메뉴 건너뛰기




Volumn 8, Issue 12, 2013, Pages

Identification of cytotoxic T lymphocyte epitopes on swine viruses: Multi-epitope design for universal T cell vaccine

Author keywords

[No Author keywords available]

Indexed keywords

EPITOPE; LEUKOCYTE ANTIGEN; PEPTIDE VACCINE; VIRUS VACCINE;

EID: 84892937198     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0084443     Document Type: Article
Times cited : (18)

References (34)
  • 1
    • 79551495241 scopus 로고    scopus 로고
    • Induction of foot-and-mouth disease virus-specific cytotoxic T cell killing by vaccination
    • doi: 10.1128/CVI.00417-10. PubMed: 21177918
    • Patch JR, Pedersen LE, Toka FN, Moraes M, Grubman MJ et al. (2011) Induction of foot-and-mouth disease virus-specific cytotoxic T cell killing by vaccination. Clin Vaccine Immunol 18: 280-288. doi: 10.1128/CVI.00417-10. PubMed: 21177918.
    • (2011) Clin Vaccine Immunol , vol.18 , pp. 280-288
    • Patch, J.R.1    Pedersen, L.E.2    Toka, F.N.3    Moraes, M.4    Grubman, M.J.5
  • 2
    • 83055188639 scopus 로고    scopus 로고
    • T-cell-inducing vaccines - What's the future
    • doi:10.1111/j.1365-2567.2011.03517.x. PubMed: 22044118
    • Gilbert SC (2012) T-cell-inducing vaccines - what's the future. Immunology 135: 19-26. doi:10.1111/j.1365-2567.2011.03517.x. PubMed: 22044118.
    • (2012) Immunology , vol.135 , pp. 19-26
    • Gilbert, S.C.1
  • 3
    • 80053585941 scopus 로고    scopus 로고
    • A human multi-epitope recombinant vaccinia virus as a universal T cell vaccine candidate against influenza virus
    • doi:10.1371/journal.pone.0025938. PubMed: 21998725
    • Goodman AG, Heinen PP, Guerra S, Vijayan A, Sorzano CO et al. (2011) A human multi-epitope recombinant vaccinia virus as a universal T cell vaccine candidate against influenza virus. PLOS ONE 6: e25938. doi:10.1371/journal.pone. 0025938. PubMed: 21998725.
    • (2011) PLOS ONE , vol.6
    • Goodman, A.G.1    Heinen, P.P.2    Guerra, S.3    Vijayan, A.4    Sorzano, C.O.5
  • 4
    • 80052891501 scopus 로고    scopus 로고
    • Cross-protective peptide vaccine against influenza A viruses developed in HLA-A*2402 human immunity model
    • doi: 10.1371/journal.pone.0024626. PubMed: 21949735
    • Ichihashi T, Yoshida R, Sugimoto C, Takada A, Kajino K (2011) Cross-protective peptide vaccine against influenza A viruses developed in HLA-A*2402 human immunity model. PLOS ONE 6: e24626. doi: 10.1371/journal.pone.0024626. PubMed: 21949735.
    • (2011) PLOS ONE , vol.6
    • Ichihashi, T.1    Yoshida, R.2    Sugimoto, C.3    Takada, A.4    Kajino, K.5
  • 5
    • 84877038783 scopus 로고    scopus 로고
    • Examination of influenza specific T cell responses after influenza virus challenge in individuals vaccinated with MVA-NP+M1 vaccine
    • doi:10.1371/journal.pone.0062778. PubMed: 23658773
    • Powell TJ, Peng Y, Berthoud TK, Blais ME, Lillie PJ et al. (2013) Examination of influenza specific T cell responses after influenza virus challenge in individuals vaccinated with MVA-NP+M1 vaccine. PLOS ONE 8: e62778. doi:10.1371/journal.pone.0062778. PubMed: 23658773.
    • (2013) PLOS ONE , vol.8
    • Powell, T.J.1    Peng, Y.2    Berthoud, T.K.3    Blais, M.E.4    Lillie, P.J.5
  • 6
    • 78049520061 scopus 로고    scopus 로고
    • Induction of a cross-reactive CD8(+) T cell response following foot-and-mouth disease virus vaccination
    • doi:10.1128/JVI.01545-10. PubMed: 20861264
    • Guzman E, Taylor G, Charleston B, Ellis SA (2010) Induction of a cross-reactive CD8(+) T cell response following foot-and-mouth disease virus vaccination. J Virol 84: 12375-12384. doi:10.1128/JVI.01545-10. PubMed: 20861264.
