Characterization and specificity of the linear epitope of the enterovirus 71 VP2 protein
Tóm tắt
Enterovirus 71 (EV71) has emerged as a major causative agent of hand, foot and mouth disease in the Asia-Pacific region over the last decade. Hand, foot and mouth disease can be caused by different etiological agents from the enterovirus family, mainly EV71 and coxsackieviruses, which are genetically closely related. Nevertheless, infection with EV71 may occasionally lead to high fever, neurologic complications and the emergence of a rapidly fatal syndrome of pulmonary edema associated with brainstem encephalitis. The rapid progression and high mortality of severe EV71 infection has highlighted the need for EV71-specific diagnostic and therapeutic tools. Monoclonal antibodies are urgently needed to specifically detect EV71 antigens from patient specimens early in the infection process. Furthermore, the elucidation of viral epitopes will contribute to the development of targeted therapeutics and vaccines. We have identified the monoclonal antibody 7C7 from a screen of hybridoma cells derived from mice immunized with the EV71-B5 strain. The linear epitope of 7C7 was mapped to amino acids 142-146 (EDSHP) of the VP2 capsid protein and was characterized in detail. Mutational analysis of the epitope showed that the aspartic acid to asparagine mutation of the EV71 subgenogroup A (BrCr strain) did not interfere with antibody recognition. In contrast, the serine to threonine mutation at position 144 of VP2, present in recently emerged EV71-C4 China strains, abolished antigenicity. Mice injected with this virus strain did not produce any antibodies against the VP2 protein. Immunofluorescence and Western blotting confirmed that 7C7 specifically recognized EV71 subgenogroups and did not cross-react to Coxsackieviruses 4, 6, 10, and 16. 7C7 was successfully used as a detection antibody in an antigen-capture ELISA assay. Detailed mapping showed that the VP2 protein of Enterovirus 71 contains a single, linear, non-neutralizing epitope, spanning amino acids 142-146 which are located in the VP2 protein's E-F loop. The S/T(144) mutation in this epitope confers a loss of VP2 antigenicity to some newly emerged EV71-C4 strains from China. The corresponding monoclonal antibody 7C7 was used successfully in an AC-ELISA and did not cross-react to coxsackieviruses 4, 6, 10, and 16 in immunofluorescence assay and Western blots. 7C7 is the first monoclonal antibody described, that can differentiate Coxsackievirus 16 from Enterovirus 71.
Tài liệu tham khảo
Pallansch M, Roos R: Enterovirus: polioviruses, coxsackieviruses, echoviruses and newer enteroviruses. In Fields Virology. Edited by: Knipe DM HP, Griffin DE, et al. 2007, Philadelphia: Lippincott Williams & Wilkins, 839-893. 5
Schmidt NJ, Lennette EH, Ho HH: An apparently new enterovirus isolated from patients with disease of the central nervous system. J Infect Dis. 1974, 129: 304-309. 10.1093/infdis/129.3.304.
WHO: Outbreak of hand, foot and mouth disease in Sarawak Cluster of deaths among infants and young children. Wkly Epidemiol Rec. 1997, 72: 211-212.
Simmonds P, Welch J: Frequency and dynamics of recombination within different species of human enteroviruses. J Virol. 2006, 80: 483-493. 10.1128/JVI.80.1.483-493.2006.
Brown BA, Oberste MS, Alexander JP, Kennett ML, Pallansch MA: Molecular epidemiology and evolution of enterovirus 71 strains isolated from 1970 to 1998. J Virol. 1999, 73: 9969-9975.
Herrero LJ, Lee CS, Hurrelbrink RJ, Chua BH, Chua KB, McMinn PC: Molecular epidemiology of enterovirus 71 in peninsular Malaysia, 1997-2000. Arch Virol. 2003, 148: 1369-1385. 10.1007/s00705-003-0100-2.
McMinn P, Lindsay K, Perera D, Chan HM, Chan KP, Cardosa MJ: Phylogenetic analysis of enterovirus 71 strains isolated during linked epidemics in Malaysia, Singapore, and Western Australia. J Virol. 2001, 75: 7732-7738. 10.1128/JVI.75.16.7732-7738.2001.
Singh S, Chow VT, Chan KP, Ling AE, Poh CL: RT-PCR, nucleotide, amino acid and phylogenetic analyses of enterovirus type 71 strains from Asia. J Virol Methods. 2000, 88: 193-204. 10.1016/S0166-0934(00)00185-3.
Ding NZ, Wang XM, Sun SW, Song Q, Li SN, He CQ: Appearance of mosaic enterovirus 71 in the 2008 outbreak of China. Virus Res. 2009, 145: 157-161. 10.1016/j.virusres.2009.06.006.
