The University of North Carolina at Chapel Hill

Other

US Government As Represented by the Secretary of the Army

North Carolina State University

Medical Research Council

Children's Hospital

Alphavax, Incorporated

University of Cape Town Observatory

Robert E Johnston

Nancy L Davis

Jonathan F Smith

Peter M Pushko

Michael D Parker

George Ludwig

Dennis A Simpson

Ronald I Swanstrom

Gene H MacDonald

Franziska B Grieder

Joseph M Thompson

Sergey A Dryga

Jenny P-y Ting

Robert A Olmsted

Philip R Johnson, Jr

Ian Caley

Philip R Johnson

Maureen Maughan

Carolyn Williamson

Paula M Keith

Lynn Morris

Salim Abdol Karim

Mark T Heise

Dennis Simpson

Brian K Martin

Loretta V Willis

Kelly Young Poe

Christy K Jurgens

Jenny P-Y Ting

The present invention provides viral adjuvants for enhancing an immune response to an immunogen. In particular embodiments, the viral adjuvant is an alphavirus adjuvant or a Venezuelan Equine Encephalitis viral adjuvant. Also provided are compositions comprising the viral adjuvant and an immunogen, and pharmaceutical formulations comprising the viral adjuvant or compositions of the invention in a pharmaceutically acceptable carrier. Further provided are methods of producing an immune response against an immunogen in a subject comprising administering the immunogen and a viral adjuvant of the invention to the subject.

Viral adjuvants

The present invention provides novel self-replicating and self-propagating chimeric viral vectors and chimeric virus particles comprising a modified genome of a carrier RNA virus packaged within structural proteins of a second virus. Also provided are pharmaceutical formulations comprising the chimeric viral vectors and virus particles and methods of inducing an immune response by administration of the same to a subject.

Live virus vaccines

Embodiments of the invention provide processes for the selection of HIV-1 subtype (clade) C isolates, selected HIV-1 subtype C isolates, their genes and modifications and derivatives thereof for use in prophylactic and therapeutic vaccines to produce proteins and polypeptides for the purpose of eliciting protection against HIV infection or disease. A process for the selection of HIV subtype isolates comprises the steps of isolating viruses from recently infected subjects; generating a consensus sequence for at least part of at least one HIV gene by identifying the most common codon or amino acid among the isolated viruses; and selecting the isolated virus or viruses with a high sequence identity to the consensus sequence. HIV-1 subtype C isolates, designated Du422, Du 151 and Du 179 (assigned Accession Numbers 01032114, 00072724 and 00072725, respectively, by the European Collection of Cell Cultures) are also provided.

Nucleic acids encoding modified South African HIV-1 subtype C gag proteins

The present invention provides compositions and methods for delivering a nucleotide sequence to a cell using an alphavirus vector that is complexed with an enhancing antibody that specifically binds to the alphavirus vector. Venezuelan Equine Encephalitis vectors are preferred. The cell may be a cell in vitro or in vivo. Alternatively, the cell may be removed from a subject, administered the alphavirus vector ex vivo and then administered to a subject. Antigen-presenting cells are preferred, with dendritic cells being more preferred. Also provided are methods of producing an immune response in a subject, e.g., for producing an immune response against an antigen associated with a pathogen or for immunotherapy of cancer of tumors.

Antibody dependent enhancement of Venezuelan Equine Encephalitis vector infection

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

Provided herein are alphavirus vectors derived from South African Arbovirus No. 86 (S.A.AR86) comprising attenuating mutations and methods of making the same. Also provided are improved viral vectors and helper constructs comprising a S.A.AR86 capsid enhancer sequence. The present invention also provides S.A.AR86 replicon and helper constructs comprising an alphavirus capsid enhancer sequence. Further provided are methods of administering an alphavirus vector comprising a heterologous nucleotide sequence (preferably encoding an immunogen or a therapeutic polypeptide) according to the invention to a cell or subject. In preferred embodiments, the alphavirus vector delivers the heterologous nucleotide sequence to the cells of the bone, bone marrow, and/or bone-associated connective tissue.

Vectors derived from South African Arbovirus No. 86

The present invention provides methods of preventing and/or treating cancers (including tumors). In one preferred embodiment, the invention is practiced to induce regression of an existing cancer or tumor and/or to prevent metastasis and/or to prevent growth of metastatic nodules. In other preferred embodiments, the invention may be used as a prophylaxis to prevent the development of primary cancers through a childhood or adult vaccination program against specific tumor antigens for cancers with high incidences. In an alternate preferred embodiment, the present invention provides methods of establishing an immune response against a universal artificial tumor antigen through a childhood or adult vaccine program, thus providing a long-term immune response that can be utilized at any point to treat any cancer which develops later in life. The present invention also provides cancer and tumor cells stably expressing an artificial antigen, preferably an artificial cell-surface antigen.

Methods and modified cells for the treatment of cancer

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

Growing community of inventors

Chapel Hill, NC, United States of America

Robert E Johnston