WO2013087945A2 - Influenza virus reassortment - Google Patents

Influenza virus reassortment Download PDF

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Publication number
WO2013087945A2
WO2013087945A2 PCT/EP2013/054227 EP2013054227W WO2013087945A2 WO 2013087945 A2 WO2013087945 A2 WO 2013087945A2 EP 2013054227 W EP2013054227 W EP 2013054227W WO 2013087945 A2 WO2013087945 A2 WO 2013087945A2
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Prior art keywords
virus
influenza
strain
segments
seq
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PCT/EP2013/054227
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English (en)
French (fr)
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WO2013087945A3 (en
Inventor
Pirada Suphaphiphat
Peter Mason
Bjoern Keiner
Philip Ralph DORMITZER
Heidi Trusheim
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Novartis AG
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Novartis AG
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Priority to CA2813723A priority Critical patent/CA2813723A1/en
Priority to AU2013205478A priority patent/AU2013205478B9/en
Priority to EP13712156.2A priority patent/EP2820126B1/en
Priority to CN201380000396.3A priority patent/CN103608453B/zh
Priority to ES13712156.2T priority patent/ES2628301T3/es
Priority to JP2013548864A priority patent/JP6054883B2/ja
Application filed by Novartis AG filed Critical Novartis AG
Priority to US13/909,013 priority patent/US9422528B2/en
Publication of WO2013087945A2 publication Critical patent/WO2013087945A2/en
Publication of WO2013087945A3 publication Critical patent/WO2013087945A3/en
Anticipated expiration legal-status Critical
Priority to US15/210,485 priority patent/US10245312B2/en
Priority to US16/272,018 priority patent/US20200000909A1/en
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/145—Orthomyxoviridae, e.g. influenza virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5252—Virus inactivated (killed)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16121—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16151—Methods of production or purification of viral material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16161—Methods of inactivation or attenuation

