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WO2011092199A1 - Protéines d'enveloppe du virus xmrv présentant une mutation au niveau de leur domaine immunosuppresseur - Google Patents

Protéines d'enveloppe du virus xmrv présentant une mutation au niveau de leur domaine immunosuppresseur Download PDF

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Publication number
WO2011092199A1
WO2011092199A1 PCT/EP2011/051062 EP2011051062W WO2011092199A1 WO 2011092199 A1 WO2011092199 A1 WO 2011092199A1 EP 2011051062 W EP2011051062 W EP 2011051062W WO 2011092199 A1 WO2011092199 A1 WO 2011092199A1
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Prior art keywords
seq
xmrv
vector
mutated
mlv
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Inventor
Thierry Heidmann
Géraldine SCHLECHT-LOUF
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Centre National de la Recherche Scientifique CNRS
Institut Gustave Roussy (IGR)
Universite Paris Sud
Viroxis SAS
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Centre National de la Recherche Scientifique CNRS
Institut Gustave Roussy (IGR)
Universite Paris Sud
Viroxis SAS
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Publication of WO2011092199A1 publication Critical patent/WO2011092199A1/fr
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    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • 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
    • A61K38/00—Medicinal preparations containing peptides
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
    • C12N2740/13022—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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
    • C12N2740/13061—Methods of inactivation or attenuation

Definitions

  • the present invention relates to mutated ENV proteins and their use as vaccine.
  • Retroviruses are viruses, the genome of which is made up of RNA. These viruses are unique in possessing an enzyme that enables synthesis from this RNA of a DNA molecule capable of integrating into the DNA of a host cell. The retrovirus then utilizes the cell machinery to replicate. HIV is one of the best-known retroviruses. Oncogenic retroviruses (or oncoretroviruses) are cancer-causing viruses. Numerous oncoretroviruses are associated with animal diseases.
  • HTLV and XMRV retroviruses
  • XMRV retrovirus Xenotropic murine leukemia virus-related virus belongs to the virus family Retro viridae and the genus gammaretro virus. It has a single-stranded RNA genome that replicates through a DNA intermediate. Its name refers to its close relationship with the murine leukemia viruses ("MuLVs"). The genome, approximately 8100 nucleotides in length, is 95% identical with several endogenous retroviruses of mice, and is 93-94% identical with several exogenous mouse viruses.
  • XMRV genomic sequences have been published to date. These sequences are almost identical, an unusual finding as retroviruses replicate their genomes with relatively low fidelity, leading to divergent viral sequences in a single host organism.
  • XMRV was first described in 2006 and has been isolated from human biological samples. Several reports have associated the virus with prostate cancer, and later with chronic fatigue syndrome (CFS) but other reports do not find an association. It has not yet been established whether XMRV is a cause of disease.
  • XMRV protein has been found in tumour-associated but nonmalignant stromal cells, but in one study is was not found in the actual prostate cancer cells, raising the possibility that the virus may indirectly support tumorigenesis [McLaughlin-Drubin ME, Munger K (2008) Biochimica et Biophysica Acta 1782 (3): 127-50]. However, in another study, XMRV proteins and nucleic acids were found in malignant cells [Aloia, et al. (2010). Cancer research, 70: 10028-10033].
  • WO2006/110589 discloses the use of the complete virus for treating prostate cancer. However, this document never demonstrates that wild type virus can be efficiently used for the vaccination of patients.
  • One aim of the invention is to provide a new mutated attenuated virus, having enhanced vaccinal efficiency.
  • Another aim of the invention is to provide a new pharmaceutical composition for treating pathologies.
  • the present invention relates to a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the present invention is based on the unexpected observation made by the Inventors that mutation in the immunosuppressive domain of XMRV ENV protein both
  • the minimal mutation according to the invention is a substitution of the E at position 14 of SEQ ID NO : 1 by R, H or K.
  • a given protein is said to possess an immunosuppressive property, if it is liable to inhibit the immune system of an organism in which it is present.
  • the immunosuppressive property of said given protein can be measured by following the general procedure described in Mangeney & Heidmann (1998) Proc. Natl. Acad. Sci. U.S.A. 95: 14920-5 and Mangeney et al. (2001) J. Gen. Virol. 82:2515-8.
  • the immunosuppressive property of a given protein can also be characterized by its immunosuppression index [(Ap ro tein-A non e)/A none ]. If the immunosuppression index of a given protein is positive then the given protein is said to be immunosuppressive, and if its immunosuppression index is equal to zero or negative, the given protein is said to have essentially no immunosuppressive activity.
  • the inhibiting of the immunosuppressive property of a given protein yields a protein with substantially no immunosuppressive activity, that is having an immunosuppression index equal to zero or negative.
  • the invention relates to a mutated XMRV ENV protein according as defined above, in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • the Inventors have demonstrated that the advantageous mutated XMRV ENV protein, harbouring the above mentioned properties, should be mutated both:
  • the above mentioned mutated XMRV ENV proteins have substantially lost, or completely lost, their immunosuppressive properties, and have a substantially enhanced, or substantially increased antigenicity compared to the antigenicity of the corresponding wild type XMRV ENV protein.
  • the above mentioned mutated XMRV ENV proteins have substantially lost, or lost, their immunosuppressive properties, and have a substantially enhanced, or increased antigenicity compared to the antigenicity of the corresponding wild type XMRV ENV protein.
  • the antigenicity of said mutated XMRV ENV proteins is at least 5 fold, preferably at least 10 fold higher compared to the antigenicity of the wild type XMRV ENV protein.
  • the invention relates to the mutated XMRV ENV previously defined, wherein said two mutations are such that
  • Advantageous mutated XMRV ENV proteins are proteins wherein: - E which is at the position 14 of SEQ ID NO : 1 is substituted by R, and A which is at the position 20 of SEQ ID NO: 1 is substituted by F,
  • the invention relates to the mutated XMRV ENV previously defined, wherein said two mutations are such that
  • the invention relates to the mutated XMRV ENV previously defined, wherein wild type XMRV ENV protein consists of the amino acid sequences chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1), and SEQ ID NO : 8, (ACY30462.1).
  • wild type XMRV ENV protein consists of the amino acid sequences chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83
  • the invention relates to the mutated XMRV ENV previously defined, wherein said mutated XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 9, SEQ ID NO : 10, SEQ ID NO : 11, SEQ ID NO : 12, SEQ ID NO : 13, SEQ ID NO : 14, SEQ ID NO : 15, SEQ ID NO : 16, SEQ ID NO : 17, SEQ ID NO : 18, SEQ ID NO : 19, SEQ ID NO : 20, SEQ ID NO : 21, SEQ ID NO : 22 and SEQ ID NO : 23.
  • the invention also relates to a nucleic acid molecule coding for mutated XMRV ENV protein as defined above.
  • the invention also relates to a nucleic acid molecule coding for mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLCAA), said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the invention relates to the nucleic acid molecule as defined above, in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • the invention relates to the nucleic acid molecule as defined above, wherein said two mutations are such that
  • the invention relates to the nucleic acid molecule as defined above, wherein said two mutations are such that
  • the invention relates to the nucleic acid molecule as defined above, wherein wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1), and SEQ ID NO : 8, (ACY30462.1).
  • wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1), and
  • the invention relates to the nucleic acid molecule as defined above coding for a mutated XMRV ENV protein consisting of the amino acid sequence chosen among the group comprising: SEQ ID NO : 9, SEQ ID NO : 10, SEQ ID NO : 11, SEQ ID NO : 12, SEQ ID NO : 13, SEQ ID NO : 14, SEQ ID NO : 15, SEQ ID NO : 16, SEQ ID NO : 17, SEQ ID NO : 18, SEQ ID NO : 19, SEQ ID NO : 20, SEQ ID NO : 21, SEQ ID NO : 22 and SEQ ID NO : 23.
  • the invention also relates to a vector comprising a nucleic acid molecule as defined above, coding for a mutated XMRV ENV as defined above.
  • the invention also relates to a vector comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the vector previously defined, coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the vector previously defined, coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the vector previously defined, coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the vector previously defined, coding for a mutated XMRV ENV protein consisting of the amino acid sequence chosen among the group comprising: SEQ ID NO : 9, SEQ ID NO : 10, SEQ ID NO : 1 1, SEQ ID NO : 12, SEQ ID NO : 13, SEQ ID NO : 14, SEQ ID NO : 15, SEQ ID NO : 16, SEQ ID NO : 17, SEQ ID NO : 18, SEQ ID NO : 19, SEQ ID NO : 20, SEQ ID NO : 21, SEQ ID NO : 22 and SEQ ID NO : 23.
  • the invention relates to the vector previously defined, further comprising nucleic acid sequences allowing the expression of said nucleic acid molecule, as defined above, said nucleic acid molecule coding for a mutated XMRV ENV protein as defined above.
