MXPA06001572A - Vaccine immunotherapy for immune suppressed patients - Google Patents

Abstract

A method of immunotherapy to treat cancer or a synergistic anti-cancer treatment by administering an effective amount of a natural cytokine mixture (NCM), an effective amount of cyclophosphamide (CY), or an effective amount of indomethacin (INDO), wherein the NCM, CY, or INDO are administered singly or in communications thereof. An anti-metastatic treatment method by promoting differentiation and maturation of immature dendritic cells in a lymph node;allowing presentation thereof;and preventing development of metastasis. A method of using an NCM as a diagnostic skin test for predicting treatment outcome. A method of pre-treating dendritic cells (DC) and a method of treating monocyte defects characterized by sinus histiocytosis or a negative NCM skin test. Compositions and methods for elicting an immune response to endogenous or exogenous tumor antigens.

Description

INMU OTERAPIA WITH VACCINE FOR IMMUNODEPRIMID PATIENTS BACKGROUND OF THE INVENTION 1. TECHNICAL FIELD The present invention relates to vaccine therapy for cancer patients. More specifically, the present invention relates to an immunotherapy with vaccine, which immunizes cancer patients, who have immunosuppression, to peptides or tumor proteins both endogenous and exogenous. 2. BACKGROUND TECHNIQUE It has become increasingly evident that human cancers have antigens, which, if reacted by host immune systems, lead to tumor regression. These antigens have been defined by both serological and cellular immune procedures. This has led to the definition of both B and T cell epitopes (Sahin U, and co-workers Curr Opin Im unol 9: 709-715, 1997; Van der Eynde, B, and collaborators Curr Opin Immunol 9: 684-693, 1997; Wang RF, et al., Immunologic Reviews 170: 85-100, 1999). Based on these results, it has become a goal of cancer immunotherapists to induce tumor regressions. However, historically, successful efforts have been sporadic and generally less in frequency and magnitude. A fundamental problem in the effort to immunize cancer patients is that the tumor bearing state is associated with immunosuppressive mechanisms derived from both the tumor and the altered immune system of the host (Kavanaugh DY, et al., Hematol-Oncol Clinics of North Amer 10 (4): 927-951, 1996), thus making immunization difficult and even impossible on a consistent basis. The suppression or immune depletion implies a reduced capacity of the immune system to respond. Such suppression can be induced by drug or by disease. The condition can be induced by drug by treatment, induced by viruses as in AIDS, or induced by a disease state such as cancer. The immune system in this condition is effectively deactivated or turned off. A variety of tumor immunization strategies have been developed. Without such an approach, all of these strategies are complex and deviate significantly from conventional immunization strategies used for infectious diseases (Weber J. Tumor, Medscape Anthology 3: 2, 2000). Such a tumor immunization strategy involves Theratope (R), a Sialyl TN polyacarid mucin antigen conjugated to tip-lane hemocyanin and administered with Detox (R) mycobacterial adjuvant and low dose cyclophosphamide (Maclean GD, et al. unother E phasis Tumor Immunol 19 (4): 309-316, 1996). However, the use of this vaccine in patients with metastatic breast and ovarian cancer has produced greater chemical responses in a low percentage of patients. A larger response means greater than 50% tumor reduction. Gene therapy has also been attempted using an adenovirus construct as an expression vector for genes expressing Papilloma virus peptide 16, has been used for the immunization of patients with cervical cancer and has produced major clinical responses in a low percentage of patients (Borysiewickz LK, et al., Lancet 347: 1524-1527, 1996). Dendritic cell-mediated therapy has also been attempted, wherein the dendritic cells were pulsed with oligopeptide fragments of prostate-specific antigens (PSA). Prostate-specific membrane antigen (PSAM) has been used in patients with metastatic prostate cancer with greater clinical responses in a low percentage of patients (Sanda MG, et al. Urology 52: 2, 1999; Murphy GP, et al., The prostate, 38: 43-78, 1999). Additionally, autologous tumors have been used with low-dose cyclophosphamide and BCG to immunize patients with cancer with malignant melanoma. However, few chemical responses were reported (Mastrangelo M J et al., Seminars in Oncology 23 (6): 773-781, 1996). Another strategy attempted included the use of MAGE antigens with a variety of vaccine adjuvants. Again, this has produced few, if any, responses in patients with malignant melanoma (personal communication Thierry Boon). Several patents of Doyle et al. (U.S. Patent Nos. 5,503,841; 5,800,810; 6,060,068; 5,643,565; 5,100,664) disclose methods for increasing the immune response in patients using Interleukin 2- (IL-2). This method is disclosed for use in response to infectious diseases and primarily works by using known antigens that are immunogenic. Limited applicability was demonstrated. As disclosed above, cancer treatment is known to require different procedures. To date, treatment with IL-2 has shown minor effects in two cancers, renal cell and malignant melanoma. (response ratios less than 20%). It is generally considered ineffective in cancer of the head and neck and neck of the squamous cell and in prostate cancer. Consequently, it is not approved for these uses. Therefore it would not be within the skill of one in the art to apply the method of the Doyle et al. Patents for the use of small peptides in the treatment of cancer. It is important to contrast the prevention with known "classical" antigens of complex structure and high molecular weights in healthy patients against the treatment (without general success) with antigens or tumor peptides in immunosuppressed patients (without success in general). The first is easy and the current viral vaccines of the inventors testify their effectiveness. The latter is almost impossible on a routine basis despite 30 years of intense effort. It is important that this invention relates to, but not exclusively to, immunizing with the endogenous peptide processed and presented by dendritic cells or administered endogenously to an environment (lymphoma node) where the dendritic cells have been prepared and can be effectively presented to T cells. This objective is considered by many immunologists to be insurmountable. The peptides are too small to be effective immunogens, their half-lives are short since they are frequently unmutated self antigens to which the patient is immunologically tolerant and the gain of a response is of importance to induce self immunity. In several of the above strategies, cell and / or tumor immunity to tumor-associated antigens has been induced (Weber J. Tumor, Medscape Anthology 3: 2, 2000; Macean GD, et al., J. Immunother Emphasis Tumor immunol 19 (4): 309-316, 1996; Borysiewickz LK, et al., Lancet 347: 1524-1527, 1996; Sanda MG et al., Urology 52: 2, 1999). This is especially so in association with tumor regression. However, the success rate of such treatments is negligible and inconsistent (< 30%). Therefore it would be useful to develop a consistent and effective method to immunize patients with cancer. BRIEF DESCRIPTION OF THE INVENTION According to the present invention, there is provided an immunotherapy method for treating cancer by administering an effective amount of a natural cytokine mixture.

