Yingfeng Lei

Abstract Old World orthohantaviruses, including Hantaan virus (HTNV), cause hemorrhagic fever with renal syndrome (HFRS) in Eurasia. Available inactivated vaccines often induce low neutralizing antibodies and short-term protection. We evaluated nucleic acid vaccines expressing a prefusion-stabilized HTNV glycoprotein in female BALB/c mice.

All animal experiments were reviewed and approved by the Animal Care and Use Committee of the Laboratory Animal Center, Air Force Medical University. The number of Animal Experimental Ethical Inspection is 20220109. And all experiments were carried out in accordance with the recommendations set out in the Guide for the Care and Use of Laboratory Animals. The JEV-P3 strain was propagated in the brains of 3-day-old inbred BALB/C suckling mice and titrated by conventional plaque assay.

In order to determine whether the JEV genome contains m6A modifications, we extracted RNA from mock- or JEV-infected neuro2a cells at at 24 hpi and 48 hpi, respectively, and subjected it to m6A-specific antibody immunoprecipitation followed by high-throughput sequencing (m6A-seq). We used a stringent peak calling method (false discovery rate[FDR] < 0.01) and sequenced the methylome of JEV with very high depth, leading to the identification of statistically significant peaks.