Use of recombinant fusion proteins and monoclonal antibodies to define linear and discontinuous antigenic sites on the dengue virus envelope glycoprotein

Mégret, Françoise; Hugnot, Jean‐Philippe; Falconar, Andrew K.; Gentry, Mary K.; Morens, David M.; Murray, Julie M.; Schlesinger, Jacob J.; Wright, P.J.; Young, Paul R.; Regenmortel, M.H.V. Van; Denis, Vincent

doi:10.1016/0042-6822(92)90450-4

Cited by 112 publications

(81 citation statements)

References 43 publications

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“…Plaque-reduction assays, performed on monkey Vero cells, have shown that mAb4E11 neutralizes the four serotypes of DENV with varying efficacies (Thullier et al, 1999). ELISAs using truncated recombinant gpE proteins as antigens have shown that the epitope of mAb4E11 is included within the ED3 domain (Megret et al, 1992;Thullier et al, 1999). Selection of peptides from a random library by the method of phage display has shown that residues 306-314 of gpE from the DENV1 serotype (gpE.DEN1) constitute a mimotope of the mAb4E11 epitope (Thullier et al, 2001).…”

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confidence: 99%

“…In particular, many antibodies that are specific for ED3 are neutralizing (Crill & Roehrig, 2001;Roehrig, 2003). Of note, their epitopes are generally sensitive to elimination of the disulfide bond (Roehrig et al, 2004).Monoclonal antibody mAb4E11 was raised against DENV1.Immunofluorescence experiments on cells of the mosquito Aedes albopictus, infected with various flaviviruses, and Western blot experiments on their total extracts have shown that mAb4E11 recognizes exclusively the viruses of the dengue group, and not other flaviviruses (Megret et al, 1992). Plaque-reduction assays, performed on monkey Vero cells, have shown that mAb4E11 neutralizes the four serotypes of DENV with varying efficacies (Thullier et al, 1999).…”

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confidence: 99%

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Mapping to completeness and transplantation of a group-specific, discontinuous, neutralizing epitope in the envelope protein of dengue virus

Lisova

Hardy

Petit

et al. 2007

Journal of General Virology

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Dengue is caused by a taxonomic group of four viruses, dengue virus types 1-4 (DENV1-DENV4). A molecular understanding of the antibody-mediated protection against this disease is critical to design safe vaccines and therapeutics. Here, the energetic epitope of antibody mAb4E11, which neutralizes the four serotypes of DENV but no other flavivirus, and binds domain 3 (ED3) of their envelope glycoprotein, was characterized. Alanine-scanning mutagenesis of the ED3 domain from serotype DENV1 was performed and the affinities between the mutant domains and the Fab fragment of mAb4E11 were measured. The epitope residues (307-312, 387, 389 and 391) were at the edges of two distinct b-sheets. Four residues constituted hot spots of binding energy. They were aliphatic and contributed to form a hydrophobic pocket (Leu308, Leu389), or were positively charged (Lys307, Lys310). They may bind the diversity residues of mAb4E11, H-Trp96-Glu97. Remarkably, cyclic residues occupy and block the hydrophobic pocket in all unrelated flaviviruses. Transplanting the epitope from the ED3 domain of DENV into those of other flaviviruses restored affinity. The epitope straddles residues of ED3 that are involved in virulence, e.g. Asn/Asp390. These results define the epitope of mAb4E11 as an antigenic signature of the DENV group and suggest mechanisms for its neutralization potency. INTRODUCTIONDengue is a re-emerging viral disease. It is caused by four types of virus, dengue virus types 1-4 (DENV1-DENV4), which belong to the species Dengue virus in the genus Flavivirus (Heinz et al., 2000). They are transmitted by Aedes mosquitoes and infect between 50 million and 100 million persons each year. The disease generally takes a mild form, dengue fever, but severe forms, dengue haemorragic fever and dengue shock syndrome, have recently become epidemic (Gubler, 2002).The immune response against an infection by DENV involves both a humoral component, in the form of neutralizing antibodies, and a cell-mediated component (Guzman & Kouri, 2002). The preferential reactivation of the memory B cells that correspond to a primary infection, and an antibody-dependent enhancement of infection, might constitute triggering mechanisms of the severe forms during a secondary infection by a different viral serotype (Halstead, 2003; Mongkolsapaya et al., 2003). A molecular understanding of the events that lead to antibody neutralization, enhancement or escape is critical to the development of efficient and secure vaccines and therapeutics. DENV1-DENV4 are enveloped RNA viruses, like all flaviviruses. The structures of the whole virus and of the envelope glycoprotein E (gpE) have been solved by electron cryomicroscopy and X-ray crystallography (Modis et al., 2003(Modis et al., , 2005 Zhang et al., 2003). Ninety dimers of gpE cover the surface of the virus. Each monomer comprises three ectodomains, ED1-ED3, and a transmembrane segment. ED2 includes the dimerization interface, a glycosylation site and the peptide of fusion with the cellular membrane. ED3 is continuou...

