A METHOD OF INDUCING AN ANTI-TUMOR RESPONSE AGAINST A LUNG METASTASIS IN A MELANOMA PATIENT

Abstract: The present invention is directed to a method of inducing an anti-tumor response against a metastatic melanoma by administering an effective amount of a composition comprising at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten-modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response such as for example regression of a melanoma. The metastatic melanoma treated according to the present invention includes metastatic melanoma which is limited to the lung and is preferably a small lung metastasis. The invention is further directed to a melanoma cell, an isolated melanoma cell membrane, a peptide isolated from such cell or membrane and a T cell having the property of inducing an anti-tumor response, a composition containing such cell, membrane, peptide, T cell or combination thereof, as well as the methods for theirisolation and preparation.

Description: A METHOD OF INDUCING AN ANTI-TUMOR RESPONSE AGAINST A LUNG METASTASIS IN A MELANOMA PATIENT CROSS-REFERENCE TO RELATED APPLICATION This application claims priority pursuant to 35 U.S.C. §119 based upon Provisional Patent Application Serial No.60/081,256 filed April 9, 1998, the entire disclosure of which is hereby incorporated by reference. REFERENCE TO GOVERNMENT GRANTS The invention described herein was made in the course of work under a grant or award from the US Public Health Services NTH grant no. CA29348. The United States Government may have certain rights in this invention. BACKGROUND OF THE INVENTION In the 1960's, certain theories that tumor cells bear specific antigens (TS A) not present on normal cells, and that the immune response to these antigens might help reject a tumor, were advanced. It was later suggested that the immune response to TSA could be increased by introducing new immunological determinants on cells. Mitchison, Transplant. Proc, 1970, 2, 92. Such determinants, although not known, were termed "helper determinants." At the time, speculations were made that compounds such as, for example, a hapten, a protein, a viral coat antigen, a transplantation antigen, or a xenogenous cell antigen could be introduced into a population of tumor cells to act as helper determinants. Clinically, the hope was that an immunologic reaction would occur against the helper determinant, as a consequence of which the reaction to the accompanying TSA would be increased, and tumor cells, which would otherwise be tolerated, destroyed. Fujiwara et al., J. Immunol., 1984, 132, 1571 showed that mouse tumor cells conjugated with the hapten trinitrophenyl (TNP), could induce systemic immunity against unmodified tumor cells in a murine system, provided that the mice were first sensitized to the hapten in the absence of hapten-specific suppressor T cells. Spleen cells from treated mice completely and specifically prevented the growth of tumors in untreated recipient animals. Flood et al, J. Immunol., 1987, 138, 3573 showed that mice immunized with a TNP- conjugated, ultraviolet light-induced "regressor" tumor were able to reject a TNP-conjugated "progressor" tumor that was otherwise non-immunologic. Moreover, these mice were subsequently resistant to challenge with unconjugated "progressor" tumor. In another experimental system, Fujiwara et al, J. Immunol, 1984, 133, 510 demonstrated that mice sensitized to trinitrochlorobenzene (TNCB) after cyclophosphamide pretreatment could be cured of large (10 mm) tumors by in situ haptenization of tumor cells; subsequently, these animals were specifically resistant to challenge with unconjugated tumor cells. However, these results could not show whether haptenized cells isolated from a human spontaneous tumor would be effective in immunotherapy and be capable of inducing tumor regression. The existence of T cells which cross-react with unmodified tissues has recently been demonstrated. Weltzien and coworkers have shown that class I MHC -restricted T cell clones generated from mice immunized with TNP-modified syngeneic lymphocytes respond to MHC-associated, TNP-modified "self peptides. Ortmann, B., et al., J. Immunol., 1992, 148, 1445. In addition, it has been established that immunization of mice with TNP-modified lymphocytes results in the development of splenic T cells that exhibit secondary proliferative and cytotoxic responses to TNP-modified cells in vitro. Shearer, G. M. Eur. J. Immunol, 1974, 4, 527. The common denominator of these experiments is sensitization with hapten in a milieu in which suppressor cells are not induced. Spleen cells from cyclophosphamide pretreated, TNCB-sensitized mice exhibited radioresistant "amplified helper function," i.e., they specifically augmented the in vitro generation of anti-TNP cytotoxicity. Moreover, once these amplified helpers had been activated by in vitro exposure to TNP-conjugated autologous lymphocytes, they were able to augment cytotoxicity to unrelated antigens as well, including tumor antigens (Fujiwara et al., 1984). Flood et al., (1987), supra, showed that this amplified helper activity was mediated by T cells with the phenotype Lyt " l + , Lyt ~ 2 " , L3T4 + , I and speculated that these cells were contrasuppressor cells, a new class of immunoregulatory T cell. Immunotherapy of patients with melanoma had shown that administration of cyclophosphamide, at high dose (1000 mg/M 2 ) or low dose (300 mg/M 2 ), three days before sensitization with the primary antigen keyhole limpet hemocyanin markedly augments the acquisition of delayed type hypersensitivity to that antigen (Berd et al., Cancer Res., 1982, 42, 4862; Cancer Res., 1984, 44, 1275). Low dose cyclophosphamide pretreatment allows patients with metastatic melanoma to develop delayed type hypersensitivity to autologous melanoma cells in response to injection with autologous melanoma vaccine (Berd et al., Cancer Res., 1986, 46, 2572; Cancer Invest., 1988, 6, 335). Cyclophosphamide administration results in reduction of peripheral blood lymphocyte non-specific T suppressor function (Berd et al., Cancer Res., 1984, 44, 5439; Cancer Res., 1987, 47, 3317), possibly by depleting CD4+, CD45R+ suppressor inducer T cells (Berd et al, Cancer Res., 1988, 48, 1671). The anti-tumor effects of this immunotherapy regimen appear to be limited by the excessively long interval between the initiation of vaccine administration and the development of delayed type hypersensitivity to the tumor cells (Berd et al, Proc. Amer. Assoc. Cancer Res., 1988, 29, 408 (#1626)). Therefore, there remains a need to increase the therapeutic efficiency of such a vaccine to make it more immunogenic. Most tumor immunologists now agree that infiltration of T lymphocytes into the tumor mass is a prerequisite for tumor destruction by the immune system. Consequently, a good deal of attention has been focused on what has become known as "TIL" therapy, as pioneered by Dr. Stephen Rosenberg at NCI. Dr. Rosenberg and others have extracted from human cancer metastases the few T lymphocytes that are naturally present, and greatly expanded their numbers by culturing them in vitro with interleukin 2 (IL2), a growth factor for T lymphocytes. Topalian et al., J. Clin. Oncol, 1988, 6, 839. However this therapy has not been very effective because the injected T cells are limited in their ability to "home" to the tumor site. Human melanomas are believed to express unique surface antigens recognizable by T lymphocytes. Old, L. 1, CancerRes., 1981, 47, 361 ; Van der Bruggen, P., et