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

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.