  • the invention relates to the vector previously defined, further comprising nucleic acid sequences allowing the expression of at least one another XMRV protein.
  • the invention relates to the vector previously defined, in association with at least one another vector as defined above comprising nucleic acid sequences allowing the expression of at least one another XMRV protein.
  • XMRV proteins can be GAG, PRO and POL XMRV proteins.
  • the invention relates to the vector previously defined, further comprising nucleic molecule coding for XMRV GAG protein. In one advantageous embodiment, the invention relates to the vector previously defined, further comprising nucleic acid molecule coding for XMRV POL protein.
  • the invention relates to the vector previously defined, further comprising nucleic acid molecule coding for XMRV PRO protein.
  • the invention relates to the vector previously defined, further comprising a first nucleic acid molecule coding for XMRV GAG protein, and a second nucleic acid molecule coding for XMRV POL protein.
  • the invention relates to the vector previously defined, further comprising a first nucleic acid molecule coding for XMRV GAG protein, and a second nucleic acid molecule coding for XMRV PRO protein. In one advantageous embodiment, the invention relates to the vector previously defined, further comprising a first nucleic acid molecule coding for XMRV PRO protein, and a second nucleic acid molecule coding for XMRV POL protein.
  • the invention relates to the vector previously defined, further comprising a first nucleic acid molecule coding for XMRV GAG protein, a second nucleic acid molecule coding for XMRV PRO protein, and a third nucleic acid molecule coding for XMRV POL protein.
  • the invention relates to the vector previously defined, said vector being a viral vector, in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a viral vector in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the invention relates to the vector previously defined, in association with at least one another vector, said another vector being a viral vector, in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector, comprising nucleic acid sequences allowing the expression of at least one another XMRV protein.
  • XMRV proteins can be GAG, PRO and POL XMRV proteins.
  • Canarypox vector as described in Poulet, H., et al (2003) Veterinary Record; 153:141-145 can be used as vector according to the invention.
  • Adenovirus vectors as described in Bayer, W., et al. (2008) Vaccine 26, 716-726, and in particular Bayer, W., et al. (2010) J. Virol. 84, 1967-1976, can be used as vector according to the mvention.
  • the invention relates to the vector previously defined, said vector being a viral vector, wherein in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for mutated XMRV ENV protein according to the invention.
  • nucleic acid molecule coding for another XMRV protein, which is not ENV protein is not ENV protein.
  • the invention relates to the vector previously defined, said vector being a viral vector, wherein in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for mutated XMRV ENV protein according to the invention.
  • nucleic acid molecule coding for GAG XMRV protein is Measles vector as described in Combredet, C, et ai. 2003. J. Virol. 77 (21), 11546-1 1554 and Guerbois, M., (2009) Virology 388 191-203, and canarypox vector as described in Poulet, H., et ai. (2003) Veterinary Record; 153:141-145.
  • the invention relates to the vector previously defined, said vector being a viral vector, wherein in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for mutated XMRV ENV protein according to the invention.
  • the invention relates to the vector previously defined, said vector being a viral vector, wherein in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for POL XMRV protein is provided.
  • the invention relates to the vector previously defined, said vector being a viral vector, wherein in particular a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • a pox vector chosen among a fowlpox, a canarypox, or a MVA (modified vaccinia virus Ankara) vector, an adenoviral vector, a measles vector, or a CMV (cytomegalovirus) vector.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for GAG XMRV protein nucleic acid molecule coding for GAG XMRV protein
  • nucleic acid molecule coding for PRO XMRV protein comprises:
  • nucleic acid molecule coding for mutated XMRV ENV protein according to the invention.
  • nucleic acid molecule coding for POL XMRV protein is provided.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for mutated XMRV ENV protein according to the invention.
  • nucleic acid molecule coding for PRO XMRV protein nucleic acid molecule coding for PRO XMRV protein
  • nucleic acid molecule coding for POL XMRV protein is provided.
  • the vector mentioned above comprises:
  • nucleic acid molecule coding for GAG XMRV protein nucleic acid molecule coding for GAG XMRV protein
  • nucleic acid molecule coding for PRO XMRV protein nucleic acid molecule coding for PRO XMRV protein
  • nucleic acid molecule coding for POL XMRV protein is provided.
  • the invention also relates to a composition, in particular a pharmaceutical composition, or a vaccinal composition, preferably in association with a pharmaceutically acceptable carrier or vehicle, comprising:
  • a mutated XMRV ENV protein as defined above, or
  • the invention also relates to a composition, in particular a pharmaceutical composition, or a vaccinal composition, as defined above, for its use as medicine or drug or vaccine.
  • the appropriate dosage of the composition of the invention can be adapted as a function of various parameters, in particular the mode of administration; the composition employed; the age, health, and weight of the host organism; the nature and extent of symptoms; kind of concurrent treatment; the frequency of treatment; and/or the need for prevention or therapy. Further refinement of the calculations necessary to determine the appropriate dosage for treatment is routinely made by a practitioner, in the light of the relevant circumstances.
  • a composition based on vector plasmids may be administered in doses of between 10 ⁇ g and 20 mg, advantageously between 100 ⁇ g and 2 mg.
  • a protein composition may be administered in one or more doses of between 10 ng and 20 mg, advantageously a dosage from about 0.1 ⁇ g to about 2 mg of the therapeutic protein per kg body weight. The administration may take place in a single dose or a dose repeated one or several times after a certain time interval.
  • the invention also relates to a composition, in particular a pharmaceutical composition, or a vaccinal composition, preferably in association with a pharmaceutically acceptable carrier or vehicle, comprising a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • a "pharmaceutically acceptable vehicle” is intended to include any and all carriers, solvents, diluents, excipients, adjuvants, dispersion media, coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like, compatible with pharmaceutical administration.
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, comprising a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA),
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, comprising a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA),
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, wherein wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1), and SEQ ID NO : 8, (ACY30462.1).
  • wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, wherein said mutated XMRV ENV protein consists of the amino acid sequence chosen among the group comprising SEQ ID NO : 9, SEQ ID NO : 10, SEQ ID NO : 11, SEQ ID NO : 12, SEQ ID NO : 13, SEQ ID NO : 14, SEQ ID NO : 15, SEQ ID NO : 16, SEQ ID NO : 17, SEQ ID NO : 18, SEQ ID NO : 19, SEQ ID NO : 20, SEQ ID NO : 21, SEQ ID NO : 22 and SEQ ID NO : 23.
  • the invention also relates to a composition, in particular a pharmaceutical composition, or a vaccinal composition, preferably in association with a pharmaceutically acceptable carrier or vehicle, comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO: 1, (LQNRRGLDILFLKEGGLC AA) , said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO: 1, (LQNRRGLDILFLKEGGLC AA) ,
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO: 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO: 1, (LQNRRGLDILFLKEGGLC AA) ,
  • the invention relates to the composition, in particular a pharmaceutical composition, defined above, wherein wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1), and SEQ ID NO : 8, (ACY30462.1).
  • wild type XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 2, (YP_512363.1), SEQ ID NO : 3, (ABB83229.1), SEQ ID NO : 4, (ACY30457.1), SEQ ID NO : 5, (ABM47429.1), SEQ ID NO : 6, (ACY3046.1), SEQ ID NO : 7, (ABB83226.1
  • the invention relates to the composition defined above, wherein said mutated XMRV ENV protein consists of the amino acid sequence chosen among the group comprising: SEQ ID NO : 9, SEQ ID NO : 10, SEQ ID NO : 11, SEQ ID NO : 12, SEQ ID NO : 13, SEQ ID NO : 14, SEQ ID NO : 15, SEQ ID NO : 16, SEQ ID NO : 17, SEQ ID NO : 18, SEQ ID NO : 19, SEQ ID NO : 20, SEQ ID NO : 21, SEQ ID NO : 22 and SEQ ID NO : 23.
  • the invention also relates to the use of
  • a mutated XMRV ENV protein as defined above, or
  • nucleic acid coding for a mutated XMRV ENV protein as defined above, or
  • nucleic acid molecule coding for a mutated XMRV ENV protein as defined above, or
  • the invention also relates to a composition
  • a composition comprising:
  • a mutated XMRV ENV protein as defined above, or
  • nucleic acid coding for a mutated XMRV ENV protein as defined above, or a vector comprising nucleic acid molecule coding for a mutated XMRV ENV protein, as defined above, or
  • the invention also relates to a method for the prevention or treatment of XMRV infection, comprising the administration in a person in a need thereof of a therapeutically effective amount of a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the invention relates to the method for the prevention or treatment of XMRV infection, as defined above, comprising the administration in a person in a need thereof of a therapeutically effective amount of a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations , said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA),
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • the invention relates to the method for the prevention or treatment of XMRV infection, as defined above, comprising the administration in a person in a need thereof of a therapeutically effective amount of a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the invention relates to the method for the prevention or treatment of XMRV infection, as defined above, comprising the administration in a person in a need thereof of a therapeutically effective amount of a nucleic acid coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations , said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • the invention also relates to a method for the prevention or treatment of XMRV infection, comprising the administration in a person in a need thereof of a therapeutically effective amount of vector comprising a nucleic acid molecule coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents at least one mutation , said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKEGGLC AA) ,
  • said at least one mutation being such that E which is at the position 14 of SEQ ID NO : 1 is substituted by R, H or K.