al., Science, 1991, 254, 1643; Mukherji, B., et al., J. Immunol, 1986, 136, 1888; and Anichini, A., et al, J. Immunol, 1989, 142, 3692. However, immunotherapeutic approaches prior to work done by the present inventor had been limited by the difficulty of inducing an effective T cell-mediated response to such antigens in vivo. In the late 1960's and early 1970's, the research group of R. Powles at St. Barthlomew's Hospital in England conducted a series of studies of vaccine treatment of acute myelogenous leukemia (AML) patients after chemotherapy-induced remission (Powles, 1974; Powles etal, 1977). They used allogeneic AML cells with BCG as an adjuvant. Several trials were performed, all with small sample sizes (N=10-15). There was some prolongation of survival using a combination of chemotherapy and immunotherapy compared to chemotherapy alone, but no prolongation of relapse- free survival. No serious toxicity was observed; autoimmunity (e.g., toxicity to normal bone marrow) was not seen. In retrospect, there were a number of technical problems with these trials: 1) allogeneic, rather than autologous, leukemia cells were used; 2) the dose of leukemia cells in the vaccine was excessive (up to 10 9 cells/dose); 3) the BCG dose was very high and BCG administration was separated by time and location from the leukemia cell vaccine; and 4) the vaccine was administered while the patients were receiving cytotoxic drugs (maintenance or consolidation chemotherapy). Use of conventional therapies to treat human cancer has been generally unsuccessful. Administration of vaccine compositions also failed to reliably induce the development of cell-mediated immunity until the work of the present inventors. Hence, there is a continued need in the art for novel methods of treating cancer, and particularly cancer metastasis. Applicants have now discovered an effective method for inducing an anti-tumor response, and particularly tumor regression, in patients suffering from metastatic melanoma. SUMMARY OF THE INVENTION The present invention is directed to a method of inducing an anti-tumor response against a melanoma by administering an effective amount of a composition comprising at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten-modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response such as for example regression of a melanoma. The melanoma treated according to the present invention includes metastatic melanoma which may be lung, lymph node or subcutaneous metastasis, and is preferably a lung metastasis. The lung metastasis to be treated according to the invention is preferably a small lung metastasis. In one preferred embodiment of the invention, a melanoma metastasis is localized to the lung of the treated mammal. The invention is further directed to a melanoma cell, an isolated melanoma cell membrane, a peptide isolated from such cell or membrane, a T cell having a property of inducing an anti-tumor response, a composition containing such cell, membrane, peptide, T cell or combinations thereof, as well as the methods for their isolation and preparation. The melanoma cell or membrane, which may be hapten modified, may be syngeneic or allogeneic. The syngeneic melanoma cell or membrane may be autologous. The melanoma cell membrane is preferably a tumor cell plasma membrane. Accordingly, in one aspect, the present invention is directed to a method of inducing an anti-tumor response against a melanoma metastasis by administering to a mammal, preferably a human, a composition comprising a therapeutically effective amount of at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten-modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti -tumor response. The present invention also relates to inducing at least one of the following anti- tumor responses: tumor necrosis, tumor regression, tumor inflammation, tumor infiltration by activated T lymphocytes, stable disease and prolongation of patient survival. In another aspect, the invention relates to a method of inducing an anti-tumor response against a lung metastasis, which is preferably a complete or a partial regression of the metastasis, and/or prolongation of survival. In yet another aspect, the present invention relates to an isolated mammalian, preferably human, melanoma cell or membrane modified with a hapten, the peptides isolated from such cells and membranes, and T cells capable of mediating an anti-tumor response, as well as compositions thereof. In yet another aspect, the invention provides for a vaccine composition and dosage forms containing a therapeutically effective amount of a mammalian, preferably human, melanoma cell or membrane modified with a hapten, the peptides isolated from such cells and membranes, T cells, or combinations thereof adapted for administration to a mammal which suffers from a metastatic melanoma, preferably lung metastasis. In another aspect of the invention, the composition contains an adjuvant, such as, for example, Bacillus Calmette-Guerin (BCG), QS-21 , detoxified endotoxin and cytokines such as interleukin-2, interleukin-4, gamma interferon (IFN-γ), interleukin- 12, interleukin- 15 and GM-CSF. Brief Description of the Drawings Figure 1 A represents Patient #1 - chest x-ray, July, 1991,^re-vαcc e: shows multiple small lung nodules, best seen in left lower lobe Figure IB represents Patient #1 - chestx-ray, September, 1991, pre- vaccine: shows increased size of multiple lung nodules, best seen in left lower lobe Figure 1C represents Patient #1 - chest x-ray, January, 1992, post-vaccine: shows regression of multiple lung nodules Figure ID represents Patient # 2 - CT, January, 199 ', pre-vaccine: 1 cm diameter lung metastasis left lung adjacent to aorta Figure IE represents Patient #2 - CT, November, 1997, post-vaccine: regression of lung metastasis noted on D Figure IF represents Patient #3 - CT, April, 1996, pre-vaccine: approximately 2 cm diameter metastasis in right lower lobe adjacent to hear boarder. Figure 1 G represents Patient #3 - CT, April, 1998, post-vaccine: regression of approximately 2 cm diameter metastasis noted on F Figure 1H represents Patient # 4 - CT, January, 1997 ', pre-vaccine: approximately 1 cm diameter metastasis in periphery of right lower lobe Figure II represents Patient #4 - CT, January, 1998, post-vaccine: regression of metastasis noted in H DETAILED DESCRIPTION OF THE INVENTION All patents, patent applications and references cited herein are hereby incorporated by reference. In case of inconsistencies, the present disclosure governs. The present invention is directed to a method of inducing an anti-tumor response against a melanoma, preferably a lung metastasis, by administering an effective amount of a composition comprising at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten- modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response such as, for example, regression of a metastasis. The cells, membranes, peptides and T cells of the invention have the property of inducing at least one of the following anti-tumor responses: tumor necrosis, tumor regression, tumor inflammation, tumor infiltration by activated T lymphocytes, delayed-type hypersensitivity response, and prolongation of patient survival. The cells, membranes, peptides, and compositions thereof are capable of eliciting T lymphocytes that have a property of infiltrating a mammalian tumor, eliciting an inflammatory immune response to a mammalian tumor, eliciting a delayed-type hypersensitivity response to a mammalian tumor and or