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Journal of Virology, June 1999, p. 4738-4747, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Dengue Virus Structural Differences That Correlate with Pathogenesis

Katrin C. Leitmeyer,1 David W. Vaughn,2 Douglas M. Watts,3 Rosalba Salas,4 Iris Villalobos , de Chacon,5 Celso Ramos,6 and Rebeca Rico-Hesse1,*

Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas 78227-53011; Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, D.C. 20307-51002; Naval Medical Research Institute Detachment, Unit 3800, APO AA 340323; Instituto Nacional de Higiene "Rafael Rangel," Ministerio de Sanidad y Assistencia Social, Caracas,4 and Servicio de Epidemiologia, Hospital Central de Maracay, La Floresta, Maracay, Aragua,5 Venezuela; and CISEI, Instituto Nacional de Salud Publica, 62508 Cuernavaca, Morelos, Mexico6

Received 1 February 1999/Accepted 12 March 1999

The understanding of dengue virus pathogenesis has been hampered by the lack of in vitro and in vivo models of disease. The study of viral factors involved in the production of severe dengue, dengue hemorrhagic fever (DHF), versus the more common dengue fever (DF), have been limited to indirect clinical and epidemiologic associations. In an effort to identify viral determinants of DHF, we have developed a method for comparing dengue type 2 genomes (reverse transcriptase PCR in six fragments) directly from patient plasma. Samples for comparison were selected from two previously described dengue type 2 genotypes which had been shown to be the cause of DF or DHF. When full genome sequences of 11 dengue viruses were analyzed, several structural differences were seen consistently between those associated with DF only and those with the potential to cause DHF: a total of six encoded amino acid charge differences were seen in the prM, E, NS4b, and NS5 genes, while sequence differences observed within the 5' nontranslated region (NTR) and 3' NTR were predicted to change RNA secondary structures. We hypothesize that the primary determinants of DHF reside in (i) amino acid 390 of the E protein, which purportedly alters virion binding to host cells; (ii) in the downstream loop (nucleotides 68 to 80) of the 5' NTR, which may be involved in translation initiation; and (iii) in the upstream 300 nucleotides of the 3' NTR, which may regulate viral replication via the formation of replicative intermediates. The significance of four amino acid differences in the nonstructural proteins NS4b and NS5, a presumed transport protein and the viral RNA polymerase, respectively, remains unknown. This new approach to the study of dengue virus genome differences should better reflect the true composition of viral RNA populations in the natural host and permit their association with pathogenesis.

* Corresponding author. Mailing address: Department of Virology and Immunology, Southwest Foundation for Biomedical Research, P.O. Box 760549, San Antonio, TX 78245-0549. Phone: (210) 258-9681. Fax: (210) 258-9776. E-mail: rricoh{at}icarus.sfbr.org.

Journal of Virology, June 1999, p. 4738-4747, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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