  • the invention also relates, in one another advantageous embodiment, to the method for the prevention or treatment of XMRV infection, as defined above comprising the administration in a person in a need thereof of a therapeutically effective amount of a vector comprising a nucleic acid coding for a mutated XMRV ENV protein in which the immunosuppressive domain of the wild type XMRV ENV protein presents two mutations, said immunosuppressive domain of the wild type XMRV ENV protein consisting of the amino acid sequence SEQ ID NO : 1, (LQNRRGLDILFLKE GGLC AA) ,
  • a which is at the position 20 of SEQ ID NO: 1, is such that it ensures that the structure of the viral XMRV ENV protein is conserved.
  • compositions mentioned above can be administered in a host via different routes: intraperitoneal (i.p.), subcutaneous (s.c), intradermal (i.d.), intramuscular (i.m.) or intravenous (i.v.) injection, oral administration and intranasal administration or inhalation.
  • the method of the invention can be carried out in conjunction with one or more conventional therapeutic modalities (e.g. radiation, chemotherapy and/or surgery).
  • therapeutic modalities e.g. radiation, chemotherapy and/or surgery.
  • the use of multiple therapeutic approaches provides the patient with a broader based intervention.
  • the method of the invention can be preceded or followed by a surgical intervention.
  • radiotherapy e.g. gamma radiation.
  • Those skilled in the art can readily formulate appropriate radiation therapy protocols and parameters which can be used (see for example Perez and Brady, 1992, Principles and Practice of Radiation Oncology, 2nd Ed. JB Lippincott Co; using appropriate adaptations and modifications as will be readily apparent to those skilled in the field).
  • Figures 1A-D Characterization of the fusogenic and IS activities of F-MLV Env and generation of a fusion-positive immunosuppression-negative specific mutant.
  • Figure 1A is a schematic representation of the F-MLV Env with the surface (SU) and transmembrane (TM) subunits, the furin cleavage site, the hydrophobic fusion peptide, the transmembrane anchor, and the IS domain with its peptide sequence indicated; the E>R and A>F substitutions generated in the single or double mutants are positioned.
  • SU surface
  • TM transmembrane
  • Figure IB represents the infectivity of Mo-MLV virions pseudotyped with F-MLV Env, wild-type (first column), single mutant (E14R second column, A20F third column), or double-mutant (DM fourth column).
  • Y axis is a logarithmic representation of the infectivity (ffu/mL).
  • Figure 1C represents the w vivo immunosuppressive (IS) activity of WT (first column) and DM (second column) F-MLV Env.
  • Y axis represents the immunosuppressive index.
  • Y- axis represents the number of virus in supernatant (RNA copy/mL), and X- axis represents hours after infection.
  • Figures 2A and B represent the requirement of the immunosuppression-positive domain for F-MLV propagation in immunocompetent mice.
  • Each circle corresponds to one individual mouse, with the lines connecting the geometric means. No viral RNA was detected in PBS-injected control mice.
  • Figure 2A represents results obtained with untreated mice.
  • Figure 2B represents results obtained with mice that were X-ray-irradiated (5 Gray) 2 days before infection.
  • Figure 3 Cellular targets of the Env-mediated immunosuppression effect, assayed by immune cell depletions in vivo. Serum viral loads after injection of WT (circles) or DM (squares) F-MLV were measured in untreated (closed symbols) or NK.1.1 -depleted (open symbols) Swiss (A) or Swiss-Nude (B) mice, or in CD8-depleted (closed symbols) or CD8- depleted plus NK1.1 -depleted (open symbols) Swiss mice (C). The data correspond to the mean +/- SD for five mice, and are representative of 2-7 independent experiments.
  • Figure 4A represents a curve showing the protection of mice immunized with DM F-MLV and challenged with WT F-MLV.
  • Swiss mice were injected with 8 x 10 8 RNA copies of DM F-MLV (white and grey circles) or injected with PBS (black circles), and the absence of viremia was checked four weeks later (day -35).
  • mice were challenged (indicated by arrow) with 4 x 10 7 RNA copies of WT F-MLV (grey and black circles) or injected with PBS (white circles), and post-challenge sera were collected at the indicated time points.
  • Each circle corresponds to one individual mouse, with the lines connecting the geometric means of 8 mice; data are representative of more than (>) 3 independent experiments, with in all cases no significant departure from full control of viremia by the vaccinated mice.
  • Y- axis represents the viral load (RNA copies/mL) and X- axis represents the days after infection
  • Figure 4B represents graphs comparing immunogenicity (B-cell responses) of WT and DM UV-inactivated F-MLV.
  • C57B1/6 mice were injected thrice at one week interval with 10 9 RNA copies of UV-inactivated WT or DM F-MLV in the presence of 50 ⁇ g of CpG ODN.
  • mice One week after the last injection, mice were blood sampled and serially diluted serum was used to detect TM-specific (left) and Gag-specific (right) IgG by ELISA. Results are the mean +/- SD of five mice, and are representative of 3 independent experiments.
  • Figure 4C represents graphs comparing immunogenicity (T-cell responses) of WT and DM UV-inactivated F-MLV.
  • C57B1/6 mice were injected thrice at one week interval with 10 9 RNA copies of UV-inactivated WT or DM F-MLV in the presence of 50 ⁇ g of CpG ODN.
  • mice were sacrificed and splenocytes restimulated in vitro for 72 h in the presence or absence of the I-A b restricted HI 9 Env peptide (left) or K b -restricted Gag peptide (right).
  • Specific IFN- ⁇ secretion by CD4 or CD8 T cells was detected in culture supernatants by standardized sandwich ELISA.
  • Figures 5A to G represents the ex vivo viremia of WT and DM GFP-marked F-MLV.
  • GFP marked F-MLV virions were generated by transient transfection of 293T cells with p57- IRESgfp plasmids, and the filtered supematants were used to infect NIH/3T3 cells under conditions similar to those in Figure 1.
  • Figure 5A represents phase-contrast microscopy visualization GFP-expressing cells infected with no virus (mock transfected 293T-cell supernatant) 4 days postinfection.
  • Figure 5B represents phase-contrast microscopy visualization GFP-expressing cells infected with DM GFP-marked F-MLV, 4 days postinfection.
  • Figure 5C represents phase-contrast microscopy visualization GFP-expressing cells infected with WT GFP-marked F-MLV 4 days postinfection.
  • Figure 5D represents fluorescence microscopy visualization GFP-expressing cells infected with no virus (mock transfected 293T-cell supernatant) 4 days postinfection.
  • Figure 5E represents fluorescence microscopy visualization GFP-expressing cells infected with DM GFP-marked F-MLV, 4 days postinfection.
  • Figure 5F represents fluorescence microscopy visualization GFP-expressing cells infected with WT GFP-marked F-MLV 4 days postinfection.
  • Figure 5G represents a curve comparing in vitro propagation kinetics of WT (black circles) and DM (gray circles) GFP-marked F-MLV.
  • the percentages of GFP+ infected NIH/3T3 cells were determined by flow cytometry using a FACScalibur flow cytometer (BD Biosciences) at days 1 , 4, and 6 postinfection (mean ⁇ SD of three independent experiments).
  • Y-axis represents the percentage of infected cells, and X-axis represnts the days post infection.
  • FIG. 6 Control injection of anti-NKl .l antibodies into NK1.1- Swiss mice does not allow mutant F-MLV to propagate.
  • Upper Phenotyping of NK1.1+ and NK1.1- Swiss mice as visualized by flow cytometry on blood samples using anti-NKl . l and anti-NKG2A,C,E antibodies.
  • PE means phycoerythrin.
  • Figure 7 Natural Treg cells do not affect F-MLV viremia.
  • Swiss mice were injected with 100 ⁇ g of anti-CD25 IgG on day -1 (Materials and Methods) and were infected at day 0 with 4 ⁇ 10 7 copies of WT or DM F-MLV.
  • Figures 8A to D represent curves analysing the antiviral antibody responses to WT and DM F-MLV [and control (PBS)].
  • Swiss mice were i.v. inoculated as in Figure 2 with 2 x 10 7 copies of WT or DM F-MLV, and serum was harvested 8 weeks postinfection.