Definitions

  • Reference 3 reports that a reassortant influenza virus containing a PB1 gene segment from A/Texas/1/77, the HA and NA segments from A/New Caledonia/20/99, a modified PA segment derived from A/Puerto Rico/8/34 and the remaining viral segments from A/Puerto Rico/8/34 shows increased growth in cells.
  • the reassortant influenza A virus comprises backbone segments from two, three, four or five donor strains
  • one or two of the donor strains may provide more than one of the backbone segments of the reassortant influenza A virus.
  • the reassortant influenza A virus cannot comprise more than six backbone segments. Accordingly, for example, if one of the donor strains provides five of the viral segments, the reassortant influenza A virus can only comprise backbone segments from a total of two different donor strains.
  • the invention also provides a reassortant influenza A virus comprising at least one backbone viral segment from a donor strain, wherein the donor strain is selected from the group consisting of 105p30 and P 8-X.
  • the at least one backbone viral segment is the PA segment it may have a sequence having at least 95% or at least 99% identity with a sequence selected from the group consisting of SEQ ID NOs: 9 and 17.
  • the at least one backbone viral segment is the PB1 segment, it may have a sequence having at least 95% or at least 99% identity with a sequence 5 selected from the group consisting of SEQ ID NOs 10 and 18.
  • the method may comprise the steps of (i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza A virus wherein the backbone viral segments are from two or more influenza strains and the PB1 and PB2 segments are from the same donor strain; and (ii) culturing the culture host in order to produce reassortant virus and optionally (iii) purifying the virus obtained in step (ii).
  • the invention also provides a method of preparing a reassortant influenza A virus of the invention comprising steps of (i) introducing into a culture host one or more expression construct(s) which encode(s) the viral segments required to produce an influenza A virus wherein one or more backbone viral segment(s) is/are from a 105p30 and/or a PR8-X influenza strain and wherein at least one viral segment is derived from a second influenza strain; and (ii) culturing the culture host in order to produce reassortant virus and optionally (iii) purifying the virus obtained in step (ii).
  • the invention provides an expression system comprising one or more expression construct(s) comprising the vRNA encoding segments of an influenza A virus wherein the expression construct(s) encode(s) the backbone viral segments from two or more influenza donor strains.
  • the expression construct(s) may encode the PB1 and PB2 segments from the same donor strain.
  • the invention also encompasses reassortant viruses which contain viral segments from more than one, for example two or three different, donor strain(s) wherein at least one viral segment, preferably not HA, is derived from the P 8-X or 105p30 influenza strains.
  • reassortant influenza viruses will typically contain the HA and/or NA segment from a vaccine strain.
  • the further donor strain(s) can be any donor strain including the donor strains of the invention.
  • some of the viral segments may be derived from the A/PR/8/34 or AA/6/60 (A/Ann Arbor/6/60) influenza strains.
  • Reassortants containing viral segments from the AA/6/60 strain may be advantageous, for example, where the reassortant virus is to be used in a live attenuated influenza vaccine.
  • Strains which can be used as vaccine strains include strains which are resistant to antiviral therapy ⁇ e.g. resistant to oseltamivir [6] and/or zanamivir), including resistant pandemic strains [7].
  • Reassortants of the donor strains of the invention wherein the HA and/or NA segment has been changed to another subtype can also be used.
  • the HI influenza subtype of the 105p30 or PR8-X strain may be changed, for example, to a H3 or H5 subtype.
  • the invention encompasses an influenza A virus which comprises one, two, three, four, five, six or seven viral segments from the 105p30 or PR8-X strains of the invention and a HA segment which is not of the HI subtype.
  • the reassortant donor strains may further comprise an NA segment which is not of the Nl subtype. Suitable techniques for reassorting the donor strains will be evident to those of skill in the art.
  • Expression constructs used in the expression systems of the invention may be uni-directional or bidirectional expression constructs. Where more than one transgene is used in the methods (whether on the same or different expression constructs) it is possible to use uni-directional and/or bi-directional expression.
  • An expression construct may be a vector, such as a plasmid or other episomal construct.
  • Such vectors will typically comprise at least one bacterial and/or eukaryotic origin of replication.
  • the vector may comprise a selectable marker which allows for selection in prokaryotic and/or eukaryotic cells. Examples of such selectable markers are genes conferring resistance to antibiotics, such as ampicillin or kanamycin.
  • the vector may further comprise one or more multiple cloning sites to facilitate cloning of a DNA sequence.
  • expression constructs of the invention can be introduced into host cells using any technique known to those of skill in the art.
  • expression constructs of the invention can be introduced into host cells by employing electroporation, DEAE-dextran, calcium phosphate precipitation, liposomes, microinjection, or microparticle-bombardment.
  • the invention utilises virus produced according to the method to produce vaccines.
  • the vaccine may comprise whole virion, split virion, or purified surface antigens (for influenza, including hemagglutinin and, usually, also including neuraminidase).
