The University of North Carolina at Chapel Hill

Other

US Government As Represented by the Secretary of the Army

North Carolina State University

Nancy L Davis

Robert E Johnston

Jonathan F Smith

Peter M Pushko

Michael D Parker

George Ludwig

Dennis A Simpson

Franziska B Grieder

Sergey A Dryga

Ronald I Swanstrom

Robert A Olmsted

Ian Caley

Maureen Maughan

Paula M Keith

Loretta V Willis

The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least alphavirus one structural protein not encoded by the first helper RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell. Preferably, the helper cell also includes a replicon RNA encoding an alphavirus packaging sequence and an inserted heterogeneous RNA.

Alphavirus RNA replicon systems

The present invention provides methods and compositions comprising a population of alphavirus replicon particles comprising two or more isolated nucleic acids selected from 1) an isolated nucleic acid encoding an env gene product or an immunogenic fragment thereof of a human immunodeficiency virus, 2) an isolated nucleic acid encoding a g&agr;g gene product or an immunogenic fragment thereof of a human immunodeficiency virus, wherein the g&agr;g gene product or immunogenic fragment thereof is modified to inhibit formation of virus-like particles containing the g&agr;g gene product or the immunogenic fragment thereof and their release from a cell, and 3) an isolated nucleic acid encoding a pol gene product or an immunogenic fragment thereof of a human immunodeficiency virus, wherein the pol gene product or immunogenic fragment thereof is modified to inhibit protease, integrase, RNase H and/or reverse transcriptase activity, and wherein the nucleic acids are each contained within a separate alphavirus replicon particle.

Alphavirus vectors and virosomes with modified HIV genes for use in vaccines

The present invention provides a method of delivering immunogenic or therapeutic proteins to bone marrow cells using alphavirus vectors. The alphavirus vectors disclosed herein target specifically to bone marrow tissue, and viral genomes persist in bone marrow for at least three months post-infection. No or very low levels of virus were detected in quadricep, brain, and sera of treated animals. The sequence of a consensus Sindbis cDNA clone, pTR339, and infectious RNA transcripts, infectious virus particles, and pharmaceutical formulations derived therefrom are also disclosed. The sequence of the genomic RNA of the Girdwood S.A. virus, and cDNA clones, infectious RNA transcripts, infectious virus particles, and pharmaceutical formulations derived therefrom are also disclosed.

System for the in vivo delivery and expression of heterologous genes in the bone marrow

The present invention provides a helper cell for expressing an infectious, replication defective, alphavirus particle in an alphavirus-permissive cell. The helper cell includes (a) a first helper RNA encoding (i) at least one alphavirus structural protein, and (ii) not encoding at least one alphavirus structural protein; and (b) a second helper RNA separate from the first helper RNA, the second helper RNA (i) not encoding the alphavirus structural protein encoded by the first helper RNA, and (ii) encoding the at least one alphavirus structural protein not encoded by the first helper RNA. Preferably, the helper cell is co-transfected with a replicon RNA encoding an alphavirus packaging segment and an inserted heterogeneous RNA, such that all of the alphavirus structural proteins assemble together into alphavirus particles in the cell, with said replicon RNA packaged therein.

System for the in vivo delivery and expression of heterologous genes in

A method of protecting a subject against a disease comprises administering a recombinant Venezuelan Equine Encephalitis (VEE) virus to the subject in an effective immunogenic amount, with the VEE virus containing a heterologous DNA segment, and with the heterologous DNA segment comprising a promoter operable in the subject operatively associated with a DNA encoding an immunogenic protein or peptide effective for protecting the subject from the disease. Preferred promoters are VEE 26S subgenomic promoters, and preferred immunogens are viral immunogens. Novel attenuating mutations useful in carrying out the invention are also disclosed.

Method of inducing an immune response with a live Venezuelan Equine

The present invention provides a recombinant DNA comprising a cDNA coding for an infectious South African Arbovirus No. 86 (S.A.AR86) virus RNA transcript and a heterologous promoter positioned upstream from the cDNA and operatively associated therewith. The present invention also provides an infectious RNA transcript encoded by the cDNA, and infectious attenuated viral particles containing the RNA transcript encoded by the cDNA.

cDNA clone for South African Arbovirus No. 86

Novel attenuating mutations of Venezuelan Equine Encephalitis (VEE) are disclosed. Further aspects of the invention include an infectious VEE virus transcript encoded by cDNA clones, infectious VEE virus particles, and pharmaceutical formulations containing such infectious particles. Also disclosed are recombinant VEE virus containing a heterologous RNA segment.

Attenuating mutations in Venezuelan Equine Encephalitis virus

A DNA comprises a cDNA clone coding for an infectious Venezuelan Equine Encephalitis Virus RNA transcript and a heterologous promoter sequence positioned upstream from the cDNA clone and operatively associated therewith. A method of making a live attenuated Togavirus useful as a vaccine, and cDNA clones which code for attenuated Togaviruses, is also disclosed.

Growing community of inventors

Chapel Hill, NC, United States of America

Nancy L Davis