  • Figure 8A represents ELISA results obtained with Serially diluted serum to detect SU specific IgG. Each symbol represents one individual mouse with the bars corresponding to the means, and results are representative of three independent experiments. Y- axis represents amount of IgG (ng/niL).
  • Figure 8B represents ELISA results obtained with Serially diluted serum to detect TM specific IgG. Each symbol represents one individual mouse with the bars corresponding to the means, and results are representative of three independent experiments. Y- axis represents amount of IgG (ng/niL).
  • Figure 8C represents ELISA results obtained with Serially diluted serum to detect TM specific IgG. Each symbol represents one individual mouse with the bars corresponding to the means, and results are representative of three independent experiments. Y- axis represents amount of IgG (arbitrary units; a.u.).
  • Figure 8D represents assay for F-MLV neutralizing antibodies in the collected sera. Neutralizing antibodies were tested for their ability to inhibit infection of NIH/ 3T3 target cells by F-MLV. Results for serum of WT ( ⁇ ) or DM (o) F-MLV-infected mice are expressed as percentage of integrated proviral copies relative to controls with serum from mock-infected (with PBS) mice. Each symbol corresponds to the mean of triplicates ⁇ SD for each individual mouse. Y-axis represents F-MLV provirus integration in percent, and X- axis indicates the sera dilutions.
  • Figures 9 A and B represent functional immunosuppressive domains in human retrovirus Envs.
  • Figure 9A represents sequence alignment of the F-MLV, XMRV, and HTLV-1 retrovirus and of the syncytin-1 (Syn- 1) ectodomains and identification of the two residues controlling their IS activity.
  • Figure 9B represents histograms showing the differential antibody responses induced by WT and DM recombinant ectodomains.
  • Recombinant proteins corresponding to the 64 amino acids (as shown in figure 9A) of the WT or DM TM ectodomain for F-MLV, XMRV, and Syn-1 and to the 84 amino acids of the WT or DM TM ectodomain fused with the MBP protein for HTLV-1 were i.v. injected three times with a 1-week interval into Swiss mice (50 ⁇ g per injection).
  • mice were blood-sampled and IgG levels were determined by ELISA using plates coated with the appropriate WT ectodomain protein for F-MLV, XMRV, and Syn-1 or with MBP-LacZ for HTLV-1. IgG levels are expressed relative to that of the WT ectodomain set as unity for retroviruses (and to that of the DM ectodomain for syncytin-1). Results are representative of 2-5 independent experiments. Similar results were observed when ELISA plates were coated with the DM ectodomain proteins from F-MLV, XMRV, and Syn-1. Y- axis represents amount of antibodies expressed in arbitrary units (a.u).
  • MCA205 tumor cells were transduced with pDFG expression vectors encoding either the WT or DM version of secreted F-MLV, XMRV, HTLV-1, and Syn-1 ectodomains.
  • the tumor rejection assay was performed as described in Materials and Methods.
  • Persistent viruses and notably retroviruses, have developed diverse strategies to subvert the host antiviral response, which allow them to escape the innate and adaptive immune system, by directly affecting and/or inducing specific sets of immune cells, including NK, CD8 T or regulatory T (Treg) cells [lannello A, et al. (2006) J Leukoc Biol 79:16-35; Schneider-Schaulies S and Dittmer U (2006) J Gen Virol 87:1423-38]. To characterize -and possibly counteract - the mechanisms by which retroviruses escape immune rejection, the Inventors thought that identification of the viral effector protein(s) associated with this activity was an unavoidable pre-requisite.
  • the Friend Virus is a complex of mouse retroviruses composed of the non-pathogenic replication-competent Friend Murine Leukemia Virus (F-MLV) and a pathogenic replication- defective spleen focus-forming virus (SFFV).
  • F-MLV non-pathogenic replication-competent Friend Murine Leukemia Virus
  • SFFV pathogenic replication- defective spleen focus-forming virus
  • This mutant allowed us to unambiguously demonstrate that i) a functional ISD is absolutely required for viremia in vivo, in immunocompetent mice, ii) the ISD inhibits both the innate and adaptive arms of the immune response, and iii) the mutations within the ISD resulting in loss of its IS activity are associated with enhanced immunogenicity of the viral antigens.
  • the Inventors also identified functional ISDs in other retroviruses, namely the human T cell leukemia virus (HTLV) and the xenotropic MLV related virus (XMRV) initially discovered in human prostate tumors [ Urisman A, et al. (2006) PLoS Pathog 2:e25], which therefore become, owing to the present mouse model and experimental results, definite targets for therapeutic antiviral approaches, as well as optimized vaccine antigens when adequately mutated.
  • HTLV human T cell leukemia virus
  • XMRV xenotropic MLV related virus
  • the MLV retroviral Env comprises within its TM subunit a sequence that the Inventors have previously delineated as responsible for its IS activity.
  • the 20 aa sequence shown in Figure 1A corresponds to that of both Mo- and F-MLV.
  • MPMV Mason-Pfizer monkey virus
  • the extent of “immunosuppression” can be quantified by an index based on tumor size, a positive index indicating that env expression facilitates tumor growth, as a consequence of its IS activity, a null or negative index pointing to no effect or even enhanced rejection, respectively (the latter may be explained by a stimulation of the immune response of the host against the new foreign antigen, represented by a non-immunosuppressive Env, expressed at the surface of the tumor cells).
  • a stimulation of the immune response of the host against the new foreign antigen represented by a non-immunosuppressive Env, expressed at the surface of the tumor cells.
  • F-MLV Env is immunosuppression-positive and, remarkably, the doubly mutated, but still functional, Env becomes immunosuppression-negative.
  • the Inventors then replaced the WT env gene by its non-immunosuppressive DM counterpart in the F-MLV proviral molecular clone 57 [Sitbon M, et al. (1990) J Virol 64:2135-40], and produced ex vivo each type of retroviral particles.
  • Viral particles were generated upon transfection of 293T cells with the WT or DM p57 plasmid and infection of NIH/3T3 producer cells with the harvested cell supernatant.
  • Virus yields for either plasmid were similar as measured by a quantitative RT-PCR assay of viral RNA in the NIH/3T3 producer cell supernatants.
  • both viruses display the same propagation kinetics in an in vitro infection assay in NIH/3T3 cells ( Figure ID; see also Figures 5 for IRES-g p-marked viruses), confirming that F-MLV DM Env is fully functional and that the introduced mutations have no effect on viral replication.
  • WT F-MLV first developed a high viremia in all animals during the primary infection phase (peak at day 7 after virus injection), followed by the establishment of a persistent infection with viremia reaching a plateau.
  • the non-immunosuppressive DM F-MLV was absent (or barely detectable in 4 out of 10 mice) at day 7, and undetectable 14 days after injection (as similarly observed when using even higher doses of virus, see Figure 4).
  • mice immunocompromised by 5 -Gray X-ray irradiation prior to infection displayed similar viral loads when injected with either WT or DM F-MLV, at least during the first 2 weeks of infection (Figure 2B), thus excluding that DM F-MLV is deficient in any step of its in vivo replication, consistent with the in vitro data ( Figure IB and Figure ID).
  • DM F-MLV was eliminated when the mouse immune system recovered from irradiation, with no more viremia detected 4 weeks post-infection.
  • NK cells Role of NK cells in virus control.
  • the Inventors searched for immune effectors responsible for rejection of the mutant F-MLV, reasoning that since these effectors were not able to eliminate WT F-MLV, they should be -directly or indirectly- the target of Env-driven immunosuppression. Due to the rapid "control" of DM F-MLV ( Figure 2A), the Inventors suspected a role for innate immune effectors, and therefore tested the possible involvement of NK cells, known for their antiviral activity, upon in vivo cell depletion. As not all the animals from the outbred Swiss mouse strain were found to express the NKl.
  • NKl .l + mice were first sorted based on their blood cell NKl .l expression, before being treated with the anti- NK1.1 antibody.
  • NK1.1 + cell depletion enabled the propagation of DM F-MLV, with viral loads similar to those observed with WT F-MLV when assayed 5 days post-infection.
  • this effect was not observed upon treatment of NKl . l -negative Swiss mice by the anti-NKl .l antibody, with no viremia detected for DM F-MLV (see Figure 6).
  • an NK1.1 + cell subset most probably NK cells, efficiently and rapidly eliminates DM F-MLV, but fails to block WT virus.
  • propagation of DM F-MLV in the antibody-treated mice was transient and the virus was completely eliminated by 7 days after infection, despite maintenance of the NK-depleted state. This observation indicates that a second mechanism for virus clearance, independent of the NKl .l + cells, takes over at a later time to eradicate the mutant virus, with a kinetics compatible with the establishment of adaptive immunity.
  • NK1.1 + cells responsible for early rejection of DM F-MLV are NK, and not NKT cells, the latter requiring the thymus to differentiate [Benlagha K, et al. (2002) Science 296:553-5].
