Development and characterization of non-glycosylated E and NS1 mutant viruses as a potential candidate vaccine for West Nile virus

Whiteman, Melissa C.; Li, Li; Wicker, Jason A.; Kinney, Richard M.; Huang, Claire Y.‐H.; Beasley, David W.C.; Chung, Kyung Min; Diamond, Michael S.; Solomon, Tom; Barrett, Alan D.T.

doi:10.1016/j.vaccine.2009.10.112

Cited by 74 publications

(77 citation statements)

References 49 publications

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“…(A) BHK21 cells were infected with WNV expressing wildtype NS1 (MOI of 0.1) or NS1 CHO null (MOI of 3) and subjected to Western blotting with 8NS1 MAb under reducing conditions. A higher MOI was required for the CHO null WNV because of a growth defect of this virus, as reported previously (62). The difference in NS1 size of the CHO null mutant is due to the absence of N-linked glycans.…”

Section: Resultssupporting

confidence: 70%

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A Short N-Terminal Peptide Motif on Flavivirus Nonstructural Protein NS1 Modulates Cellular Targeting and Immune Recognition

Youn¹,

Cho

Fremont

et al. 2010

J Virol

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Flavivirus NS1 is a versatile nonstructural glycoprotein, with intracellular NS1 functioning as an essential cofactor for viral replication and cell surface and secreted NS1 antagonizing complement activation. Even though NS1 has multiple functions that contribute to virulence, the genetic determinants that regulate the spatial distribution of NS1 in cells among different flaviviruses remain uncharacterized. Here, by creating a panel of West Nile virus-dengue virus (WNV-DENV) NS1 chimeras and site-specific mutants, we identified a novel, short peptide motif immediately C-terminal to the signal sequence cleavage position that regulates its transit time through the endoplasmic reticulum and differentially directs NS1 for secretion or plasma membrane expression. Exchange of two amino acids within this motif reciprocally changed the cellular targeting pattern of DENV or WNV NS1. For WNV, this substitution also modulated infectivity and antibody-induced phagocytosis of infected cells. Analysis of a mutant lacking all three conserved N-linked glycosylation sites revealed an independent requirement of N-linked glycans for secretion but not for plasma membrane expression of WNV NS1. Collectively, our experiments define the requirements for cellular targeting of NS1, with implications for the protective host responses, immune antagonism, and association with the host cell sorting machinery. These studies also suggest a link between the effects of NS1 on viral replication and the levels of secreted or cell surface NS1.

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Section: Resultssupporting

confidence: 70%

“…We evaluated this phenotype as previous studies suggested that loss of N-linked glycans on NS1 affects infectivity of different flaviviruses (15,48,62). WNV NS1 has three N-linked glycans (amino acids 130, 175, and 207), whereas DENV NS1 has two (amino acids 130 and 207).…”

Section: Vol 84 2010 Genetic Basis Of Flavivirus Ns1 Cell Targetingmentioning

confidence: 99%

A Short N-Terminal Peptide Motif on Flavivirus Nonstructural Protein NS1 Modulates Cellular Targeting and Immune Recognition

Youn¹,

Cho

Fremont

et al. 2010

J Virol

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“…While WNV vaccines are in various stages of clinical development, it is unclear when one or more of these might be approved for human use. Approaches include inactivated WNV vaccines (44), E glycoprotein conjugated to virus-like particles (45), live attenuated WNV vaccines (46,47), a single-cycle replicon vaccine (48), a plasmid DNA vaccine (49), and chimeric viral vectors expressing the key WNV prM and E proteins on the backbone of either a canary pox virus (50), dengue virus (51), influenza virus (52), or yellow fever virus (53). Among the most advanced candidates in human development is Chimerivax-WN02, a live attenuated chimeric vaccine based on the yellow fever virus 17D clone.…”

Section: Discussionmentioning

confidence: 99%

An Inactivated Cell Culture Japanese Encephalitis Vaccine (JE-ADVAX) Formulated with Delta Inulin Adjuvant Provides Robust Heterologous Protection against West Nile Encephalitis via Cross-Protective Memory B Cells and Neutralizing Antibody

Petrovsky

Larena

Siddharthan

et al. 2013

J Virol

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e West Nile virus (WNV), currently the cause of a serious U.S. epidemic, is a mosquito-borne flavivirus and member of the Japanese encephalitis (JE) serocomplex. There is currently no approved human WNV vaccine, and treatment options remain limited, resulting in significant mortality and morbidity from human infection. Given the availability of approved human JE vaccines, this study asked whether the JE-ADVAX vaccine, which contains an inactivated cell culture JE virus antigen formulated with Advax delta inulin adjuvant, could provide heterologous protection against WNV infection in wild-type and ␤2-microglobulindeficient (␤2m ؊/؊ ) murine models. Mice immunized twice or even once with JE-ADVAX were protected against lethal WNV challenge even when mice had low or absent serum cross-neutralizing WNV titers prior to challenge. Similarly, ␤2m ؊/؊ mice immunized with JE-ADVAX were protected against lethal WNV challenge in the absence of CD8 ؉ T cells and prechallenge WNV antibody titers. Protection against WNV could be adoptively transferred to naive mice by memory B cells from JE-ADVAX-immunized animals. Hence, in addition to increasing serum cross-neutralizing antibody titers, JE-ADVAX induced a memory Bcell population able to provide heterologous protection against WNV challenge. Heterologous protection was reduced when JE vaccine antigen was administered alone without Advax, confirming the importance of the adjuvant to induction of cross-protective immunity. In the absence of an approved human WNV vaccine, JE-ADVAX could provide an alternative approach for control of a major human WNV epidemic.

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“…Investigations include live attenuated vaccines, recombinant subunit vaccines, vectorized vaccines, DNA vaccines with constructs that express the WNV E protein, live recombinant vaccines, and an attenuated strain based on nonglycosylated E and mutant NS1 proteins (15,235). A neutralizing, WNV-specific monoclonal antibody, E16 (MGAWN1), which penetrates the CNS in animal models, produced neutralizing antibodies in phase I trials (15).…”

Section: Therapeuticsmentioning

confidence: 99%

West Nile Virus: Biology, Transmission, and Human Infection

et al. 2012

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Development and characterization of non-glycosylated E and NS1 mutant viruses as a potential candidate vaccine for West Nile virus

Cited by 74 publications

References 49 publications

A Short N-Terminal Peptide Motif on Flavivirus Nonstructural Protein NS1 Modulates Cellular Targeting and Immune Recognition

A Short N-Terminal Peptide Motif on Flavivirus Nonstructural Protein NS1 Modulates Cellular Targeting and Immune Recognition

An Inactivated Cell Culture Japanese Encephalitis Vaccine (JE-ADVAX) Formulated with Delta Inulin Adjuvant Provides Robust Heterologous Protection against West Nile Encephalitis via Cross-Protective Memory B Cells and Neutralizing Antibody

West Nile Virus: Biology, Transmission, and Human Infection

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