Lentiviral vector based immunological compounds against malaria

Patent metadata

Field	Value
Publication number	US-9822153-B2
Application number	US-201514799633-A
Country	US
Kind code	B2
Filing date	Jul 15, 2015
Priority date	May 3, 2010
Publication date	Nov 21, 2017
Grant date	Nov 21, 2017

Abstract

The invention relates to lentiviral vector particles pseudotyped with a determined heterologous viral envelope protein or viral envelope proteins originating from a RNA virus and which comprise in its genome at least one recombinant polynucleotide encoding at least one polypeptide(s) carrying epitope(s) of an antigen of a Plasmodium parasite capable of infecting a mammalian host. The lentiviral vector particles are used in order to elicit an immunological response against malaria parasites.

First claim

The invention claimed is:

1. A lentiviral vector particle (i) which is pseudotyped with at least one determined heterologous viral envelope protein originating from a RNA virus and (ii) which comprises in its genome at least one recombinant polynucleotide encoding at least one polypeptide comprising at least one epitope of a pre-erythrocytic stage antigen of a Plasmodium parasite capable of infecting a mammalian host.

2. The lentiviral vector according to claim 1 which is a replication-incompetent HIV-based vector particle.

3. The lentiviral vector particle according to claim 1 wherein the at least one recombinant polynucleotide comprises a nucleic acid sequence encoding a polypeptide(s) of an antigen from the circumsporozoite protein (CSP).

4. The lentiviral vector particle according to claim 1 , wherein the at least one recombinant polynucleotide has a mammalian codon optimized nucleotide sequence.

5. The lentiviral vector particle according to claim 1 , wherein the at least one recombinant polynucleotide encodes at least a polypeptide of the CSP antigen, said polypeptide being devoid of the glycosylphosphatidylinositol (GPI)-anchoring motif of said CSP.

6. The lentiviral vector particle according to claim 1 , which is an integration-deficient vector particle.

7. The lentiviral vector particle according to claim 1 , which is an integration-competent vector particle.

8. The lentiviral vector particle according to claim 1 , which is pseudotyped with at least one viral transmembrane glycosylated (G) envelope protein(s) of a Vesicular Stomatitis Virus (VSV) chosen from the group consisting of VSV-G protein(s) of the Indiana strain, VSV-G protein(s) of the New Jersey strain, VSV-G protein(s) of the Cocal strain, VSV-G protein of the Isfahan strain, VSV-G protein(s) of Chandipura strain, VSV-G protein(s) of Pyri strain and VSV-G protein(s) of the SVCV strain.

9. The lentiviral vector particle according to claim 1 , recovered from mammalian cells co-transfected with:
a) a vector plasmid comprising lentiviral cis-active sequences necessary for packaging, reverse transcription, and transcription; a functional lentiviral DNA flap element; and a polynucleotide of a truncated mammalian codon-optimized sequence of the circumsporozoite (cs) gene of a Plasmodium parasite, under the control of regulatory expression sequences;
b) a VSV-G envelope expression plasmid comprising a polynucleotide encoding a VSV-G envelope protein or envelope proteins under the control of regulatory expression sequences; and
c) an encapsidation plasmid comprising lentiviral gag-pol coding sequences suitable for the production of integration-competent vector particles or modified gag-pol coding sequences suitable for the production of integration-deficient vector particles, wherein said gag-pol sequences are from the same lentivirus sub-family as the DNA flap element, wherein said gag-pol or modified gag-pol sequences are under the control of regulatory expression sequences.

10. The lentiviral vector particles according to claim 1 recovered from a stable cell line transfected with
a) a vector plasmid comprising lentiviral cis-active sequences necessary for packaging, reverse transcription, and transcription; a functional lentiviral DNA flap element; and a polynucleotide of a truncated mammalian codon-optimized sequence of the cs gene of a Plasmodium parasite, under the control of regulatory expression sequences;
b) a VSV-G envelope expression plasmid comprising a polynucleotide encoding a VSV-G envelope protein or envelope proteins under the control of regulatory expression sequences; and
c) an encapsidation plasmid comprising lentiviral gag-pol coding sequences suitable for the production of integration-competent vector particles or modified gag-pol coding sequences suitable for the production of integration-deficient vector particles, wherein said gag-pol sequences are from the same lentivirus sub-family as the DNA flap element, wherein said gag-pol or modified gag-pol sequences are under the control of regulatory expression sequences.

11. The lentiviral particles according to claim 1 , which comprises in its genome lentiviral-based sequences devoid of functional lentiviral genes, wherein said lentiviral-based sequence comprise:
cis-active sequences necessary for packaging, reverse transcription, and transcription and a functional lentiviral DNA flap element, wherein said cis-acting sequences comprise at least one modification selected from:
a) the 3′ long terminal repeat (LTR) sequence from the lentiviral genome is truncated and devoid of the enhancer of the U3 region;
b) the 3′ LTR sequence from the lentiviral genome is truncated and devoid of the U3 region or partly deleted in the U3 region; and
c) the U3 region of the LTR5′ is replaced by a non lentiviral U3 region or by a promoter suitable to drive tat-independent primary transcription.

12. A combination of lentiviral vector particles for separate administration to a mammalian host, which comprises:
(i) a lentiviral vector particle according to claim 1 , pseudotyped with at least one first determined heterologous viral envelope protein; and
(ii) provided separately from lentiviral vector particles in (i), a lentiviral vector particle according to claim 1 , pseudotyped with at least one second determined heterologous viral envelope protein;
wherein said first and second at least one viral envelope protein do not sero-neutralize with each other and are suitable for in vivo transduction of mammalian cells.

13. A combination of lentiviral vector particles according to the claim 12 , wherein said first and second at least one viral envelope proteins are selected from:
VSV-G of Indiana strain and VSV-G of New Jersey strain;
wherein one or both of said first and second envelope proteins are modified versions of native VSV-G of Indiana strain or/and VSV-G of New Jersey strain;
a chimeric VSV-G protein wherein at least one of the following domains is from an Indiana strain: the export determinant YTDIE (amino acids 501 to 505 of SEQ ID NO: 77), the cytoplasmic tail, the transmembrane domain and the cytoplasmic domain; and
the first at least one viral envelope protein is either VSV-G of Indiana strain or VSV-G of New Jersey strain and the second at least one viral envelope protein is selected from the group consisting of VSV-G protein(s) of the Cocal strain, VSV-G protein of the Isfahan strain, VSV-G protein(s) of Chandipura strain, VSV-G protein(s) of Pyri strain and VSV-G protein(s) of the SVCV strain.

14. A composition comprising a lentiviral vector particle according to claim 1 and a physiologically acceptable vehicle.

15. The composition of claim 14 , further comprising an adjuvant and/or an immunostimulant.

16. The lentiviral vector particle of claim 10 , wherein the functional lentiviral DNA flap element and the lentiviral gag-pol sequences are from HIV-1.

17. The lentiviral vector particle of claim 11 , wherein the lentiviral-based sequences are from HIV-1.

Assignees

• Charneau Pierre
• Coutant Frederic Philippe
• Pasteur Institut

Inventors

• Charneau Pierre
• Coutant Frederic Philippe

Classifications

• A61P37/04

  Immunostimulants · CPC title

• C12N2740/16043

  viral genome or elements thereof as genetic vector · CPC title

• A61P33/06

  Antimalarials · CPC title

• A61K39/015 Primary

  Hemosporidia antigens, e.g. Plasmodium antigens · CPC title

• C12N2740/16023

  Virus like particles [VLP] · CPC title