(NCM) including, but not limited to, IL-1, IL-2, IL-6, IL-8, IL-12, IFN-α, TNF-α, GM-CSF, G-CSF, recombinant the same and combinations thereof. In addition, the present invention provides a method of immunotherapy for treating cancer by administering an effective amount of cyclophosphamide.

(CY) and an effective amount of indomethacin (INDO). Various methods of anti-cancer treatment are also provided wherein the administration of an effective amount of CY occurs in conjunction with an effective amount of a nonsteroidal anti-inflammatory drug (NSAID) that includes, but is not limited to, indomethacin (INDO) , ibuprofen, celecoxib (Celebrex®), rofecoxib (Vioxx®), CoxII inhibitors, and combinations thereof. More specifically, the present invention provides a method of immunotherapy for treating cancer by administering an effective amount of a CY in combination with an effective amount of INDO and an effective amount of IFN-α, IL-2, IL-1, and TNF- to. Additionally, the present invention provides an immunotherapy method for treating cancer by administering an effective amount of a CY in combination with an effective amount of INDO and an effective amount of recombinant IL-2, recombinant IFN-α, recombinant TFN-a, and recombinant IL-1. The present invention further provides a synergistic anti-cancer treatment by administering an effective amount of CY and INDO in combination with an NCM described herein. In addition, the present invention provides a method of anti-metastatic treatment by promoting the differentiation and maturation of mature dendritic cells in a lymphoma node.; allowing the presentation by the mature dendritic cells resulting from the antigen to T cells to gain immunization of the T cells to the antigen; and preventing the development of metastasis. Alternatively, the present invention provides an anti-metastatic method by unblocking immunization at a lymphoma node; and by generating systematic immunity. The present invention also provides a skin test and a method of pretreatment of dendritic cells (DC) by applying an effective amount of CY and INDO in combination with an NCM described herein. The present invention further provides a method for treating monocyte defects characterized by sinus histiocytosis or a negative skin test of NCM by applying an effective amount of CY and INDO in combination with an NCM described herein. Finally, the present invention provides compositions and methods for inducing an immune response to endogenous or exogenous tumor antigens. BRIEF DESCRIPTION OF THE DRAWINGS Other advantages of the present invention are readily appreciated as they come to be understood by reference to the following detailed description when considered in connection with the accompanying drawings in which: Figure 1 is a bar graph showing the size of the lymphoma node in controls, and cancer controls or IRX-2 populations (NCM) treated with squamous cell head and neck cancer (H &S; NSC); Figure 2 shows two bar graphs, one showing the area of the T cell and the second showing density in the controls and controls of squamous cancer of the head and neck and patients treated with NCM (IRX-2); Figure 3 shows two bar graphs showing an area of B cell and follicles in the three treatment groups; Figure 4 shows a comparison of other cells and sinus histocytosis in the three treatment groups; Figure 5 is a graph showing an adjustment diagram of the B & T and Cancer B &T; Figure 6 is a graph illustrating the survival percentage of patients treated in forty-eight months; Figure 7 is a graph illustrating the survival of the full and partial responders compared to the minor and non-responders; Figure 8 is a graph illustrating the relationship of the pathology index for survival; Figure 9 is a graph showing the ratio of lymphocyte infiltration for survival; Figure 10 is a graph illustrating the i percent survival (dose response) of patients treated in forty-four months, where "x" is equal to approximately 100 lU / mL of IL-2; Figure 11 is a graph of the dendritic cell illustrating the effects of NCM treatment on activated mature dendritic cells; and Figure 12 is a graph illustrating the peptide specific DTH response of mice immunized with conjugate and adjuvants, where the response is indicated as swelling in mm for individual mice (dots) and for the average (bar), the adjuvant is list on the x-axis, na? ve indicates the non-immunized mice and all other mice were immunized with Ovalbumin conjugates of PSMA peptide except where indicated (KLH). DETAILED DESCRIPTION OF THE INVENTION In general, the present invention provides methods for treating patients using immunotherapy with vaccine wherein the patients are immunosuppressed or immunosuppressed. For immunosuppressed, it is proposed that the patient has reduced cellular immunity and thus the impaired ability to respond to new antigens. More specifically, in the blood, T lymphocyte counts are reduced and / or the function of these cells is impaired, as is shown, for example, by the PHA proliferation assay. By "adjuvant" a composition is proposed with the ability to increase the immune response to a particular antigen. To be effective, an adjuvant must be delivered at or near the antigen site. Such ability is manifested by a significant increase in immune mediated protection. The increase in immunity is typically manifested by a significant (usually greater than 10 fold) increase in the titer of the antibody raised to the antigen. The increase in cellular immunity can be measured by a positive skin test, a cytotoxic T cell analysis, ELISPOT analysis for dlFN or IL-2, or infiltration of the T cell into the tumor (as described below). For "antigen associated with tumor", an analogous protein or peptide (which was previously shown to work by pulsing the ex vivo dendritic cell) or another equivalent antigen is proposed. This may include, but is not limited to PSMA peptides, MAGE peptides (Sahin U, et al., Curr Opin Immunol 9: 