Vaccine candidates generated by codon and codon pair deoptimization of enterovirus A71 protect against lethal challenge in mice

ABSTRACTZika virus (ZIKV) infection during the large epidemics in the Americas is related to congenital abnormities or fetal demise. To date, there is no vaccine, antiviral drug, or other modality available to prevent or treat Zika virus infection. Here we designed novel live attenuated ZIKV vaccine candidates using a codon pair deoptimization strategy. Three codon pair-deoptimized ZIKVs (Min E, Min NS1, and Min E+NS1) werede novosynthesized and recovered by reverse genetics and contained large amounts of underrepresented codon pairs in the E gene and/or NS1 gene. The amino acid sequence was 100% unchanged. The codon pair-deoptimized variants had decreased replication fitness in Vero cells (Min NS1 ≫ Min E > Min E+NS1), replicated more efficiently in insect cells than in mammalian cells, and demonstrated diminished virulence in a mouse model. In particular, Min E+NS1, the most restrictive variant, induced sterilizing immunity with a robust neutralizing antibody titer, and a single immunization achieved complete protection against lethal challenge and vertical ZIKV transmission during pregnancy. More importantly, due to the numerous synonymous substitutions in the codon pair-deoptimized strains, reversion to wild-type virulence through gradual nucleotide sequence mutations is unlikely. Our results collectively demonstrate that ZIKV can be effectively attenuated by codon pair deoptimization, highlighting the potential of Min E+NS1 as a safe vaccine candidate to prevent ZIKV infections.IMPORTANCEDue to unprecedented epidemics of Zika virus (ZIKV) across the Americas and the unexpected clinical symptoms, including Guillain-Barré syndrome, microcephaly, and other birth defects in humans, there is an urgent need for ZIKV vaccine development. Here we provided the first attenuated versions of ZIKV with two important genes (E and/or NS1) that were subjected to codon pair deoptimization. Compared to parental ZIKV, the codon pair-deoptimized ZIKVs were mammal attenuated and preferred insect to mammalian cells. Min E+NS1, the most restrictive variant, induced sterilizing immunity with a robust neutralizing antibody titer and achieved complete protection against lethal challenge and vertical virus transmission during pregnancy. More importantly, the massive synonymous mutational approach made it impossible for the variant to revert to wild-type virulence. Our results have proven the feasibility of codon pair deoptimization as a strategy to develop live attenuated vaccine candidates against flaviviruses such as ZIKV, Japanese encephalitis virus, and West Nile virus.

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A Polysaccharide Purified From Ganoderma lucidum Acts as a Potent Mucosal Adjuvant That Promotes Protective Immunity Against the Lethal Challenge With Enterovirus A71

Frontiers in Immunology ◽

10.3389/fimmu.2020.561758 ◽

2020 ◽

Vol 11 ◽

Author(s):

Yu-Li Lin ◽

Chiaho Shih ◽

Pei-Yun Cheng ◽

Chiao-Li Chin ◽

An-Ting Liou ◽

...

Keyword(s):

Ganoderma Lucidum ◽

Protective Immunity ◽

Lethal Challenge ◽

Mucosal Adjuvant ◽

Enterovirus A71

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Extensive genomic recoding by codon-pair deoptimization selective for mammals is a flexible tool to generate attenuated vaccine candidates for dengue virus 2

Virology ◽

10.1016/j.virol.2019.09.003 ◽

2019 ◽

Vol 537 ◽

pp. 237-245 ◽

Cited By ~ 1

Author(s):

Charles B. Stauft ◽

Yutong Song ◽

Oleksandr Gorbatsevych ◽

Petraleigh Pantoja ◽

Idia V. Rodriguez ◽

...

Keyword(s):

Dengue Virus ◽

Flexible Tool ◽

Vaccine Candidates ◽

Attenuated Vaccine ◽

Codon Pair

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Protection from lethal challenge in a neonatal mouse model by circulating recombinant form coxsackievirus A16 vaccine candidates

Journal of General Virology ◽

10.1099/vir.0.063560-0 ◽

2014 ◽

Vol 95 (5) ◽

pp. 1083-1093 ◽

Cited By ~ 29

Author(s):

Jingliang Li ◽

Junliang Chang ◽

Xin Liu ◽

Jiaxin Yang ◽

Haoran Guo ◽

...

