Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC)

Translated title of the contribution: Production and characterization of monoclonal antibodies to a bovine herpesvirus type 1 strain defective on the glycoprotein C

Evandro Reinoldo Winkelmann, Letícia Frizzo Da Silva, Sandra Vanderli Mayer, Ketty Cristina Mazzutti, Rudi Weiblen, Eduardo Furtado Flores

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Most monoclonal antibodies (MAbs) already produced against bovine herpesvirus type 1 (BoHV-1) react with glycoprotein C (gC), an abundant and immunodominant antigen present on the viral envelope. In order to obtain MAbs with other protein specificities, antigens of a BoHV-1 gC-negative strain were used to immunize BALB/c mice. After fusion and selection of 54 HAT-resistant hybridomas, three clones have been obtained (1F1, 2H4 and 4D7) that secrete IgG2a antibodies reacting to BoHV-1 antigens. These MAbs reacted with viral antigens in immunofluorescence (IFA) and immunoperoxidase (IPX) in dilutions up to 1:640 (hybridoma supernatants) and 1:20.000 (ascitis fluid). The three MAbs showed a wide spectrum of reactivity, recognizing antigens of 14 herpesviruses isolated from respiratory or genital disease (supposedly BoHV-1) and with 17 isolates of neurological disease (likely BoHV-5) and displayed varied levels of neutralizing activity against all these viruses. The protein specificity could not be demonstrated directly as none of the MAbs bound to viral antigens in Western blot. On the other hand, the three MAbs reacted with cells infected with a BoHV-5 strain defective in glycoproteins gE and gI, demonstrating they are directed to other viral proteins. By exclusion (gC, gE and gI) and due to their neutralizing activity, these MAbs are probably directed to conserved epitopes on other envelope glycoproteins harboring neutralizing epitopes: gB or gD. In this sense, besides being useful for diagnosis purposes, these MAbs may be very useful for mapping neutralizing epitopes in these glycoproteins.

Original languagePortuguese
Pages (from-to)1066-1072
Number of pages7
JournalCiencia Rural
Volume37
Issue number4
StatePublished - Jul 2007
Externally publishedYes

Fingerprint

Bovine Herpesvirus 1
glycoproteins
monoclonal antibodies
Monoclonal Antibodies
cattle
neutralization
Bovine herpesvirus 5
epitopes
Glycoproteins
hybridomas
Viral Antigens
viral antigens
Hybridomas
antigens
Antigens
Epitopes
Epitope Mapping
Immunodominant Epitopes
cattle diseases
glycoprotein gC, herpes simplex virus type 1

Keywords

  • BoHV-1
  • BoHV-5
  • Bovine herpesvirus
  • Monoclonal antibodies
  • Virus neutralization

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Animal Science and Zoology
  • veterinary(all)

Cite this

Winkelmann, E. R., Da Silva, L. F., Mayer, S. V., Mazzutti, K. C., Weiblen, R., & Flores, E. F. (2007). Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC). Ciencia Rural, 37(4), 1066-1072.

Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC). / Winkelmann, Evandro Reinoldo; Da Silva, Letícia Frizzo; Mayer, Sandra Vanderli; Mazzutti, Ketty Cristina; Weiblen, Rudi; Flores, Eduardo Furtado.

In: Ciencia Rural, Vol. 37, No. 4, 07.2007, p. 1066-1072.

Research output: Contribution to journalArticle

Winkelmann, ER, Da Silva, LF, Mayer, SV, Mazzutti, KC, Weiblen, R & Flores, EF 2007, 'Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC)', Ciencia Rural, vol. 37, no. 4, pp. 1066-1072.
Winkelmann ER, Da Silva LF, Mayer SV, Mazzutti KC, Weiblen R, Flores EF. Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC). Ciencia Rural. 2007 Jul;37(4):1066-1072.
Winkelmann, Evandro Reinoldo ; Da Silva, Letícia Frizzo ; Mayer, Sandra Vanderli ; Mazzutti, Ketty Cristina ; Weiblen, Rudi ; Flores, Eduardo Furtado. / Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC). In: Ciencia Rural. 2007 ; Vol. 37, No. 4. pp. 1066-1072.
@article{2482c7778a394bd980a50ddeb4b61a09,
title = "Produ{\cc}{\~a}o e caracteriza{\cc}{\~a}o de anticorpos monoclonais contra uma cepa do herpesv{\'i}rus bovino tipo 1 defectiva na glicoprote{\'i}na C (gC)",
abstract = "Most monoclonal antibodies (MAbs) already produced against bovine herpesvirus type 1 (BoHV-1) react with glycoprotein C (gC), an abundant and immunodominant antigen present on the viral envelope. In order to obtain MAbs with other protein specificities, antigens of a BoHV-1 gC-negative strain were used to immunize BALB/c mice. After fusion and selection of 54 HAT-resistant hybridomas, three clones have been obtained (1F1, 2H4 and 4D7) that secrete IgG2a antibodies reacting to BoHV-1 antigens. These MAbs reacted with viral antigens in immunofluorescence (IFA) and immunoperoxidase (IPX) in dilutions up to 1:640 (hybridoma supernatants) and 1:20.000 (ascitis fluid). The three MAbs showed a wide spectrum of reactivity, recognizing antigens of 14 herpesviruses isolated from respiratory or genital disease (supposedly BoHV-1) and with 17 isolates of neurological disease (likely BoHV-5) and displayed varied levels of neutralizing activity against all these viruses. The protein specificity could not be demonstrated directly as none of the MAbs bound to viral antigens in Western blot. On the other hand, the three MAbs reacted with cells infected with a BoHV-5 strain defective in glycoproteins gE and gI, demonstrating they are directed to other viral proteins. By exclusion (gC, gE and gI) and due to their neutralizing activity, these MAbs are probably directed to conserved epitopes on other envelope glycoproteins harboring neutralizing epitopes: gB or gD. In this sense, besides being useful for diagnosis purposes, these MAbs may be very useful for mapping neutralizing epitopes in these glycoproteins.",
keywords = "BoHV-1, BoHV-5, Bovine herpesvirus, Monoclonal antibodies, Virus neutralization",
author = "Winkelmann, {Evandro Reinoldo} and {Da Silva}, {Let{\'i}cia Frizzo} and Mayer, {Sandra Vanderli} and Mazzutti, {Ketty Cristina} and Rudi Weiblen and Flores, {Eduardo Furtado}",
year = "2007",
month = "7",
language = "Portuguese",
volume = "37",
pages = "1066--1072",
journal = "Ciencia Rural",
issn = "0103-8478",
publisher = "Universidade Federal de Santa Maria",
number = "4",