709-715, 1997; Wang RF, et al., Immunology Reviews 170: 85-100, 1999), Papilloma virus peptides (E6 and E7), MAGE fragments, NY ESO-1 and other similar antigens. Previously, these antigens were not considered to be effective in treating patients based either on their size, that is, they are very small or that they were previously thought not to have immunogenic properties (ie, auto antigens). By "NCM", it is proposed as a mixture of natural cytokine, as defined and set forth in U.S. Patent No. 5,632,983 and 5,698,194. The NCM may include recombinant cytokines. Briefly, the NCM is prepared in the continuous presence of a 4-aminoquinolone antibiotic and with the continuous or pulsed presence of a mitogen, which in the preferred embodiment is PHA. T lymphopenia (low T cell levels in the blood) is a diagnostic feature of cellular immune deficiency, while the impaired function of existing lymphocytes is the other characteristic. There is no generally accepted (clinically approved) way to treat T lymphopenia. Bone marrow transplants (tympanium transplantations + -) have been used in cases of severe combined immunodeficiency (SCID-congenital), irradiation or induced chemotherapy). The recombinant IL2 has been treated in AIDS with some effect due to a lot of toxicity. There are two ways to make new T cells to try to correct T lymphocytopenia. One way, as in rIL-2 therapy, expands T cells in the periphery, that is, memory T cells (CD45RO) (blood, node of lymphoma and vessel). The other involves the processing in the thymus of the new T cells of the precursors derived from the bone marrow, this happens naturally in children but not in adults. These new cells are called "thymic migrans" recently and have the surface marker of the "na? Ve" T cells, that is, CD45RA. NCM therapy (Tymosin or the plus) results in the production of these cells. New T as well as expands pre-existing memory T cells. More specifically, the present invention relates to immunization to provide an immune response to antigens, which is administered either endogenously or exogenously. Such antigens in the past may have been believed to be immunogenic while others used in the present invention may have been previously thought to be non-immunogenic. Any antigen can be used with the present invention. Examples of such antigens are EADPTGHSY (melanoma) of the MAGE-1 protein, EVDPIGHLY (lung carcinoma) of MAGE-3, EVDPIGHLY (lung carcinoma) of MAGE-3, and many others.

(See Bellone et al., Immunology Today, Vol. 20, No. 10, p 457-462 (1999)). The present invention is directed towards the effect of antigen processing in general; therefore, any antigen can be used with the present invention. The present invention can be extended to all forms of tumor antigens and haptens that include peptides and / or carbohydrates. The present invention can be extended to areas of applicability as in the AIDS virus vaccine in HIV + patients; another difficulty for driving situations; kidney transplants, aged individuals and the like. The present invention utilizes several general derivative method steps to obtain immunization in subjects where such immunization was previously believed to be impossible. More specifically, the present invention provides a method for overcoming immune depression by inducing the production of T cells, na? Ve. The term "na? Ve" T cells is proposed for recently produced T cells, even in adults, where these T cells have not yet been exposed to the antigen. Such T cells at this non-specific stage are not yet capable of becoming specific in presentation by a mature dendritic cell having antigen, such as tumor peptide, exposed therein. Thus, the present invention fills or generates new T cells. In general this is achieved by mixing a mixture of natural cytokine (NCM). The NCM includes, but is not limited to, IL1, IL2, IL6, IL8, IL10, IL12, IFNd, TNFa, G-, and GM-CSF, recombinants thereof, and combinations thereof. The quantity and proportions of these constituents are detailed below. Preferably, approximately 150-600 units of IL2 are contained in the NCM. In any modality, it is preferred that the NCM be injected around the lymphatics draining into the regional lymphoma nodes to an injury, such as a tumor and other persistent lesions that are treated. Peritumoral injection has been associated with the small response, a progression and thus contraindicated. A (10) day injection schedule is optimal and a (20) day injection protocol, while clinically effective, tends to reduce the TH1 response and shifts towards a less desirable TH2 response as measured by the lymphoid infiltration in the Cancer. Bilateral injections are also effective. Where radical neck dissection has occurred, contralateral injection is effective.

It is preferable to block the endogenous suppression of T cells, as is caused by several cancer lesions. Blocking is effected by the co-delivery of low dose cyclophosphamide (CY) and a non-steroidal anti-inflammatory drug (NSAID). The NSAID of choice is indomethacin (INDO). While INDO is the most effective in the NSAID, it is also arguably the most toxic. NSAIDs, Cox II, Vioxx® and Celebrex® are also less effective. Ibuprofen was effective, but histological responses were characteristic of a TH2 mediated response before THL, this is less desirable. The side effects of NSAIDs will be treated aggressively with proton inhibitors and an analogous E of prostaglandin. Zinc and multi-vitamins are useful agents to help the restoration of T-cell immunity. Treatment with anti-suppression and zinc without NSM is effective. In summary, the minimum regimen is the perilymphatic treatment with the NCM combined with the counter-suppression using CY and an NSAID. The alternative regimen is the previously mentioned regimen also includes zinc and vitamins, possibly including the addition of selenium. The preferable dosage of Zinc is 50 to 75 mg. A standard multivitamin can be administered. Zinc can be an available gluconate.