    • (2010) J Virol , vol.84 , pp. 12375-12384
    • Guzman, E.1    Taylor, G.2    Charleston, B.3    Ellis, S.A.4
  • 7
    • 33751259349 scopus 로고    scopus 로고
    • DNA immunization with 2C FMDV non-structural protein reveals the presence of an immunodominant CD8+, CTL epitope for Balb/c mice
    • doi:10.1016/j.antiviral.2006.07.002. PubMed: 16890298
    • Barfoed AM, Rodriguez F, Therrien D, Borrego B, Sobrino F et al. (2006) DNA immunization with 2C FMDV non-structural protein reveals the presence of an immunodominant CD8+, CTL epitope for Balb/c mice. Antiviral Res 72: 178-189. doi:10.1016/j.antiviral.2006.07.002. PubMed: 16890298.
    • (2006) Antiviral Res , vol.72 , pp. 178-189
    • Barfoed, A.M.1    Rodriguez, F.2    Therrien, D.3    Borrego, B.4    Sobrino, F.5
  • 8
    • 48649108828 scopus 로고    scopus 로고
    • Identification of immunodominant T-cell epitopes present in glycoprotein 5 of the North American genotype of porcine reproductive and respiratory syndrome virus
    • doi:10.1016/j.vaccine.2008.06.047. PubMed: 18590788
    • Vashisht K, Goldberg TL, Husmann RJ, Schnitzlein W, Zuckermann FA (2008) Identification of immunodominant T-cell epitopes present in glycoprotein 5 of the North American genotype of porcine reproductive and respiratory syndrome virus. Vaccine 26: 4747-4753. doi:10.1016/j.vaccine.2008.06.047. PubMed: 18590788.
    • (2008) Vaccine , vol.26 , pp. 4747-4753
    • Vashisht, K.1    Goldberg, T.L.2    Husmann, R.J.3    Schnitzlein, W.4    Zuckermann, F.A.5
  • 9
    • 84860320829 scopus 로고    scopus 로고
    • The utility and limitations of current web-available algorithms to predict peptides recognized by CD4 T cells in response to pathogen infection
    • doi:10.4049/jimmunol.1103640. PubMed: 22467652
    • Chaves FA, Lee AH, Nayak JL, Richards KA, Sant AJ (2012) The utility and limitations of current web-available algorithms to predict peptides recognized by CD4 T cells in response to pathogen infection. J Immunol 188: 4235-4248. doi:10.4049/jimmunol.1103640. PubMed: 22467652.
    • (2012) J Immunol , vol.188 , pp. 4235-4248
    • Chaves, F.A.1    Lee, A.H.2    Nayak, J.L.3    Richards, K.A.4    Sant, A.J.5
  • 10
    • 0036184702 scopus 로고    scopus 로고
    • Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus
    • PubMed: 11842250
    • Armengol E, Wiesmüller KH, Wienhold D, Büttner M, Pfaff E et al. (2002) Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus. J Gen Virol 83: 551-560. PubMed: 11842250.
    • (2002) J Gen Virol , vol.83 , pp. 551-560
    • Armengol, E.1    Wiesmüller, K.H.2    Wienhold, D.3    Büttner, M.4    Pfaff, E.5
  • 11
    • 84858708796 scopus 로고    scopus 로고
    • Characterisation of vaccine-induced, broadly cross-reactive IFN-gamma secreting T cell responses that correlate with rapid protection against classical swine fever virus
    • doi: 10.1016/j.vaccine.2012.02.029. PubMed: 22366027
    • Graham SP, Haines FJ, Johns HL, Sosan O, La Rocca SA et al. (2012) Characterisation of vaccine-induced, broadly cross-reactive IFN-gamma secreting T cell responses that correlate with rapid protection against classical swine fever virus. Vaccine 30: 2742-2748. doi: 10.1016/j.vaccine.2012.02.029. PubMed: 22366027.
