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Titolo:
Construction of a recombinant adeno-associated virus (rAAV) vector expressing murine interleukin-12 (IL-12)
Autore:
Paul, D; Qazilbash, MH; Song, KM; Xu, H; Sinha, BK; Liu, J; Cowan, KH;
Indirizzi:
NCI, NIH, Med Branch, Bethesda, MD 20892 USA NCI Bethesda MD USA 20892NCI, NIH, Med Branch, Bethesda, MD 20892 USA NEI, NIH, Bethesda, MD 20892 USA NEI Bethesda MD USA 20892NEI, NIH, Bethesda, MD 20892 USA NHLBI, NIH, Hematol Branch, Bethesda, MD 20892 USA NHLBI Bethesda MD USA 20892 , NIH, Hematol Branch, Bethesda, MD 20892 USA
Titolo Testata:
CANCER GENE THERAPY
fascicolo: 2, volume: 7, anno: 2000,
pagine: 308 - 315
SICI:
0929-1903(200002)7:2<308:COARAV>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
CYTOKINE GENE-THERAPY; T RESPONSES INVITRO; WILD-TYPE P53; ADENOASSOCIATED VIRUS; IN-VIVO; ANTITUMOR IMMUNITY; HUMAN-LYMPHOCYTES; TUMOR-REGRESSION; COLON-CARCINOMA; NK CELLS;
Keywords:
adeno-associated virus; interleukin-12 vector; gene transfer;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
59
Recensione:
Indirizzi per estratti:
Indirizzo: Cowan, KH NCI, NIH, Med Branch, 10-12N-226, Bethesda, MD 20892 USA NCI 10-12N-226 Bethesda MD USA 20892 226, Bethesda, MD 20892 USA
Citazione:
D. Paul et al., "Construction of a recombinant adeno-associated virus (rAAV) vector expressing murine interleukin-12 (IL-12)", CANC GENE T, 7(2), 2000, pp. 308-315

Abstract

IL-12 is a heterodimeric cytokine that is known to induce tumor regressionand long-term antitumor immunity. Recombinant adeno-associated virus (rAAV) vectors are advantageous for gene therapy in that they lack pathogenicityin humans, infect dividing as well as nondividing cells, and show a broad range of infectivity. We constructed an rAAV vector expressing interleukin-12 (IL-12) for cancer immunotherapy studies in a mouse model by inserting murine IL-12 (mIL-12) p35 and p40 cDNAs into the plasmid pRep4 and insertingthe encephalomyocarditis virus internal ribosomal entry site between the p35 and p40 cDNAs. The mIL-12 expression cassette containing the Rous sarcoma virus promoter and a simian virus 40 polyadenylation signal was subclonedinto the AAV plasmid p008Sub/NeoR which contains two AAV inverted terminalrepeat sequences and the NeoR gene driven by the thymidine kinase promoter. rAAV virions (10(4) infectious particles/ml) were generated by cotransfection of rAAV-mIL-12 and a helper plasmid (pAAV/Ad) into 293 cells previously infected with adenovirus 5. After infection of DG fibroblasts with rAAV-mIL-12, G418-resistant clones were isolated. Each of the 1D D6 clones isolated produced up to 5.2 ng/10(6) cells/48 hours of mIL-12 as determined by enzyme-linked immunosorbent assay. Induction of interferon-gamma, enhanced lymphocyte proliferation, and cytotoxicity assays confirmed biologically functional IL-12 production by the vector. This is the First report indicating that an rAAV vector expresses mIL-12, which can be used to model the effects of mIL-12 alone and/or in combination with Other antitumor agents.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 16/07/20 alle ore 06:04:32