Generation and characterization of Japanese encephalitis virus specific monoclonal antibodies.

Abstract

Monoclonal antibodies (MoAbs) specifically against Japanese encephalitis virus (JEV) were generated by fusion of immunized mouse spleen cells with NS-1 myeloma cells. Nakayama-NIH (Na) and three Taiwan local strains of JEV, i.e., TL isolated from a patient's brain in 1965, NT109 (JE7) isolated from Cx. tritaeniorhynchus in 1985, and RP9, a plaque purified clone of NT109, were used in the immunization. The specificities of moAbs were determined by immunoprecipitation and western blotting, using JEV-infected cell lysates. They were confirmed by the same methods using recombinant JEV proteins as antigens. From Na immunization, 4 anti-E, 3 anti-NS1 and 3 anti-NS3 moAbs were generated. Seventeen anti-E, three anti-NS1 and three anti-NS3 specific moAbs were generated from mice immunized with Taiwan local JEV strains. Overall 21 anti-E, 6 anti-NS1, and 6 anti-NS3 moAbs were produced and characterized. The isotypes of these moAbs were also determined and described. Interestingly, a majority of the moAbs generated for RP9 were IgG1 isotype. In conclusion, 33 moAbs specific to JEV were generated and characterized, and some of these anti-JEV moAbs were made against Taiwan local isolates. These moAbs provide a powerful tool to study JEV, especially the antigenic properties of Taiwan's local strains.