    • (2012) Vaccine , vol.30 , pp. 2742-2748
    • Graham, S.P.1    Haines, F.J.2    Johns, H.L.3    Sosan, O.4    La Rocca, S.A.5
  • 12
    • 0029583635 scopus 로고
    • Classical swine fever virus-specific cytotoxic T lymphocytes and identification of a T cell epitope
    • doi: 10.1099/0022-1317-76-12-3039
    • Pauly T, Elbers K, Konig M, Lengsfeld T, Saalmuller A et al. (1995) Classical swine fever virus-specific cytotoxic T lymphocytes and identification of a T cell epitope. J Gen Virol 76 (12): 3039-3049. doi: 10.1099/0022-1317-76- 12-3039.
    • (1995) J Gen Virol , vol.76 , Issue.12 , pp. 3039-3049
    • Pauly, T.1    Elbers, K.2    Konig, M.3    Lengsfeld, T.4    Saalmuller, A.5
  • 13
    • 79951645476 scopus 로고    scopus 로고
    • Potent CD8+ T-cell immunogenicity in humans of a novel heterosubtypic influenza A vaccine, MVA-NP+M1
    • doi:10.1093/cid/ciq039. PubMed: 21148512
    • Berthoud TK, Hamill M, Lillie PJ, Hwenda L, Collins KA et al. (2011) Potent CD8+ T-cell immunogenicity in humans of a novel heterosubtypic influenza A vaccine, MVA-NP+M1. Clin Infect Dis 52: 1-7. doi:10.1093/cid/ciq039. PubMed: 21148512.
    • (2011) Clin Infect Dis , vol.52 , pp. 1-7
    • Berthoud, T.K.1    Hamill, M.2    Lillie, P.J.3    Hwenda, L.4    Collins, K.A.5
  • 14
    • 84862219175 scopus 로고    scopus 로고
    • Preliminary assessment of the efficacy of a T-cell-based influenza vaccine, MVA-NP+M1, in humans
    • doi: 10.1093/cid/cis327. PubMed: 22441650
    • Lillie PJ, Berthoud TK, Powell TJ, Lambe T, Mullarkey C et al. (2012) Preliminary assessment of the efficacy of a T-cell-based influenza vaccine, MVA-NP+M1, in humans. Clin Infect Dis 55: 19-25. doi: 10.1093/cid/cis327. PubMed: 22441650.
    • (2012) Clin Infect Dis , vol.55 , pp. 19-25
    • Lillie, P.J.1    Berthoud, T.K.2    Powell, T.J.3    Lambe, T.4    Mullarkey, C.5
  • 15
    • 70349331592 scopus 로고    scopus 로고
    • Prediction of T cell epitopes for the utility of vaccine development from structural proteins of dengue virus variants using in silico methods
    • Somvanshi P, Seth PK (2009) Prediction of T cell epitopes for the utility of vaccine development from structural proteins of dengue virus variants using in silico methods. Indian Journal of Biotechnology 8: 193-198.
    • (2009) Indian Journal of Biotechnology , vol.8 , pp. 193-198
    • Somvanshi, P.1    Seth, P.K.2
  • 16
    • 77956210124 scopus 로고    scopus 로고
    • Immunogenicity of novel Dengue virus epitopes identified by bioinformatic analysis
    • doi:10.1016/j.virusres.2010.07.014. PubMed: 20638434
    • Sánchez-Burgos G, Ramos-Castañeda J, Cedillo-Rivera R, Dumonteil E (2010) Immunogenicity of novel Dengue virus epitopes identified by bioinformatic analysis. Virus Res 153: 113-120. doi:10.1016/j.virusres.2010.07. 014. PubMed: 20638434.
    • (2010) Virus Res , vol.153 , pp. 113-120
    • Sánchez-Burgos, G.1    Ramos-Castañeda, J.2    Cedillo-Rivera, R.3    Dumonteil, E.4
  • 17
    • 81255147901 scopus 로고    scopus 로고
    • Identification of potential human respiratory syncytial virus and metapneumovirus T cell epitopes using computational prediction and MHC binding assays
    • doi:10.1016/j.jim.2011.08.004. PubMed: 21854782
    • Rock MT, McKinney BA, Yoder SM, Prudom CE, Wright DW et al. (2011) Identification of potential human respiratory syncytial virus and metapneumovirus T cell epitopes using computational prediction and MHC binding assays. J Immunol Methods 374: 13-17. doi:10.1016/j.jim.2011.08.004. PubMed: 21854782.
