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DNA-vaccines based on constructs derived from the genomes of human and animal pathogens

a technology of human and animal genomes, applied in the field of dnavaccines based on constructs derived from the genomes of human and animal pathogens, can solve the problems of non-selective expression of pathogen's genes, and achieve the effects of minimal number of molecular modifications of vectors and/or inserted polynucleotides, easy growth and manipulation, and enhanced transcription

Inactive Publication Date: 2007-10-11
POWDER JECT VACCINES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This approach elicits a robust and protective immune response, including both humoral and cell-mediated immunity, potentially offering better protection than traditional vaccines by delivering a broader array of antigens and their regulatory elements, similar to natural infection.

Problems solved by technology

In contrast, the use of heterologous promoters in previously described expression library immunization would result in non-selective expression of the pathogen's genes.

Method used

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  • DNA-vaccines based on constructs derived from the genomes of human and animal pathogens
  • DNA-vaccines based on constructs derived from the genomes of human and animal pathogens
  • DNA-vaccines based on constructs derived from the genomes of human and animal pathogens

Examples

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Effect test

example 1

Nucleic Acid Immunization Using HSV-2 Cosmids

[0112] In order to assess the specificity and effectiveness of nucleic acid immunization using DNA vaccine cosmids containing HSV-2 genomic DNA, the following studies were carried out.

A. Cosmid Preparation

[0113] HSV-2 Cosmids:

[0114] HSV-2 genomic DNA was obtained by infecting Vero cells (ATCC # CCL-81) with HSV-2 strain MS (ATCC # VR-54T) and isolating genomic DNA 10′ from the infected cells. Cosmids containing fragments of the HSV-2 genome were generated by digesting HSV-2 genomic DNA, as described above, with the restriction enzyme EcoRI (New England Biolabs). As shown in FIG. 1, the HSV genome has been mapped and contains at least 10 EcoR1 sites, indicated in the figure with arrows. The digested DNA was ligated into cosmids using the SuperCos 1 Cosmid Vector Kit from Stratagene following the manufacturers instructions. Positive cosmids were first identified from the fragment sizes generated by an EcoRI digest of purified cosmid DN...

example 2

Nucleic Acid Immunization Using HSV-2 Plasmids

A. Plasmid Construction

[0124] HSV-2 genomic DNA was obtained by infecting VERO cells (ATCC #CCL-81) with HSV-2 strain MS (ATCC #VR-54T) and isolating genomic DNA from purified viral particles. PCR was performed using the genomic DNA to amplify segments for cloning into a plasmid vector.

[0125] The primer gB2, having the following sequence: CGC GTC TAG AAA CGT TCG CGA CCA CGG GTG AC (SEQ ID NO: 3) and primer gB3, having the following sequence: CGC GTC TAG ATG ATG GGG TCC CGC TAA CTC GC (SEQ ID NO: 4), which correspond to sequences at 58160 and 52930 of the HSV-2 genome, respectively, were used to amplify a product of approximately 5200 bp by PCR. A second PCR product was generated with using primer gB2 (SEQ ID NO: 3) and primer gB4, having the following sequence: CGC GTC TAG ACC TTC ATG ACC GCG CTG GTC CT (SEQ ID NO: 5) which correspond to sequences at 58160 and 49670, respectively, of the HSV-2 genome. This produced a product of appro...

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Abstract

Methods of eliciting an immune response in a subject by administering one or more large genomic DNA fragments are provided. Also provided are methods of identifying sequences encoding antigenic polypeptides. Also provided are vaccine compositions comprising one or more large genomic DNA fragments.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is related to U.S. provisional application Ser. No. 60 / 163,297, filed 3 Nov. 1999, from which priority is claimed pursuant to 35 U.S.C. §119(e)(1) and which application is incorporated herein by reference in its entirety.TECHNICAL FIELD [0002] The present invention relates generally to vaccine compositions and methods of use thereof. More particularly, the invention pertains to eliciting an immune response in a subject by administering one or more large genomic DNA fragments to the subject. BACKGROUND [0003] Vaccines which induce a cell-mediated immune response are emerging as important strategies in combating parasites, autoimmune disorders, allergic diseases and cancers. Conventional vaccination strategies generally involve administration of either “live” or “dead” vaccines. Ertl et al. (1996) J. Immunol. 156:3579-3582. The so-called live vaccines include attenuated microbes and recombinant molecules based on a living ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/245A61K39/25A61P31/22
CPCA61K39/245C12N2710/16634A61K2039/53A61K39/12A61K2039/545A61P31/22
Inventor SWAIN, WILLIAM F.ROBERTS, LEE K.PAYNE, LENDON G.BRAUN, RALPH P.
Owner POWDER JECT VACCINES INC
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