{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T05:55:07Z","timestamp":1791179707828,"version":"4.1.0"},"reference-count":53,"publisher":"Microbiology Society","issue":"9","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2004,9,1]]},"abstract":"<jats:p>The first flavivirus chimera encoding dengue 4 virus (D4) PrM and E structural proteins in a Japanese encephalitis virus (JEV) backbone was successfully generated using the long-PCR based cDNA-fragment stitching (LPCRcFS) technique, demonstrating the technique's applicability for rapid preparation of flavivirus chimeras. The JEV\/D4 chimera multiplied at levels equal to JEV and D4 in the mosquito cell line C6\/36, while in a mouse neuronal cell line (N2a) JEV replicated efficiently, but JEV\/D4 and D4 did not. In mouse challenge experiments, JEV\/D4 showed a lack of neuroinvasiveness similar to D4 when inoculated intraperitoneally, but demonstrated attenuated neurovirulence (LD<jats:sub>50<\/jats:sub>=3\u00b717\u00d710<jats:sup>4<\/jats:sup>\u2005f.f.u.) when inoculated intracranially. It was also noted that mice receiving intraperitoneal challenge with JEV\/D4 possessed D4-specific neutralization antibody and in addition clearly showed resistance to JEV intraperitoneal challenge (at 100\u00d7LD<jats:sub>50<\/jats:sub>). This suggests that immunity to anti-JEV non-structural protein(s) offers protection against JEV infection<jats:italic>in vivo<\/jats:italic>. Dengue secondary infection was also simulated by challenging mice pre-immunized with dengue 2 virus, with D4 or JEV\/D4. Mice showed higher secondary antibody response to challenge with JEV\/D4 than to D4, at 210\u200a000 and 37\u200a000 averaged ELISA units, respectively. Taken together, aside from demonstrating the LPCRcFS technique, it could be concluded that the PrM and E proteins are the major determinant of neuroinvasiveness for JEV. It is also expected that the JEV\/D4 chimera with its pathogenicity in mice and atypical immune profile, could have applications in dengue prophylactic research,<jats:italic>in vivo<\/jats:italic>efficacy assessment of dengue vaccines and development of animal research on models of dengue secondary infection.<\/jats:p>","DOI":"10.1099\/vir.0.80120-0","type":"journal-article","created":{"date-parts":[[2004,8,9]],"date-time":"2004-08-09T15:02:31Z","timestamp":1092063751000},"page":"2503-2513","source":"Crossref","is-referenced-by-count":15,"title":["Fusion PCR generated Japanese encephalitis virus\/dengue 4 virus chimera exhibits lack of neuroinvasiveness, attenuated neurovirulence, and a dual-flavi immune response in mice"],"prefix":"10.1099","volume":"85","author":[{"given":"Edward Gitau Matumbi","family":"Mathenge","sequence":"first","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria del Carmen","family":"Parquet","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasutomo","family":"Funakoshi","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seiji","family":"Houhara","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pooi Fong","family":"Wong","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akitoyo","family":"Ichinose","sequence":"additional","affiliation":[{"name":"Central Laboratory, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Futoshi","family":"Hasebe","sequence":"additional","affiliation":[{"name":"CREST, Japan Science and Technology Corporation, Saitama 332-0012, Japan"},{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shingo","family":"Inoue","sequence":"additional","affiliation":[{"name":"CREST, Japan Science and Technology Corporation, Saitama 332-0012, Japan"},{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kouichi","family":"Morita","sequence":"additional","affiliation":[{"name":"CREST, Japan Science and Technology Corporation, Saitama 332-0012, Japan"},{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki City 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"crossref","first-page":"3794","DOI":"10.1128\/JVI.68.6.3794-3802.1994","article-title":"NS2B-3 proteinase-mediated processing in the yellow fever virus structural region: in vitro and in vivo studies","volume":"68","author":"Amberg","year":"1994","journal-title":"J Virol"},{"key":"R2","doi-asserted-by":"crossref","first-page":"4317","DOI":"10.1016\/S0264-410X(03)00487-0","article-title":"Mutations which enhance the replication of dengue virus type 4 and an antigenic chimeric dengue virus type 2\/4 vaccine candidate in Vero cells","volume":"21","author":"Blaney","year":"2003","journal-title":"Vaccine"},{"key":"R3","doi-asserted-by":"crossref","first-page":"10342","DOI":"10.1073\/pnas.88.22.10342","article-title":"Construction of intertypic chimeric dengue viruses by substitution of structural protein genes","volume":"88","author":"Bray","year":"1991","journal-title":"Proc Natl Acad Sci U S A"},{"key":"R4","first-page":"135","article-title":"Enzyme-linked immunosorbent assay (ELISA) on Japanese encephalitis virus. 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