}

TY - JOUR

T1 - Produção e caracterização de anticorpos monoclonais contra uma cepa do herpesvírus bovino tipo 1 defectiva na glicoproteína C (gC)

AU - Winkelmann, Evandro Reinoldo

AU - Da Silva, Letícia Frizzo

AU - Mayer, Sandra Vanderli

AU - Mazzutti, Ketty Cristina

AU - Weiblen, Rudi

AU - Flores, Eduardo Furtado

PY - 2007/7

Y1 - 2007/7

N2 - Most monoclonal antibodies (MAbs) already produced against bovine herpesvirus type 1 (BoHV-1) react with glycoprotein C (gC), an abundant and immunodominant antigen present on the viral envelope. In order to obtain MAbs with other protein specificities, antigens of a BoHV-1 gC-negative strain were used to immunize BALB/c mice. After fusion and selection of 54 HAT-resistant hybridomas, three clones have been obtained (1F1, 2H4 and 4D7) that secrete IgG2a antibodies reacting to BoHV-1 antigens. These MAbs reacted with viral antigens in immunofluorescence (IFA) and immunoperoxidase (IPX) in dilutions up to 1:640 (hybridoma supernatants) and 1:20.000 (ascitis fluid). The three MAbs showed a wide spectrum of reactivity, recognizing antigens of 14 herpesviruses isolated from respiratory or genital disease (supposedly BoHV-1) and with 17 isolates of neurological disease (likely BoHV-5) and displayed varied levels of neutralizing activity against all these viruses. The protein specificity could not be demonstrated directly as none of the MAbs bound to viral antigens in Western blot. On the other hand, the three MAbs reacted with cells infected with a BoHV-5 strain defective in glycoproteins gE and gI, demonstrating they are directed to other viral proteins. By exclusion (gC, gE and gI) and due to their neutralizing activity, these MAbs are probably directed to conserved epitopes on other envelope glycoproteins harboring neutralizing epitopes: gB or gD. In this sense, besides being useful for diagnosis purposes, these MAbs may be very useful for mapping neutralizing epitopes in these glycoproteins.

AB - Most monoclonal antibodies (MAbs) already produced against bovine herpesvirus type 1 (BoHV-1) react with glycoprotein C (gC), an abundant and immunodominant antigen present on the viral envelope. In order to obtain MAbs with other protein specificities, antigens of a BoHV-1 gC-negative strain were used to immunize BALB/c mice. After fusion and selection of 54 HAT-resistant hybridomas, three clones have been obtained (1F1, 2H4 and 4D7) that secrete IgG2a antibodies reacting to BoHV-1 antigens. These MAbs reacted with viral antigens in immunofluorescence (IFA) and immunoperoxidase (IPX) in dilutions up to 1:640 (hybridoma supernatants) and 1:20.000 (ascitis fluid). The three MAbs showed a wide spectrum of reactivity, recognizing antigens of 14 herpesviruses isolated from respiratory or genital disease (supposedly BoHV-1) and with 17 isolates of neurological disease (likely BoHV-5) and displayed varied levels of neutralizing activity against all these viruses. The protein specificity could not be demonstrated directly as none of the MAbs bound to viral antigens in Western blot. On the other hand, the three MAbs reacted with cells infected with a BoHV-5 strain defective in glycoproteins gE and gI, demonstrating they are directed to other viral proteins. By exclusion (gC, gE and gI) and due to their neutralizing activity, these MAbs are probably directed to conserved epitopes on other envelope glycoproteins harboring neutralizing epitopes: gB or gD. In this sense, besides being useful for diagnosis purposes, these MAbs may be very useful for mapping neutralizing epitopes in these glycoproteins.

KW - BoHV-1

KW - BoHV-5

KW - Bovine herpesvirus

KW - Monoclonal antibodies

KW - Virus neutralization

UR - http://www.scopus.com/inward/record.url?scp=34447290828&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34447290828&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:34447290828

VL - 37

SP - 1066

EP - 1072

JO - Ciencia Rural

JF - Ciencia Rural

SN - 0103-8478

IS - 4

ER -