  • Chemical means for inactivating a virus include treatment with an effective amount of one or more of the following agents: detergents, formaldehyde, ⁇ -propiolactone, methylene blue, psoralen, carboxyfullerene (C60), binary ethylamine, acetyl ethyleneimine, or combinations thereof.
  • Non-chemical methods of viral inactivation are known in the art, such as for example UV light or gamma irradiation.
  • the methods of the invention may also be used to produce live vaccines.
  • live vaccines are usually prepared by purifying virions from virion-containing fluids.
  • the fluids may be clarified by centrifugation, and stabilized with buffer (e.g. containing sucrose, potassium phosphate, and monosodium glutamate).
  • buffer e.g. containing sucrose, potassium phosphate, and monosodium glutamate.
  • influenza virus vaccines include Medlmmune's FLUMISTTM product
  • Vaccine compositions may include preservatives such as thiomersal or 2-phenoxyethanol. It is preferred, however, that the vaccine should be substantially free from (i.e. less than 5 ⁇ g/ml) mercurial material e.g. thiomersal-free [31,41]. Vaccines containing no mercury are more preferred. An a-tocopherol succinate can be included as an alternative to mercurial compounds [31]. Preservative-free vaccines are particularly preferred.
  • Preferred amounts of surfactants are: polyoxyethylene sorbitan esters (such as Tween 80) 0.01 to 1%, in particular about 0.1 %; octyl- or nonylphenoxy polyoxyethanols (such as Triton X-100, or other detergents in the Triton series) 0.001 to 0.1 %, in particular 0.005 to 0.02%; polyoxyethylene ethers (such as laureth 9) 0.1 to 20 %, preferably 0.1 to 10 % and in particular 0.1 to 1% or about 0.5%.
  • polyoxyethylene sorbitan esters such as Tween 80
  • octyl- or nonylphenoxy polyoxyethanols such as Triton X-100, or other detergents in the Triton series
  • polyoxyethylene ethers such as laureth 9
  • the emulsion may also include a 3-de-O- acylated monophosphoryl lipid A (3d MPL).
  • 3d MPL 3-de-O- acylated monophosphoryl lipid A
  • Another useful emulsion of this type may comprise, per human dose, 0.5-10 mg squalene, 0.5-11 mg tocopherol, and 0.1 -4 mg polysorbate 80 [52] e.g. in the ratios discussed above.
  • the vial is preferably made of a glass or plastic material.
  • the vial is preferably sterilized before the composition is added to it.
  • vials are preferably sealed with a latex-free stopper, and the absence of latex in all packaging material is preferred.
  • the vial may include a single dose of vaccine, or it may include more than one dose (a 'multidose' vial) e.g. 10 doses.
  • Preferred vials are made of colourless glass.
  • the immune response raised by these methods and uses will generally include an antibody response, preferably a protective antibody response.
  • Methods for assessing antibody responses, neutralising capability and protection after influenza virus vaccination are well known in the art. Human studies have shown that antibody titers against hemagglutinin of human influenza virus are correlated with protection (a serum sample hemagglutination-inhibition titer of about 30-40 gives around 50% protection from infection by a homologous virus) [62].
  • Antibody responses are typically measured by hemagglutination inhibition, by microneutralisation, by single radial immunodiffusion (S ID), and/or by single radial hemolysis (SRH). These assay techniques are well known in the art.
  • an oseltamivir or zanamivir compound in the 7 days prior to receiving the vaccine, people with egg allergies and people travelling abroad.
  • the vaccines are not suitable solely for these groups, however, and may be used more generally in a population. For pandemic strains, administration to all age groups is preferred.
  • compositions of the invention satisfy 1 , 2 or 3 of the CPMP criteria for efficacy.
  • these criteria are: (1) >70% seroprotection; (2) >40% seroconversion; and/or (3) a GMT increase of >2.5-fold.
  • elderly (>60 years) these criteria are: (1) >60% seroprotection; (2) >30% seroconversion; and/or (3) a GMT increase of >2-fold.
  • Treatment can be by a single dose schedule or a multiple dose schedule. Multiple doses may be used in a primary immunisation schedule and/or in a booster immunisation schedule. In a multiple dose schedule the various doses may be given by the same or different routes e.g.
  • uninfected MDCK cells are plated at a density of 1.8xl0 4 cells/well in 96 well plates in 100 ⁇ of DMEM with 10% FCS. The next day, medium is aspirated and cells are infected with viruses in a volume of 50 ⁇ 1 (viruses diluted in DMEM + 1% FCS). The cells are incubated at 37°C until the next day.
  • MDCK cells are infected with 105p30 and PR8-X at a moi of 10 "3 and samples are taken at several time points after infection. The titre is determined by a FFA assay. The results show that 105p30 grows even faster in MDCK cells compared to PR8-X ( Figure 6).
  • reassortant influenza strains are produced which contain backbone numbers 17 and 19 and the HA and NA segments from either a second H3 influenza (strain 1) virus or a pandemic HI influenza virus (strain 3).
  • a second H3 influenza strain 1
  • a pandemic HI influenza virus strain 3
  • the equivalent wildtype H3 strain 2 influenza virus
  • a reassortant influenza virus comprising the same HA and NA segments and all backbone segments from PR8-X
  • pandemic HI influenza virus a reassortant influenza virus comprising the same HA and NA segments and all backbone segments from PR8-X is used.
  • SEQ ID NO: 7 (HA, A/New Caledonia/20/1999)
  • SEQ ID NO: 8 (NA, A/New Caledonia/20/1999)
  • SEQ ID NO: 31 (PB2, A/New Caledonia/20/1999)
  • SEQ ID NO: 38 (NP, A/Wisconsin/67/2005)

