Direct injection of a recombinant retroviral vector induces human immunodeficiency virus-specific immune responses in mice and nonhuman primates

Irwin, M J; Laube, Lisa S.; Lee, V; Austin, Mark; Chada, Sunil; Anderson, C G; Townsend, Kay; Jolly, Douglas J.; Warner, John F.

doi:10.1128/jvi.68.8.5036-5044.1994

Cited by 52 publications

(26 citation statements)

References 40 publications

(29 reference statements)

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“…This PCR product was inserted into the pBluescript II (SKϩ) and was designated pBluescript II (SKϩ) HBe-c and verified by dideoxynucleotide sequencing to be correct (23,42). The HBeAg coding region from pBluescript II (SKϩ) was inserted into the N2-based Moloney murine leukemia virus backbone at the XhoI and ClaI sites (3,10). This construct encoded the HBeAg gene followed by the neomycin phosphotransferase II (NEO) gene being driven by an internal simian virus 40 promoter.…”

Section: Construction Of Vectors Expressing Hbcag and Hbeagmentioning

confidence: 99%

“…The VPCL-producing LHBc-NEO(6A3) was generated from the same G418-resistant pool that produced the LHBc(3A4) retrovirus vector. A negative control, the N2 ␤-gal vector, was described previously (10).…”

Section: Construction Of Vectors Expressing Hbcag and Hbeagmentioning

confidence: 99%

“…Direct in vivo administration of recombinant retrovirus vectors encoding human immunodeficiency virus Env and Rev proteins can induce MHC-restricted CD8 ϩ CTL in mice, rhesus monkeys, and baboons with high efficiency (10,15). Since recombinant retrovirus vectors can prime CTL responses to novel antigens efficiently, we were interested in extending this approach to the HBcAg and HBeAg to ask whether retrovirus vectors encoding these proteins are able to induce cell-mediated responses.…”

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confidence: 99%

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Characterization of CD8+ cytotoxic T-lymphocyte responses after genetic immunization with retrovirus vectors expressing different forms of the hepatitis B virus core and e antigens

Townsend¹,

Sällberg²,

O'Dea³

et al. 1997

J Virol

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Cytotoxic T-lymphocyte (CTL) activity appears to play an important role in resolving hepatitis B virus (HBV) infection, and the ability to induce such responses remains an important goal for developing effective immunotherapeutics. A panel of recombinant retrovirus vectors expressing different forms of the HBV core antigen (HBcAg) or e antigen (eAg) were found to induce antigen-specific major histocompatibility complexrestricted CTL responses in both mice and macaques. In addition, a novel retrovirus vector expressing an HBcAg-neomycin phosphotransferase II (HBc-Neo) fusion protein [LHBc-NEO(6A3)], which allows the measurement of the anti-Neo antibody response as a means of directly tracking biological activity of the vector, was generated. Doses greater than 10 7 CFU were necessary to induce CTL responses in H-2 k mice. Intramuscular injections with 10 8 CFU of the LHBc-NEO(6A3) retrovirus vector into rhesus monkeys induced HBc/eAgspecific antibody production and CD8 ؉ CTLs. The CTL response from one of the two responder rhesus monkeys was directed against a 9-residue peptide, GELMTLATW, at positions 63 to 71 of the HBc/eAg sequence. The CTL response is long lived, being detectable as late as 16 weeks after immunization, and can be boosted upon reimmunization. The potent ability of recombinant retrovirus vectors to induce HBcAg-and eAg-specific CTL responses may prove beneficial as a therapeutic treatment for chronic hepatitis B infection.

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Section: Construction Of Vectors Expressing Hbcag and Hbeagmentioning

confidence: 99%

Section: Construction Of Vectors Expressing Hbcag and Hbeagmentioning

confidence: 99%

mentioning

confidence: 99%

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Characterization of CD8+ cytotoxic T-lymphocyte responses after genetic immunization with retrovirus vectors expressing different forms of the hepatitis B virus core and e antigens

Townsend¹,

Sällberg²,

O'Dea³

et al. 1997

J Virol

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“…The vector titers were determined by serial dilution and colony-forming unit (CFU) assay. 26 The DA/β-gal producer cell line has been described. 26 For some experiments, a control vector identical to the HSV-TK retroviral vector but containing the human γ-interferon (γ−IFN) cDNA 31 in place of HSV-TK cDNA, was used.…”

Section: Retroviral Vector Constructmentioning

confidence: 99%

Ablation of Tumor Cells In Vivo by Direct Injection of HSV‐Thymidine Kinase Retroviral Vector and Ganciclovir Therapy