stimulating T lymphocytes in vitro. The melanoma cells, membranes, peptides and T cells of the invention and compositions thereof may be used for treating melanoma in a mammal, preferably a human, including treating metastatic and primary melanoma. Metastatic melanomas may include lymph node, lung and subcutaneous metastasis. Stage I, II, III, or IV cancer may be treated with the preparations, compositions and methods of the present invention, preferably stages III and IV. The lung metastasis to be treated according to the invention is preferably a small lung metastasis. For purposes of the present invention, a "small" lung metastasis is less than about 2 cm in diameter, preferably less than about 1.5 cm in diameter and most preferably less than about 1 cm in diameter. In one preferred embodiment of the invention, a melanoma metastasis in a mammal is localized to the lung of said mammal. Lung metastases to be treated according to the present invention may be single or multiple nodules. In one embodiment, the present invention is used to treat domestic animals such as, for example, members of feline, canine, equine and bovine families. It will be understood that any disclosure in this specification with respect to use of isolated melanoma cells equally applies to use of melanoma cells, membranes, peptides, T cells or to a combination thereof. Preparations of the Invention and Compositions thereof Melanoma cells, membranes, peptides and T cells are herein collectively referred to as the preparations of the invention. Furthermore, a term "extract" may be used to collectively refer to disrupted melanoma cells, isolated melanoma cell membranes and melanoma cell peptides. The isolated melanoma cell, membrane or peptide of the present invention are prepared from mammalian, preferably human, melanoma cells. The source of melanoma cells may be lung, lymph node or subcutaneous tumor masses including metastatic masses. In one embodiment of the invention, these materials are isolated from a melanoma of an animal from a feline, canine, equine or bovine family. T cells of the invention are isolated from tumor masses, such as for example metastatic lung and lymph node masses, and may be expanded in vitro. For the purposes of the present invention, a melanoma cell is intended to include both whole and disrupted melanoma cells. The melanoma cells for use in the vaccine composition of the invention, as well as those melanoma cells from which membranes and peptides are isolated, may be live, attenuated, or killed cells. Melanoma cells which do not grow and divide after administration into the subject such that they are substantially in a state of no growth can be used in the present invention. Such cells are preferred if they are administered to the patient. As used herein, the phrase "cells in a state of no growth" means live, attenuated or killed cells, cells in GO phase, whole or disrupted (or both whole and disrupted), i.e., cells that do not divide in vivo. Conventional methods of suspending cells in a state of no growth are known to skilled artisans and may be useful in the present invention. For example, cells may be irradiated prior to use such that they do not grow and divide. Melanoma cells may be irradiated, for example at 2500 R. Melanoma cell membranes and peptides may be isolated from either melanoma cells in a no growth state or melanoma cells that are capable of growing and dividing in vivo. Preferably, in the latter case, the melanoma cell membrane or peptide preparation is not contaminated with melanoma cells that are capable of dividing in vivo. Melanoma cells, membranes and peptides are isolated from the melanoma cells which may be of lymph node, lung and subcutaneous origin. Preferably, the melanoma cells originate from the same subject who is to be treated. The melanoma cells are preferably syngeneic (e.g. autologous). To be defined as "syngeneic," the melanoma cell need not be completely (i.e., 100 %) genetically identical to either the tumor cell or the non-tumor, somatic cell of the treated patient. Genetic identity of the MHC molecules between the tumor cell and the patient is generally sufficient. Additionally, there may be genetic identity between a particular antigen on the melanoma cell and an antigen present on the patient's tumor cells. Genetic identity may be determined according to the methods known in the art. For purposes of the present invention, a melanoma cell that has been genetically altered (using for example recombinant DNA technology) to become genetically identical with respect to, for example, the particular MHC molecules of the patient and/or the particular antigen on the patient's cancer cells is also within the meaning of the term "syngeneic" melanoma cell. However, such cells may also be referred to as "MHC-identical" or "MHC-compatible." Melanoma cells from mammals of the same species that differ genetically, such as allogeneic cells, may also be used for the preparation of melanoma cell membranes and peptides of the invention. The melanoma cells may be, and are not limited to, cells dissociated from biopsy specimens or from tissue culture. Membranes isolated from allogeneic cells and stem cells are also within the scope of the present invention. Melanoma cell membranes may include all cellular membranes, such as outer membrane, nuclear membranes, mitochondrial membranes, vacuole membranes, endoplasmic reticular membranes, golgi complex membranes, and lysosome membranes. In one embodiment of the invention greater than about 50% of the membranes are melanoma cell plasma membranes. Preferably, greater than about 60% of the membranes consist of melanoma cell plasma membranes, with greater than about 70% being more preferred, 80% being even more preferred, 90% being even more preferred, 95% being even more preferred, and 99% being most preferred. Preferably, the isolated membranes are substantially free of nuclei and cells. For example, a membrane preparation is substantially free of nuclei or cells if it contains less than about 100 cells and/or nuclei in about 2 x 10 8 cell equivalents (c.e.) of membrane material. A cell equivalent is that amount of membrane isolated from the indicated number of cells. An isolated melanoma cell membrane which is substantially free of cells and/or nuclei may contain lymphocytes and/or lymphocyte membranes. Preferably, the isolated melanoma cell membranes are the outer cell membranes, i.e., melanoma cell plasma membranes. The membrane preparation of the invention may contain the entire outer membrane or a fraction thereof. An isolated membrane of the invention containing a fraction of the outer membrane contains at least an MHC molecule fraction and/or a heat shock protein fraction of the outer membrane. The size of membrane fragments is not critical. The isolated melanoma cells as well as melanoma cell membranes may be modified, for example, with a hapten. Such modified melanoma cells and membranes have at least one of the following properties: (i) eliciting T lymphocytes that infiltrate the tumor of a treated mammal, (ii) eliciting an inflammatory immune response against the tumor of the mammal, and (iii) eliciting a delayed-type hypersensitivity response to the tumor of the mammal. Modified tumor cell membranes and cells also have the property of stimulating T cells in vitro. The peptides of the present invention may be isolated from a hapten modified melanoma cell or membrane. The peptides of the present invention may be hapten-modified. For the purpose of the present