Double-copy bicistronic retroviral vector platform for gene therapy and tissue engineering: application to melanoma vaccine development

Wiznerowicz, Maciej; Fong, Andrew Z. C.; Maćkiewicz, Andrzej; Rg, Hawley

doi:10.1038/sj.gt.3300500

Cited by 28 publications

(12 citation statements)

References 14 publications

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“…44 Others have placed promoter-transgene cassettes in the 3′LTR to achieve double-copy vectors. 18,[45][46][47][48] Our studies, therefore, have important implications for future design of vectors with inserts within the 3′LTR, given the usefulness of chromatin insulator elements, customized lineage-specific LTR vectors or double-copy vectors.…”

Section: Discussionmentioning

confidence: 97%

Mechanism of Reduction in Titers From Lentivirus Vectors Carrying Large Inserts in the 3′LTR

et al. 2009

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Self-inactivating (SIN) lentiviruses flanked by the 1.2-kb chicken hypersensitive site-4 (cHS4) insulator element provide consistent, improved expression of transgenes, but have significantly lower titers. The mechanism by which this occurs is unknown. Lengthening the lentiviral (LV) vector transgene cassette by an additional 1.2 kb by an internal cassette caused no further reduction in titers. However, when cHS4 sequences or inert DNA spacers of increasing size were placed in the 3'-long terminal repeat (LTR), infectious titers decreased proportional to the length of the insert. The stage of vector life cycle affected by vectors carrying the large cHS4 3'LTR insert was compared to a control vector: there was no increase in read-through transcription with insertion of the 1.2-kb cHS4 in the 3'LTR. Equal amount of full-length viral mRNA was produced in packaging cells and viral assembly/packaging was unaffected, resulting in comparable amounts of intact vector particles produced by either vectors. However, LV vectors carrying cHS4 in the 3'LTR were inefficiently processed following target-cell entry, with reduced reverse transcription and integration efficiency, and hence lower transduction titers. Therefore, vectors with large insertions in the 3'LTR are transcribed and packaged efficiently, but the LTR insert hinders viral-RNA (vRNA) processing and transduction of target cells. These studies have important implications in design of integrating vectors.

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Section: Discussionmentioning

confidence: 97%

Mechanism of Reduction in Titers From Lentivirus Vectors Carrying Large Inserts in the 3′LTR

et al. 2009

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“…Furthermore, several investigators have shown that bicistronic vectors can be easily generated for adenoviral as well as other types of vectors. [37][38][39] Consequently, one can envision the use of a cell autonomous marker such as GFP in conjunction with a stem cell gene (eg SCL, Gata-1, PU.1, etc) so that biological changes can be monitored and characterized for specific cells. In addition, the capacity of adenoviral vectors to transduce non-dividing cells is of critical importance for the analysis of hematopoietic stem cells.…”

Section: Discussionmentioning

confidence: 99%

Genetic manipulation of primitive leukemic and normal hematopoietic cells using a novel method of adenovirus-mediated gene transfer

et al. 1999

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“…Plasmid construction DCCMV ± TNF vector was generated by blunt end ligation of human TNF cDNA, excised as EcoRI fragment from the MSCV ±TNF retroviral vector (kindly provided by Dr. Robert G. Hawley, Holland Laboratories, Rockville, MA ) , into a filled SalI site of DCCMV 21 between the hCMV-IE promoter and the EMCV IRES -Neo cassette.…”

Section: Micementioning

confidence: 99%

Antitumor effects of the combination therapy with TNF-α gene–modified tumor cells and interleukin 12 in a melanoma model in mice

et al. 2000

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In the present study, TNF -gene ± transduced B78 melanoma cells ( B78 / TNF ) were used as a vaccine and combined with interleukin ( IL ) -12 in the treatment of B78 melanoma -bearing mice. The combined administration of genetically modified melanoma cells and IL -12 induced specific protective antitumor immunity resulting in a decreased rate of the tumor take following a rechallenge with parental B78 cells. When used therapeutically, intratumoral injections of irradiated B78 / TNF melanoma cells and IL -12 exerted strong antitumor effects and led to complete regression of established tumors in 50% of mice. Injections of irradiated B78 / TNF cells alone did not influence tumor development and IL -12 itself significantly delayed tumor growth but without curative effect. FACS analysis of parental B78 melanoma cells and its B78 / TNF genetically modified variant showed that a proportion of cells of both cell lines expressed B7 -1 ( CD80 ) costimulatory molecule and that the expression of this molecule was increased during incubation with IFN -. Moreover, IFN -markedly augmented expression of major histocompatibility class ( MHC ) class I and II molecules on B78 / TNF cells that were primarily MHC class I and II negative with no substantial effect on MHC -negative parental B78 melanoma. IFN -also synergized in cytostatic / cytotoxic effects with TNF -against B78 melanoma in vitro. Lymphocyte depletion studies in vivo showed reduction of the antitumor response in mice treated with anti -NK monoclonal antibodies ( mAbs ) as well as in mice treated with anti -CD4 + anti -CD8 mAbs. The results suggest that, when used therapeutically, IL -12 and a vaccine containing TNF -gene ± transduced tumor cells may reciprocally augment their overall antitumor effectiveness by facilitating development of systemic antitumor immunity and by stimulating local effector mechanisms of the tumor destruction. Cancer Gene Therapy ( 2000 ) 7, 1581 ± 1590

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Double-copy bicistronic retroviral vector platform for gene therapy and tissue engineering: application to melanoma vaccine development

Cited by 28 publications

References 14 publications

Mechanism of Reduction in Titers From Lentivirus Vectors Carrying Large Inserts in the 3′LTR

Mechanism of Reduction in Titers From Lentivirus Vectors Carrying Large Inserts in the 3′LTR

Genetic manipulation of primitive leukemic and normal hematopoietic cells using a novel method of adenovirus-mediated gene transfer

Antitumor effects of the combination therapy with TNF-α gene–modified tumor cells and interleukin 12 in a melanoma model in mice

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