Terapia genica anti HIV-1: effetto di mutanti transdominanti negativi di TAT sulla replicazione e riattivazione del virus dalla latenza

Dottorato di ricerca in biotecnologie. A.a 1993-94. Coordinatore Giuseppe Barbanti Brodano, tutore Roberto ManservigiConsiglio Nazionale delle Ricerche - Biblioteca Centrale - P.le Aldo Moro, 7, Rome; Biblioteca Nazionale Centrale - Piazza Cavalleggeri, 1, Florence / CNR - Consiglio Nazionale delle RichercheSIGLEITItal

Comunicazione orale, Keynote speaker: Herpes Simplex Vector Expressing Hiv1 Tat: A New Strategy For Vaccine Development

Recent major breakthroughs in the field of HSV-1 technology authorize and support the use of HSV-1 based vectors as vaccine vectors against HSV-1 infection or for the delivery of foreign antigens. One of the major obstacles in developing effective vaccines against intracellular pathogens is the incomplete understanding on the mechanisms that contribute to effective immune responses. The identification of a new class of adjuvants, expressed by HSV vectors, able to direct and broad antigen presentation by the major histocompatibility complexes (MHC) can be used to achieve a stronger effect or a more potent skewing of immune responses. The focus of our laboratory is to develop vaccines able to activate and modulate effective immune responses not only against HSV but also against other intracellular pathogens such as Mycobacterium tuberculosis (Mtb). In this context, we have investigated, whether HIV-1 Tat protein co-expressed with HSV-1 vector antigens, may act as a molecule capable to induce such broad and protective immunity against intracellular pathogens. The concept of Tat as a candidate adjuvant has derived from in vitro and in vivo evidences indicating that the biologically active clade B Tat protein possesses several immunomodulatory features making it an attractive adjuvant for other antigens and suggesting that it can be exploited for vaccination strategies and therapeutic interventions aimed at modulating antigen-specific immune responses in different types of human diseases. In recent studies we have demonstrated that an attenuated replication competent HSV vector expressing Tat, as immunostimulatory molecule, is immunogenic and confers protection in mice challenged with a lethal dose of wild-type HSV1. Moreover, in preliminary experiments using a non-replicative HSV vector expressing Mtb antigens and Tat we observed that Tat can function as a potent adjuvant that modulates the CTL epitope hierarchy and broadens the epitope-specific T cell responses of heterologous antigens. Our results suggest that HSV vector backbone co-expressing an adjuvant molecule is able to broaden epitope responses and that this can be a new concept for a future vaccine against intracellular pathogens

Herpes Simplex Virus Type 1-Derived Recombinant and Amplicon Vectors

Herpes simplex virus type 1 (HSV-1) is a human pathogen whose lifestyle is based on a long-term dual interaction with the infected host, being able to establish both lytic and latent infections. The virus genome is a 153 kbp double-stranded DNA molecule encoding more than 80 genes. The interest of HSV-1 as gene transfer vector stems from its ability to infect many different cell types, both quiescent and proliferating cells, the very high packaging capacity of the virus capsid, the outstanding neurotropic adaptations that this virus has evolved, and the fact that it never integrates into the cellular chromosomes, thus avoiding the risk of insertional mutagenesis. Two types of vectors can be derived from HSV-1, recombinant vectors and amplicon vectors, and different methodologies have been developed to prepare large stocks of each type of vector. This chapter summarizes (1) the two approaches most commonly used to prepare recombinant vectors through homologous recombination, either in eukaryotic cells or in bacteria, and (2) the two methodologies currently used to generate helper-free amplicon vectors, either using a bacterial artificial chromosome (BAC)-based approach or a Cre/loxP site-specific recombination strategy

Recombinant herpes simplex virus and uses therefor

(WO200592374) Provided is a replication-deficient HSV encoding a heterologous antigen, and a vaccine comprising the HSV.The vaccine may be directed to viruses, intracellular bacteria or tumours.Also provided is an anticancer vaccine comprising replication-deficient HSV encoding an angiogenesis inhibitor, a cytokine and a suicide gene.A challenge model for screening candidate antigens is also provided, as is an UL13 mutant HSV vector.Further provided are HSV vectors for expression of Hex A and NTFs, and methods comprising said HSV

