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Parapoxvirus vectors

a parapoxvirus and vector technology, applied in the field ofviral immunotherapy, can solve the problems of a major obstacle to effective immunotherapy, and achieve the effect of improving the cytokine/chemokine profil

Pending Publication Date: 2021-01-07
TRANSGENE SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention is about a new type of virus called pseudocowpoxvirus (PCPV) that has been altered to contain foreign nucleic acid. This foreign nucleic acid can be inserted into the genome of the PCPV and can be used to treat or prevent diseases caused by a pathogenic organism or an unwanted cell division. The PCPV can also be engineered to have reduced viral functions or to remove disease-related proteins. The invention also includes a method for generating the PCPV and a composition containing the PCPV for therapeutic use.

Problems solved by technology

Indeed, diseased cells have evolved potent immunosuppressive mechanisms for eluding the immune system, posing a major obstacle to effective immunotherapy.

Method used

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  • Parapoxvirus vectors
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Examples

Experimental program
Comparison scheme
Effect test

example 1

f Secreted Cytokines / Chemokines in Infected Human Peripheral Blood Mononuclear Cells (PBMCs)

[0175]Human peripheral blood mononuclear cells (PBMCs) were isolated from whole human blood of two healthy donors (EFS, Etablissement de Strasbourg; donor A and B). PBMCs were isolated by density gradient centrifugation using Ficoll-Paque PLUS (GE HealthCare) and Blood Separation Tubes (Greiner) and stored in liquid nitrogen.

[0176]Frozen aliquots of PBMCs were thawed in RPMI supplemented with 10% Fetal calf serum (FSC) and dispatched in 24 well plates (5·105 cells in 500 μL / well). After 5 to 6 hours, cells were infected with the different viruses (see below) at multiplicities of infection (MOI) between 10−3 and 10. After overnight incubation (16 hours), cells were scraped, cells and supernatant were transferred in Eppendorf tubes and centrifuged 10 min at 300 g. Supernatant was isolated and frozen at −80° C. Cytokine and chemokine profiles were quantified by a Multiplex approach in the supern...

example 2

PCPV on Human Monocyte-Derived Dendritic Cells, M2 Macrophages and Myeloid-Derived Suppressor Cells (MDSCs)

[0180]Type I interferons like IFN-alpha are key players in immunotherapy of cancer as well as altered immunosuppressive tumor environment and improved adaptive anti-tumor responses against vaccine encoded and / or tumor-presented antigens as described e.g. by Parker et al. (2016, Nat Rev Cancer 16(3): 131-44) and Zitvogel et al. (2015, Nat Rev Immunol. 15(7): 405-14) To address these aspects, we looked in vitro in human primary immune cells at the activation of human monocyte-derived dendritic cells (moDC) and at the effect on immunosuppressive cell populations (re-programming of immunosuppressive cells like M2 macrophages and MDSCs) upon PCPV treatment. Preclinical studies on innate and adaptive immunity were also probed in murine syngeneic tumor models.

[0181]Stimulation of moDCs.

[0182]To probe the activation of antigen-presenting cells (APCs), we worked with monocyte-derived de...

example 3

ion of Recombinant PCPV and Anti-Tumor Properties

[0192]The interest for PCPV as viral backbone was evaluated by generating recombinant PCPV encoding tumor antigens and testing them in tumor control experiments.

[0193]Recombinant PCPV vectors were constructed from the wild-type strain TJS (ATTC, VR-634). PCPV, as all the Parapoxvirus, lack genes present or conserved in other poxviruses. These comprises homologues of most poxviral genes likely involved in nucleotide metabolism, including homologues of ribonucleotide reductase (RR), thymidine kinase (TK), guanylate kinase and thymidylate kinase. Therefore, the locus TK generally used for generation of recombinant vaccinia virus could not be used for introduction of recombinant gene in PCPV. However, it was shown that recombinant ORF virus could be generated by insertion of the transgene in the non-essential VEGF gene (Rziha et al., 2000, J. Biotechnol. 83(1-2): 137-145). This locus was therefore evaluated for the generation of recombina...

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Abstract

The present invention is in the field of viral immunotherapy. The invention provides new pseudocowpox (PCPV) viruses, in particular recombinant PCPV, composition thereof as well as their therapeutic use for preventing or treating diseases, and, notably, proliferative diseases like cancers and restenosis and infectious diseases such as chronic ones. The present invention also provides methods for generating and amplifying such a PCPV and a method for eliciting or stimulating and / or re-orienting an immune response using such a PCPV. More specifically, the invention provides an alternative to the existing poxvirus vectors such as MVA (Modified Virus Ankara) and may be largely used for the therapeutic vaccination.

Description

TECHNICAL FIELD OF THE INVENTION[0001]The present invention is in the field of viral immunotherapy. The invention provides new pseudocowpox (PCPV) viruses, in particular recombinant PCPV, composition thereof as well as their therapeutic use for preventing or treating diseases, and, notably, proliferative diseases like cancers and restenosis and infectious diseases. The present invention also provides methods for generating and amplifying such a PCPV and a method for eliciting or stimulating and / or re-orienting an immune response using such a PCPV. More specifically, the invention provides an alternative to the existing poxvirus vectors such as MVA (Modified Virus Ankara) and may be largely used for the therapeutic vaccination.BACKGROUND ART[0002]Immunotherapy seeks to boost the host's immune system to help the body to eradicate pathogens and abnormal cells. Widely used in traditional vaccination, immunotherapy is also being actively investigated as a potential modality for treating ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/00A61K39/29C07K14/53C07K16/28C12N15/86
CPCA61K39/00117A61K39/292C07K14/53C12N2710/20043C12N15/86A61K2039/5256C12N2710/20034C07K16/2878A61K39/12A61K2039/5254C12N2710/24243
Inventor RITTNER, KAROLAREMY, CHRISTELLETHIOUDELLET, CHRISTINE
Owner TRANSGENE SA
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