Establishment of a panel of in-house polyclonal antibodies for the diagnosis of enterovirus infections
The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin‐embedded tissues play an im...
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| Published in | Neuropathology Vol. 35; no. 2; pp. 107 - 121 |
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| Main Authors | , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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Blackwell Publishing Ltd
01.04.2015
Wiley Subscription Services, Inc John Wiley and Sons Inc |
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| Abstract | The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin‐embedded tissues play an important role in recognizing infectious agents in tissue samples, six in‐house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1 protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C. Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross‐reacted with non‐enteroviruses within the family Picornaviridae. However, all cross‐reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non‐structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally‐infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in‐house anti‐enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus. |
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| AbstractList | The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin-embedded tissues play an important role in recognizing infectious agents in tissue samples, six in-house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1 protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C. Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross-reacted with non-enteroviruses within the family Picornaviridae. However, all cross-reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non-structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally-infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in-house anti-enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus. The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin-embedded tissues play an important role in recognizing infectious agents in tissue samples, six in-house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1 protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C. Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross-reacted with non-enteroviruses within the family Picornaviridae. However, all cross-reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non-structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally-infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in-house anti-enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus.The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin-embedded tissues play an important role in recognizing infectious agents in tissue samples, six in-house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1 protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C. Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross-reacted with non-enteroviruses within the family Picornaviridae. However, all cross-reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non-structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally-infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in-house anti-enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus. The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin‐embedded tissues play an important role in recognizing infectious agents in tissue samples, six in‐house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71 , one was raised against the recombinant VP1 protein of coxsackievirus B3 , and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C . Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross‐reacted with non‐enteroviruses within the family Picornaviridae . However, all cross‐reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non‐structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally‐infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71 , coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in‐house anti‐enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B , echoviruses, enterovirus A71 and poliovirus. |
