Study on the Porcinophilic Foot-and-Mouth Disease Virus I. Production and Characterization of Monoclonal Antibodies against VP1

Abstract

ABSTRACT. Monoclonal antibodies (MAbs) reported here were produced against the porcinophilic foot-and-mouth disease virus (FMDV) that caused the devastating swine disease on 1997 in Taiwan. A panel (25) of MAbs were found to react with VP1 of O/Taiwan/97 (O/ 97) by ELISA with various potencies. The biological identities of these VP1 reacting MAbs, such as neutralization activity, isotype and capability to distinguish between two serotype O FMDVs, O/97 and O/Taiwan/KM1/99 (O/99), were further analyzed. Eleven out of the total eighteen O/97 neutralizing MAbs were able to neutralize heterologous O/99. Eight O/97 neutralizing and five non-neutralizing MAbs could differentiate two serotype O FMDVs by immunofluorescence assay (IFA) implied that these thirteen MAbs recognized O/ 97 specific epitope(s). Furthermore, reactivities of the VP1 reacting MAbs with a 29 amino acids synthetic peptide (P29) representing the βG-βH loop of VP1 were analyzed by ELISA and fourteen were found positive. MAb clone Q10E-3 reacting strongest with VP1 and P29, neutralizing both but not differentiating two serotype O viruses suggested that the antibody binding site might involve the RGD motif and its C terminal conserved region on βG-βH loop. MAbs with diverse characters presented in this study were the first raised against porcinophilic FMDV. The complete set of MAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV. KEY WORDS: ELISA, foot-and-mouth disease virus, monoclonal antibody, neutralizing activity, VP1.J. Vet. Med. Sci. 68(8): 859-864, 2006 All cloven-hoofed species are susceptible to foot-andmouth disease (FMD) that is considered the most contagious animal disease. The outbreaks of FMD in Argentina, Europe, Asia, South Africa and Uruguay [8] have brought to world attention the devastating effects of the disease in a naïve population and economic costs of control and eradication. The swine-adapted strain of FMD virus which occurred in Taiwan in 1997 killed more than 4 million pigs, hit related industries heavily and made Taiwan losing FMD free status [27]. To regain the free status, an eradication campaign has been implemented since then.The FMD virus, O/Taiwan/97 (O/97), produced high morbidity and mortality in swine but did not affect cattle [5]. The distinct host tropism was demonstrated that the mutated 3A coding region of O/97 played a major role in the attenuation of this virus in bovine [3]. Besides, limited biologic character of this porcinophilic virus was known. With the high specificity of antigen (Ag) recognition, monoclonal antibody (MAb) has been applied on the field of determination of antigenic epitope, diagnostic development, vaccine research and studies of viral pathogenesis for years. Therefore, production of MAbs against the porcinophilic FMDV would be beneficial to both basic and applied researches.This paper described the production of a panel of MAbs against porcinophilic O/97 strain FMDV. Their reactivities with VP1 were determi...

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