  • NK cells per se are sufficient to eradicate a non- immunosuppressive F-MLV both at the early stage of infection, and also at a later stage, when the virus is then additionally cleared by a T cell-dependent mechanism.
  • the Inventors chose to inject UV-inactivated F-MLV in inbred C57B1/6 mice, in order to (i) work with the same amounts of antigen for WT or DM F-MLV and (ii) analyze specific T cell responses against well defined immunodominant H-2 b - restricted antigenic peptides [Chen W, et al. (1996) J Virol 70:7773-82 ; Iwashiro M, et al. (1993) J Virol 67:4533-42].
  • Figure 4 shows that injection of the non-immunosuppressive DM F-MLV reproducibly induced significantly higher responses than its wild type counterparts, for both anti-TM and anti-Gag IgG and for IFN- ⁇ secretion by CD4 and CD8 T cells, suggesting that Env-mediated immunosuppression impairs, at least quantitatively, the induction of anti-viral responses.
  • DM F-MLV Env could be introduced back into the complete F- MLV cloned provirus, resulting in a mutant F-MLV retrovirus that disclosed infectivity and replication efficiency in vitro identical to that of WT virus, as assayed on NIH/3T3 cells with either the "basic" virus or an IRES-g p-marked version of the virus.
  • Full conservation of the functionality of DM F-MLV was further ascertained by the in vivo infection assay performed on immunocompromised X-ray- irradiated mice, which disclosed viremia identical to that of the wild-type virus, at least in the initial stages of infection.
  • the physiological impact of the IS function carried by a retroviral Env could then be unambiguously and specifically determined.
  • the Inventors could demonstrate that the non- immunosuppressive DM F-MLV is unable to propagate in untreated, immunocompetent mice, under conditions where such mice become persistently infected by WT F-MLV.
  • lack of viral propagation of DM F-MLV in vivo is not dependent on some "mechanical" defect of the virus but actually depends on loss of the Env-associated IS function, viremia being undistinguishable from that of WT virus in the X-irradiated mice assayed in parallel.
  • Treg cell inhibition or depletion is not sufficient for virus elimination, [Zelinskyy G, et al. (2006) Eur J Immunol 36:2658-70; Zelinskyy G, et al. (2009) Blood 114:3199-207; He H, et al. (2004) J Virol. 78:11641-7].
  • Treg cells -either natural or induced- are most probably not primary effectors of Env-mediated immunosuppression.
  • DC Dendritic cells
  • mice and cells were cultured in Swiss (N-tropic F-MLV permissive), Balb/c and C57BL/6 mice, 6-10 weeks old, were from Janvier (Laval, France). Swiss-Nude mice were from Institut Gustave Roussy breeding center. 293T (CRL11268), HeLa (CCL2), NIH/3T3 (CRL-1658), and MCA205 cells [Suzuki T, et al. (1995) J. Exp. Med. 182:477-486] were cultured in DMEM with 10 % FCS, streptomycin (100 ⁇ g/mL) and penicillin (100 units/mL). Mouse splenocytes were cultured in RPMI with 10 % FCS, antibiotics as above and 5 x 10 "5 M ⁇ -mercaptoethanol.
  • phCMV-envF-MLV was constructed by inserting the F-MLV env gene retrieved by PCR from p57 ([Sitbon M, et al. (1990) J Virol 64:2135-40]; gift from Dr Mougel) using primers 1 and 2 (see SI text for primer sequences) and digested with Xhol and Mlul, into phCMV-envHERV-T [Blaise S, et al. (2003) Proc Natl Acad Sci U S A 100:13013-8], digested with the same enzymes.
  • Each mutant derivative was constructed by a three- fragment ligation of phCMV-envF-MLV opened by Clal/Avrll, and two PCR products generated with primer pairs 3-4 and 5-6 for the E14R mutation, 3-7 and 6-8 for the A20F mutation, and 2-3 and 6-8 for the E14R+A20F mutation, restricted with Clal and Avrll.
  • the p57 DM F-MLV was constructed by inserting the BstZl lI/BsmI fragment of the phCMV-envF-MLV double mutant into p57 digested with the same enzymes.
  • the pDFG retroviral vector expressing the WT or DM F-MLV Env (and hygromycin resistance) for stable transduction of MCA205 cells was constructed by inserting the BspEI/MluI fragment from phCMV-envF-MLV into pDFG- MoTMl (10) digested with AgEI/MluI.
  • the bacterial expression vectors for WT and DM F- MLV Env ectodomains were constructed by inserting a PCR fragment, generated with WT or DM phCMV-envF-MLV as a template and primers 9-10 and digested with Ncol/Xhol, into pET28(+)b (Novagen) digested with the same enzymes.
  • Mo-MLV virions pseudotyped with WT or DM F-MLV Env were produced as in [Blaise S, et al. (2004) J Virol 78:1050-4] by cotransfecting 7.5 x 10 5 293T cells with 0.55 ⁇ g of phCMV-envF-MLV, 1.75 ⁇ g of a Mo-MLV gag-pol vector [Blaise S, et al. (2004) J Virol 78:1050-4] and 1.75 ⁇ g of a LacZ-marked defective retroviral vector (pMFGsnls/acZ), by calcium phosphate precipitation (Invitrogen).
  • pMFGsnls/acZ LacZ-marked defective retroviral vector
  • viral supernatants were harvested, filtered through 0.45 ⁇ m-pore-size membranes, and 5-500 ⁇ of supernatant supplemented with 8 ⁇ g polybrene/mL were transferred onto NIH/3T3 target cells (seeded in 24-well plates at 10 4 cells per well the day before infection).
  • Viral titers were measured by X-Gal staining 3 days post infection, and expressed as lacZ ffu/mL of viral supernatant.
  • MLV virions containing the WT or DM F-MLV env-expressing pDFG retroviral vector were produced by transfecting 293T cells with 1.75 ⁇ g of the pDFG vector plus non-retroviral expression vectors for the MLV proteins (0.55 ⁇ g for MLV env and 1.75 ⁇ g for MLV gag- pol, ref. 33). Released particles were then used to transduce MCA205 cells (5 x 10 5 cells).
  • Tumor area was determined by measuring perpendicular tumor diameters thrice weekly, and extent of "immunosuppression" quantified by an index based on tumor size: (A env - A none )/A none , where Ae nv and A none are the mean areas at the peak of growth of tumors from mice injected with env-expressing or control cells, respectively.
  • F-MLV production and assay 7.5 x 10 5 293T cells were transfected with 4 ⁇ g of WT or DM p57 DNA using calcium phosphate transfection (Invitrogen). Cell supernatants were collected 48 h later, filtered through 0.45 ⁇ m-pore-size membranes, supplemented with 8 ⁇ g/mL polybrene and used to infect 5 x 10 5 NIH/3T3 cells. Infected cells were cultured for 4 additional days, expanded, and supernatants collected from day 4 to 8 post-infection. Viral particles were concentrated by ultracentrifugation, resuspended in PBS, and frozen for further use.
  • Viral RNA from 2 ⁇ of concentrated virus or 20 ⁇ of cell supernatant or 20 ⁇ of mouse serum was extracted using the RNAeasy microkit (QIAgen), reverse-transcribed using the MoMLV reverse transcription kit (Applied Biosystems) with random hexamers as primers, and the cDNA was quantified by real-time PCR using the Platinum SYBR Green qPCR kit (Invitrogen) and primers CTCAGGGAGCAGCGGGA (SEQ ID NO : 24) and TAGCTTAAGTCTGTTCCAGGCAGTG (SEQ ID NO : 25). Only values >10 4 RNA copies/mL are significant.
  • PK136, YTS169 and PC61 hybridomas producing depleting antibodies against the NK1.1, CD8cc and CD25 antigens, respectively, were from Dr L. Zitvogel. Ascites were produced in Nude mice and antibodies were purified by FPLC using protein A columns. NK cell depletions were performed by intraperitoneal injection of 300 ⁇ g of anti-NKl . l antibodies at day -3, 0, 3, 7 and 10 post-infection for Swiss mice. For Nude mice, two additional injections were performed at day 13 and 17.
  • CD8 T cell depletions were performed by injecting 100 ⁇ g of anti-CD8 antibodies at day -1, 0, 5 and 10 post-infection.
  • CD25 depletions were performed by intraperitoneal injection of 100 ⁇ g of anti-CD25 antibodies the day before infection. Depletions were checked at day 5 post-infection by quantifying NK, CD8 and CD25 cells in mouse blood by flow cytometry, using the 20d5 anti- NKG2A/C/E (Serotec), 53-6.7 anti-CD8 (Miltenyi Biotec), and 3C7 anti-CD25 (BD Biosciences) antibodies, respectively.