Chumakov M, Voroshilova M, Shindarov L, Lavrova I, Gracheva L, Koroleva G, Vasilenko S, Brodvarova I, Nikolova M, Gyurova S, et al: Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria. Arch Virol. 1979, 60: 329-340. 10.1007/BF01317504.
Nagy G, Takatsy S, Kukan E, Mihaly I, Domok I: Virological diagnosis of enterovirus type 71 infections: experiences gained during an epidemic of acute CNS diseases in Hungary in 1978. Arch Virol. 1982, 71: 217-227. 10.1007/BF01314873.
King AMQ, Brown F, Christian P, Hovi T, Hyypia T, et al: Picornaviridae Virus Taxonomy Seventh Report of the International Committee for the Taxonomy of Viruses pp 657-673. 2000, New York: Academic, 657-673.
Mertens T, Pika U, Eggers HJ: Cross antigenicity among enteroviruses as revealed by immunoblot technique. Virology. 1983, 129: 431-442. 10.1016/0042-6822(83)90181-2.
Chang LY, Lin TY, Huang YC, Tsao KC, Shih SR, Kuo ML, Ning HC, Chung PW, Kang CM: Comparison of enterovirus 71 and coxsackie-virus A16 clinical illnesses during the Taiwan enterovirus epidemic, 1998. Pediatr Infect Dis J. 1999, 18: 1092-1096. 10.1097/00006454-199912000-00013.
Chan KP, Goh KT, Chong CY, Teo ES, Lau G, Ling AE: Epidemic hand, foot and mouth disease caused by human enterovirus 71, Singapore. Emerg Infect Dis. 2003, 9: 78-85.
Lum LC, Wong KT, Lam SK, Chua KB, Goh AY, Lim WL, Ong BB, Paul G, AbuBakar S, Lambert M: Fatal enterovirus 71 encephalomyelitis. J Pediatr. 1998, 133: 795-798. 10.1016/S0022-3476(98)70155-6.
Centers for Disease Control T: The activity of human enterovirus in Taiwan between 1998 and 2006. 2009, [http://www.cdc.gov.tw/public/Attachment/7121014355971.pdf]
Chang LY, Lin TY, Hsu KH, Huang YC, Lin KL, Hsueh C, Shih SR, Ning HC, Hwang MS, Wang HS, Lee CY: Clinical features and risk factors of pulmonary oedema after enterovirus-71-related hand, foot, and mouth disease. Lancet. 1999, 354: 1682-1686. 10.1016/S0140-6736(99)04434-7.
McMinn PC: An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev. 2002, 26: 91-107. 10.1111/j.1574-6976.2002.tb00601.x.
Chow M, Newman JF, Filman D, Hogle JM, Rowlands DJ, Brown F: Myristylation of picornavirus capsid protein VP4 and its structural significance. Nature. 1987, 327: 482-486. 10.1038/327482a0.
Chang HW, Liu CC, Lin MH, Ho HM, Yang YT, Chow YH, Chong P, Sia C: Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates. J Virol Methods. 2011, 173: 189-195. 10.1016/j.jviromet.2011.02.003.
Oberste MS, Maher K, Kilpatrick DR, Pallansch MA: Molecular evolution of the human enteroviruses: correlation of serotype with VP1 sequence and application to picornavirus classification. J Virol. 1999, 73: 1941-1948.
Tan CS, Cardosa MJ: High-titred neutralizing antibodies to human enterovirus 71 preferentially bind to the N-terminal portion of the capsid protein VP1. Arch Virol. 2007, 152: 1069-1073. 10.1007/s00705-007-0941-1.
Foo DG, Alonso S, Phoon MC, Ramachandran NP, Chow VT, Poh CL: Identification of neutralizing linear epitopes from the VP1 capsid protein of Enterovirus 71 using synthetic peptides. Virus Res. 2007, 125: 61-68. 10.1016/j.virusres.2006.12.005.
Oberste MS, Maher K, Kilpatrick DR, Flemister MR, Brown BA, Pallansch MA: Typing of human enteroviruses by partial sequencing of VP1. J Clin Microbiol. 1999, 37: 1288-1293.
Minor PD, Ferguson M, Evans DM, Almond JW, Icenogle JP: Antigenic structure of polioviruses of serotypes 1, 2 and 3. J Gen Virol. 1986, 67 (Pt 7): 1283-1291.
Fiore L, Ridolfi B, Genovese D, Buttinelli G, Lucioli S, Lahm A, Ruggeri FM: Poliovirus Sabin type 1 neutralization epitopes recognized by immunoglobulin A monoclonal antibodies. J Virol. 1997, 71: 6905-6912.