In order to maximize the clinical response and for the greatest increase in the survival rate, the degree and type of lymphocyte filtration is important. Lymphocyte: infiltration of granulocyte or macrophage of a 90:10 ratio is optimal. The infiltration of T and / or B cells is preferably diffuse and intense and not peripheral. Light infiltration of less than 20% is not associated with a robust clinical response. The reduction and fragmentation of the tumor in the histological samples is preferred in the reflection of a good response. The key to changes in the lymphoma node for good response involves at least five (5) aspects. Enlargement of the lymphoma node is preferred and not only the reversal of the induced reduction in tumor size but above all it increases in size compared to normal. The areas of cell T and B indicate an immunization. Sinus histocytosis (SH) is thought to be the accumulation of mature dendritic cells, which have tumor antigens ingested and processed but are not able to mature and present these tumor peptides to naïve T cells capable of stimulating THl and TH2 cells which lead to the T cell and B cells of cytotoxin. The inversion of SH is preferred. Thus, the present invention is provided for the immunization of unbind at a regional lymphoma node by promoting the differentiation and maturation of immature dendritic cells at a regional lymph node and thus allowing presentation by mature dendritic cells resulting from peptides. small, generally nine amino acids in length to T cells to gain immunization of T cells. Additionally, induction of mature dendritic cells is required. Finally, the mobilization of peripheral blood T lymphocytes in T lymphocytopenic patients is desired in the presence of induction of na? Ve T cells capable of responding to dendritic cells displaying endogenous tumor peptides. (See, Sprent, et al. Science, Vol 293, July 13, 2001, pgs 254-248) '. In view of the foregoing, the key mechanistic characteristics of the present invention are the in vivo maturation of dendritic cells that result in the effective presentation of peptide antigen. Based on the examples presented below, increases in non-compromised T cells na? Ve positive CD45 RA have been found. With the antigen, this leads to the clonal expansion of the T and B cells, creating immunity in the patient. The resulting infiltration in tumors by hematogenous spread leads to the robust destruction of the tumor. The result as it is found in the data right away, increases to survival due to immunological memory. (See, Sprent et al., Cited above). It is logically predicted that exogenously provided extracted or synthetic tumor peptides provided (see, Bellone et al. Cited above) can be delivered at the distal or co-primed regional or distal lymphoma node and provide tumor-specific antigen T cells, with or without B cells. Three examples are shown below. In view of the above, it can be concluded that the action of the NCM plus the other agents is useful for any of the tumor antigens (proteins and peptides, synthetic and endogenous). Many of these peptides are not normally immunogenic and only when presented by an activated, mature dendritic cell will they be effective in the immunization of natural naïve T cells. Thus, the appearance of an immune T cell means, de facto, that a dendritic cell has been made or allowed to act appropriately. Also de facto, the activation and maturation of the dendritic cell is going to be considered a key factor in cancer immunodeficiency as well as well-known defects in T cells such as decreased number and function with anergy and putative apoptosis. With reference more specifically to the protocol and medicament supplied in accordance with the present invention, the invention uses the NCM to immunize patients, such as patients with cancer as well as patients with other lesions or disease conditions that produce antigens. More specifically, the present invention utilizes a method for increasing the immune response of cancer patients to a cancer by administering an effective amount of a composition containing therein the NCM and a tumor associated antigen, wherein the NCM acts as a adjuvant to produce the immune response. The antigen associated with tumor may be either an endogenously processed tumor peptide preparation resident in the regional nodes of cancer patients or in conjunction with a tumor antigen preparation administered exogenously at or near these nodes. Tumor peptides as well as antigens are included herein even though the peptides are not expected to be immunogenic where protein antigens associated with tumors would be more likely since they are thus complete. In the preferred embodiment, the composition of the present invention involves administration of the NCM plus a tumor-associated specific antigen, as defined below with low doses of CY, a cyclooxygenase inhibitor, and other similar compounds that have been shown to increase additionally the effects of the composition of the present invention. According to the present invention, an NCM that has been previously shown to be effective and promote T cell development and function in aged immunosuppressed mice is provided. In the administration of this NCM to immunosuppressed patients with cancer in the head and neck, it was shown in this application for the first time that a mobilization of the T lymphocytes in the blood of cancer patients treated with the NCM produces an increase in the cells T na? Ve, immature supporting both CD2 and CD45 R ?. This is one of the first demonstrations that human adults can generate natural T cells. Previous references: Mackall et al. (New England Journal of Medicine (1995), Vol. 332, pp.143-149) and a review by Mackall (Stem Cells 2000, Vol. 18. pp. 10.18) discuss the inability to generate New T cells in adults but not in children, and discusses the problem of trying to fill T cells followed by cancer chemotherapy and / or radiotherapy. In general, there is a dogma that new T cells do not regenerate in the adult human. However, after transplantation of bone marrow for intensive chemotherapy, it has been shown that new T cells can be generated in the adult. Non-molecular therapy to date has been able to achieve this, although lymphocyte counts have been achieved with prolonged and intense therapy with recombinant interleukin-2 in patients infected with HIV. These have not been clearly demonstrated to be thymus-derived T cells and are presumably an expansion of pre-existing peripheral T cells. Previously, Cortesina and colleagues used a natural IL-2, perilymphatically in patients with cancer in the head and neck and induced severe tumor regressions (Cortesina G, et al., Cancer 62: 2482-2485,1988) with some infiltration of the tumor with leukocytes (Valente G, et al, MODEM Pathol 3 (6): 702-708, 1990). The non-treatable recurrences occurred and the response was classified as non-specific and without memory and thus not immunological (Cortesina G, and collaborators Br J Cáncer 69: 572-577, 1994). Repeated attempts to confirm the initial observations with the IL-2 recoiabinant were substantially unsuccessful (Hadden JW, In 'I J Immunopharmacol 11/12: 629-644, 1997). The method of the present invention involves using a NCM with local perilymphatic injections or other injections that are known to those skilled in the art to provide sufficient localization of the immunotherapy compounds. In the preferred embodiment, the injections