| Author | Sato, Yuko Suzuki, Tadaki Nagata, Noriyo Shimizu, Hiroyuki Nakajima, Noriko Iwata-Yoshikawa, Naoko Iwasaki, Takuya Yamashita, Teruo Minagawa, Hiroko Kotani, Osamu Koike, Satoshi Sata, Tetsutaro Taguchi, Fumihiro Hasegawa, Hideki |
| AuthorAffiliation | 1 Department of Pathology National Institute of Infectious Diseases Aichi Japan 2 Department of Virology II National Institute of Infectious Diseases Aichi Japan 4 Neurovirology Project Tokyo Metropolitan Institute of Medical Science Aichi Japan 5 Department of Microbiology and Medical Zoology Aichi Prefectural Institute of Public Health Aichi Japan 3 Department of Virology and Viral Infections Faculty of Veterinary Medicine Nippon Veterinary and Life Science University Aichi Japan |
| AuthorAffiliation_xml | – name: 2 Department of Virology II National Institute of Infectious Diseases Aichi Japan – name: 5 Department of Microbiology and Medical Zoology Aichi Prefectural Institute of Public Health Aichi Japan – name: 4 Neurovirology Project Tokyo Metropolitan Institute of Medical Science Aichi Japan – name: 1 Department of Pathology National Institute of Infectious Diseases Aichi Japan – name: 3 Department of Virology and Viral Infections Faculty of Veterinary Medicine Nippon Veterinary and Life Science University Aichi Japan |
| Author_xml | – sequence: 1 givenname: Osamu surname: Kotani fullname: Kotani, Osamu organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 2 givenname: Naoko surname: Iwata-Yoshikawa fullname: Iwata-Yoshikawa, Naoko organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 3 givenname: Tadaki surname: Suzuki fullname: Suzuki, Tadaki organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 4 givenname: Yuko surname: Sato fullname: Sato, Yuko organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 5 givenname: Noriko surname: Nakajima fullname: Nakajima, Noriko organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 6 givenname: Satoshi surname: Koike fullname: Koike, Satoshi organization: Neurovirology Project, Tokyo Metropolitan Institute of Medical Science, Aichi, Japan – sequence: 7 givenname: Takuya surname: Iwasaki fullname: Iwasaki, Takuya organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 8 givenname: Tetsutaro surname: Sata fullname: Sata, Tetsutaro organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 9 givenname: Teruo surname: Yamashita fullname: Yamashita, Teruo organization: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Aichi, Japan – sequence: 10 givenname: Hiroko surname: Minagawa fullname: Minagawa, Hiroko organization: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Aichi, Japan – sequence: 11 givenname: Fumihiro surname: Taguchi fullname: Taguchi, Fumihiro organization: Department of Virology and Viral Infections, Faculty of Veterinary Medicine, Nippon Veterinary and Life Science University, Aichi, Japan – sequence: 12 givenname: Hideki surname: Hasegawa fullname: Hasegawa, Hideki organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan – sequence: 13 givenname: Hiroyuki surname: Shimizu fullname: Shimizu, Hiroyuki organization: Department of Virology II, National Institute of Infectious Diseases, Aichi, Japan – sequence: 14 givenname: Noriyo surname: Nagata fullname: Nagata, Noriyo email: nnagata@niid.go.jp organization: Department of Pathology, National Institute of Infectious Diseases, Aichi, Japan |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25263613$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1089_mab_2019_0031 crossref_primary_10_1016_j_idcr_2021_e01059 crossref_primary_10_1371_journal_pone_0148184 crossref_primary_10_1128_JVI_00864_16 crossref_primary_10_1093_jnen_nlz124 crossref_primary_10_1186_s12879_021_06858_2 crossref_primary_10_1097_INF_0000000000004052 |
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| Copyright | 2014 Japanese Society of Neuropathology 2014 Japanese Society of Neuropathology. Copyright © 2015 Japanese Society of Neuropathology |
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| Keywords | paraffin-embedded tissue viral encephalitis immunohistochemistry enterovirus polyclonal antibody |
