang E, Leal G, Forrester NL, Plante K, Rossi SL, Partidos CD, Adams AP, Seymour RL, Weger J, Borland EM, Sherman MB, Powers AM, Osorio JE, Weaver SC. 2012. Attenuation of Chikungunya virus vaccine strain 181/clone 25 is determined by two amino acid substitutions in the E2 envelope glycoprotein. J. Virol. 86:6084-6096. http://dx.doi.org/10.1128/JVI.06449-11.
60. Hoarau JJ, Jaffar Bandjee MC, Krejbich Trotot P, Das T, Li-Pat-Yuen G, Dassa B, Denizot M, Guichard E, Ribera A, Henni T, Tallet F, Moiton MP, Gauzere BA, Bruniquet S, Jaffar Bandjee Z, Morbidelli P, Martigny G, Jolivet M, Gay F, Grandadam M, Tolou H, Vieillard V, Debre P, Autran B, Gasque P. 2010. Persistent chronic inflammation and infection by Chikungunya arthritogenic alphavirus in spite of a robust host immune response. J. Immunol. 184:5914-5927. http://dx.doi.org/10.4049/jimmunol.0900255.
61. Chow A, Her Z, Ong EK, Chen JM, Dimatatac F, Kwek DJ, Barkham T, Yang H, Renia L, Leo YS, Ng LF. 2011. Persistent arthralgia induced by Chikungunya virus infection is associated with interleukin-6 and granulocyte macrophage colony-stimulating factor. J. Infect. Dis. 203:149-157. http://dx.doi.org/10.1093/infdis/jiq042.
62. Rohatgi A, Corbo JC, Monte K, Higgs S, Vanlandingham DL, Kardon G, Lenschow DJ. 2014. Infection of myofibers contributes to the increased pathogenicity during infection with an epidemic strain of Chikungunya virus. J. Virol. 88:2414-2425. http://dx.doi.org/10.1128/JVI.02716-13.
63. + T cells during Chikungunya virus infection in mice. J. Immunol. 190:259-269. http://dx.doi.org/10.4049/jimmunol.1202177.
64. Silva LA, Khomandiak S, Ashbrook AW, Weller R, Heise MT, Morrison TE, Dermody TS. 2014. A single-amino-Acid polymorphism in chikungunya virus E2 glycoprotein influences glycosaminoglycan utilization. J. Virol. 88:2385-2397. http://dx.doi.org/10.1128/JVI.03116-13.
65. Nakaya HI, Gardner J, Poo YS, Major L, Pulendran B, Suhrbier A. 2012. Gene profiling of Chikungunya virus arthritis in a mouse model reveals significant overlap with rheumatoid arthritis. Arthritis Rheum. 64:3553-3563. http://dx.doi.org/10.1002/art.34631.
66. Long KM, Whitmore AC, Ferris MT, Sempowski GD, McGee C, Trollinger B, Gunn B, Heise MT. 2013. Dendritic cell immunoreceptor regulates Chikungunya virus pathogenesis in mice. J. Virol. 87:5697-5706. http://dx.doi.org/10.1128/JVI.01611-12.