take place in the neck but can be applied in other locations as required by the disease to be treated. These clinical regressions induced treatment in a high percentage of patients who also showed improved recurrence-free survival (Hadden JW et al, Arch Otoaringol Head Neck Surg. 120: 395-403, 1994; Meneses A et al., Arch Pathol Lab. Med. 122: 447-454, 1998); Barrera J et al., Arch Otolaryngol Head Neck Surg 126: 345-351, 2000; Whiteside et al., Cancer Res. 53: 564-5662, 1993). Whiteside et al. (Cancer Res. 53: 5654-5662, 1993) observed that in head and neck cancer, the tumor injection of recombinant interleukin-2 produced a T-cell lymphocyte infiltrate, but without significant clinical responses. Peritumor injection of Multikine (Celsci Website) (in combination with perilymphatic injection in up to 150 patients resulted in significant tumor responses, ie greater than 50% tumor reduction in only 11 patients, making its response ratio less than 10% in contrast to the degree In addition, they observed 50% of non-responders where the applicants have only observed 20% .The peritumoral and intratumoral injection can be associated with the progression of the disease even in patients who initially had had a positive response to the NCM protocol, thus nullifying its benefit.Patitutional injection is thus contraindicated and excluded as part of the present invention.This has led to the interpretation that the tumor is not the immunization site and the present application presents the documentation that the regional lymphoma node is the immunization site, then the analysis of the data revealed the regional lymphoma nodes, which indicated that the regional lymphoma node is the immunization site for postulated tumor antigens (Figure 14 and 18). With the identification of a number of different tumor antigens, this has been an intricate question over the last decade that gave the presence of such antigens, they have not been effectively employed in immunization protocols. Sporadic positive examples have been reported, but for the most part, the data are negative. The problem of antigen presentation has been focused in the last decade and the dendritic cell has emerged as a critical player in the presentation of tumor-derived peptides. See DeLaugh and Lotts, Current Opinion Immunology, 2000, Vol. 12, pp. 583-588; Banchereau and colboradores, Annual Reviews of Immulology, (2000), Vol. 18, pp. 767-811; also Albert et al., Nature, Vol. 392, pp. 86-89 (1998). In summary, in order for tumor antigens to be appropriately antigenic, they must come from an apoptotic rather than from a necrotic tumor cell (Albert, Nature, 39 2: 86-87, 1997). They need to be captured by mature dendritic cells that have the morphology of large histocytes. These immature dendritic cells process the antigen (endosymes, phagocytosis and digestion) and develop into mature dendritic cells, which show the fragments of the peptide (in general nine amino acids) of the antigen digested in the MHC cleft for presentation to the T cells. The T cells in order to respond must have the antigen presented by them in the MHC cleft plus the various co-stimulatory signs. Researchers, such as Murphy et al., 1999, have used dendritic cells generated in culture and then pulsed with tumor antigens and have achieved a small degree of success in immunizing patients against prostate-specific membrane antigen peptides. Unfortunately, this procedure for pulsing dendritic cells is difficult to manage and has been rather inefficient. In the present invention, the Applicants have shown that the cells present in the sinuses of the lymphoma node, which accumulate in cancer, are cells of the dendritic cell class and that they follow the in vivo treatment with the NCM protocol of the present invention. invention, these cells disappear and the antigen eventually becomes immunogenic to the T cells. They are able to respond to the tumor. Thus, one aspect of this invention is capable of generating a micro-environment in the regional lymphoma node, which allows the processing and presentation of the effective antigen. Immunization, which derives the results in the T cells are able to circulate to the lesion and destroy tumors, is de facto demonstration of the processing of the appropriate antigen by dendritic cells. Additionally, none of the patients treated with the NCM developed distant metastasis, which is expected in up to 15% clinically and up to 50% pathologically. This indicates that a systematic immunity rather than just a local immunity has been induced by the treatment. This is a drastic improvement over the compositions in the prior art, because the prior art compositions at best were inconsistently effective against metastatic disease. The ability of the composition of the present invention to create systematic immunity allows more effective and efficient treatment of a patient. In addition, the magnitude of the systematic response enables an individual to be administered smaller doses without limiting the effectiveness of the treatment and without toxicity. Literature (Hadden JW, Int'I J Immunopharmacol 11/12: 629-644, 1997; Hadden JW. Immunology and immunotherapy of breast cancer: An update: Int'I J Immunopharmacol 21: 79-101, 1999) has indicated that for both SSC and adenocarcinomas, the two largest types of cancer, the regional lymphoma nodes reflect tumor-related abnormalities, including sinus histocytosis, lymphoid depletion and frequently the presence of lymphocytes associated with anergic tumors (able to react to tumor cells with expansion and recovery ex vivo using IL-2). Then, with the metastases, lymphoid exhaustion and depressive function occur. Additionally, cervical lymphoma nodes not implicated in such patients have shown a reduction in average size and an increase in sinus histocytosis associated with head and neck cancers. (See, Figures 1-4). Specifically related to the composition, the composition of the present invention involves the mixture of the natural cytokine plus either the antigen associated with the endogenous or exogenous tumor. Additionally, low dose of CY, cyclooxygenase inhibitors, zinc and other similar compounds have been shown to further increase the effects of the composition of the present invention. Immunization for treatment of patients with cellular immune deficiency associated with cancer, HIV infection, aging, renal transplants and other such deficiencies can be achieved with the composition of the present invention. The present invention has numerous modalities. In one embodiment, the present invention provides an immunotherapy method for treating cancer by administering an effective amount of an NCM that includes cytokines including, but not limited to, IL-1, IL-2, IL-6, IL-8. , IL-12, IFN-d, TNF-a GM-CSF, recombinants thereof and combinations thereof. The above method also includes administering from 75 to 500 units of the IL-2 equivalence, where administration preferably occurs bilaterally in the lymphatics draining into the lymphoma nodes. Alternatively, administration may occur unilaterally. The NCM is administered for at least one to ten days and has approximately twenty days. In a preferred embodiment, administration occurs bilaterally and for approximately 10 days. The NCM can be administered before surgery or radiation therapy. Alternatively, the NCM can be administered during tumor recurrence. In addition, an effective amount of CY can be administered to the NCM.