  • Recombinant proteins, peptides and oligonucleotides were produced as in [Mangeney M, et al. (2007) Proc Natl Acad Sci U S A 104:20534-9] using BL21 (DE3) E.coli cells (Stratagene) and pET28(+)b-derived expression vectors (see Plasmids).
  • Recombinant WT and DM TM subunit ectodomains and Gag protein were soluble and were purified on HiTrap Chelating HP columns (Amersham). WT and DM ectodomains were further purified through a Superdex 75 HR10/30 column (Amersham) to isolate the - major- trimeric form.
  • the synthetic GagL 85 _ 9 3 (CCLCLTVFL (SEQ ID NO: 57)) peptide corresponding to the immunodominant Derestricted CD8 + T cell epitope from F-MLV Gag [Chen W, et al. (1996) J Virol 70:7773-82] and the synthetic H19-Envi 22 -i4i (EPLTSLTPRCNTAWNRLKL (SEQ ID NO: 58)) peptide corresponding to the immunodominant I-A b -restricted CD4 + T cell epitope from F-MLV gp70 [Iwashiro M, et al. (1993) J Virol 67:4533-42] were from Sequentia (Evry, France).
  • CpG oligonucleotides ODN 1826; TCCATGACGTTCCTGACGTT (SEQ ID NO : 26), CpG ODN) and control ODN (ODN 1982; TCCAGGACTTCTCTCAGGTT (SEQ ID NO : 27)) were synthesized by Proligo (France).
  • Anti-F-MLV antibodies detection and IFN- ⁇ assay IgG levels in serially diluted mouse sera were assayed by indirect ELISA using MaxiSorp microplates (Nunc, Denmark) coated with recombinant Gag or TM subunit ectodomains (2 ⁇ g/mL), anti-mouse IgG antibodies conjugated to HRP (Amersham), and BD-Opteia revelation kit (Sigma). Mouse anti-Hiss antibody (QIAgen) was used for standardization.
  • p57-lKES-gfp plasmids are derived from the F-MLV containing p57 plasmid (see above) by introducing an IKES-gfp sequence 3' to the F-MLV env gene.
  • Both the WT and the immunosuppression-negative DM versions were constructed, by first introducing Sail and Mlul restriction sites 3' to the env gene (via a three-fragment ligation between WT or DM p57 opened by Xhol and Bsml, and two PCR products generated with primer pairs S1-S2 and S3- S4).
  • PCR amplification was performed using primer pair S7-S8 and WT and DM phCMV-envF-MLV plasmids as templates. Sfil/Mlul digested PCR fragments were then introduced into the pDFG vector opened with the same enzymes.
  • the XMRV Env ectodomain encoding DNA sequence was generated by ligation of 3 sets of paired 70-75 oligomers with 4 nt cohesive ends for each oligomer, including a Sfil and Mlul restriction site for the external fragments.
  • the ligation product was introduced into the Sfil/Mlul opened pDFG vector.
  • This plasmid was used as a template for PCR amplification of the XMRV Env ectodomain with primer pair SI 1-S12 to introduce Ncol and Xhol restriction sites and the PCR fragment was introduced into the Ncol/Xhol digested pET28 plasmid.
  • pCMV-HTLVenv was a gift from C. Pique.
  • the DM pCMV-HTLVenv mutant was generated by triple ligation of PCR fragments generated with primer pairs S15-S16 and S17-S18, digested with Kpnl and Nsil respectively, and the opened Kpnl-Nsil restricted pCMV- HTLVenv.
  • the WT and DM HTLV Env ectodomains were then PCR-amplified with primer pair S19-S20, digested with Sfi-Mlu, and introduced into pDFG opened with the same enzymes.
  • coli MBP were constructed by a three- fragment ligation of pMal-c2x opened with Bglll and Hindlll, a Bglll and Pstl cut PCR fragment generated with primer pair S21-22 using pMal-c2x as a template, and a Pstl and Hindlll cut PCR fragment generated with primer pair S23-24 using pCMV- HTLVenv as a template.
  • These vectors encode a 84-residues long HTLV ectodomain fused to the C-terminus of MBP through a tri-alanine linker, identical to the fusion protein that has been crystallized [Jonjic S, et al.
  • the empty pMal-c2x vector encodes the 85-residues long alpha subunit of E. coli ⁇ -galactosidase fused to the C- terminus of MBP, and was used as a control protein.
  • Neutralizing antibodies were tested by their ability to inhibit infection of NIH/3T3 target cells by F-MLV, as described for another retrovirus in Langhammer et al. [Malim MH, Emerman M (2008) Cell Host Microbe 3:388-98] . 50 of F-MLV (2 x 10 6 copies/mL) were incubated with serial dilutions of heat-inactivated serum for 45 min at 37°C and then transferred to NIH/3T3 cells seeded at 5000 cells per well into 96-well microplates.
  • Cianciolo G Copeland TD, Orozlan S, Snyderman R (1985) Inhibition of lymphocyte proliferation by a synthetic peptide homologous to retroviral envelope proteins.

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Abstract

La présente invention concerne une protéine d'enveloppe mutée du virus XMRV dans laquelle le domaine immunosuppresseur de la protéine d'enveloppe du virus XMRV de phénotype sauvage présente au moins une mutation, ledit domaine immunosuppresseur de la protéine d'enveloppe du virus XMRV de phénotype sauvage étant constitué de la séquence d'acides aminés SEQ ID NO : 1 (LQNRRGLDILFLKEGGLCAA), et ladite mutation faisant que E en position 14 de SEQ ID NO : 1 est remplacé par R, H ou K.
PCT/EP2011/051062 2010-01-26 2011-01-26 Protéines d'enveloppe du virus xmrv présentant une mutation au niveau de leur domaine immunosuppresseur Ceased WO2011092199A1 (fr)

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WO2013050048A3 (fr) * 2011-10-07 2013-07-18 Skau Aps Identification et atténuation des domaines immunosuppresseurs dans des protéines de fusion de virus à arn enveloppés
WO2020043908A1 (fr) 2017-09-01 2020-03-05 Inprother Aps Vaccin destiné à être utilisé dans la prophylaxie et/ou le traitement d'une maladie
WO2024218400A1 (fr) 2023-04-21 2024-10-24 Inprother Aps Expression améliorée d'antigènes affichés en surface

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005095442A1 (fr) * 2004-03-30 2005-10-13 Institut Gustave Roussy Sequence polypeptidique impliquee dans la modulation de l'effet immunosuppressif de proteines virales
WO2006103562A2 (fr) * 2005-03-30 2006-10-05 Centre National De La Recherche Scientifique (Cnrs) Retrovirus endogene et proteines codees par un gene env en tant que cible pour le traitement du cancer
WO2006110589A2 (fr) 2005-04-07 2006-10-19 The Cleveland Clinic Foundation Gammaretrovirus associe au cancer

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005095442A1 (fr) * 2004-03-30 2005-10-13 Institut Gustave Roussy Sequence polypeptidique impliquee dans la modulation de l'effet immunosuppressif de proteines virales
WO2006103562A2 (fr) * 2005-03-30 2006-10-05 Centre National De La Recherche Scientifique (Cnrs) Retrovirus endogene et proteines codees par un gene env en tant que cible pour le traitement du cancer
WO2006110589A2 (fr) 2005-04-07 2006-10-19 The Cleveland Clinic Foundation Gammaretrovirus associe au cancer

Non-Patent Citations (92)

* Cited by examiner, † Cited by third party
Title
