Antigenic analysis of Japanese encephalitis virus by using monoclonal antibodies

Abstract

Hybridoma cells were produced by fusing P3X63Ag8.653 mouse myeloma cells with spleen cells from BALB/c mice immunized with Japanese encephalitis (JE) virus, Nakayama-RFVL strain. The resulting 26 clones produced hemagglutination inhibition antibodies against the homologous strain. The hemagglutination inhibition reactivity of each clone was tested against six flaviviruses: JE, Murray Valley encephalitis (MVE), Egypt 101 strain of West Nile (WN), St. Louis encephalitis (SLE), Russian spring summer encephalitis, and dengue type 1. The 26 monoclonal antibodies fell into four groups: 14 JE species-specific antibodies, 6 antibodies reactive to JE and MVE viruses, 3 antibodies to three or four viruses in the JE-MVE-WN-SLE subgroup, and 3 antibodies to all six flaviviruses. Furthermore, antigenic comparison of 27 strains of JE virus was carried out by using five JE species-specific monoclonal antibodies. Of these, 24 strains were isolated in various parts of Japan, and 3 strains came from Southeast Asia. In reactivity, the 27 strains were classified into at least four antigenic groups. The results showed that the Nakayama-Yakken strain is a mutant strain which lacks the Nakayama strain-specific antigen and that the recently isolated strains are immunologically different from Nakayama and JaGAr 01 strains. One clone (NARMA 13) produced a JE species-specific antibody which showed almost the same titer against 26 JE virus strains, whereas one clone (NARMA 5) produced a Nakayama strain-specific antibody which reacted only to the Nakayama-RFVL and Nakayama-Yoken strains.

Effect of Cytokines on Japanese Encephalitis Virus Production by Human Monocytes

“…The number of infected monocytes was determined by immunoperoxidase staining using horseradish peroxidase (HRPO)-labeled F(ab')2 fragment of NARMA 7 (JE species-specific monoclonal antibody) (14) at 48 hr after infection as described previously (15) . The infected monocytes were air-dried on glass slides and fixed with acetone at -20 C for 10 min, and reacted with HRPO-F (ab') 2 fragment of NARMA 7 at 37 C for 1 hr.…”

Antigenic and Genetic Variations among Japanese Encephalitis Virus Strains Belonging to Genotype 1

“…Since *Address correspondence to Dr . Ayub Ali, Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-then, antigenic, biochemical and genetic differences among JE virus strains have been demonstrated by crossneutralization tests using post-vaccination human sera (3), polyclonal or monoclonal antibody reactivities (1,5,13,14,16,17,(31)(32)(33)49) and T1 oligonucleotide fingerprints of the 42S genome RNA (4,26). Recently, Chen et al (9,10) identified four genotypes of JE virus by analyzing 240 nucleotides from the prM gene region, and tried to explain the transmission, evolution, epidemiology and possible pathogenesis of JE virus.…”

Studies on the Antigenic Structure of Japanese Encephalitis Virus Using Monoclonal Antibodies

“…However, the immunological classification of strains of JE virus is not yet clear. In the previous study (13), we produced 26 monoclonal antibodies (NARMA 1 to 26) that reacted with the Nakayama-RFVL strain of JE virus in hemagglutinationinhibition (HI) tests. These antibodies fell into four groups: JE species-, subgroup-, and genus-specific antibodies and antibodies reactive with JE and Murray Valley encephalitis (MVE) viruses.…”