Howard

Kalthoff

Fong³

1999

Annals of the New York Academy of Sciences

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The introduction of therapeutic genes into proliferating tumor cells in vivo by direct intralesional injection of retroviral vectors can provide an effective and valuable approach for the treatment of a variety of solid tumor types. Efficient transduction of tumor cells in situ by direct injection was demonstrated using a retroviral vector containing the beta-galactosidase (beta-gal) gene. Ablation therapy in vivo was demonstrated using a retroviral vector containing the Herpes simplex virus thymidine kinase gene (HSV-TK) to deliver the TK gene into the murine colorectal tumor cell line CT26. Ablation of CT26 tumor cells in situ was achieved by directly injecting high-titer HSV-TK retroviral vector preparations into the site of tumor cell inoculation followed by intraperitoneal (i.p.) delivery of ganciclovir (GCV). This gene therapy strategy demonstrated a markedly lower rate of tumor progression, with several complete regressions, compared to animals in control groups. We also demonstrated that resistance to subsequent challenges with unmodified CT26 cells and an enhanced cellular immune response is associated with tumor regression in immunocompetent animals. Our results demonstrate the feasibility of direct in situ administration of HSV-TK retroviral vectors for the treatment of cancer and suggest that a cellular immune response may be elicited by this therapy.

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“…The final formulation was approximately 25 mM tromethamine, pH 7.4, 60 mM NaCl, 1 mg/ml arginine, 5 mg/ml HSA, and 50 mg/ml lactose. Vector preparations were shown to be free of replication competent retrovirus by hygromycin marker rescue assay of vector, or by Mus dunni/producer cell co-cultivated as described [42]. The viruses used for these studies had a titer of 1-3 × 10 8 cfu/ml.…”

Section: Vectorsmentioning

confidence: 99%

Retroviral gene transfer into cord blood stem/progenitor cells using purified vector stocks

et al. 1998

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Cord blood (CB) progenitor/stem cells (P/SC) are ideal targets for early gene therapy in individuals prenatally diagnosed with genetic disorders. Most retroviral transduction protocols were developed using adult peripheral blood stem cells (PBSC) and bone marrow (BM). Less is known about retroviral transduction of CB P/SC. We examined how timing, multiplicity of infection (MOI), and polycations in the transduction media affect transduction efficiency. Rates of transduction were determined in recently isolated CD34+ enriched CB cells and in colonies derived after various times in liquid cultures (LC). CB mononuclear cells (MNC) were separated by ficoll-hypaque centrifugation and enriched for CD34+ cells. Purity was assessed by flow cytometry. Transduction were performed with clinical-grade retroviral stocks at MOIs of 1-20. Transduction was performed with fetal bovine serum (FBS) or autologous plasma, IL-3, GM-CSF, IL-6, and SCF. The retroviral vector contained LacZ and neomycin resistance (neo) reporter genes. Transduction was determined by X-gal stain and by PCR amplification of the reporter genes. No drug selection was used. Twenty-five experiments were done. CB volumes ranged from 35-150 ml. MNC and CD34+ cell counts ranges were: 0.14-840 x 10(6) and 0.1-4.2 x 10(6), respectively. Transduction efficiency in liquid cultures ranged from 4-63%. Higher rates were seen using MOI > or = 10, 2 microg/ml polybrene, and 10% autologous CB plasma. In colonies, transduction rates were 63 to 72% by PCR and 32% by X-gal staining. In LTC-IC derived colonies, transduction was 7% by PCR. Short incubations of CD34+ CB cells with purified retroviral stocks, polybrene, and autologous sera result in high transduction rates of committed progenitors and moderately low efficiencies of transduction of LTC-IC in the absence of drug selection.

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Direct injection of a recombinant retroviral vector induces human immunodeficiency virus-specific immune responses in mice and nonhuman primates

Cited by 52 publications

References 40 publications

Characterization of CD8+ cytotoxic T-lymphocyte responses after genetic immunization with retrovirus vectors expressing different forms of the hepatitis B virus core and e antigens

Characterization of CD8+ cytotoxic T-lymphocyte responses after genetic immunization with retrovirus vectors expressing different forms of the hepatitis B virus core and e antigens

Ablation of Tumor Cells In Vivo by Direct Injection of HSV‐Thymidine Kinase Retroviral Vector and Ganciclovir Therapy

Retroviral gene transfer into cord blood stem/progenitor cells using purified vector stocks

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