Buttinelli G, Donati V, Ruggeri FM, Joki-Korpela P, Hyypia T, Fiore L: Antigenic sites of coxsackie A9 virus inducing neutralizing monoclonal antibodies protective in mice. Virology. 2003, 312: 74-83. 10.1016/S0042-6822(03)00182-X.
Frimann TH, Barfoed AM, Aasted B, Kamstrup S: Vaccination of mice with plasmids expressing processed capsid protein of foot-and-mouth disease virus-importance of dominant and subdominant epitopes for antigenicity and protection. Vaccine. 2007, 25: 6191-6200. 10.1016/j.vaccine.2007.06.002.
Joki-Korpela P, Roivainen M, Lankinen H, Poyry T, Hyypia T: Antigenic properties of human parechovirus 1. J Gen Virol. 2000, 81: 1709-1718.
Alho A, Marttila J, Ilonen J, Hyypia T: Diagnostic potential of parechovirus capsid proteins. J Clin Microbiol. 2003, 41: 2294-2299. 10.1128/JCM.41.6.2294-2299.2003.
Abed Y, Wolf D, Dagan R, Boivin G: Development of a serological assay based on a synthetic peptide selected from the VP0 capsid protein for detection of human parechoviruses. J Clin Microbiol. 2007, 45: 2037-2039. 10.1128/JCM.02432-06.
Wu Y, Yeo A, Phoon MC, Tan EL, Poh CL, Quak SH, Chow VT: The largest outbreak of hand; foot and mouth disease in Singapore in 2008: The role of enterovirus 71 and coxsackievirus A strains. Int J Infect Dis. 2010, 14: e1076-e1081. 10.1016/j.ijid.2010.07.006.
Meng T, Kolpe AB, Kiener TK, Chow VT, Kwang J: Display of VP1 on the Surface of Baculovirus and Its Immunogenicity against Heterologous Human Enterovirus 71 Strains in Mice. PLoS One. 2011, 6: e21757-10.1371/journal.pone.0021757.
WHO: A guide to clinical management and public health response for hand, foot and mouth disease. 2011
Lum LC, Wong KT, Lam SK, Chua KB, Goh AY: Neurogenic pulmonary oedema and enterovirus 71 encephalomyelitis. Lancet. 1998, 352: 1391-
Tan EL, Yong LL, Quak SH, Yeo WC, Chow VT, Poh CL: Rapid detection of enterovirus 71 by real-time TaqMan RT-PCR. J Clin Virol. 2008, 42: 203-206. 10.1016/j.jcv.2008.01.001.
Sherry B, Mosser AG, Colonno RJ, Rueckert RR: Use of monoclonal antibodies to identify four neutralization immunogens on a common cold picornavirus, human rhinovirus 14. J Virol. 1986, 57: 246-257.
Usherwood EJ, Nash AA: Lymphocyte recognition of picornaviruses. J Gen Virol. 1995, 76 (Pt 3): 499-508.
Pulli T, Lankinen H, Roivainen M, Hyypia T: Antigenic sites of coxsackievirus A9. Virology. 1998, 240: 202-212. 10.1006/viro.1997.8908.
Lea S, Hernandez J, Blakemore W, Brocchi E, Curry S, Domingo E, Fry E, Abu-Ghazaleh R, King A, Newman J, et al: The structure and antigenicity of a type C foot-and-mouth disease virus. Structure. 1994, 2: 123-139. 10.1016/S0969-2126(00)00014-9.
Rossmann MG, Arnold E, Erickson JW, Frankenberger EA, Griffith JP, Hecht HJ, Johnson JE, Kamer G, Luo M, Mosser AG, et al: Structure of a human common cold virus and functional relationship to other picornaviruses. Nature. 1985, 317: 145-153. 10.1038/317145a0.
Liu CC, Chou AH, Lien SP, Lin HY, Liu SJ, Chang JY, Guo MS, Chow YH, Yang WS, Chang KH, et al: Identification and characterization of a cross-neutralization epitope of Enterovirus 71. Vaccine. 2011, 29: 4362-4372. 10.1016/j.vaccine.2011.04.010.
Miao LY, Pierce C, Gray-Johnson J, DeLotell J, Shaw C, Chapman N, Yeh E, Schnurr D, Huang YT: Monoclonal antibodies to VP1 recognize a broad range of enteroviruses. J Clin Microbiol. 2009, 47: 3108-3113. 10.1128/JCM.00479-09.
Yokoyama WM: Production of monoclonal antibodies. Curr Protoc Cell Biol. 2001, 16 (16): 1-