Recombinant expression vectors comprising a human codon-optimized marburg virus (MARV) angola glycoprotein gene insert and method of immunization employing said vector
Inventors
Sullivan, Nancy • Chakrabarti, Bimal • Yang, Zhi-Yong • Pau, Maria Grazia • Goudsmit, Jaap • Nabel, Gary
Assignees
HEALTH AND HUMAN SERVICES GOVERNMENT OF United States, THE, Secretary of, Department of • US Department of Health and Human Services
Publication Number
US-8101739-B2
Publication Date
2012-01-24
Expiration Date
2025-09-27
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
The invention is related to a nucleic acid molecule comprising a polynucleotide encoding a modified filovirus glycoprotein (GP) having at least one amino acid change located in a relatively conserved region of said GP that decreases in vitro cytotoxicity and retains immunogenicity when compared to in vitro cytotoxicity and immunogenicity of a wild type filovirus GP, and related modified filovirus GPs, plasmid DNAs, recombinant viruses, adenoviruses, pharmaceutical compositions, vaccine compositions, antibodies that are specifically reactive with the modified filovirus GPs, and related methods of making and using the same.
Core Innovation
The invention relates to nucleic acid molecules encoding modified filovirus glycoproteins (GP) with at least one amino acid change in a relatively conserved region of the GP that decreases in vitro cytotoxicity while retaining immunogenicity compared to the wild type filovirus GP. These modifications include single amino acid substitutions such as E71D or G102A in Ebola Zaire GP or corresponding positions in other filovirus strains. The invention also encompasses plasmid DNAs, recombinant viruses—including adenoviruses—pharmaceutical and vaccine compositions comprising these nucleic acid molecules or modified GPs, and antibodies specifically reactive with the modified GPs, alongside methods for making and using these components.
The problem addressed arises from the cytotoxic effects exhibited by filovirus GP when expressed in cultured human cells, which disrupts cellular function and vascular integrity and is linked to Ebola virus pathogenesis. Previous deletions, such as the mucin-like domain, while removing cytotoxicity, led to loss of many T- and B-cell epitopes and decreased vaccine efficacy. The invention solves this problem by identifying specific point mutations that abolish GP in vitro cytopathicity but retain immunogenicity and protective efficacy, providing a safer and effective antigen for vaccine development against filovirus infections.
Further, the invention determines that deletion of the transmembrane domain eliminates cytopathicity but reduces protective efficacy significantly, while single amino acid substitutions at conserved sites preserve membrane anchoring and vaccine efficacy. Optimized recombinant adenovirus vectors encoding these modified GPs induce protective immune responses at lower doses in nonhuman primate models, and protective immunity can be achieved with GP alone without the nucleoprotein (NP) antigen, simplifying the vaccine composition.
Claims Coverage
The patent claims encompass isolated nucleic acid molecules, plasmid DNAs, recombinant viruses, adenoviruses, immunogenic compositions comprising modified Marburg virus GP sequences, and methods of inducing or boosting immune responses against Marburg virus antigens in primates using these genetic constructs.
Isolated nucleic acid molecule comprising the Marburg/Angola GP/h sequence
An isolated nucleic acid molecule having the specific SEQ ID NO:5 that encodes the modified Marburg virus Angola strain glycoprotein.
Plasmid DNA construct with CMV promoter and Marburg GP insert
An isolated plasmid DNA comprising the cytomegalovirus immediate early enhancer promoter, HTLV-1 R region, and the Marburg/Angola GP/h insert of SEQ ID NO:5, enabling expression of the modified GP.
Recombinant virus comprising Marburg GP insert
An isolated recombinant virus that includes the Marburg/Angola GP/h nucleic acid insert of SEQ ID NO:5 for use in immune response induction.
Adenovirus vector encoding modified Marburg GP
An isolated adenovirus containing the Marburg/Angola GP/h insert of SEQ ID NO:5 designed for vaccination applications.
Immunogenic composition comprising nucleic acid molecule
An immunogenic composition containing the nucleic acid molecule of SEQ ID NO:5 in an immunogenically effective dose.
Method of boosting immune response using recombinant virus
A method of boosting an immune response to a Marburg virus antigen in a primate by administering the recombinant virus comprising SEQ ID NO:5, enhancing a previously primed immune response.
Method of inducing immune response using nucleic acid molecule
A method of inducing an immune response to a Marburg virus antigen in a primate by administering the isolated nucleic acid molecule of SEQ ID NO:5.
Prime-boost immunization method with nucleic acid molecule and recombinant virus
A method comprising priming with the nucleic acid molecule of SEQ ID NO:5 followed by boosting with a composition comprising the antigen or a recombinant virus encoding the antigen to induce a stronger immune response.
The claims collectively cover nucleic acid sequences encoding a human codon-optimized modified Marburg virus glycoprotein, expression constructs including plasmids and adenoviral vectors harboring these sequences, immunogenic compositions containing them, and methods for inducing or boosting immune responses in primates by administering these nucleic acid molecules or recombinant viruses.
Stated Advantages
Modified filovirus glycoproteins exhibit decreased in vitro cytotoxicity while retaining immunogenicity compared to wild type, enhancing safety of vaccines.
Single amino acid mutations eliminate cytopathicity without reducing protective efficacy, improving vaccine design.
Recombinant adenoviral vectors encoding the modified GP confer protection at lower doses than previously used, enabling more efficient immunization.
Protection can be achieved with GP alone without the nucleoprotein (NP), simplifying vaccine formulation.
Adenoviral vectors are replication-deficient, non-replicating, safe, can be manufactured in high yields, and are well tolerated in vivo.
Documented Applications
Use as vaccines or immunogenic compositions to induce or boost immune responses against filoviruses, specifically Ebola and Marburg viruses in primates.
Prophylactic vaccination of humans or non-human primates to prevent filovirus (Ebola or Marburg virus) infection and disease.
Use of recombinant adenovirus vectors expressing the modified filovirus GP for accelerated or prime-boost immunization regimens to elicit cellular and humoral immunity.
Interested in licensing this patent?