invention, peptides are compounds of two or more amino acids. Peptides will preferably be of low molecular weight, of about 1,000 kD to about 10,000 kD, more preferably of about 1,000 to about 5,000. The peptide may preferably be about 8 to about 20 amino acids, in addition the peptide may be haptenized. Peptides may be isolated from the cell surface, cell interior, or any combination of the two locations. The extract may be particular to type of cancer cell (versus normal cell). The peptide of the present invention includes and is not limited to a peptide which binds to the major histocompatibility complex or to a cell surface-associated protein such as a heat shock protein. This peptide may be a protein encoded by cancer oncogenes or mutated anti-oncogenes. Both individual peptides and small peptide-containing fractions of a melanoma cell or membrane are within the scope of the present invention. A small peptide-containing fraction is that fraction of melanoma peptides which has the ability to stimulate T cells and contains peptides that are isolated together in a particular purification step. For example, a pool of samples eluted from an HPLC column may represent such a small peptide-containing fraction. The useful peptides of the invention have a property of stimulating T lymphocytes. The ability of peptides to stimulate T cells can be identified by using standard assays, such as by measuring uptake of labelled nucleotides by T cells or by measuring production of cytokines such as and not limited to gamma interferon, tumor necrosis factor (TNF), and IL-2. Allogeneic melanoma cell membranes and peptides isolated from allogeneic melanoma cells may also be used in the methods of the present invention with syngeneic (e.g. autologous) antigen presenting cells. This approach permits immunization of a patient with melanoma cell membranes or peptides originating from a source other than the patient's own melanoma. Syngeneic antigen-presenting cells process allogeneic membranes or peptides such that the patient's cell-mediated immune system may respond to them. A melanoma cell, membrane or peptide (modified or un-modified) as referred to in this specification includes any form in which such preparation may be stored or administered such as, for example, resuspended in a diluent, as a pellet, frozen or lyophilized. Mammalian T cells capable of mediating regression of a tumor or another specific immune response directed against a tumor (as evidenced, for example, by T cell expansion in vitro, or T cell cytotoxicity) are also within the scope of the present invention. The T cell may be elicited in vivo by immunization of the patient with a composition comprising haptenized cells of the same tumor type or may be produced from such T cells by cloning in vitro. In one preferred embodiment, the isolated human T cell expresses a Vβ receptor, which may be Vβl, Vβ5, Vβl3, or Vβl4. The T cells to be used in the method of the present invention, may be cytotoxic T lymphocytes (CTL), or more generally, tumor infiltrating lymphocytes (TIL), i.e., a type of effector lymphocyte associated with cell mediated immunity directed against the tumor. The T cells isolated from a patient may be CD8+ T cells and MHC class I specific. The cells, membranes, peptides and T cells of the invention may be employed in the methods of the invention singly or in combination with other compounds, including and not limited to other compositions of the invention. Accordingly, cells, membranes, peptides or T cells may be used alone or co-administered. For purposes of the present invention, co- administration includes administration together and consecutively. Further, the preparations of the invention may be co-administered with other compounds including and not limited to cytokines such as interleukin-2, interleukin-4, gamma interferon (LFN-γ), interleukin- 12, interleukin- 15 and GM-CSF. They may also be used in conjunction with other cancer treatments including and not limited to chemotherapy, radiation, antibodies, and antisense oligonucleofides. However, it is the advantage of the present invention that it can be useful alone as a cancer treatment, such that the need for additional therapies is unnecessary. A composition of the present invention may contain the isolated melanoma cell, membrane, peptide, T cell of the invention (modified or unmodified) or a combination thereof and a pharmaceutically acceptable carrier or diluent, such as and not limited to Hanks solution, saline, phosphate-buffered saline, sucrose solution, and water. In general, the pharmaceutically-acceptable carrier is selected with regard to the intended route of administration and the standard pharmaceutical practice. The proportional ratio of active ingredient to carrier naturally depends on the chemical nature, solubility, and stability of the compositions, as well as the dosage contemplated and can be optimized using common knowledge in the art. In one preferred embodiment of the invention, a composition of the invention is a vaccine composition containing an effective amount of an isolated hapten-modified melanoma cell, membrane, peptide, T cell or a combination thereof. For purposes of this disclosure, "an effective amount" is the amount necessary to achieve a desired result. For example, in a method for inducing an anti -tumor response, "an effective amount" means the amount of melanoma cell, membrane, peptide or T cell that has the property of causing at least one of the following: tumor necrosis, tumor regression, tumor inflammation, tumor infiltration by activated T lymphocytes, and prolongation of patient survival. Similarly, in a method for stimulating T cells in vitro, "an effective amount" is that amount of cells, membranes or peptides that results in T cell stimulation. The vaccine composition may contain, for example, at least 10 4 melanoma cells per dose, preferably at least 10 5 cells, and most preferably at least 10 6 cells. A dose is that amount ofthe vaccine composition that is administered in a single administration. With respect to melanoma membranes, the vaccine composition may contain, for example, at least 10 4 c.e. of isolated membranes per dose, preferably at least 10 5 c.e., and most preferably at least 10 6 c.e. In one embodiment, the vaccine composition contains from about 10 5 to about 2.5 x 10 7 cells, c.e. membranes, or a combination thereof per dose, more preferably about 5 x 10 6 cells/c.e. In another embodiment, the vaccine composition contains melanoma cell peptides isolated from about 10 5 to about 2.5 x 10 7 cells, c.e. membranes, or a combination thereof per dose, more preferably from about 5 x 10 6 cells/c.e. The amount ofthe tumor cells, membranes or peptides ofthe invention to be used generally depends on such factors as the affinity ofthe compounds for cancerous cells, the amount of cancerous cells present and the solubility of the composition. Dosages may be set with regard to weight and clinical condition of the patient. A vaccine composition of the invention may be packaged in a dosage form suitable for intradermal, intravenous, intraperitoneal, intramuscular, and subcutaneous administration. Alternatively, the dosage form may contain the isolated preparations ofthe invention thereof to be reconstituted at the time of the administration with, for example, a suitable diluent. Hapten The melanoma cells, melanoma cell membranes and melanoma peptides ofthe present invention may be used as modified, unmodified, or a combination of modified and unmodified. For purposes ofthe present invention, modified includes and is not limited to modification with a hapten. Any small molecule that does not alone induce an immune response (but that enhances immune response against another molecule to which it is conjugated or otherwise attached) may function as a hapten. Generally, the molecule used should have less than about 1 ,000 mw. A variety of haptens are known in the art such as for example: TNP (Kempkes et al, J. Immunol 1991 147:2461); phosphorylcholine (Jang et al, Eur. J. Immunol. 1991 21 : 1303); nickel (Pistoor et al. , J. Invest. Dermatol. 1995 105:92); arsenate (Nalefski and Rao, J. Immunol. 1993 750:3806). Generally, haptens suitable for use in the present invention have the property of binding to a hydrophilic amino acid (such as for example lysine). Hapten can be conjugated to a cell via ε-amino groups of lysine or -COOH groups. Additionally, hapten that can bind to hydrophobic amino acids such as tyrosine and histidine via diazo coupling can also be used. Examples of haptens suitable for use in the present invention are: dinitrophenyl, trinitrophenyl, N-iodoacetyl-N'-(5-sulfonic 1 -naphthyl) ethylene diamine, trinitrobenzenesulfonic acid, fluorescein isothiocyanate, arsenic acid benzene isothiocyanate, trinitrobenzenesulfonic acid, phosphorylcholine, sulfanilic acid, arsanilic acid, dinitrobenzene- S-mustard (Nahas and Leskowitz, Cellular Immunol. 198054:241) and combinations thereof. In view ofthe present disclosure, a skilled artisans would be able to choose haptens for use in the present invention. For example, haptens can be routinely tested using a delayed type hypersensitivity (DTH) test. Adjuvant In one embodiment, the melanoma cell, melanoma cell membrane, peptide or T cell is administered with an immunological adjuvant. The adjuvant has the property of augmenting an immune response to the preparations ofthe present invention. Representative examples of adjuvants are BCG, or the synthetic adjuvant, QS-21 comprising a homogeneous saponin purified from the bark of Quillaja saponaria, Corynebacterium parvum (McCune et al, Cancer 1979 43:1619), saponins in general, detoxified endotoxin and cytokines such as interleukin-2, interleukin-4, gamma interferon (IFN-γ), interleukin- 12, interleukin- 15, GM- CSF and combinations thereof. It will be understood that the adjuvant may be subject to optimization. In other words, the skilled artisan may use routine experimentation to determine the most optimal adjuvant to use. Methods of Making the Preparations of the Invention The melanoma cells for use in the present invention may be prepared as follows. Tumors are processed as described by Berd et al. (1986), supra, Sato, et al. (1997), U.S. Patent No. 5,290,551, and applications U.S. Serial Nos. 08/203,004, 08/479,016, 08/899,905, 08/942,794, or corresponding PCT application PCT/US96/09511 , each of which is incorporated herein by reference in its entirety. Briefly, the cells are extracted by dissociation, such as for example by enzymatic dissociation with collagenase and DNase, by mechanical dissociation in a blender, by teasing with tweezers, using mortar and pestle, cutting into small pieces using a scalpel blade. Melanoma cell membranes are prepared from melanoma cells by disrupting the cells using, for example, hypotonic shock, mechanical dissociation and enzymatic dissociation, and separating various cell components by centrifugation. Briefly, the following steps may be used: lysing tumor cells, removing nuclei from the lysed tumor cells to obtain nuclei-free tumor cells, obtaining substantially pure membranes free from cells and nuclei, and subjecting the tumor cell membranes to a hapten to obtain hapten-modified tumor cell membranes. Membrane isolation may be conducted in accordance with the methods of Heike et al. In one embodiment ofthe invention, intact cells and nuclei may be removed by consecutive centrifugation until membranes are substantially free of nuclei and cells, as determined microscopically. For example, lysed cells may be centrifuged at low speed, such as for example, at about 500-2,000 g for about five minutes. The separation procedure may be such that less than about 100 cells and or nuclei remain in about 2 x 10 8 cell equivalents (c.e.) of membrane material. The postnuclear supernatant containing membranes may be pelleted by ultracentrifugation, for example at about 100,000 g for about 90 minutes, for example. The pellet contains total membranes. Membranes may be resuspended, for example, in about 8% sucrose, 5 mM Tris, pH 7.6 and frozen at about -80°C until use. Any diluent may be used, preferably one that acts as a stabilizer. To determine the quality of membrane preparation (about 6 x 10 7 c.e. membranes) may be cultured under standard cell culture conditions. Cell colonies should not develop and cells or nuclei should not be detected by light microscopy. Modification ofthe prepared cells or membranes with DNP or another hapten may be performed by known methods, e.g. by the method of Miller and Claman, J. Immunol. , 1976, 117, 1519, incorporated herein by reference in its entirety, which involves a 30 minute incubation of tumor cells or membranes with a hapten under sterile conditions, followed by washing with sterile saline. The hapten-modification may be confirmed by flow cytometry using a monoclonal anti-hapten antibody. The dissociated cells or isolated membranes may be used fresh or stored frozen, such as in a controlled rate freezer or in liquid nitrogen until needed. The cells and membranes are ready for use upon thawing. Preferably, the cells or membranes are thawed shortly before they are to be administered to a patient. For example, on the day that a patient is to be skin tested or treated, the cells or membranes may be thawed. Optionally, the cells or membranes may be washed, and optionally irradiated to 2500 R. They may be washed again and then suspended in Hanks balanced salt solution without phenol red. Allogeneic melanoma cell membranes may be prepared as described above. However, prior to administration to a subject they are co-incubated with syngeneic (e.g. autologous) antigen presenting cells. Syngeneic antigen-presenting cells process allogeneic membranes such that the patient's cell-mediated immune system may respond to them. This approach permits immunization of a patient with melanoma cell membranes originating from a source other than the patient's own tumor. Allogeneic melanoma cell membranes are incubated with antigen-presenting cells for a time period varying from about several hours to about several days. The membrane-pulsed antigen presenting cells are then washed and injected into the patient. Antigen-presenting cells may be prepared in a number of ways including for example the methods of Grabbe et al., 1995 and Siena et al., 1995. Briefly, blood is obtained, for example by venipuncture or by leukapheresis, from the patient to be immunized. Alternatively, bone marrow may be obtained. From any of these sources, mononuclear leukocytes are isolated by gradient centrifugation. The leukocytes may be further purified by positive selection with a monoclonal antibody to the antigen, CD34. The purified leukocytes may be cultured and expanded in tissue culture medium (for example, RPMI-1640 supplemented with serum, such as fetal calf serum, pooled human serum, or autologous serum). Alternatively, serum-free medium may be used. To stimulate the growth of antigen- presenting cells, cytokines may be added to the culture medium. Cytokines include and are not limited to granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin 4 (IL4), TNF (tumor necrosis factor), interleukin 3 (IL3), FLT3 ligand and granulocyte colony stimulating factor (G-CSF). The antigen-presenting cells isolated and expanded in culture may be dendritic cells, monocytes, macrophages, and Langerhans cells, for example. The peptides of the invention may be isolated from cells according to an established technique of Rotzschke et al, Nature, 1990, 348, 252, the disclosure of which is hereby incorporated by reference in its entirety. The cells are treated with a weak acid, such as and not limited to trifluoroacetic acid. The cells are then centrifuged and the supernatant is saved. Compounds having a molecular weight greater than 5,000 are removed from the supernatant by gel filtration (G25 Sepharose, Pharmacia). The remainder ofthe supernatant is separated on a reverse-phase HPLC column (Superpac Pep S, Pharmacia LKB) in 0.1%. trifluoroacetic acid (TFA) using a gradient of increasing acetonitrile concentration; flow rate 1 ml/min, fraction size 1 ml. Fractions containing small peptides are obtained by HPLC according to the method of Sambrook et al, Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989), concentrated, and frozen. The fractions are screened for immunological activity by allowing them to bind to autologous B lymphoblastoid cells which are then tested for ability to stimulate melanoma- specific T lymphocytes. T cells are isolated from biopsies by known techniques, such as preparation of single cell suspension, filtration, depletion of monocytes and isolation of a subset expressing a particular TCR type by causing that subset to expand in the presence of a TCR-subtype specific antibody and/or in the presence of IL-2 and/or in the presence of a superantigen. The T cells of interest may be expanded in vitro using methods known in the art. Specifically, T lymphocytes may be prepared from tumors as follows. Single cell suspensions may be prepared from tumors by digestion with a mixture of 0.14% collagenase, 0.03%) DNase and optionally 2.5 U/ml hyaluronidase (Sigma Chemical CO., St. Louis, MO) for 3 hours at room temperature. The cells may be filtered through a layer of no. 100 nylon mesh then washed and resuspended in buffer, e.g., Hanks Buffered Saline. The mixture of cells may be depleted of monocytes by panning ofthe mixture over plastic dishes in a final volume of 2ml RPMI- 1640 supplemented with 10% pooled human serum and cultured for one week. T cells can be expanded by exposure to antibodies and or an to immunostimulatory cytokine (such as, IL-2) and/or superantigens as disclosed e.g., inPCT/US93/05213. Activity ofthe T cells may be measured after four to five weeks of in vitro stimulation. T cells can also be purified using Dynabeads (DYNAL, Lake Success, New York) coated with various antibodies, e.g., anti-BV14 or anti-CD8+, to enhance the degree and speed of purification of the T cells prior to or after expansion ofthe T cells. Alternative methods of T cells expansion in vitro involve use of superantigens or monoclonal antibodies to a T cell receptor expressed by the tumor infiltrating cells or use of an immunostimulatory cytokines such as TNF, gamma interferon, and an interleukin (IL-1, IL-2, IL-12 etc.). Methods of Inducing an Anti-tumor Response The present invention is directed to a method of inducing an anti-tumor response against a melanoma by administering an effective amount of a composition comprising at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten-modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response such as for example regression of a melanoma. The melanoma treated according to the present invention includes metastatic melanoma which may be lung, lymph node or subcutaneous metastasis, and is preferably a lung metastasis. The lung metastasis to be treated according to the invention is preferably a small lung metastasis. In one preferred embodiment of the invention, a melanoma metastasis in a mammal is limited to the lung of said mammal. Primary cancers may also be treated according to the invention. Stage I, II, III, or IV cancer may be also be treated according to the invention, and preferably stages III and IV. In one embodiment ofthe invention, domestic animals may be treated. In one preferred embodiment, a mammal, preferably a human, having a single or multiple lung metastases is treated. Small metastases are particularly suited for treatment according to the present invention. The size of such metastases may be about 2 cm in diameter, preferably less than about 1.5 cm, and most preferably less than about 1 cm. An anti-tumor response resulting from such a treatment may be a partial or a complete regression ofthe metastatic tumor or a stable disease. A "complete" regression indicates about 100% regression for a period of at least one month, more preferably for a period of at least three months. A "partial" regression indicates more than about 50 % regression for a period of at least one month, more preferably for a period of at least three months. A "stable" disease indicates a condition in which there is no significant growth of the tumor after the vaccine treatment. Another anti-tumor response that may be observed upon following the treatment ofthe invention is prolongation of survival. Prior to administration ofthe vaccine composition ofthe invention, the subject may be immunized to the hapten used to modify tumor cells and membranes by applying it to the skin. For example, dinitrofluorobenzene (DNFB) may be used to immunize the patient against DNP. Subsequently (about two weeks later, for example), the subject may be injected with a preparation of the invention. In one embodiment ofthe invention, the patient is not immunized prior to administration ofthe vaccine. A pharmaceutically acceptable amount of a low-dose cyclophosphamide or another low-dose chemotherapy may be administered preceding the administration of the composition. A haptenized vaccine composition may optionally be followed by administration of a pharmaceutically acceptable amount of a non-haptenized composition. A non-haptenized composition may also be administered in accordance with the methods of the present invention. The composition may be administered (such as by reinjection) for a total of at least three and preferably at least six treatments. In one embodiment, the total number of administrations (including the initial administration) may be eight, and in another embodiment may be ten. The vaccination schedule may be designed by the attending physician to suit the particular subject's condition. The vaccine injections may be administered, for example, every week, every 2 weeks, or every 4 weeks. A booster vaccine may be administered. Preferably, one or two booster vaccines are administered. The booster vaccine may be administered, for example, after about six months or about one year after the initial administration. The present invention may be used following conventional treatment for cancer, such as surgery. Excised tumors or collected tumor cells may be used to prepare tumor cells and membranes as described above. The preparations of the invention may be administered by any suitable route, including inoculation and injection, for example, intradermal, intravenous, intraperitoneal, intramuscular, and subcutaneous. There may be multiple sites of administration per each vaccine treatment. For example, the vaccine composition may be administered by intradermal injection into at least two, and preferably three, contiguous sites per administration. In one embodiment ofthe invention, the vaccine composition is administered on the upper arms or legs. The effectiveness of the vaccine may be improved by administering various biological response modifiers. These agents work by directly or indirectly stimulating the immune response. Biological response modifiers ofthe present invention include and are not limited to interleukin- 12, interleukin- 15 and gamma interferon. In one embodiment, IL12 is given following each vaccine inj ection. Administration of IL 12 to patients with inflammatory responses may cause the T lymphocytes within the tumor mass to proliferate and become more active. The increased T cell numbers and functional capacity leads to immunological destruction and regression ofthe tumors. The modified tumor cells, membranes and peptides each have the property of stimulating T cells. "Stimulation" for purposes of the present invention refers to inducing proliferation of T cells as well as production of cytokines by T cells in vitro. Proliferation of T cells may be detected and measured by the uptake of modified nucleotides, such as and not limited to 3 H thymidine, I25 IUDR (iododeoxyuridine); and dyes such as 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) which stain live cells. In addition, production of cytokines such as and not limited to IFNγ, tumor necrosis factor (TNF), and IL-2 may be useful in exhibiting T cell proliferation. Production of cytokines may be detected and measured using tests well known in the art. Cytokine production should be above the background level, which is generally above 25 picograms/ml, and is preferably above 100 picograms/ml. The following non-limiting examples further illustrate the invention. EXAMPLES Example 1 : Sixteen patients suffering from melanoma and having lung metastases were treated according to the present invention. All treated patients had measurable lung metastases. For purposes of the present invention, the term "measurable" lung metastasis refers to a metastasis that is visible on an X ray. Melanoma cells were isolated from lymph node and lung metastases and prepared in accordance with the methods set forth above. The cells were obtained by dissociation of metastatic masses enzymatically (with collagenase and DNAse). The cells were conjugated to DNP as described herein. Some ofthe patients were sensitized to DNP by topical application of 5 % dinitrochlorobenzene to the upper arm. The first vaccine was preceded by a low dose cyclophosphamide (CY). On day 0, the patients were given cyclophosphamide 300 Mg/M 2 i.v. Three days later, they were injected intradermally with a vaccine containing 2.5 X 10 6 to 25 X 10 6 autologous, cryopreserved, irradiated (2500 R) tumor cells mixed with BCG. Mo$t patients were treated every week for 6 treatments. The patients were compared to their condition prior to treatment with the vaccine. The patients treated with other cancer therapies prior to the vaccine study were removed from such treatments at least two months prior to starting the vaccine study. Accordingly, the patients were untreated beginning the vaccine study. Partial responses (PR) (documented by computerized tomography) were observed in 3/16 patients, and a fourth patient has exhibited ongoing tumor regression with <50% regression to date (stable disease). The details ofthe PR's were as follows: Patient 1: 90% regression of multiple (>50) small (5-10 mm diameter) lung nodules; Patient 2: 75% regression of a solitary 1 cm diameter nodule; and Patient 3: complete regression of a 1 cm nodule with partial regression of two accompanying nodules. All responses were in patients whose metastases were limited to the lung. Tumor regression was not evident in any responder until 4-6 months after beginning vaccine treatment. The survival ofthe 4 patients with PR or stable disease is 34.5, 8.5+, 9.2+, and 17+ months, respectively. Accordingly, a DNP-melanoma cell vaccine can induce radiographically documented regression of melanoma metastases. Small lung metastases may be particularly susceptible to immunological destruction. Example 2: Tumor masses for the preparation of a vaccine composition ofthe invention may be obtained from lymph node, lung or subcutaneous metastatic masses and processed as previously described. Briefly, cells may be extracted by enzymatic dissociation with collagenase and DNase and by mechanical dissociation. Cell membranes may be isolated as described in this specification, and frozen in a controlled rate freezer, and stored in liquid nitrogen until needed. On the day that a patient is to be treated, the membranes may be thawed, washed, and resuspended in Hanks balanced salt solution without phenol red. Modification with DNP may be performed by the method of Miller and Claman (1976). This involves a 30 minute incubation of melanoma cells with dinitrofluorobenzene (DNFB) under sterile conditions, followed by washing with sterile saline. Melanoma patients having small multiple lung metastases and melanoma patients with metastases localized to the lung may be treated as follows. The vaccine composition may contain a minimum of 2.5x 10 6 c.e. trypan-blue-excluding melanoma cell membranes, and a maximum of 7.5x10 6 c.e. melanoma cell membranes, suspended in 0.2 ml Hanks solution. Each vaccine treatment may consist of three injections into contiguous sites. The freeze-dried material may be reconstituted with 1 ml sterile water or phosphate buffered saline, pH 7.2 (PBS). Appropriate dilutions may be made in sterile buffered saline. Then 0.1 ml may be drawn up and mixed with the vaccine just before injection. The first and second vaccines may be mixed with 0.1 ml of a 1 : 10 dilution of Tice BCG ("Tice- 1 "). BCG is a Tice strain (substrain ofthe Pasteur Institute strain) obtained from Organon Teknika Corporation (Durham, NC). The third and fourth vaccines may be mixed with 0.1 ml of a 1:100 dilution ("Tice-3"). The fifth and sixth and booster vaccines may be mixed with 0.1 ml of a 1:1000 dilution ("Tice-5"). The ideal vaccine reaction is an inflammatory papule with no more than small (< 5mm) central ulceration. Skin testing may be performed by the intradermal injection of 0.1 ml of test material on the forearm, and DTH is assessed at 48h by measuring the mean diameter of induration. The following materials may be tested: 1) 1x10^ autologous melanoma cell membranes umnodified and modified with DNP; both enzymatically-dissociated (TCE) and mechanically-dissociated (TCM) melanoma cells may be used; 2) 3x10^ autologous peripheral blood lymphocytes unmodified and modified with DNP; 3) Hanks solution; and 4) PPD-intermediate strength. Also, contact sensitivity to DNFB may be tested by applying 200 μg DNFB to the skin ofthe ventral surface ofthe upper arm and examining the area for a circle of induration at 48 hours. The full battery of DTH tests may be performed following the six week course of vaccine administration. Pre-treatment DTH testing may be limited to DNP-modified melanoma cell membranes, PPD, and diluent. This strategy is designed to avoid: 1) sensitizing patients to DNP-modified lymphocytes and 2) tolerizing patients by injection of unmodified melanoma cells. All patients may have blood collected for separation and cryopreservation of lymphocytes and serum each time skin-testing is performed. Periodically, these may be tested for: response to autologous cancer cells, as measured by proliferation, cytokine