Keyword(s):

Mouse Model ◽

Vaccine Development ◽

Lethal Dose ◽

Neonatal Mouse ◽

Recombinant Form ◽

Control Group ◽

Coxsackievirus A16 ◽

Lethal Challenge ◽

Vaccine Candidates

Circulating coxsackievirus A16 (CA16) is a major cause of hand, foot and mouth disease (HFMD) in South-east Asia. At present, there is no vaccine against CA16. Pathogenic animal models that are sensitive to diverse circulating CA16 viruses would be desirable for vaccine development and evaluation. In this study, we isolated and characterized several circulating CA16 viruses from recent HFMD patients. These CA16 viruses currently circulating in humans were highly pathogenic in a newly developed neonatal mouse model; we also observed and analysed the pathogenesis of representative circulating recombinant form CA16 viruses. An inactivated CA16 vaccine candidate, formulated with alum adjuvant and containing submicrogram quantities of viral proteins, protected neonatal mice born to immunized female mice from lethal-dose challenge with a series of CA16 viruses. Further analysis of humoral immunity showed that antibody elicited from both the immunized dams and their pups could neutralize various lethal viruses by a cytopathic effect in vitro. Moreover, viral titres and loads in the tissues of challenged pups in the vaccine group were far lower than those in the control group, and some were undetectable. This lethal-challenge model using pathogenic CA16 viruses and the vaccine candidates that mediated protection in this model could be useful tools for the future development and evaluation of CA16 vaccines.

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Purified Inactivated Zika Vaccine Candidates Afford Protection against Lethal Challenge in Mice

Scientific Reports ◽

10.1038/s41598-018-34735-7 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 9

Author(s):

Whitney R. Baldwin ◽

Jill A. Livengood ◽

Holli A. Giebler ◽

Janae L. Stovall ◽

Karen L. Boroughs ◽

...

Keyword(s):

Lethal Challenge ◽

Vaccine Candidates

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Synonymous Deoptimization of Foot-and-Mouth Disease Virus Causes AttenuationIn Vivowhile Inducing a Strong Neutralizing Antibody Response

Journal of Virology ◽

10.1128/jvi.02167-15 ◽

2015 ◽

Vol 90 (3) ◽

pp. 1298-1310 ◽

Cited By ~ 25

Author(s):

Fayna Diaz-San Segundo ◽

Gisselle N. Medina ◽

Elizabeth Ramirez-Medina ◽

Lauro Velazquez-Salinas ◽

Marla Koster ◽

...

Keyword(s):

Antibody Response ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Live Attenuated Vaccine ◽

Vaccine Candidates ◽

Attenuated Vaccine ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Codon Pair

ABSTRACTCodon bias deoptimization has been previously used to successfully attenuate human pathogens, including poliovirus, respiratory syncytial virus, and influenza virus. We have applied a similar technology to deoptimize the capsid-coding region (P1) of foot-and-mouth disease virus (FMDV). Despite the introduction of 489 nucleotide changes (19%), synonymous deoptimization of the P1 region rendered a viable FMDV progeny. The resulting strain was stable and reached cell culture titers similar to those obtained for wild-type (WT) virus, but at reduced specific infectivity. Studies in mice showed that 100% of animals inoculated with the FMDV A12 P1 deoptimized mutant (A12-P1 deopt) survived, even when the animals were infected at doses 100 times higher than the dose required to cause death by WT virus. All mice inoculated with the A12-P1 deopt mutant developed a strong antibody response and were protected against subsequent lethal challenge with WT virus at 21 days postinoculation. Remarkably, the vaccine safety margin was at least 1,000-fold higher for A12-P1 deopt than for WT virus. Similar patterns of attenuation were observed in swine, in which animals inoculated with A12-P1 deopt virus did not develop clinical disease until doses reached 1,000 to 10,000 times the dose required to cause severe disease in 2 days with WT A12. Consistently, high levels of antibody titers were induced, even at the lowest dose tested. These results highlight the potential use of synonymous codon pair deoptimization as a strategy to safely attenuate FMDV and further develop live attenuated vaccine candidates to control such a feared livestock disease.IMPORTANCEFoot-and-mouth disease (FMD) is one of the most feared viral diseases that can affect livestock. Although this disease appeared to be contained in developed nations by the end of the last century, recent outbreaks in Europe, Japan, Taiwan, South Korea, etc., have demonstrated that infection can spread rapidly, causing devastating economic and social consequences. The Global Foot-and-Mouth Disease Research Alliance (GFRA), an international organization launched in 2003, has set as part of their five main goals the development of next-generation control measures and strategies, including improved vaccines and biotherapeutics. Our work demonstrates that newly developed codon pair bias deoptimization technologies can be applied to FMD virus to obtain attenuated strains with potential for further development as novel live attenuated vaccine candidates that may rapidly control disease without reverting to virulence.