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PCT/EP2013/054227 2012-03-02 2013-03-02 Influenza virus reassortment Ceased WO2013087945A2 (en)

Priority Applications (9)

Application Number Priority Date Filing Date Title
AU2013205478A AU2013205478B9 (en) 2012-03-02 2013-03-02 Influenza virus reassortment
EP13712156.2A EP2820126B1 (en) 2012-03-02 2013-03-02 Influenza virus reassortment
CN201380000396.3A CN103608453B (zh) 2012-03-02 2013-03-02 流感病毒重配
ES13712156.2T ES2628301T3 (es) 2012-03-02 2013-03-02 Recombinación de virus de gripe
JP2013548864A JP6054883B2 (ja) 2012-03-02 2013-03-02 インフルエンザウイルスの再集合
CA2813723A CA2813723A1 (en) 2012-03-02 2013-03-02 Influenza virus reassortment
US13/909,013 US9422528B2 (en) 2012-03-02 2013-06-03 Influenza virus reassortment
US15/210,485 US10245312B2 (en) 2012-03-02 2016-07-14 Influenza virus reassortment
US16/272,018 US20200000909A1 (en) 2012-03-02 2019-02-11 Influenza virus reassortment

Applications Claiming Priority (4)

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US201261605922P 2012-03-02 2012-03-02
US61/605,922 2012-03-02
US201261685766P 2012-03-23 2012-03-23
US61/685,766 2012-03-23

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WO2013087945A3 WO2013087945A3 (en) 2013-11-14

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CN (1) CN103608453B (enExample)
AU (1) AU2013205478B9 (enExample)
CA (1) CA2813723A1 (enExample)
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Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014115104A1 (en) * 2013-01-23 2014-07-31 Novartis Ag Influenza virus reassortment
RU2605317C1 (ru) * 2015-06-04 2016-12-20 Федеральное государственное автономное образовательное учреждение высшего образования "Новосибирский национальный исследовательский государственный университет" (Новосибирский государственный университет, НГУ) АДАПТИРОВАННЫЕ ПАНДЕМИЧЕСКИЕ ШТАММЫ ВИРУСА ГРИППА A/Tomsk/273/2010-MA1(H1N1pdm09), A/Tomsk/273/2010-MA2(H1N1pdm09) И A/Tomsk/273/2010-MA3(H1N1pdm09) ДЛЯ ОЦЕНКИ ДЕЙСТВИЯ ПРОТИВОВИРУСНЫХ ПРЕПАРАТОВ (ВАРИАНТЫ)
WO2016207853A2 (en) 2015-06-26 2016-12-29 Seqirus UK Limited Antigenically matched influenza vaccines
WO2017040203A1 (en) * 2015-08-28 2017-03-09 Yoshihiro Kawaoka Generation of infectious influenza viruses from virus-like particles
US10030231B2 (en) 2013-01-23 2018-07-24 Seqirus UK Limited Influenza virus reassortment
EP3764098A1 (en) 2015-07-07 2021-01-13 Seqirus UK Limited Influenza potency assays
US11384339B2 (en) 2011-08-26 2022-07-12 Wisconsin Alumni Research Foundation (Warf) Influenza viruses with mutant PB2 gene segment as live attenuated vaccines
US11633469B2 (en) 2016-07-15 2023-04-25 The Research Foundation For Microbial Diseases Of Osaka University Reassortant influenza virus production method
US11802273B2 (en) 2014-06-20 2023-10-31 Wisconsin Alumni Research Foundation (Warf) Mutations that confer genetic stability to additional genes in influenza viruses
US11807872B2 (en) 2019-08-27 2023-11-07 Wisconsin Alumni Research Foundation (Warf) Recombinant influenza viruses with stabilized HA for replication in eggs
US11851648B2 (en) 2019-02-08 2023-12-26 Wisconsin Alumni Research Foundation (Warf) Humanized cell line
US12076387B2 (en) 2010-03-23 2024-09-03 Wisconsin Alumni Research Foundation (Warf) Vaccines comprising mutant attenuated influenza viruses
US12122807B2 (en) 2019-05-01 2024-10-22 Wisconsin Alumni Research Foundation (Warf) Influenza virus replication for vaccine development
US12144857B2 (en) 2018-08-20 2024-11-19 Wisconsin Alumni Research Foundation (Warf) Vectors for eliciting immune responses to non-dominant epitopes in the hemagglutinin (HA) protein
US12251436B2 (en) 2017-10-25 2025-03-18 Wisconsin Alumni Research Foundation (Warf) Recombinant influenza viruses with stabilized HA for replication in eggs
US12290562B2 (en) 2020-03-25 2025-05-06 Wisconsin Alumni Research Foundation (Warf) Recombinant multivalent influenza viruses
US12343390B2 (en) 2018-08-07 2025-07-01 Wisconsin Alumni Research Foundation (Warf) Recombinant biologically contained filovirus vaccine
US12365880B2 (en) 2020-01-24 2025-07-22 The University Of Tokyo Recombinant influenza viruses with stabilized NA
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