Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv

a technology of immunogenic composition and plasmid, which is applied in the field of plasmids and immunogenic compositions, can solve the problems of limiting the expression of multiple genes from one vector backbone in a single target cell, raising safety and technical issues, and difficult to achieve optimal expression of all encoded genes

Inactive Publication Date: 2007-08-16
WYETH LLC
View PDF14 Cites 17 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0036] In a particular embodiment, the present invention provides an immunogenic composition for inducing an immune response to HIV in a vertebrate host, where the immunogenic composition comprises five plasmids as described above, and where each plasmid further comprises a selectable marker selected from the group con

Problems solved by technology

While, viral regulatory elements are advantageous for use in plasmid DNA vectors, the use of unmodified viral vectors to express the relevant genes may raise safety and technical issues not encountered with plasmid DNA.
Current plasmid DNA designs, however, limit the expression of multiple genes from one vector backbone in a single target cell.
When cells must be co-transfected with multiple plasmids, it is difficult to achieve optimal expression of all encoded genes, especially when the plasmid is being used in vivo.
None of the existing plasmid designs have solved the problem of providing a DNA plasmid suitable for expressing more than two independent open reading frames in hu

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
  • Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
  • Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv

Examples

Experimental program
Comparison scheme
Effect test

example 1

Selection and Modification of HIV Genes

[0179] One of skill in the art would appreciate that sequence information from many viruses and bacteria is available in the art. More particularly, sequence information can be used to clone genes for use in expressing polypeptides in plasmids of the invention. Information on many sequences from HIV and other pathogens is available from the HIV sequence database at the Los Alamos National Laboratory and the National Center for Biotechnology Information at the United States National Library of Medicine, (8600 Rockville Pike, Bethesda, Md. 20894).

[0180] In one embodiment of the invention, the following HIV genes were selected for inclusion into a single exemplary DNA plasmid expressing most of the HIV genome: gag gene from the HXB2 isolate and the pol gene from the HXB2 isolate. The complete HXB2 sequence is listed in the GenBank computer database under the accession number K03455. The nef, tat and vif genes were derived from the NL4-3 isolate....

example 2

Construction of Single, Double and Triple Transcriptional Unit Plasmids

[0191] The plasmids discussed in these examples are set forth in Tables 1 and 2.

[0192] A triple transcriptional unit expression cassette was constructed by using a variety of components in a circular double stranded DNA plasmid. See FIG. 1. The first component was a first transcriptional unit for expressing polypeptides in eukaryotic cells, composed of the simian cytomegalovirus (SCMV) promoter, a cloning site and bovine growth hormone (BGH) poly-A signal. The second component is a second transcriptional unit for expressing polypeptides in eukaryotic cells, which consists of human cytomegalovirus (HCMV) immediate early promoter, a cloning site and the SV40 polyadenylation (polyA) signal. Separating the first and second transcriptional units is spacer region 1. The third component is a third transcriptional unit for expressing polypeptides in eukaryotic cells and is composed of the Herpes simplex virus Lap1 prom...

example 3

Triple Transcriptional Unit Plasmid Containing Six HIV Genes

[0193] As a demonstration of the use of the three transcriptional unit plasmid DNA vectors, a plasmid vector capable of co-expressing three eukaryotic open reading frames was created. The three transcriptional unit plasmid DNA vector was created by inserting the following selected genes encoding HIV-1 antigens into the triple transcriptional unit expression cassette described in Example 2. All cloning techniques were performed following conventional procedures (Sambrook et al. 1989).

[0194] First, an HIV-1 gag-pol fusion gene was inserted into the PmeI-XhoI cloning site between the SCMV and BGH poly-A sites of the first transcriptional unit. The gag gene was derived from the HXB2 isolate, and, similarly, the pol gene was also derived from the HXB2 isolate. The complete HXB2 sequence is listed in the GenBank computer database under the accession number K03455. One of skill in the art would understand that other HIV-1 gag an...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Dielectric strengthaaaaaaaaaa
Dielectric strengthaaaaaaaaaa
Volumeaaaaaaaaaa
Login to view more

Abstract

The invention provides a DNA plasmid comprising: (a) a first transcriptional unit comprising a nucleotide sequence that encodes a first polypeptide operably linked to regulatory elements including a first promoter and a first polyadenylation signal; (b) a second transcriptional unit comprising a nucleotide sequence that encodes a second polypeptide operably linked to regulatory elements including a second promoter and a second polyadenylation signal; (c) a third transcriptional unit comprising a nucleotide sequence that encodes a third polypeptide operably linked to regulatory elements including a third promoter and a third polyadenylation signal; and wherein said first, said second and said third promoters are each derived from different transcriptional units; and wherein said first, said second and said third polyadenylation signals are each derived from different transcriptional units. The invention further relates to immunogenic compositions for inducing an immune response to HIV comprising combinations of two, three, or four plasmids, where each plasmid is expressing a defined antigen, which may be a single antigen or a fusion of two or three antigens.

Description

FIELD OF THE INVENTION [0001] This invention relates to plasmids, immunogenic compositions and methods to improve prophylactic and therapeutic immune responses to antigens. BACKGROUND OF THE INVENTION [0002] Immunization using plasmid DNA-based immunogenic compositions is a powerful tool that is useful for developing approaches to prevent or treat infectious diseases or in the treatment of ongoing disease processes. Plasmid DNA immunization has been extensively tested in animal models where it has been found to be effective in inducing both cellular and humoral immune responses against a wide variety of infectious agents and tumor antigens. See Donnelly J J, et al., Ann. Rev. Immunol.; 15: 617-48 (1997); Iwasaki A, et al., J Immunol 158 (10): 4591-601 (1997); Wayne, C. L. and Bennett M., Crit. Rev. Immunol., 18: 449-484 (1998). [0003] An important advantage of plasmid DNA immunization is that genes can be cloned, modified and positioned into a potential plasmid DNA expression vector...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K39/21A61K48/00C12N15/867
CPCA61K39/21A61K2039/55538A61K2039/54A61K2039/545C07K14/005C12N15/85C12N2740/16122C12N2740/16134C12N2740/16222C12N2740/16234C12N2740/16322C12N2740/16334C12N2800/107C12N2830/00C12N2840/20C12N2840/60A61K2039/53A61K39/12A61P31/18A61P37/04C12N15/86C12N15/63
Inventor SIDHU, MANINDER K.ELDRIDGE, JOHN H.EGAN, MICHAELISRAEL, ZIMRA
Owner WYETH LLC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap