Compositions and methods for treatment of neoplastic disease

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...

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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.

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