Herpes Simplex Virus Type 1 (HSV-1HSV-1 )-Derived Recombinant Vectorsvectors for Gene Transfergene transfer and Gene Therapygene therapy

Herpes simplex virus type 1 (HSV-1 ) is a human pathogen whose lifestyle is based on a long-term dual interaction with the infected host, being able to establish both lytic and latent infections. The virus genome is a 153-kilobase pair (kbp) double-stranded DNA molecule encoding more than 80 genes. The interest of HSV-1 as gene transfer vector stems from its ability to infect many different cell types, both quiescent and proliferating cells, the very high packaging capacity of the virus capsid , the outstanding neurotropic adaptations that this virus has evolved, and the fact that it never integrates into the cellular chromosomes, thus avoiding the risk of insertional mutagenesis . Two types of vectors can be derived from HSV-1, recombinant vectors and amplicon vectors, and different methodologies have been developed to prepare large stocks of each type of vector. This chapter summarizes the approach most commonly used to prepare recombinant HSV-1 vectors through homologous recombination , either in eukaryotic cells or in bacteria.Herpes simplex virus type 1 (HSV-1) is a human pathogen whose lifestyle is based on a long-term dual interaction with the infected host, being able to establish both lytic and latent infections. The virus genome is a 153-kilobase pair (kbp) double-stranded DNA molecule encoding more than 80 genes. The interest of HSV-1 as gene transfer vector stems from its ability to infect many different cell types, both quiescent and proliferating cells, the very high packaging capacity of the virus capsid , the outstanding neurotropic adaptations that this virus has evolved, and the fact that it never integrates into the cellular chromosomes, thus avoiding the risk of insertional mutagenesis. Two types of vectors can be derived from HSV-1, recombinant vectors and amplicon vectors, and different methodologies have been developed to prepare large stocks of each type of vector. This chapter summarizes the approach most commonly used to prepare recombinant HSV-1 vectors through homologous recombination , either in eukaryotic cells or in bacteria

Use of replication deficient hsv-1 as a vaccine vector for the deli vary of hiv-1 tat antigen

USE - For the treatment and/or prophylaxis of a condition associated with the virus, or the condition optionally being a disease condition. Also for the treatment of a condition associated with a DNA virus selected from order Caudovirales, including families Myoviridae, Podoviridae, Siphoviridae; order Herpesvirales, including families Alloherpesvirdae, Herpersviridae, and Malacoherpesviridae; order Ligamenvirales, including families Lipothrixviridae, and Rudiviridae; and families Adenovirdae, Ampullaviridae, Ascoviridae, Asfarviridae, Baculoviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Fuselloviridae, Globuloviridae, Guttaviridae, Hytrosaviridae, Iridoviridae, Miminviridae, Nimaviridae, Papillomaviridae, Phycodnaviridae, Plasmaviridae, Polydnaviruses, Polyomaviridae, Poxviridae, and Tectiviridae (preferably Herpesvirales, especially Herpesviridae, and the virus is herpes simplex virus type 1, or HSVI). The virus is an avirulent form selected from replication-deficient forms of the virus, attenuated forms of the virus, and forms of the virus modified, to prevent integration of the viral genome into the host genome (claimed). ADVANTAGE - The HIV1 Tat expressed by replication-competent HSV1 vectors not only does not dysregulate the immune system against HSV infection, but increases and broadens the Th1-like and CTL responses against HSV1 immunodominant and subdominant T cell epitopes, as well as eliciting detectable immunoglobulin G (IgG) responses. A similar attenuated HSV1 recombinant vector without Tat (HSV1-LacZ) induces lower T cell responses, which are directed only against the immunodominant epitopes, and fails to generate measurable IgG serum responses. Immunization with the present vaccines is effective, and the present vaccines are capable of protecting BALB/c and C57BL/6 mice against HSV lesions and death after challenge with a lethal dose of wild type HSV1 virus

Herpetic vectors inhibiting angiogenesis and inducing cell suicide in gliomas

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Enhanced tumor cell killing in the presence of ganciclovir by herpes simplex virus type 1 vector-directed coexpression of human tumor necrosis factor-alpha and herpes simplex virus thymidine kinase

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