In addition, an effective amount of an NSAID can be administered where the NSAID can be, but is not limited to INDO, Ibuprofen, celecoxib (Celebrex®), rofecoxib (Vioxx®), CoxII inhibitors, combinations thereof, and any other NSAIDs known to those skilled in the art. In another embodiment of the present invention, there is provided an immunotherapy method for treating cancer by administering an effective amount of CY and an effective amount of INDO. Another embodiment of the present invention provides a method of synergistic anti-cancer treatment by administering an effective amount of a CY and an effective amount of an NSAID, wherein the NSAID can be, but is not limited to INDO, Ibuprofen, celecoxib (Celebrex ®), rofecoxib (Vioxx®), CoxII inhibitors, combinations thereof, and the like. Another embodiment of the present invention provides an immunotherapy method for treating cancer by administering an effective amount of CY in combination with an effective amount of INDO and an effective amount of IFN-d IL-2, IL-1, and TNF-α. An additional embodiment is directed to a method of immunotherapy for treating cancer by administering an effective amount of CY in combination with an effective amount of INDO and an effective amount of the recombinant IL-2, recombinant IFN-d, recombinant TFN-a, and recombinant IL-1. A synergistic anti-cancer treatment is also provided by the present invention, wherein the treatment includes the steps of administering an effective amount of CY and INDO in combination with an NCM. The NCM may include, but is not limited to, IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM-CSF, G-CSF, recombinant the same, combinations thereof, and any other similar cytokine known to those skilled in the art. A method of anti-metastatic treatment is another embodiment of the present invention wherein the method includes steps to promote the differentiation and maturation of immature dendritic cells in a lymphoma node, allowing presentation by mature dendritic cells resulting from an antigen. to T cells to gain immunization of the T cells to the antigen, and prevent the development of metastasis. An additional modality is provided for an anti-metastatic method that includes the steps to unblock immunization in a lymphoma node, and 'generate systematic immunity. This method also includes the stage to prevent the development of metastasis. Other embodiments of the present invention provide a method for using a natural cytokine mixture as a skin diagnostic test to predict the effect of treatment by administering a NCM intracutaneously and determining a response to the NCM within 24 hours, wherein a negative skin test indicates insensitivity to "the NCM and predicts the failure of patients to respond to surgery with or without radiotherapy." Another modality provides a method for pretreatment of dendritic cells (DC) by applying an effective amount of CY and INDO in combination with a NCM that includes cytokines such as, but not limited to, IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM-CSF, G -CSF, recombinants thereof, combinations thereof, and any other similar cytokine known to those skilled in the art The present invention provides a method for treating monocyte defects characterized by the histiocytosis of breast or a negative skin test of NCM by applying an effective amount of CY and INDO in combination with an NCM. The NCM includes but is not limited to IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof, combinations of them, and any other similar cytokine known to those skilled in the art. In addition, the present invention provides various methods for inducing an immune response to endogenous or exogenous tumor antigens by administering an effective amount of an NCM that includes several cytokines including, but not limited to, IL-1, IL-2, IL -6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof, combinations thereof. Another embodiment of the present invention provides administering the NCM described above and an effective amount of CY to induce an immune response to endogenous or exogenous tumor antigens. In still another embodiment of the present invention, the method for inducing an immune response to endogenous and exogenous tumor antigens occurs by administering an effective amount of an NCM; an effective amount of CY; and an effective amount of INDO, wherein the NCM includes cytokines such as, but are not limited to IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM -CSF, G-CSF, recombinants thereof, combinations thereof. The present invention also provides a composition for inducing an immune response to endogenous or exogenous tumor antigens. The composition includes an effective amount of NCM, wherein the NCM includes cytokines such as, but not limited to, IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF- a, GM-CSF, G-CSF, recombinants thereof, combinations thereof. In a further embodiment, the composition includes an effective amount of CY. In still another embodiment, the composition further includes an effective amount of INDO. For any of the above modalities, the following administration details and / or protocols for treatment are used. Administration and protocols for treatment: Supply of Gene Products / Synthetic Antigens: The compounds of the present invention (including NCM), and antigens and exogens are administered and dosed to achieve optimal immunization, taking into account the clinical condition of the patient, individual, place and method of administration, administration program, patient's age, sex, body weight. The "pharmaceutically effective amount" for purposes herein is determined by such considerations as are known in the art. The amount must be effective to achieve reduction, fragmentation and infiltration of the tumor, protection of survival or faster recovery, or improvement or elimination of symptoms. In the method of the present invention, the compounds of the present invention can be administered in various ways. It should be noted that they can be administered as the compound or as a pharmaceutically acceptable salt and can be administered alone or as an active ingredient in combination with pharmaceutically acceptable carriers, diluents, adjuvants and vehicles. The compounds can be administered intra- or subcutaneously, either peri or intralinfatically, intranodally or intrasplenically or intramuscularly, intreperitoneally and intratotaxically. Implants of the compounds can also be useful. The patient being treated is a warm-blooded animal and, in particular, mammals that include man. Pharmaceutically acceptable carriers, diluents, adjuvants and carriers as well as implant carriers generally refer to a non-toxic, inert solid or liquid fillers, diluents or encapsulating material that do not react with the active ingredients of the invention.

The doses may be single dose or multiple doses over a period of several days. When the compound of the present invention is administered, it is generally formulated in an injectable unit dosage form (solution, suspension, emulsion). Pharmaceutical formulations suitable for injection include sterile aqueous solutions or dispersions and sterile powders for reconstitution into sterile injectable solutions or dispersions. It may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Non-aqueous vehicles such as cottonseed oil, sesame oil, olive oil, soybean oil, corn oil, sunflower oil, or peanut oil and esters, such as isopropyl myristate, can also be used as solvent systems for composite compositions. Additionally, various additives that increase the stability, sterility and isotonicity of the compositions can be added, including antimicrobial preservatives, antioxidants, chelating agents and regulatory suspensions. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. In many cases, it is desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of the dosage form can be caused by the use of delaying adsorption agents, for example, aluminum monostearate and gelatin. According to the present invention, however, any vehicle, diluent or additive used would have to be compatible with the compounds. The peptides can be polymerized or conjugated to carriers such as human serum albumin as is well known in the art. Sterile injectable solutions can be prepared by incorporating the compounds used in the practice of the present invention in the required amount of the appropriate solvent with several of the ingredients, as desired. A pharmacological formulation of the present invention can be administered to the patient in an injectable formulation containing any compatible carrier, such as various carriers, additives and diluents; or the compounds used in the present invention can be administered parenterally to the patient in the form of subcutaneous slow release implants or targeted delivery systems such as monoclonal antibodies, vectored delivery, iontophoretic, polymer matrices, liposomes and microspheres. Examples of delivery system useful in the present invention include: U.S. Patent Nos. 5,225,182; 5,169,383; 5,167,616; 4,959,217; 4,925,678; 4,487,603; 4,486,194; 4,447,233; 4,447,224; 4,439,196; and 4,475,196. Many other implants such as delivery systems and modules are well known to those skilled in the art. The foregoing provides a protocol for using the NCM as an adjuvant to immunize patients against tumor antigens, either autologous or as defined proteins or peptides. The antigen preparations to be used: Bn cancer: 1) PSMA peptides (9) obtained commercially Prostate 2) Fragments MAGE 1 & 3 & MAGE & NY ESO-1 Melanoma, obtained from Ludwig Inst. Of Immunol. H & NSC 3) Papilloma Virus E6 & E7 obtained Cervical SCC commercially The commercial route of antigen administration is preferably the neck because it is accessible and it contains > 30% of the bodies of lymphoma nodes and systematic immunity can be contemplated for the result.