release, and cytotoxicity. Patients may be evaluated for metastatic disease before vaccine therapy begins. After the end ofthe first six weeks of vaccine therapy, evaluations may be performed every three months. Evaluations may continue through year two, every four months in year three, and every six months thereafter. Patients who are responders at the 1 year evaluation may receive a final booster injection of vaccine. Then their condition may be followed without further treatment. An efficacy study to determine whether DNP-vaccine prolongs relapse-free and/or total survival in these patients may also be conducted. Survival parameters (Kaplan-Meier method) may be measured. Example 3: Four patients that responded to the vaccine of the invention as described in Example 1 , were continued to be monitored to assess the effectiveness ofthe vaccine. Three patients (Patients 2, 3 and 4 in Example 1) received a total of 6 vaccines, a vaccine per week, with a booster vaccine at six and 12 months from the first vaccine. One patient (Patient 1) received weekly vaccines for 12 weeks without booster vaccine. Two patients had complete remission and two patients had partial remission. Patient 1 (of Example 1) had a partial remission for a duration of 12 months. Patient 2 (of Example 1) had partial remission for a period of 8 months. Patient 3 (of Example 1) had a complete remission for a period of 29 months, and the fourth patient (of Example 1) had a complete remission in excess of 27 months. The term "complete remission" indicates that all detectable tumors were completely gone. The term "partial remission" generally indicates that there was at least a 50% decrease in the diameter of the tumors. However, in the present case, with respect to Patient 1 and Patient 2, about 90% decrease in the diameter ofthe tumors was observed. The results are represented in Figures A-I. REFERENCES Anichini, A., et al., J. Immunol, 1989, 142(10):3692 Andreasen, R.B., et al., Scand. J. 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Claims: WHAT IS CLAIMED IS: A method of inducing an anti-tumor response against a melanoma metastasis in a mammal comprising administering to said mammal a composition comprising a therapeutically effective amount of at least one of the following: (i) a hapten-modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten- modified melanoma cell membrane, (iii) a peptide isolated from said hapten- modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response; wherein said metastasis is localized to the lung of said mammal. 2. The method of claim 1 wherein said anti-tumor response is at least one of the following: tumor necrosis, tumor regression, tumor inflammation, tumor infiltration by activated T lymphocytes, stable disease and prolongation of patient survival. 3. The method of claim 1 wherein said anti-tumor response is a complete or a partial regression ofthe metastasis. 4. The method of claim 1 wherein said mammal is a human. 5. The method of claim 1 wherein said membrane is a syngeneic melanoma cell membrane or an allogeneic melanoma cell membrane. 6. The method of claim 5 wherein said melanoma cell membrane is allogeneic and said composition further comprises an antigen presenting cell. 7. The membrane of claim 1 wherein said membrane comprises a membrane fraction comprising an MHC molecule, a heat shock protein or a combination thereof. 8. The method of claim 1 wherein said hapten is selected from the group consisting of dinitrophenyl, trinitrophenyl, N-iodoacetyl-N'-(5-sulfonic 1 -naphthyl) ethylene diamine, trinitrobenzenesulfonic acid, fluorescein isothiocyanate, arsenic acid benzene isothiocyanate, trinitrobenzenesulfonic acid, sulfanilic acid, arsanilic acid, dinitrobenzene-S- mustard and combinations thereof. 9. The method of claim 1 wherein said hapten is dinitrophenyl. 10. The method of claim 1 wherein said composition further comprises an adjuvant. 11. The composition of claim 10 wherein said adjuvant is selected from the group consisting of Bacillus Calmette-Guerin, QS-21 , detoxified endotoxin and a cytokine. 12. The method of claim 1 wherein said administration is repeated at least six times at spaced apart intervals. 13. The method of claim 12 wherein said administration is repeated every week. 14. The method of claim 1 further comprising administering a therapeutically effective amount of cyclophosphamide prior to administration of said composition. 15. The method of claim 14 wherein said therapeutically effective amount of cyclophosphamide comprises administering a dose of about 300 mg/M 2 of cyclophosphamide prior to administration of said composition. 16. A method of inducing an anti -tumor response against a metastasis in a mammal suffering from melanoma comprising administering to said mammal a composition comprising a therapeutically effective amount of at least one ofthe following: (i) a hapten- modified syngeneic mammalian melanoma cell substantially in a no growth phase, (ii) a hapten-modified melanoma cell membrane, (iii) a peptide isolated from said hapten-modified melanoma cell or membrane and (iv) a T cell capable of mediating an anti-tumor response; wherein said metastasis is a small lung metastasis. 17. The method of claim 16 wherein said anti-tumor response is at least one ofthe following: tumor necrosis, tumor regression, tumor inflammation, tumor infiltration by activated T lymphocytes, stable disease and prolongation of patient survival. 18. The method of claim 16 wherein said anti -tumor response is a complete or a partial regression ofthe lung metastasis. 19. The method of claim 16 wherein said mammal is a human. 20. The method of claim 16 wherein said membrane is a syngeneic melanoma cell membrane or an allogeneic melanoma cell membrane. 21. The method of claim 20 wherein said melanoma cell membrane is allogeneic and said composition further comprises an antigen presenting cell. 22. The membrane of claim 16 wherein said membrane comprises a membrane fraction comprising an MHC molecule, a heat shock protein or a combination thereof. 23. The method of claim 16 wherein said hapten is selected from the group consisting of dinitrophenyl, trinitrophenyl, N-iodoacetyl-N'-(5-sulfonic 1 -naphthyl) ethylene diamine, trinitrobenzenesulfonic acid, fluorescein isothiocyanate, arsenic acid benzene isothiocyanate, trinitrobenzenesulfonic acid, sulfanilic acid, arsanilic acid, dinitrobenzene-S- mustard and combinations thereof. 24. The method of claim 16 wherein said hapten is dinitrophenyl. 25. The method of claim 16 wherein said composition further comprises an adjuvant. 26. The composition of claim 25 wherein said adjuvant is selected from the group consisting of Bacillus Calmette-Guerin, QS-21 , detoxified endotoxin and a cytokine. 27. The method of claim 16 wherein said administration is repeated at least six times at spaced apart intervals. 28. The method of claim 27 wherein said administration is repeated every week. 29. The method of claim 16 further comprising administering a therapeutically effective amount of cyclophosphamide prior to administration of said composition. 30. The method of claim 29 wherein said therapeutically effective amount of cyclophosphamide comprises administering a dose of about 300 mg/M 2 of cyclophosphamide prior to administration of said composition.

Inventors: BERD DAVID, BERD, DAVID
Applicants: BERD DAVID, UNIV JEFFERSON, BERD, DAVID, THOMAS JEFFERSON UNIVERSITY
Application year: 1999
Main classification: A61K39
IPC codes: A61K31, A61K35, A61K39, A61P35