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Development and Characterization of Recombinant Vesicular Stomatitis Virus (rVSV)-based Bivalent Vaccine Against COVID-19 Delta Variant and Influenza Virus

10.1101/2021.12.14.472657 ◽

2021 ◽

Author(s):

Zhujun Ao ◽

Maggie Jing Ouyang ◽

Titus Abiola Olukitibi ◽

Bryce Warner ◽

Robert Vendramelli ◽

...

Keyword(s):

Immune Responses ◽

Neutralizing Antibodies ◽

Influenza A ◽

Respiratory Infections ◽

Convincing Evidence ◽

Influenza Viruses ◽

Lethal Challenge ◽

Vaccine Candidates ◽

Bivalent Vaccine ◽

Wide Range

COVID-19 and influenza are both highly contagious respiratory diseases with a wide range of severe symptoms and cause great disease burdens globally. It has become very urgent and important to develop a bivalent vaccine that is able to target these two infectious diseases simultaneously. In this study, we generated three attenuated replicating recombinant VSV (rVSV) vaccine candidates. These rVSV-based vaccines co-express SARS-CoV-2 Delta variant spike protein (SP) or the receptor binding domain (RBD) and four copies of the highly conserved M2 ectodomain (M2e) of influenza A fused with the Ebola glycoprotein DC-targeting/activation domain. Animal studies have shown that immunization with these bivalent rVSV vaccines induced efficient but variable levels of humoral and cell-mediated immune responses against both SARS-CoV-2 and influenza M2e protein. Significantly, our vaccine candidates induced production of high levels of neutralizing antibodies that protected cells against SARS-CoV-2 Delta and other SP-pseudovirus infections in culture. Furthermore, vaccination with the bivalent VSV vaccine via either intramuscular or intranasal route efficiently protected mice from the lethal challenge of H1N1 and H3N2 influenza viruses and significantly reduced viral load in the lungs. These studies provide convincing evidence for the high efficacy of this bivalent vaccine to prevent influenza replication and initiate robust immune responses against SARS-CoV-2 Delta variants. Further investigation of its efficacy to protect against SARS-CoV-2 Delta variants will provide substantial evidence for new avenues to control two contagious respiratory infections, COVID-19 and influenza.

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Proteomics-Based Identification of Anchorless Cell Wall Proteins as Vaccine Candidates against Staphylococcus aureus

Infection and Immunity ◽

10.1128/iai.00617-08 ◽

2009 ◽

Vol 77 (7) ◽

pp. 2719-2729 ◽

Cited By ~ 50

Author(s):

Eva Glowalla ◽

Bettina Tosetti ◽

Martin Krönke ◽

Oleg Krut

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Survival Rates ◽

Proteome Analysis ◽

Surface Proteins ◽

Human Neutrophils ◽

Cell Wall Proteins ◽

Lethal Challenge ◽

Vaccine Candidates

ABSTRACT Staphylococcus aureus is an important human pathogen with increasing clinical impact due to the extensive spread of antibiotic-resistant strains. Therefore, development of a protective polyvalent vaccine is of great clinical interest. We employed an intravenous immunoglobulin (IVIG) preparation as a source of antibodies directed against anchorless S. aureus surface proteins for identification of novel vaccine candidates. In order to identify such proteins, subtractive proteome analysis (SUPRA) of S. aureus anchorless cell wall proteins was performed. Proteins reacting with IVIG but not with IVIG depleted of S. aureus-specific opsonizing antibodies were considered vaccine candidates. Nearly 40 proteins were identified by this preselection method using matrix-assisted laser desorption ionization—time of flight analysis. Three of these candidate proteins, enolase (Eno), oxoacyl reductase (Oxo), and hypothetical protein hp2160, were expressed as glutathione S-transferase fusion proteins, purified, and used for enrichment of corresponding immunoglobulin Gs from IVIG by affinity chromatography. Use of affinity-purified anti-Eno, anti-Oxo, and anti-hp2160 antibodies resulted in opsonization, phagocytosis, and killing of S. aureus by human neutrophils. High specific antibody titers were detected in mice immunized with recombinant antigens. In mice challenged with bioluminescent S. aureus, reduced staphylococcal spread was measured by in vivo imaging. The recovery of S. aureus CFU from organs of immunized mice was diminished 10- to 100-fold. Finally, mice immunized with hp2160 displayed statistically significant higher survival rates after lethal challenge with clinically relevant S. aureus strains. Taken together, our data suggest that anchorless cell wall proteins might be promising vaccine candidates and that SUPRA is a valuable tool for their identification.

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