Low Dose CY: Low dose CY has been used to increase cell immunity and decrease suppression by lymphocytes in mice and cancer patients (BERD D., Progerss in Clin Biol. Res 288: 449-458, 1989; BERD D, et al. Cancer Research 47: 3317-3321, 1987) and this has been used in the effective immunotherapy of cancer patients (Weber J., Medscape Anthology 3: 2, 2000, Murphy GP, Tjoa BA, Simons SJ The Prostate, 38: 43-78, 1999, Hadden JW, and co-workers, Arch Otolaryngol Head Neck Surg 120: 395-403, 1994). Zinc: Zinc deficiency is associated with improved cellular immunity and treatment with zinc is immunorestorative in mice (Hadden JW, Int'l J Immunopharmacol 17: 696-701, 1995; Saha A., et al., Int'l Immunophamacol 17: 729-734, 1995). A cyclooxygenase (COXi) inhibitor such as INDO: Cancers produce prostaglandins and induce the production of prostaglandins in host macrophages (Eden JW, The immunopharmacology of head and neck cancer: An update, Int'l J Im unophamacol 11/12: 629-644, 1997). Since prostaglandins are well known to be immunosuppressive for T cells, inhibition of PG synthesis with cyclooxygenase inhibitors is appropriate. Purification of the Recombinant Protein: Marshak et al, "Strategies for Protein Purification and Characterization, A laboratory course manual." CSHL Press, 1996. Dosage and Frequency of Antigens: 1-1000 μg, preferably 10-500; soluble form (partially polymerized or conjugated to the carrier if necessary). Administration of the dosage occurs on Day 1, Day 12 and Day 21. The Pre-Rx occurs on Day 12, Day 21, and Day 31. The site of the injections is local (ie injections in the neck) . Expected responses: 1) Tumor reduction; 2) Pathological changes of the tumor (reduction, fragmentation, lymphoid infiltration); 3) Humoral immunity to the antigen (RAI or ELISA); and / or 4) Cellular Immunity to the antigen (in vitro lymphocyte proliferation of the intracutaneous skin test, of ELISPOT ASSAY). Oligopeptides such as PSMA fragments, MAGE, E6, E7 peptides would be poorly immunogenic even if they pulsed in the dendritic cells. Thus effective immunization would not be expected to occur. Even with effective immunization, regression of the tumor would be considered surprising by this method, particularly at a distance as with the serv and prostate. The regression of metastatic disease is always a surprising event with immunotherapy. The degree and frequency of clinical responses are a factor in the effectiveness and thus the novelty of this procedure. The skin diagnostic tests are otherwise to obtain more effective immunization. Patients can be pretreated with the NCM of the present invention to induce better responses (increase of NCM and PHA skin tests and lymphocyte counts and reversal of lymphoma node abnormalities). As a result, an adjuvant strategy has been created, where the combination immunorestauración and adjuvance occurs, causes the peptides and immunogenic proteins to take place, and obtain the degree of immune response to effect the regression of the tumor at a distance that is possible. Patients can be treated by the skin for one or more tumor peptides before consideration of the protocol, 100 pg of one or more tumor peptides can be administered perilymphatically in the neck with NCM using the NCM protocol as discussed below in on day 1 and 10 of the NCM series. The combination will be repeated on day 21. In addition to the response and histology of the tumor, the immune reaction to the peptides will be monitored by repeating the skin test or by other means known in the art. The above discussion provides a real basis for the use of the present invention. The methods used with a utility of the present invention can be shown by the non-limiting examples and accompanying figures. EXAMPLES MATERIALS AND METHODS: All the stages that relate to cell culture are performed under sterile conditions. The general methods of cell immunology not described herein are performed as described in the general references for cellular immunology techniques such as Mishell and Shiigi (Selected Methods in Cellular Immunology, 1981) and are well known to those skilled in the art. Preparation of the Natural Cytokine Mixture (NCM) The white cells of lymph curdled from human blood are collected from negative donors of hepatitis virus and HIV negative. In an alternative embodiment, the animals could be the source of cells for veterinary uses. Donor cells accumulate and are placed in ficoll hypaque gradients (Pharmacy) to produce neutrophil and erythrocyte free lymphocytes. Alternative methods could be used that would result in the same population of starting lymphocytes as are known in the art. The lymphocytes are washed and distributed in the medium X vi o-10 (Whittaker Bioproducts) to the surface activated cell culture flasks for selection of the cell subsets (MICROCELLECTOR ™ T-25 Cell Culture flasks) in the which are immobilized stimulants, that is, mitogens similar to PHA. In an experiment set, the medium X vivo-15 and X vivo-20 were used as indicated. The immobilization process for the stimulants as described by the manufacturer to immobilize several substances for extraction procedures, that is, to separate cells, in the flasks. Alternatively, the lymphocytes are exposed to stimulants for example PHA for 2-4 hours, then washed three times. The cells are incubated for 24-48 hours in vivo vivo-10 medium with 80 μg / ml of ciprofloxacin (Miles Lab) a 370 in a C02 / air incubator. Alternatively, the RPMl 1640 medium could be used (Webb et al. 1973). In general, HSA is used at 0.1 to 0.5% (weight by volume). After incubation the floats are emptied and collected. Human serum albumin (HSA) can be added to further stabilize the interleukins if the HSA-free medium is used for generations. The supernatants are stored at 4 ° C to -70 ° C. Stimulation of lymphocytes: The objective was to find a way to stimulate lymphocytes to produce high levels of interleukin-2 in the absence of serum and in some way that did not produce significant amounts of PHA in the float. To do this, the PHA was immobilized in surface activated cell culture flasks for selection of cell subsets (AIS MICROCELLECTOR ™ T-25 plates) as described in the manufacturer's instructions for removal of the "extraction cell". "or