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Hansson SF, Korsgren S, Ponten F, Korsgren O. Enteroviruses and the pathogenesis of type 1 diabetes revisited: cross-reactivity of enterovirus capsid protein (VP1) antibodies with human mitochondrial proteins. J Pathol 2013; 229: 719-728. Trabelsi A, Grattard F, Nejmeddine M, Aouni M, Bourlet T, Pozzetto B. Evaluation of an enterovirus group-specific anti-VP1 monoclonal antibody, 5-D8/1, in comparison with neutralization and PCR for rapid identification of enteroviruses in cell culture. J Clin Microbiol 1995; 33: 2454-2457. Zoll J, Erkens HS, Lanke K et al. Saffold virus, a human Theiler's-like cardiovirus, is ubiquitous and causes infection early in life. PLOS Pathogen 2009; 5: e1000416. Shimizu H, Utama A, Yoshii K et al. Enterovirus 71 from fatal and nonfatal cases of hand, foot and mouth disease epidemics in Malaysia, Japan and Taiwan in 1997-1998. Jpn J Infect Dis 1999; 52: 12-15. Kallajoki M, Kalimo H, Wesslen L, Auvinen P, Hyypia T. In situ detection of enterovirus genomes in mouse myocardial tissue by ribonucleic acid probes. Lab Invest 1990; 63: 669-675. McKinney MD, Moon SJ, Kulesh DA, Larsen T, Schoepp RJ. Detection of viral RNA from paraffin-embedded tissues after prolonged formalin fixation. J Clin Virol 2009; 44: 39-42. Cree BC, Bernardini GL, Hays AP, Lowe G. A fatal case of coxsackievirus B4 meningoencephalitis. Arch Neurol 2003; 60: 107-112. Ho M, Chen ER, Hsu KH et al. An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group. N Engl J Med 1999; 341: 929-935. King A, Adams M, Lefkowitz E, Carstens E. Virus 2009; 44 2009; 47 2009; 40 2012; 167 2006; 78 2000; 48 2009; 83 1997; 350 1983; 1 1995; 33 1997; 2 2012; 12 2007; 38 2009; 2009 2000; 17 2000; 13 1995; 69 2010; 28 2004; 78 1993; 31 2012; 170 2003; 9 2008; 26 2008; 67 1999; 52 2003; 83 2011; 29 2005; 79 2012; 62 1988 2007; 125 1993; 46 2004; 85 1990; 33 2012 2013; 87 2013; 229 2010 2013; 85 1997; 67 2008; 14 2011; 30 1999; 341 2005; 43 2012; 38 1998; 133 1990; 80 2011; 8 2010; 84 1999 1990; 63 1990; 64 2010; 47 1991; 65 2006; 44 2002; 67 2005; 446 2011; 51 2000; 31 2000; 83 2009; 5 2003; 60 2012; 6 2007; 45 1987; 28 2012; 86 2012; 9 2012; 84 e_1_2_6_51_1 e_1_2_6_74_1 e_1_2_6_53_1 e_1_2_6_76_1 e_1_2_6_32_1 Klump WM (e_1_2_6_46_1) 1990; 64 e_1_2_6_30_1 e_1_2_6_72_1 Saiz JC (e_1_2_6_13_1) 1991; 65 Shimizu H (e_1_2_6_40_1) 1999; 52 e_1_2_6_36_1 e_1_2_6_59_1 e_1_2_6_11_1 e_1_2_6_34_1 e_1_2_6_17_1 e_1_2_6_15_1 e_1_2_6_38_1 e_1_2_6_57_1 e_1_2_6_62_1 e_1_2_6_64_1 e_1_2_6_43_1 e_1_2_6_20_1 e_1_2_6_41_1 e_1_2_6_60_1 e_1_2_6_9_1 e_1_2_6_5_1 e_1_2_6_7_1 Odoom JK (e_1_2_6_70_1) 2012; 12 e_1_2_6_24_1 e_1_2_6_49_1 e_1_2_6_3_1 e_1_2_6_22_1 e_1_2_6_66_1 e_1_2_6_28_1 e_1_2_6_26_1 e_1_2_6_47_1 e_1_2_6_68_1 e_1_2_6_52_1 e_1_2_6_73_1 e_1_2_6_54_1 e_1_2_6_75_1 e_1_2_6_10_1 e_1_2_6_31_1 e_1_2_6_50_1 e_1_2_6_71_1 Kallajoki M (e_1_2_6_33_1) 1990; 63 e_1_2_6_14_1 e_1_2_6_35_1 e_1_2_6_12_1 e_1_2_6_39_1 e_1_2_6_56_1 King A (e_1_2_6_18_1) 2012 e_1_2_6_16_1 e_1_2_6_37_1 e_1_2_6_63_1 Ezyaguirre EJ (e_1_2_6_19_1) 2010 e_1_2_6_42_1 e_1_2_6_65_1 e_1_2_6_21_1 e_1_2_6_61_1 Harlow E (e_1_2_6_55_1) 1999 Zwol AL (e_1_2_6_58_1) 2009; 2009 e_1_2_6_8_1 e_1_2_6_4_1 Harlow E (e_1_2_6_45_1) 1988 e_1_2_6_6_1 e_1_2_6_25_1 e_1_2_6_48_1 e_1_2_6_23_1 e_1_2_6_2_1 e_1_2_6_29_1 e_1_2_6_44_1 e_1_2_6_67_1 e_1_2_6_27_1 e_1_2_6_69_1 |
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| SubjectTerms | Animals Antibodies, Viral - immunology Antigens Capsid Proteins - immunology Coxsackievirus B3 Coxsackievirus Infections - diagnosis Coxsackievirus Infections - immunology Echovirus Infections - diagnosis Echovirus Infections - immunology Enterovirus Enterovirus - classification Enterovirus - immunology Enterovirus - isolation & purification Enterovirus Infections - diagnosis Enterovirus Infections - immunology Evaluation Studies as Topic Fluorescent Antibody Technique, Indirect Humans Immunohistochemistry Infections Medical research Mice Original paraffin-embedded tissue Picornaviridae Poliovirus Polyclonal antibodies polyclonal antibody Rodents Sensitivity and Specificity Serotyping viral encephalitis |
| Title | Establishment of a panel of in-house polyclonal antibodies for the diagnosis of enterovirus infections |
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