Generation of virus-like particles and demonstration of lipid rafts as sites of filovirus entry and budding
OwnerSina Bavari, M. Javad Aman +1inventors
InventorsSina Bavari, M. Javad Aman, Alan L. Schmaljohn
Life of this patent
Estimating the term…
Abstract
In this application is described a method for the formation of filovirus-like particles for both Ebola and Marburg and their use as a diagnostic and therapeutic agent as well as a filovirus vaccine. Also described is the association of Ebola and Marburg with lipid rafts during assembly and budding, and the requirement of functional rafts for entry of filoviruses into cells.
Claims44
A filovirus virus like particle, VLP, comprising filovirus envelope glycoprotein, GP, and filovirus matrix protein, VP40.
A filovirus VLP, produced by expressing in a cell a polynucleotide encoding filovirus envelope glycoprotein, GP, and filovirus matrix protein, VP40 such that said polynucleotide is expressed and said VLP is produced.
A method for inhibiting the association of a filovirus envelope glycoprotein GP with lipid rafts, comprising inhibiting palmitoylation at cysteine residues 670 and 672 of said GP.
A method for preventing filovirus trafficking into and out of a cell comprising disrupting lipid rafts of said cell.
A method for preventing filovirus trafficking said method comprising introducing to a cell cholesterol synthesis inhibitors.
A filovirus vaccine comprising VLP and a nucleic acid encoding an agent capable of eliciting an immune response against said filovirus.
A method for introducing an agent into a cell, comprising packaging said agent into a VLP producing a packed VLP and allowing the packed VLP to enter said cell.
A method for testing an agent involved in filovirus budding, comprising introducing said agent to a cultured cell producing filovirus VLP and monitoring the presence or absence of a change in the budding of VLP as compared to a control by measuring VLPs in supernatant of said cultured cell, wherein a reduction or increase in the number of VLP in the superntant indicates a negative or positive agent, respectively, on filovirus budding.
A method for inhibiting Ebola virus infection in a cell comprising administering to said cell lipid raft-disrupting agents.
A kit for testing agents involved in Ebola budding said kit comprising a cell producing Ebola VLPs and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
An Ebola VLP-producing cell comprising a mammalian cell expressing Ebola GP and VP40.
A kit for testing agents involved in Marburg budding said kit comprising a cell producing Marburg VLPs and ancillary reagents for detecting VLPs in the supernatant of said cells when cells are cultured.
A Marburg VLP-producing cell comprising a mammalian cell expressing Marburg GP and VP40.
An immunogenic composition comprising, in a physiologically acceptable vehicle, Ebola VLPs.
A method for stimulating an Ebola virus specific immune response, said method comprising administering to a subject an immunologically sufficient amount of Ebola VLPs in a physiologically acceptable vehicle.
An immunogenic composition comprising, in a physiologically acceptable vehicle, Marburg VLPs.
A method for stimulating a Marburg virus specific immune response, said method comprising administering to a subject an immunologically sufficient amount of Marburg VLPs in a physiologically acceptable vehicle.
Dependencies are inferred from the claim text.