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confidence: 99%

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confidence: 99%

Mapping to completeness and transplantation of a group-specific, discontinuous, neutralizing epitope in the envelope protein of dengue virus

Lisova

Hardy

Petit

et al. 2007

Journal of General Virology

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“…(i) Limited sequencing studies performed on the E gene indicated a uniform rate of random nucleotide mutations. (ii) The region encodes a protein domain relevant for viral antigenicity and pathogenicity (Mandl et al, 1989;Lobigs et al, 1990;M~gret et al, 1992). (iii) Amino acid sequence comparison between Asibi and 17D strain E proteins showed two major Ivory Coast Aedesfurcifer 199l * These strains were associated with an epidemic of YF.…”

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Geographic distribution and evolution of yellow fever viruses based on direct sequencing of genomic cDNA fragments

Lepiniec

Dalgarno

Huong

et al. 1994

Journal of General Virology

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We have compared the nucleotide sequence of an envelope protein gene fragment encoding amino acids 291 to 406 of 22 yellow fever (YF) virus strains of diverse geographic and host origins isolated over a 63 year time span. The nucleotide fragment of viral RNA was examined by direct sequencing ofa PCR product derived from complementary DNA. Alignment with the prototype Asibi strain sequence showed divergence of 0 to 21'5 % corresponding to a maximum of 5-2 % divergence in the amino acid sequence. Taking 10% nucleotide divergence as a cut-off point, the 22 YF virus strains fell into three topotypes which corresponded to different geographical areas, namely West Africa, Central-East Africa, and South America. Two subgroups were defined in West Africa, a genotypic group circulating in the

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“…Flavivirus antigens have been expressed successfully in recombinant prokaryotic and eukaryotic cell culture systems. Purified Escherichia coil fusion proteins have been described and reported to be feasible flavivirus vaccine candidates, usable diagnostic reagents, and useful for defining tlavivirus antigenic sites (12,35,75,79 Although the live virus vaccines are usually preferable to inactivated or subunit vaccines, alternative approaches to vaccinology still play an important role in disease control strategies. Proteins prepared in baculovirus expression systems have been used to prepare antigens suitable for vaccination.…”

Section: Vaccine Developmentmentioning

confidence: 99%

Insect-transmitted vertebrate viruses: Flaviviridae

Ludwig

Iacono-Connors

1993

In Vitro Cell Dev Biol - Animal

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SUMMARYThe Flaviviridae include almost 70 viruses, nearly half of which have been associated with human disease. These viruses are among the most important arthropod-borne viruses worldwide and include dengue, yellow fever, and Japanese encephalitis viruses. Morbidity and mortality caused by these viruses vary, but collectively they account for millions of encephalitis, hemorrhagic fever, arthralgia, rash, and fever cases per year. Most of the members of this family are transmitted between vertebrate hosts by arthropod vectors, most commonly mosquitoes or ticks. Transmission cycles can be simple or complex depending on the hosts, vectors, the virus, and the environmental factors affecting both hosts and viruses. Replication of virus in invertebrate hosts does not seem to result in any significant pathology, which suggests a close evolutionary relationship between virus and vector. Another example of this relationship is the ability of these viruses to grow in invertebrate ceil culture, where replication usually results in a steady state, persistent infection, often without cytopathic effect. Yields of virus from insect cell culture vary but are generally similar to yields in vertebrate cells. Replication kinetics are comparable between insect and vertebrate ceil lines, despite differences in incubation temperature. Both vertebrate and insect cell culture systems continue to play a significant role in flavivirus isolation and the diagnosis of disease caused by these agents. Additionally, these culture systems permit the study of flavivirus attachment, penetration, replication, and release from cells and have been instrumental in the production and characterization of live-attenuated vaccines. Both vertebrate and insect cell culture systems will continue to play a significant role in basic and applied flavivirus research in the future.

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Use of recombinant fusion proteins and monoclonal antibodies to define linear and discontinuous antigenic sites on the dengue virus envelope glycoprotein

Cited by 112 publications

References 43 publications

Mapping to completeness and transplantation of a group-specific, discontinuous, neutralizing epitope in the envelope protein of dengue virus

Mapping to completeness and transplantation of a group-specific, discontinuous, neutralizing epitope in the envelope protein of dengue virus

Geographic distribution and evolution of yellow fever viruses based on direct sequencing of genomic cDNA fragments

Insect-transmitted vertebrate viruses: Flaviviridae

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