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Combination Immunogene Therapy

a combination immunogene and tumor technology, applied in the field of tumor immunotherapy, can solve the problems of affecting affecting the survival rate of cancer patients, and affecting the success rate of chemotherapeutics as anticancer agents, so as to increase the immunogenicity of human tumors and effective cancer immunotherapy

Inactive Publication Date: 2010-12-16
RADIENT PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]In order to provide effective cancer immunotherapy, the art needs means to increase the im...

Problems solved by technology

However, specificity has been the major problem with conventional anticancer agents.
Success with chemotherapeutics as anticancer agents has also been hampered by the phenomenon of multiple drug resistance, resistance to a wide range of structurally unrelated cytotoxic anticancer compounds [Gerlach et al.
In addition, certain cancers are non-responsive to known chemotherapeutics agents and patients with these cancers invariably die within a short period following diagnosis (e.g., glioblastoma multiforme, recurrent metastatic melanoma, breast, lung and pancreatic cancers).
However, given the incidence of cancer, even in seemingly immunologically normal individuals, it is clear that the immune system fails to recognize and destroy all tumor cells.

Method used

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Examples

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example 1

[0104]Construction of Mono- and Bi-Cistronic Retroviral Vectors and Packaging of Recombinant Virus

[0105]Retroviral gene therapy vectors were constructed that contained the genes for GM-CSF, B7-2, GM-CSF and B7-2, or GFP gene as shown schematically in FIG. 1. FIG. 1 provides a schematic showing the map of the parental vector (pLSN), pLSNB70 (encodes human B7-2), pLSNGM1 (encodes human GM-CSF), pLSN-BG9 (encodes both B7-2 and GM-CSF) and pLSN-GFP (encodes GFP).

[0106]All genes were cloned into the polylinker region of the MLV-based pLSN plasmid [Robinson et al. (1995) Gene Therapy 2:269 and co-pending application Ser. No. 08 / 336,132]. Therapeutic genes were inserted into pLSN such that their expression was under the control of the retroviral LTR (long terminal repeat) whereas a neomycin-resistance gene was under the control of an internal SV40 promotor. The vectors lacked the gag, pol, or env genes necessary for retroviral packaging in order to render them replication incompetent. Thes...

example 2

[0128]Expression of B7-2 and GM-CSF in the Human Glioblastoma Cell Line D54MG

[0129]Viral particles containing recombinant retroviral genomes encoding either GM-CSF, B7-2, B7-2 / GM-CSF or GFP were used to transduce the human glioblastoma cell line D54MG.

[0130]a) Growth Of D54MG In Tissue Culture

[0131]The human glioblastoma cell line D54MG [obtained from Dr. D. Bigner, Duke University, Durham, N.C.; Bigner et al. (1981) J. Neuropathol. Exp. Neurol. 40:201] was cultured in Dulbucco's Modified Eagle's Media (DMEM) with 10% fetal bovine serum (Gibco), 0.2 units / ml penicillin-streptomycin solution (Sigma), and 0.2 mM glutamate at 37° C. in a humidified atmosphere containing 5% CO2.

[0132]b) Retroviral Transduction

[0133]Frozen virus stock was thawed at 37° C. Polybrene was added to the thawed virus solution at a final concentration of 4 μg / ml. Culture media was added to the virus solution to bring the final volume to 1.5 ml. D54MG cells in logarithmic growth phase in T25 flasks (approximatel...

example 3

[0145]Tumor Growth Efficiency and Human PBL Reconstitution in SCID / Beige and SCID / nod Mice

[0146]This example describes the reconstitution of SCID / nod and SCID / beige mice with human PBL and the engraftment of human tumor cells in these mouse strains.

[0147]a) Animals and Human PBL Reconstitution

[0148]For most experiments, four to five week old female C.B-17-SCID-beige mice were purchased from Taconic (Germantown, N.Y.). For the first vaccination / challenge experiment, four to five week old female SCID / nod mice were obtained from Dr. L. Pilarski (Cross Cancer Institute, University of Alberta, Edmonton, Alberta). The mice were maintained in filtered cages in a virus free environment and received cotrimoxazole in their drinking water twice per week.

[0149]Hu-PBL-SCID mouse reconstitution was carried out as previously described [Zhang et al. (1996) Proc. Natl. Acad. Sci. USA 93:14720]. Briefly, each mouse was intraperitoneally (IP) injected with 2-3×107 PBLs resuspended in 0.5 ml of Hanks' ...

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Abstract

The present invention provides to immunogene therapy protocols for the treatment of tumors. In particular, the present invention provides combinations of immune-modulating proteins that induce systemic immunity against tumors. In addition, the present invention provides humanized animal models suitable for the evaluation of anti-human tumor immunity and permit the identification of combinations of immune-modulating genes which when delivered to human tumor cells induce an effective anti-tumor response, including a systemic anti-tumor response.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. (USPASN) 10,785,577 filed Feb. 23, 2004 which is a continuation of U.S. patent application Ser. No. 09 / 826,025 filed Apr. 4, 2001, now U.S. Pat. No. 6,730,512, which is a continuation of U.S. patent application Ser. No. 08 / 838,702, filed Apr. 9, 1997, now abandoned. The entire contents of each of these applications are incorporated by reference herein.FIELD OF THE INVENTION[0002]The present invention relates to immunotherapy for the treatment of tumors. In particular the present invention provides combinations of immune-modulating proteins that induce systemic immunity against tumors and provides humanized animal models for immunogene therapy.BACKGROUND OF THE INVENTION[0003]Conventional treatment of cancer typically involves the use of chemotherapeutic agents. The father of chemotherapy, Paul Ehrlich, imagined the perfect chemotherapeutic as a “magic bullet;” such a c...

Claims

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Application Information

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IPC IPC(8): A61K38/20A61K35/00A61K38/19A61K38/21A61P35/00C12N15/63A01K67/027A61K39/00C07K14/16C12N7/04
CPCA01K67/0271A61K39/00C07K14/005C12N7/00C12N2710/10343Y02A50/41C12N2740/13043C12N2740/16034C12N2740/16322Y02A50/412C12N2710/10361A61P35/00Y02A50/30
Inventor CHANG, LUNG-JI
Owner RADIENT PHARMA
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