    • (2011) J Immunol Methods , vol.374 , pp. 13-17
    • Rock, M.T.1    McKinney, B.A.2    Yoder, S.M.3    Prudom, C.E.4    Wright, D.W.5
  • 18
    • 59449094834 scopus 로고    scopus 로고
    • NetMHCpan, a method for MHC class I binding prediction beyond humans
    • doi:10.1007/s00251-008-0341-z. PubMed: 19002680
    • Hoof I, Peters B, Sidney J, Pedersen LE, Sette A et al. (2009) NetMHCpan, a method for MHC class I binding prediction beyond humans. Immunogenetics 61: 1-13. doi:10.1007/s00251-008-0341-z. PubMed: 19002680.
    • (2009) Immunogenetics , vol.61 , pp. 1-13
    • Hoof, I.1    Peters, B.2    Sidney, J.3    Pedersen, L.E.4    Sette, A.5
  • 20
    • 17844385106 scopus 로고    scopus 로고
    • The role of the proteasome in generating cytotoxic T-cell epitopes: Insights obtained from improved predictions of proteasomal cleavage
    • doi:10.1007/s00251-005-0781-7. PubMed: 15744535
    • Nielsen M, Lundegaard C, Lund O, Ke?mir C (2005) The role of the proteasome in generating cytotoxic T-cell epitopes: insights obtained from improved predictions of proteasomal cleavage. Immunogenetics 57: 33-41. doi:10.1007/s00251-005-0781-7. PubMed: 15744535.
    • (2005) Immunogenetics , vol.57 , pp. 33-41
    • Nielsen, M.1    Lundegaard, C.2    Lund, O.K.C.3
  • 21
    • 82755161001 scopus 로고    scopus 로고
    • Porcine major histocompatibility complex (MHC) class I molecules and analysis of their peptide-binding specificities
    • doi:10.1007/s00251-011-0555-3. PubMed: 21739336
    • Pedersen LE, Harndahl M, Rasmussen M, Lamberth K, Golde WT et al. (2011) Porcine major histocompatibility complex (MHC) class I molecules and analysis of their peptide-binding specificities. Immunogenetics 63: 821-834. doi:10.1007/s00251-011-0555-3. PubMed: 21739336.
    • (2011) Immunogenetics , vol.63 , pp. 821-834
    • Pedersen, L.E.1    Harndahl, M.2    Rasmussen, M.3    Lamberth, K.4    Golde, W.T.5
  • 22
    • 84892944837 scopus 로고    scopus 로고
    • Identification of peptides from foot-and-mouth disease virus structural proteins bound by class I swine leukocyte antigen (SLA) alleles, SLA-1*0401 and SLA-2*0401
    • Pedersen LE, Harndahl M, Nielsen M, Patch JR, Jungersen G, et al. (2012) Identification of peptides from foot-and-mouth disease virus structural proteins bound by class I swine leukocyte antigen (SLA) alleles, SLA-1*0401 and SLA-2*0401. Anim Genet.
    • (2012) Anim Genet
    • Pedersen, L.E.1    Harndahl, M.2    Nielsen, M.3    Patch, J.R.4    Jungersen, G.5
  • 23
    • 33747826322 scopus 로고    scopus 로고
    • Reconstruction of a swine SLA-I protein complex and determination of binding nonameric peptides derived from the foot-and-mouth disease virus
    • doi:10.1016/j.vetimm.2006.06.002. PubMed: 16870265
    • Gao FS, Fang QM, Li YG, Li XS, Hao HF et al. (2006) Reconstruction of a swine SLA-I protein complex and determination of binding nonameric peptides derived from the foot-and-mouth disease virus. Vet Immunol Immunopathol 113: 328-338. doi:10.1016/j.vetimm.2006.06.002. PubMed: 16870265.
    • (2006) Vet Immunol Immunopathol , vol.113 , pp. 328-338
    • Gao, F.S.1    Fang, Q.M.2    Li, Y.G.3    Li, X.S.4    Hao, H.F.5
  • 24
    • 68949166049 scopus 로고    scopus 로고
    • In silico prediction and ex vivo evaluation of potential T-cell epitopes in glycoproteins 4 and 5 and nucleocapsid protein of genotype-I (European) of porcine reproductive and respiratory syndrome virus
    • doi:10.1016/j.vaccine.2009.07.029. PubMed: 19646408
    • Díaz I, Pujols J, Ganges L, Gimeno M, Darwich L et al. (2009) In silico prediction and ex vivo evaluation of potential T-cell epitopes in glycoproteins 4 and 5 and nucleocapsid protein of genotype-I (European) of porcine reproductive and respiratory syndrome virus. Vaccine 27: 5603-5611. doi:10.1016/j.vaccine.2009.07.029. PubMed: 19646408.
    • (2009) Vaccine , vol.27 , pp. 5603-5611
    • Díaz, I.1    Pujols, J.2    Ganges, L.3    Gimeno, M.4    Darwich, L.5
  • 25
    • 23844439739 scopus 로고    scopus 로고
    • Identification of classical swine fever virus protein E2 as a target for cytotoxic T cells by using mRNA-transfected antigen-presenting cells
    • doi:10.1099/vir.0.80907-0. PubMed: 16099911
    • Ceppi M, de Bruin MG, Seuberlich T, Balmelli C, Pascolo S et al. (2005) Identification of classical swine fever virus protein E2 as a target for cytotoxic T cells by using mRNA-transfected antigen-presenting cells. J Gen Virol 86: 2525-2534. doi:10.1099/vir.0.80907-0. PubMed: 16099911.
    • (2005) J Gen Virol , vol.86 , pp. 2525-2534
    • Ceppi, M.1    De Bruin, M.G.2    Seuberlich, T.3    Balmelli, C.4    Pascolo, S.5
  • 26
    • 79955977371 scopus 로고    scopus 로고
    • Identification of two internal signal peptide sequences: Critical for classical swine fever virus non-structural protein 2 to trans-localize to the endoplasmic reticulum
    • doi:10.1186/1743-422X-8-236. PubMed: 21592340
    • Guo KK, Tang QH, Zhang YM, Kang K, He L (2011) Identification of two internal signal peptide sequences: critical for classical swine fever virus non-structural protein 2 to trans-localize to the endoplasmic reticulum. Virol J 8: 236. doi:10.1186/1743-422X-8-236. PubMed: 21592340.
    • (2011) Virol J , vol.8 , pp. 236
    • Guo, K.K.1    Tang, Q.H.2    Zhang, Y.M.3    Kang, K.4    He, L.5
  • 27
    • 80455128598 scopus 로고    scopus 로고
    • Immunogenicity of recombinant classic swine fever virus CD8(+) T lymphocyte epitope and porcine parvovirus VP2 antigen coexpressed by Lactobacillus casei in swine via oral vaccination
    • doi:10.1128/CVI.05204-11. PubMed: 21940406
    • Xu Y, Cui L, Tian C, Zhang G, Huo G et al. (2011) Immunogenicity of recombinant classic swine fever virus CD8(+) T lymphocyte epitope and porcine parvovirus VP2 antigen coexpressed by Lactobacillus casei in swine via oral vaccination. Clin Vaccine Immunol 18: 1979-1986. doi:10.1128/CVI.05204-11. PubMed: 21940406.
    • (2011) Clin Vaccine Immunol , vol.18 , pp. 1979-1986
    • Xu, Y.1    Cui, L.2    Tian, C.3    Zhang, G.4    Huo, G.5
  • 28
    • 79955655632 scopus 로고    scopus 로고
    • Complex assembly, crystallization and preliminary X-ray crystallographic studies of the swine major histocompatibility complex molecule SLA-1*1502
    • doi:10.1107/S174430911100741X. PubMed: 21543863
    • Pan X, Qi J, Zhang N, Li Q, Yin C et al. (2011) Complex assembly, crystallization and preliminary X-ray crystallographic studies of the swine major histocompatibility complex molecule SLA-1*1502. Acta Crystallogr Sect F Struct Biol Cryst Commun 67: 568-571. doi:10.1107/S174430911100741X. PubMed: 21543863.
