Generation and characterization of Japanese encephalitis virus specific monoclonal antibodies.
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.
Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, R.O.C.
S H
SH
Ma
Y L
YL
Lin
Y Y
YY
Huang
C I
CI
Liu
S S
SS
Chen
H Y
HY
Chiang
L K
LK
Chen
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D004664:Encephalitis Virus, Japanese / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D020365:RNA Helicases; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins / Q000276:immunology; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000911:Antibodies, Monoclonal; D020365:RNA Helicases; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C100315:NS1 protein, Flavivirus; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases