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Recombinant Isfahan viral vectors

A technology of Isfahan, virus, applied in the field of recombinant Isfahan virus vector

Pending Publication Date: 2017-05-31
PROFECTUS BIOSCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cross-reactivity of cellular immune responses against the rVSV core protein may limit this approach

Method used

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  • Recombinant Isfahan viral vectors
  • Recombinant Isfahan viral vectors
  • Recombinant Isfahan viral vectors

Examples

Experimental program
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Effect test

Embodiment 1

[0169] Analysis and Recovery of Isfahan Virus (ISFV)

[0170] A system has been developed for the recovery of recombinant Isfahan virus (rISFV) from plasmid DNA encoding the ISFV genomic cDNA. The safety of wild-type (wt) rISFV and the attenuated variant rISFVN4ΔCT25gag1 was investigated in a highly sensitive 4- to 5-week-old NIH Swiss Webster mouse intracranial neurovirulence model. with LD with 50 wtVSV IN In contrast, unmodified rISFV wt exhibited LD 50 >10 3 PFU. with 10 4 LD 50 rVSV IN N2CT1 vs. rISFVN4ΔCT25gag1 exhibits LD 50 >10 7 . These results suggest that rISFV and VSV IN Compared to being fundamentally less pathogenic, multiple attenuation strategies (N shuffling, truncation of the G protein cytoplasmic tail) may not be required to achieve e.g. rVSV IN Similar safety and immunogenicity of the vector.

[0171] Isfahan Phylogenetic Analysis. Available sequences for Vesivirus are downloaded from GenBank. Using MUSCLE algorithm (Gouy et al., 2010, Molecul...

Embodiment 2

[0202] Construction of attenuated rISFV vectors and truncated ISFV G cytoplasmic tails using gene shuffling

[0203] Construction of rISFV-N4G5-MCS1. Plasmid pPBS-ISFV-008 contains the nucleic acid sequence 5'-N 1 -P 2 -M 3 -G 4 -L 5 -3' [antigenome of rISFV]. Subscript numbers indicate the genomic location of each ISFV gene, P (encoding phosphoprotein), M (encoding matrix protein), G (encoding attachment protein), N (encoding nucleocapsid protein) and L (encoding polymerase protein).

[0204] Plasmid pPBS-ISFV-009 contains the nucleic acid sequence 5'-MCS 1 -N 2 -P 3 -M 4 -G 5 -L 6 -3' (antigenome of rISFV with an additional transcription cassette at position 1). According to this formula, the MCS (multiple cloning site) is the empty transcription unit (TU) at position 1 immediately upstream of the ISFVN in the rISFV antigenome. Subscript numbers indicate the antigenomic position of each ISFV gene, P (encoding phosphoprotein), M (encoding matrix protein), G (enco...

Embodiment 3

[0243] animal research

[0244] A series of mouse studies were performed to investigate the relative safety and efficacy of immunogenic compositions comprising rISFV vectors expressing alphavirus proteins. Due to the known neurovirulence properties of vesicular and related viruses, mouse intracranial (IC) LD 50 Models were used for preliminary evaluation of vector safety (Olitsky et al., Journal of Experimental Medicine. 1934, 59:159–71; Frank et al., Am J Vet Res. 1945, Jan:28–38; Sabin et al., Journal of Experimental Medicine. Medicine. 1937, 66:15-34; Rao et al., Lancet. 2004, 364(9437):869-74). Efficacy was evaluated in rigorous VEEV and EEEV challenge models.

[0245] Neurovirulence of rISFV vectors. A preliminary study was performed to investigate the neurovirulence properties of unmodified rISFV and a highly attenuated variant expressing HIV-1 gag (rISFV-N4GΔCT25HIVgag1). with known LD from previous studies 50 rVSV IN The N2CT1 vector was used as a positive contr...

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Abstract

Certain embodiments are directed to recombinant vesiculovirus encoding a heterologous polynucleotide and methods of using the same.

Description

[0001] priority [0002] This application claims priority to US Provisional Patent Application No. 61 / 946,734, filed March 1, 2014, which is hereby incorporated by reference in its entirety. [0003] Statement Regarding Federally Funded Research [0004] This invention was made with government support N01-AI-30027, HHSN272201000040I and HHSN2720004 / D04 awarded by the National Institutes of Health / National Institute of Allergy and Infectious Diseases. The government has certain rights in this invention. [0005] About the sequence listing [0006] The sequence listing required by 37 CFR 1.821-1.825 is filed electronically with this application. The Sequence Listing is incorporated herein by reference. Background technique [0007] Recombinant vesicular stomatitis virus (rVSV) has been developed as a carrier platform for some human pathogens (Finke and Conzelmann.Current Topics in Microbiology and Immunology.2005,292:165-200; Jones et al.Nature Medicine.2005,11(7 ):786-90; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12C12N15/00
CPCA61K39/12C12N2760/20243C12N2770/36134C12N15/86A61K2039/545C12N2740/16234C12N2760/20222C12N2760/20262C12N7/00A61P35/00A61P37/04Y02A50/30C12N2760/20221A61K2039/5256A61K39/21C12N2760/20271C12N2740/16071C12N2770/36171C12N7/02C12N7/04C12N2740/16034C12N2760/20211
Inventor 法鲁克·纳萨尔德米特里厄斯·马塔索夫罗迪翁·V·戈尔恰科夫斯蒂芬·哈姆瑞贝卡·诺瓦克罗伯特·L·西摩约翰·H·爱尔德里奇罗伯特·B·泰什大卫·K·克拉克特丽莎·E·莱瑟姆斯科特·韦弗
Owner PROFECTUS BIOSCI
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