Low population Japanese encephalitis virus (JEV) seroprevalence in Udayapur district, Nepal, three years after a JE vaccination programme: A case for further catch up campaigns?

The live attenuated Japanese encephalitis (JE) vaccine SA14-14-2 has been used in Nepal for catch-up campaigns and is now included in the routine immunisation schedule. Previous studies have shown good vaccine efficacy after one dose in districts with a high incidence of JE. The first well-documented dengue outbreak occurred in Nepal in 2006 with ongoing cases now thought to be secondary to migration from India. Previous infection with dengue virus (DENV) partially protects against JE and might also influence serum neutralising antibody titres against JEV. This study aimed to determine whether serum anti-JEV neutralisation titres are: 1. maintained over time since vaccination, 2. vary with historic local JE incidence, and 3. are associated with DENV neutralising antibody levels. We conducted a cross-sectional study in three districts of Nepal: Banke, Rupandehi and Udayapur. Udayapur district had been vaccinated against JE most recently (2009), but had been the focus of only one campaign, compared with two in Banke and three in Rupandehi. Participants answered a short questionnaire and serum was assayed for anti-JEV and anti-DENV IgM and IgG (by ELISA) and 50% plaque reduction neutralisation titres (PRNT50) against JEV and DENV serotypes 1-4. A titre of 1:10 was considered seropositive to the respective virus. JEV neutralising antibody seroprevalence (PRNT50 1:10) was 81% in Banke and Rupandehi, but only 41% in Udayapur, despite this district being vaccinated more recently. Sensitivity of ELISA for both anti-JEV and anti-DENV antibodies was low compared with PRNT50. DENV neutralising antibody correlated with the JEV PRNT50 1:10, though the effect was modest. IgM (indicating recent infection) against both viruses was detected in a small number of participants. We also show that DENV IgM is present in Nepali subjects who have not travelled to India, suggesting that DENV may have become established in Nepal. We therefore propose that further JE vaccine campaigns should be considered in Udayapur district, and similar areas that have had fewer vaccination campaigns. Author summary In Nepal, immunisation using a live attenuated vaccine is given against Japanese encephalitis (JE), caused by the mosquito-transmitted JE virus (JEV). JE immunisation has taken place via catch-up campaigns and is now part of the routine immunisation programme. Although previous studies have shown good vaccine efficacy in areas where there is a lot of natural exposure to the virus (high endemicity), it is suggested that the efficacy may wane in areas where transmission is lower. Dengue virus (DENV) belongs to the same family and genus as JEV. Previous infection with DENV may also influence the immune response to JEV. Therefore, we conducted a cross-sectional study in Nepal to measure immunity to JE, in districts of differing historic JE incidence, and time from JE vaccination. This showed that neutralising antibody to JEV was found more frequently in districts which had been the subject of more vaccination campaigns, rather than in the most recently vaccinated district. In addition, we cannot rule out a role for natural exposure to JEV in maintaining higher antibody levels. Additionally, the study showed that previous exposure to DENV was positively associated with an immune response to JEV, though this effect was modest. We conclude that there is a need to consider further JE vaccine catch up campaigns in some areas especially given that we could detect JEV IgM, indicating ongoing transmission. We show that ELISA yielded many false negative results for exposure to JEV or vaccination, when compared with neutralising antibody. We also identified some individuals during the course of the study with DENV IgM in their blood, but with no history of travel to India. This suggests that DENV may have become established in some areas of Nepal.