ALOIA ET AL., CANCER RESEARCH, vol. 70, 2010, pages 10028 - 10033
ANDREWS DM ET AL., MOL IMMUNOL, vol. 42, 2005, pages 547 - 55
ANDREWS DM; ANDONIOU CE; SCALZO AA; VAN DOMMELEN SL; WALLACE ME ET AL.: "Cross-talk between dendritic cells and natural killer cells in viral infection", MOL IMMUNOL, vol. 42, 2005, pages 547 - 55, XP004686652, DOI: doi:10.1016/j.molimm.2004.07.040
BAYER, W. ET AL., J. VIROL., vol. 84, 2010, pages 1967 - 1976
BAYER, W. ET AL., VACCINE, vol. 26, 2008, pages 716 - 726
BENIT LAURENCE ET AL: "Identification, phylogeny, and evolution of retroviral elements based on their envelope genes", JOURNAL OF VIROLOGY, THE AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 75, no. 23, 1 December 2001 (2001-12-01), pages 11709 - 11719, XP002289562, ISSN: 0022-538X, DOI: DOI:10.1128/JVI.75.23.11709-11719.2001 *
BENLAGHA K ET AL., SCIENCE, vol. 296, 2002, pages 553 - 5
BENLAGHA K; KYIN T; BEAVIS A; TEYTON L; BENDELAC A: "A thymic precursor to the NK T cell lineage", SCIENCE, vol. 296, 2002, pages 553 - 5
BLAISE S ET AL., J VIROL, vol. 78, 2004, pages 1050 - 4
BLAISE S ET AL., J. GEN. VIROL, vol. 82, 2001, pages 1597 - 1600
BLAISE S ET AL., J. GEN. VIROL., vol. 82, 2001, pages 1597 - 1600
BLAISE S ET AL., PROC NATL ACAD SCI U S A, vol. 100, 2003, pages 13013 - 8
BLAISE S; DE PARSEVAL N; BENIT L; HEIDMANN T: "Genomewide screening for fusogenic human endogenous retrovirus envelopes identifies syncytin 2, a gene conserved on primate evolution", PROC NATL ACAD SCI USA, vol. 100, 2003, pages 13013 - 8, XP002289560, DOI: doi:10.1073/pnas.2132646100
BLAISE S; MANGENEY M; HEIDMANN T: "The envelope of Mason-Pfizer monkey virus has immunosuppressive properties", J. GEN. VIROL., vol. 82, 2001, pages 1597 - 1600, XP055052502, DOI: doi:10.1099/0022-1317-82-7-1597
BLAISE S; RUGGIERI A; DEWANNIEUX M; COSSET FL; HEIDMANN T: "Identification of an envelope protein from the FRD family of human endogenous retroviruses (HERV-FRD) conferring infectivity and functional conservation among simians", J VIROL, vol. 78, 2004, pages 1050 - 4, XP002336699, DOI: doi:10.1128/JVI.78.2.1050-1054.2004
BOWIE AG; UNTERHOLZNER L, NAT REV IMMUNOL, vol. 8, 2008, pages 911 - 22
BOWIE AG; UNTERHOLZNER L: "Viral evasion and subversion of pattern-recognition receptor signalling", NAT REV IMMUNOL, vol. 8, 2008, pages 911 - 22
CHEN W ET AL., J VIROL, vol. 70, 1996, pages 7773 - 82
CHEN W; QIN H; CHESEBRO B; CHEEVER MA: "Identification of a gag-encoded cytotoxic T-lymphocyte epitope from FBL-3 leukemia shared by Friend, Moloney, and Rauscher murine leukemia virus-induced tumors", J VIROL, vol. 70, 1996, pages 7773 - 82
CIANCIOLO G ET AL., SCIENCE, vol. 230, 1985, pages 453 - 455
CIANCIOLO G; COPELAND TD; OROZLAN S; SNYDERMAN R: "Inhibition of lymphocyte proliferation by a synthetic peptide homologous to retroviral envelope proteins", SCIENCE, vol. 230, 1985, pages 453 - 455, XP055052522
COMBRCDET, C. ET AL., J. VIROL., vol. 77, no. 21, 2003, pages 11546 - 11554
COMBREDET, C. ET AL., J. VIROL., vol. 77, no. 21, 2003, pages 11546 - 11554
DITTMER U ET AL., IMMUNITY, vol. 20, 2004, pages 293 - 303
DITTMER U; HE H; MESSER RJ; SCHIMME S; OLBRICH AR ET AL.: "Functional impairment of CD8(+) T cells by regulatory T cells during persistent retroviral infection", IMMUNITY, vol. 20, 2004, pages 293 - 303
GUERBOIS, M., VIROLOGY, vol. 388, 2009, pages 191 - 203
GUERMONPREZ P ET AL., ANNU REV IMMUNOL, vol. 20, 2002, pages 621 - 67
GUERMONPREZ P; VALLADEAU J; ZITVOGEL L; THERY C; AMIGORENA S: "Antigen presentation and T cell stimulation by dendritic cells", ANNU REV IMMUNOL, vol. 20, 2002, pages 621 - 67, XP009012587, DOI: doi:10.1146/annurev.immunol.20.100301.064828
HANSEN TH; BOUVIER M, NAT REV IMMUNOL, vol. 9, 2009, pages 503 - 13
HANSEN TH; BOUVIER M: "MHC class I antigen presentation: learning from viral evasion strategies", NAT REV IMMUNOL, vol. 9, 2009, pages 503 - 13
HE H ET AL., J VIROL., vol. 78, 2004, pages 11641 - 7
HE H; MESSER RJ; SAKAGUCHI S; YANG G; ROBERTSON SJ ET AL.: "Reduction of retrovirus-induced immunosuppression by in vivo modulation of T cells during acute infection", J VIROL., vol. 78, 2004, pages 11641 - 7
IANNELLO A ET AL., J LEUKOC BIOL, vol. 79, 2006, pages 16 - 35
IANNELLO A; DEBBECHE 0; MARTIN E; ATTALAH LH; SAMARANI S ET AL.: "Viral strategies for evading antiviral cellular immune responses of the host", JLEUKOC BIOL, vol. 79, 2006, pages 16 - 35
IWASHIRO M ET AL., J VIROL, vol. 67, 1993, pages 4533 - 42
IWASHIRO M ET AL., PROC NATL ACAD SCI USA, vol. 98, 2001, pages 9226 - 30
IWASHIRO M; KONDO T; SHIMIZU T; YAMAGISHI H; TAKAHASHI K ET AL.: "Multiplicity of virus-encoded helper T-cell epitopes expressed on FBL-3 tumor cells", J VIROL, vol. 67, 1993, pages 4533 - 42
IWASHIRO M; MESSER RJ; PETERSON KE; STROMNES IM; SUGIE T ET AL.: "Immunosuppression by CD4+ regulatory T cells induced by chronic retroviral infection", PROC NATL ACAD SCI US A, vol. 98, 17 July 2001 (2001-07-17), pages 9226 - 30, XP002488511, DOI: doi:10.1073/pnas.151174198
JONJIC S ET AL., CURR OPIN IMMUNOL, vol. 20, 2008, pages 30 - 8
JONJIC S; BABIC M; POLIC B; KRMPOTIC A: "Immune evasion of natural killer cells by viruses", CURR OPIN IMMUNOL, vol. 20, 2008, pages 30 - 8, XP022487766, DOI: doi:10.1016/j.coi.2007.11.002
LETVIN NL ET AL., J VIROL, vol. 81, 2007, pages 12368 - 74
LETVIN NL; RAO SS; DANG V; BUZBY AP; KORIOTH-SCHMITZ B ET AL.: "No evidence for consistent virus-specific immunity in simian immunodeficiency virus-exposed, uninfected rhesus monkeys", J VIROL, vol. 81, 2007, pages 12368 - 74
LO SC ET AL., PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 107, no. 36, 2010, pages 15874 - 15879
LOMBARDI VC ET AL., SCIENCE, vol. 326, no. 5952, 2009, pages 585 - 9
MALIM MH; EMERMAN M, CELL HOST MICROBE, vol. 3, 2008, pages 388 - 98
MALIM MH; EMERMAN M: "HIV-1 accessory proteins--ensuring viral survival in a hostile environment", CELL HOST MICROBE, vol. 3, 2008, pages 388 - 98
MANGENEY ET AL., J. GEN. VIROL., vol. 82, 2001, pages 2515 - 8
MANGENEY M ET AL., PROC NATL ACAD SCI U S A, vol. 104, 2007, pages 20534 - 9
MANGENEY M ET AL., PROC NATL ACAD SCI US A, vol. 104, 2007, pages 20534 - 9
MANGENEY M ET AL., PROC NATL ACAD SCI USA, vol. 104, 2007, pages 20534 - 9
MANGENEY M ET AL: "The full-length envelope of an HERV-H human endogenous retrovirus has immunosuppressive properties", JOURNAL OF GENERAL VIROLOGY, SOCIETY FOR GENERAL MICROBIOLOGY, SPENCERS WOOD, GB, vol. 82, no. 10, 1 October 2001 (2001-10-01), pages 2515 - 2518, XP002298110, ISSN: 0022-1317 *
MANGENEY M, RENARD M, SCHLECHT-LOUF G, BOUALLAGA I, HEIDMANN O, LETZELTER C, RICHAUD A, DUCOS B, HEIDMANN T.: "Placental syncytins: Genetic disjunction between the fusogenic and immunosuppressive activity of retroviral envelope proteins.", PROC NATL ACAD USA, vol. 104, no. 51, 12 December 2007 (2007-12-12), pages 20534 - 20539, XP002633415 *