    • (2011) Acta Crystallogr Sect F Struct Biol Cryst Commun , vol.67 , pp. 568-571
    • Pan, X.1    Qi, J.2    Zhang, N.3    Li, Q.4    Yin, C.5
  • 29
    • 79960630749 scopus 로고    scopus 로고
    • Characterization of antigenic regions in the porcine reproductive and respiratory syndrome virus by the use of peptide-specific serum antibodies
    • doi:10.1016/j.vaccine.2011.04.071. PubMed: 21554913
    • Vanhee M, Van Breedam W, Costers S, Geldhof M, Noppe Y et al. (2011) Characterization of antigenic regions in the porcine reproductive and respiratory syndrome virus by the use of peptide-specific serum antibodies. Vaccine 29: 4794-4804. doi:10.1016/j.vaccine.2011.04.071. PubMed: 21554913.
    • (2011) Vaccine , vol.29 , pp. 4794-4804
    • Vanhee, M.1    Van Breedam, W.2    Costers, S.3    Geldhof, M.4    Noppe, Y.5
  • 30
    • 0345352755 scopus 로고    scopus 로고
    • Bioinformatics tools for identifying class I-restricted epitopes
    • doi:10.1016/S1046-2023(02)00351-1. PubMed: 12725794
    • Martin W (2003) Bioinformatics tools for identifying class I-restricted epitopes. Methods 29: 289-298. doi:10.1016/S1046-2023(02)00351-1. PubMed: 12725794.
    • (2003) Methods , vol.29 , pp. 289-298
    • Martin, W.1
  • 31
    • 38049169092 scopus 로고    scopus 로고
    • Large-scale validation of methods for cytotoxic T-lymphocyte epitope prediction
    • doi:10.1186/1471-2105-8-424. PubMed: 17973982
    • Larsen MV, Lundegaard C, Lamberth K, Buus S, Lund O et al. (2007) Large-scale validation of methods for cytotoxic T-lymphocyte epitope prediction. BMC Bioinformatics 8: 424. doi:10.1186/1471-2105-8-424. PubMed: 17973982.
    • (2007) BMC Bioinformatics , vol.8 , pp. 424
    • Larsen, M.V.1    Lundegaard, C.2    Lamberth, K.3    Buus, S.4    Lund, O.5
  • 32
    • 80052876042 scopus 로고    scopus 로고
    • MimoPro: A more efficient Web-based tool for epitope prediction using phage display libraries
    • doi:10.1186/1471-2105-12-199. PubMed: 21609501
    • Chen WH, Sun PP, Lu Y, Guo WW, Huang YX et al. (2011) MimoPro: a more efficient Web-based tool for epitope prediction using phage display libraries. BMC Bioinformatics 12: 199. doi:10.1186/1471-2105-12-199. PubMed: 21609501.
    • (2011) BMC Bioinformatics , vol.12 , pp. 199
    • Chen, W.H.1    Sun, P.P.2    Lu, Y.3    Guo, W.W.4    Huang, Y.X.5
  • 33
    • 83155180577 scopus 로고    scopus 로고
    • Toward more accurate pan-specific MHC-peptide binding prediction: A review of current methods and tools
    • PubMed: 21949215
    • Zhang L, Udaka K, Mamitsuka H, Zhu S (2012) Toward more accurate pan-specific MHC-peptide binding prediction: a review of current methods and tools. Brief Bioinform, 13: 350-64. PubMed: 21949215.
    • (2012) Brief Bioinform , vol.13 , pp. 350-364
    • Zhang, L.1    Udaka, K.2    Mamitsuka, H.3    Zhu, S.4
  • 34
    • 77953149616 scopus 로고    scopus 로고
    • NetCTLpan: Pan-specific MHC class I pathway epitope predictions
    • doi:10.1007/s00251-010-0441-4. PubMed: 20379710
    • Stranzl T, Larsen MV, Lundegaard C, Nielsen M (2010) NetCTLpan: pan-specific MHC class I pathway epitope predictions. Immunogenetics 62: 357-368. doi:10.1007/s00251-010-0441-4. PubMed: 20379710.
    • (2010) Immunogenetics , vol.62 , pp. 357-368
    • Stranzl, T.1    Larsen, M.V.2    Lundegaard, C.3    Nielsen, M.4


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.