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Compositions and methods for treatment of neoplastic disease

a neoplastic disease and composition technology, applied in the field of compositions and methods for treating tumors and cancer, can solve the problems of inconsistent results, little or no effect, and inability to demonstrate unacceptable toxicity, and achieve the effect of high ifn production

Inactive Publication Date: 2005-05-26
TERMAN DAVID
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0034] Sickled erythrocytes are useful in the present invention since they have natural ligands for integrins expressed on tumor neovasculature which facilitates their targeting to the tumor endothelium. Sickled erythrocyte membranes acquire oxyLDL using fusigenic techniques with oxyLDL containing liposomes and apoproteins via gene transfection in the nucleated pre-reticulocyte phase. The oxyLDL and apoproteins expressed by the sickled cells facilitates targeting to oxyLDL, LOX-1 and SREC receptors present on the tumor microvasculature. These erythrocytes are also useful for carrying nucleic acids for transfection of the tumor endothelial cells in vivo. Vesicles derived from sickled erythrocytes are more rigid, prothrombotic and target the tumor microvascularture more effectively than the parent cell. They also carry oxyLDL to receptors on tumor endothelium. Likewise, vesicles, exosomes or SAg-GPI-digalctosylceramides shed from from SAg transfected tumor cells are capable of inducing potent tumoricidal responses and are useful in the present invention.
[0037] For in vivo immunization, tumor cells are transfected with nucleic acids encoding SAgs together with a carbohydrate modifying enzyme such as galactosyl transferase to produce the Gal epitope, Staphylococcal hyaluronidase, Streptococcal capsular polysaccharide, Staphylococcal erythrogenic toxin, Staphylococcal Protein A, Staphylococcal □ hemolysin, Staphylococcal coagulase, costimulants such as B7-1 and B7.2, chemoattractants and chemokines. SAgs are also cotransfected into tumor cells with gene clusters encoding the biosynthesis of highly immunogenic microbial Lipid A, membrane or capsular polysaccharides, lipoproteins and peptidoglycans. Nucleic acids are useful when transfected alone. However combinations are preferred. The cotransfection into tumor cells of the SAg-encoding nucleic acid together with the nucleic acids encoding Gal or GalCer biosynthesis is particularly useful. The cotransfection into tumor cells of the nucleic acid encoding SAg with nucleic acids encoding Staphylococcal erythrogenic toxins and hyaluronidase allows the transfected tumor cells to simulate the in vivo inflammatory activity of a Staphylococcus or leukocyte or macrophage by secreting enzymes and toxins which induce a sterile cellulitis in tumor sites.
[0041] Also provided is a tumor specific T cell or NKT cell population which is activated by SAgs or the tumor cell transfectants above to produce a population of tumor specific effector cells useful in adoptive immunotherapy. After ex vivo stimulation, the T cells or NKT cells used for adoptive immunotherapy should preferentially express CD44 which indicates that they are capable of trafficking and homing to tumor sites. Additionally, the T cell population used for ex vivo immunization is engineered to overexpress the TCR variable V□ and invariant Vα sites specific for SAg and glycosylceramide binding respectively and to produce IFN by exogenous delivery of corticosteroids or a heavy metal. A particularly useful population of therapeutic tumor specific effector T cells or NKT cells which demonstrates overexpressed CD44 together with V□ variable and Vα invariant regions and high IFN production. Also provided are methods for reactivating anergic T cells in cancer patients by transfecting nucleic acids encoding the SAg receptors to produce a T cell population which may now be stimulated with exogenous SAgs.

Problems solved by technology

However, results with these measures, while beneficial in some tumors, has had only marginal effects in many patients and little or no effect in many others, while demonstrating unacceptable toxicity.
While the initial observations of tumor killing effects with the immobilized Protein A perfusion system have been confirmed, some have obtained inconsistent results.
Second, various methods of immobilizing Protein A to solid supports have been used, sometimes resulting in loss of biological activity of the perfusion system.
Third, the plasma used for perfusion over immobilized Protein A has often been stored and treated in different ways, also resulting in occasional inactivation of the system.
Moreover, the substance(s) or factors responsible for the anti-tumor effect of this extremely complex perfusion system have not been previously defined.
First, final products are quite different.

Method used

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  • Compositions and methods for treatment of neoplastic disease
  • Compositions and methods for treatment of neoplastic disease
  • Compositions and methods for treatment of neoplastic disease

Examples

Experimental program
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example 1

Preparation of Plasmids for Making DNA Templates for any Gene of Interest and the Process Transfection

[0389] Mammalian oncogenes, and genes for oncogenic transcription factors, angiogenic factors, growth factor receptors and amplicons as well as bacterial and SAg plasmids and DNA are prepared as described in the text references. When necessary, they are modified to forms suitable for transfection into mammalian tumor cells or accessory cells using methods well described in the art. (Old R W et al., Principles of Gene Manipulation, 5th Ed., Blackwell 1994).

[0390] As a representative SAg, enterotoxin B plasmid DNA is prepared by the method of Jones C L et al., J. Bacteriology 166 29-33 (1986) and Ranelli et al., Proc. Natl. Acad. Sci. USA 82:5850-5854 (1985) using the CsCl-ethidium bromide density gradient centrifugation of cleared lysates as described (Clewell, D B et al., Proc. Natl. Acad. Sci. USA 62-1159-1166 (1969)). S. aureus chromosomal DNA was isolated as described by Betley...

example 2

Cells Transfected with Nucleic Acids Encoding SAgs

[0536] Cultured VX-2 carcinoma cells were shown to retain their tumorigenic activity after implantation into New Zealand white rabbits. Progressive tumor outgrowth was observed over a 3 week period. Nucleic acid encoding SEB isolated and characterized by Gaskill et al, J. Biol. Chem. 263:6276 (1988) and Ranelli et al., Proc. Natl Acad. Sci. U.S.A 82:5850 (1985) were used to transfect tissue cultured VX-2 carcinoma cells using transfection methodology described in Example 1. Transfectants were selected using G418 and the survival of SEB-transfected VX-2 carcinoma cells was observed. In additional experiments, attempts were made to transfect murine 205 and 207 tumor cells with nucleic acid encoding SEB(the kind gift from Dr. Saleem Khan) and Streptococcal pyrogenic exotoxin A (the kind gift of Dr. Joseph Ferretti). Successfiil transfection of murine MCA 205 and B16 cells by nucleic acids encoding SEA and SEC2 was achieved shortly ther...

example 3

Naked SAg DNA and Cells Co-Transfected with SAg DNA and with Additional Nucleic Acid Encoding Anti-Tumor Motifs or Products

[0537] Nucleic acids encoding a SAg are injected in naked or plasmid form into a host with cancer as a means of activating T cells and initiating an anti-tumor response. They may also be used as a vaccine to prevent the occurrence or recurrence of tumor in a host. Under circumstances where it is desirable to activate CD4 cells to produce a TH-1 cytokine response the nucleic acid construct used to transfect cells contains immunostimulatory sequences such as unmethylated CpG sequences. Nucleic acids encoding SAgs may be co transfected into tumor cells together with nucleic acid encoding other constituents capable of promoting an anti-tumor response. A list of possible components of nucleic acid constructs for direct administration and / or transfection of tumor cells which are administered to the host is presented in Table II.

[0538] The nucleic acid construct or c...

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Abstract

The present invention comprises compositions and methods for treating a tumor or neoplastic disease in a host, The methods employ conjugates comprising superantigen polypeptides or nucleic acids with other structures that preferentially bind to tumor cells and are capable of inducing apoptosis. Also provided are superantigen-glycolipid conjugates and vesicles that are loaded onto antigen presenting cells to activate both T cells and NKT cells. Cell-based vaccines comprise tumor cells engineered to express a superantigen along with glycolipids products which, when expressed, render the cells capable of eliciting an effective anti-tumor immune response in a mammal into which these cells are introduced. Included among these compositions are tumor cells, hybrid cells of tumor cells and accessory cells, preferably dendritic cells. Also provided are T cells and NKT cells activated by the above compositions that can be administered for adoptive immunotherapy.

Description

CROSS-REFERENCE TO RELATED DOCUMENTS [0001] The Instant application is a continuation application of U.S. application Ser. No. 09 / 650,884 filed on Aug. 30, 2000 which claims priority to provisional applicaton 60 / 151,470 filed on Aug. 30, 1999. Both of the above referenced applications are incorporated herein in their entirety by reference.BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The invention relates generally to immunotherapeutic compositions and methods for treating tumors and cancer. The methods are based on the expression of superantigen (“SAg”) alone or in combination with other molecules in transfected host cells (tumor cells, accessory cells or lymphocytes). Other therapeutic methods are based on administering T cells which are activated by cells engineered to express SAg and other immunostimulatory molecules and structures. [0004] 2. Description of the Background Art [0005] Therapy of the neoplastic diseases has largely involved the use of chemothe...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K48/00C07K14/31C07K14/315C07K14/47C07K14/705C07K14/715C12N5/08C12P21/04
CPCA61K39/0011A61K48/005A61K2039/5156A61K2039/55544C07K2319/33C07K14/3156C07K14/70503C07K14/7156C07K14/31A61P35/00A61K39/4611A61K2239/31A61K2239/38A61K39/4613A61K39/464499A61K2239/56A61K39/4632A61K39/4615A61K2239/57A61K39/4622
Inventor TERMAN, DAVID
Owner TERMAN DAVID
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