MANGENEY M; HEIDMANN T, PROC. NATL. ACAD. SCI. U. S. A., vol. 95, 1998, pages 14920 - 14925
MANGENEY M; HEIDMANN T, PROC. NATL. ACAD. SCI. U.S.A., vol. 95, 1998, pages 14920 - 14925
MANGENEY M; HEIDMANN T: "Tumor cells expressing a retroviral envelope escape immune rejection in vivo", PROC. NATL. ACAD. SCI. U. S. A., vol. 95, 1998, pages 14920 - 14925, XP002298109, DOI: doi:10.1073/pnas.95.25.14920
MANGENEY M; RENARD M; SCHLECHT-LOUF G; BOUALLAGA I; HEIDMANN 0 ET AL.: "Placental syncytins: Genetic disjunction between the fusogenic and immunosuppressive activity of retroviral envelope proteins", PROC NATL ACAD SCI USA, vol. 104, 2007, pages 20534 - 9, XP002633415, DOI: doi:10.1073/PNAS.0707873105
MANGENEY; HEIDMANN, PROC. NATL. ACAD. SCI. U.S.A., vol. 95, 1998, pages 14920 - 5
MANSFIELD K ET AL., J VIROL, vol. 82, 2008, pages 4135 - 48
MANSFIELD K; LANG SM; GAUDUIN MC; SANFORD HB; LIFSON JD ET AL.: "Vaccine protection by live, attenuated simian immunodeficiency virus in the absence of high- titer antibody responses and high-frequency cellular immune responses measurable in the periphery", J VIROL, vol. 82, 2008, pages 4135 - 48
MCLAUGHLIN-DRUBIN ME; MUNGER K, BIOCHIMICA ET BIOPHYSICA ACTA, vol. 1782, no. 3, 2008, pages 127 - 50
METZGER MJ ET AL., JOURNAL OF VIROLOGY, vol. 84, no. 4, 2010, pages 1874 - 80
MYERS L ET AL., J IMMUNOL, vol. 183, 2009, pages 1636 - 43
MYERS L; MESSER RJ; CARMODY AB; HASENKRUG KJ: "Tissue-specific abundance of regulatory T cells correlates with CD8+ T cell dysfunction and chronic retrovirus loads", J IMMUNOL, vol. 183, 2009, pages 1636 - 43
PEREZ; BRADY: "Principles and Practice of Radiation Oncology", 1992, JB LIPPINCOTT CO
POULET, H. ET AL., VETERINARY RECORD, vol. 153, 2003, pages 141 - 145
RALAINIRINA N ET AL., JLEUKOC BIOL, vol. 81, 2007, pages 144 - 53
RALAINIRINA N; POLI A; MICHEL T; POOS L; ANDRES E ET AL.: "Control of NK cell functions by CD4+CD25+ regulatory T cells", JLEUKOC BIOL, vol. 81, 2007, pages 144 - 53
RENARD M ET AL., J MOL BIOL, vol. 352, 2005, pages 1029 - 34
RENARD M ET AL., JMOL BIOL, vol. 352, 2005, pages 1029 - 34
RENARD M; VARELA PF; LETZELTER C; DUQUERROY S; REY FA ET AL.: "Crystal structure of a pivotal domain of human syncytin-2, a 40 million years old endogenous retrovirus fusogenic envelope gene captured by primates", JMOL BIOL, vol. 352, 2005, pages 1029 - 34, XP005074546, DOI: doi:10.1016/j.jmb.2005.07.058
ROBERTSON SJ ET AL., J IMMUNOL, vol. 176, 2006, pages 3342 - 9
ROBERTSON SJ; MESSER RJ; CARMODY AB; HASENKRUG KJ: "In vitro suppression of CD8+ T cell function by Friend virus-induced regulatory T cells", J IMMUNOL, vol. 176, 2006, pages 3342 - 9
ROUSE BT ET AL., IMMUNOL REV, vol. 212, 2006, pages 272 - 86
ROUSE BT; SARANGI PP; SUVAS S: "Regulatory T cells in virus infections", IMMUNOL REV, vol. 212, 2006, pages 272 - 86
SCHLECHT-LOUF G.; RENARD M.; MANGENEY M.; LETZELTER C.; RICHAUD A.; DUCOS B.; BOUALLAGA I.; HEIDMANN T.: "Retroviral infection in vivo requires an immune escape virulence factor encrypted in the envelope protein of oncoretroviruses", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 107, no. 8, 23 February 2010 (2010-02-23), pages 3782 - 3787, XP002633414 *
SCHNEIDER-SCHAULIES S; DITTMER U, J GEN VIROL, vol. 87, 2006, pages 1423 - 38
SCHNEIDER-SCHAULIES S; DITTMER U: "Silencing T cells or T-cell silencing: concepts in virus-induced immunosuppression", J GEN VIROL, vol. 87, 2006, pages 1423 - 38
SITBON M ET AL., J VIROL, vol. 64, 1990, pages 2135 - 40
SITBON M; ELLERBROK H; POZO F; NISHIO J; HAYES SF ET AL.: "Sequences in the U5- gag-pol region influence early and late pathogenic effects of Friend and Moloney murine leukemia viruses", J VIROL, vol. 64, 1990, pages 2135 - 40
SMITH RA, RETROVIROLOGY, vol. 7, no. 1, 2010, pages 112
SUZUKI T ET AL., J. EXP. MED., vol. 182, 1995, pages 477 - 486
SUZUKI T; TAHARA H; NARULA S; MOORE KW; ROBBINS PD ET AL.: "Viral interleukin 10 (IL-10), the human herpes virus 4 cellular IL-10 homologue, induces local anergy to allogeneic and syngeneic tumors", J. EXP. MED., vol. 182, 1995, pages 477 - 486, XP000979495, DOI: doi:10.1084/jem.182.2.477
URISMAN A ET AL., PLOS PATHOG, vol. 2, 2006, pages E25
URISMAN A ET AL., PLOS PATHOGENS, vol. 2, no. 3, 2006, pages E25
URISMAN A ET AL: "Identification of a novel gammaretrovirus in prostate tumors of patients homozygous for R462Q RNASEL variant", PLOS PATHOGENS, PUBLIC LIBRARY OF SCIENCE, SAN FRANCISCO, CA, US, vol. 2, no. 3, 1 March 2006 (2006-03-01), pages 211 - 225, XP002402639, ISSN: 1553-7366 *
URISMAN A; MOLINARO RJ; FISCHER N; PLUMMER SJ; CASEY G ET AL.: "Identification of a novel Gammaretrovirus in prostate tumors of patients homozygous for R462Q RNASEL variant", PLOS PATHOG, vol. 2, 2006, pages E25, XP002669943, DOI: doi:10.1371/JOURNAL.PPAT.0020025
ZELINSKYY G ET AL., BLOOD, vol. 114, 2009, pages 3199 - 207
ZELINSKYY G ET AL., EUR J IMMUNOL, vol. 36, 2006, pages 2658 - 70
ZELINSKYY G ET AL., J VIROL, vol. 79, 2005, pages 10619 - 26
ZELINSKYY G; DIETZE KK; HUSECKEN YP; SCHIMMER S; NAIR S ET AL.: "The regulatory T cell response during acute retroviral infection is locally defined and controls the magnitude and duration of the virus-specific cytotoxic T cell response", BLOOD, vol. 114, 2009, pages 3199 - 207
ZELINSKYY G; KRAFT AR; SCHIMMER S; ARNDT T; DITTMER U: "Kinetics of CD8+ effector T cell responses and induced CD4+ regulatory T cell responses during Friend retrovirus infection", EUR J IMMUNOL, vol. 36, 2006, pages 2658 - 70
ZELINSKYY G; ROBERTSON SJ; SCHIMMER S; MESSER RJ; HASENKRUG KJ ET AL.: "CD8+ T-cell dysfunction due to cytolytic granule deficiency in persistent Friend retrovirus infection", J VIROL, vol. 79, 2005, pages 10619 - 26

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WO2013050048A3 (fr) * 2011-10-07 2013-07-18 Skau Aps Identification et atténuation des domaines immunosuppresseurs dans des protéines de fusion de virus à arn enveloppés
US10961279B2 (en) 2011-10-07 2021-03-30 Isd Immunotech Aps Identification and attenuation of the immunosuppressive domains in fusion proteins of enveloped RNA viruses
WO2020043908A1 (fr) 2017-09-01 2020-03-05 Inprother Aps Vaccin destiné à être utilisé dans la prophylaxie et/ou le traitement d'une maladie
CN111630060A (zh) * 2017-09-01 2020-09-04 盈珀治疗有限公司 用于在疾病的预防和/或治疗中使用的疫苗
CN113056477A (zh) * 2017-09-01 2021-06-29 盈珀治疗有限公司 用于在疾病的预防和/或治疗中使用的疫苗
CN111630060B (zh) * 2017-09-01 2024-01-23 盈珀治疗有限公司 用于在疾病的预防和/或治疗中使用的疫苗
EP4450516A2 (fr) 2017-09-01 2024-10-23 InProTher ApS Vaccin destiné à être utilisé dans la prophylaxie et/ou le traitement d'une maladie
WO2024218400A1 (fr) 2023-04-21 2024-10-24 Inprother Aps Expression améliorée d'antigènes affichés en surface

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