pulsed in the cells followed by washing (pulse techniques). The medium used in these experiments was X vivolo (Whittaker) and was approved for administration to humans by the U.S. Food and Drug Administration for activated cell lymphokine (LAK) protocols of interleukin-2. Serum-free medium capable of supporting the proliferation of human lymphocytes similar to minimal essential medium (MEM) or RPMI-1640 (Sigma) could also be used. Initial experiments indicated that the PHA (HA-16, Murex Diagnostics Ltd., Dartford, UK) could be mobilized by the technique described by the manufacturer and that under appropriate optimal cell number conditions of 7.5-15xl06 / ml, exposure time from 24 hours to 48 hours , and PHA concentration of 25 or 50 μg / ml a high product of IL-2 in serum free floating eTL could be obtained. Additional detailed information is disclosed in International Publication Number WO 03/035004 A2 of Hadden and International Publication Number WO 02/34119 A2 of Hadden, which are hereby incorporated by reference in their entirety. Preparation of the NCM For the examples described herein, accumulated human peripheral blood clotting lymphs obtained from INCAN Blood Bank were incubated with phytohemagglutinin (Murex, Dartford UK) and washed. The cells were incubated in the serum-free medium (x-vivo 10, BioWhitaker) for twenty-four hours. The lots were prepared from six blood donors and screened by the INCAN Blood Bank for hepatitis B and C viruses, HTLV 1 and 2 and HIV. After 24 hours of culture, the cultures were centrifuged, the float was filtered through 0.2-micron filters, and the natural cytokine mixture was placed in vials. The activity of the batches of the NCM of the present invention averaged 200 U / ml of IL-2 as determined by ELISA. The bottles of the batches were stored at -70 ° until use. The cytokines were analyzed using commercial ELISA kits (Quantikine ™, R & D Systems, Inc., Minneapolis, MN) (See, - Table 1). The biological activity of the NCM of the present invention was confirmed using a murine cytotoxic T cell line (CTLL-2), which was originally developed as an indicator of biological activity of IL-2. Table I: NCM cytokine contents for five batches Patients Forty-two patients with squamous cell carcinomas stage I-IV of the head and neck entered the experiment (Hadden et al., International Immunopharmacology 3; 1073-1081 (2003)). Twenty-seven patients were treated with one or more 10-day NCM courses of the present invention. Fifteen patients received a twenty-day NCM course of the present invention at two neck sites. Four of these patients received an NCM preparation of the present invention that had low levels of cytokines (l / 4x, where these patients were excluded from the statistical analysis). Ten other patients did not meet the criteria for inclusion of the study at the time of study entry but were treated with the NCM in a compassionate manner. The compassionate treatment occurred because of a negative skin test response to the NCM, the occurrence of metastatic disease or that is not a surgical candidate. The average age of all patients was 66.0 years (range 34-86) and the male: female ratio was 4: 1. The majority of patients had stage III squamous cell carcinomas & amp; amp;; V of the larynx or oral cavity. Stage I and II patients had tongue injuries. The response was estimated based on the criteria and standard oncology tumor. Survival was estimated using the Kaplan-Meier diagrams using a computer software statistical data package (GRAPHPAD ™, San Diego, C?) Where the complete response (CR), partial response (PR) of greater than 50% was recorded. of tumor reduction, and no response or less than 25% tumor reduction. Treatment Regime Each treatment cycle lasted twenty days. Each cycle was started with an intravenous infusion of cyclophosphamide (CY) at 300 mg / m2 and indomethacin (INDO) at 25 mg orally three times a day, and zinc (65 mg of elemental zinc as sulfate) once a day. Patients also underwent the skin test with NCM at 0.1 L administered intradermally to exclude the allergic response to the components and display delayed-type hypersensitivity (DTH) responses to the NCM on day 4. Patients were treated with NCM at 1.0 mL for ten days or 2.0 L for twenty days intramuscularly administered at the insertion of the elastocleidomastoid muscle 2.0 cm below the tip of the tip. In patients who have undergone surgery, the treatment was administered in the lateral contralateral of the neck, intact next to the surgery. All patients gave their consent to the signed report to participate in the study and the study was approved by INCAN's Institutional Review Board and Research Committee as well as the Mexican Ministry of Health. Patients with any fundamental immune disorder, severe systemic disease, or patients requiring immunosuppressive therapy were excluded from the experiment. Patients who were surgical candidates were taken for surgery on day 21; Post-operative radiation therapy was administered to patients who were at high risk of recurrence (for example, those with involved nodes and / or positive surgical margins at the discretion of the surgeon and the radiation oncologist's consultation. routine was performed on all pre and post-treatment biopsy specimens.To quantify the various histological components of the tumor, a representative biopsy section containing a tumor was selected under low dust and the amount of the tumor was expressed as one percent of the tumor. Total area The remaining stroma was evaluated by the percent of the area having lymphocytes Tumors were further evaluated for the percent of the area of the specimen that was solid and the percent fragmented with the interdispersed leukocyte as infiltration as described. II: Characteristics of the Study Patient in Head and Neck Cancer Patient Number s 42 Average age (average) 65.0 (34-91) Relationship H: M 32: 8 KPS Medium (average) 100 (70-100) Treatment regime 10 days 20 20 days 22 Compassionate use 7 Low dose 4 Primary tumor Larynx 15 Tongue 8 Lined 5 Sine 3 Tonsila 3 Mouth Floor 1 Troll Retromolar 2 Mucosa Buccal 2 Lip 1 External Ear 1 Unknown Primary 1 Diagnosis Stage I 1 II 6 III 12 IV 23 EXAMPLE 1 Local perilymphatic injections in the neck that have NCM plus. low-dose CY, INDO, and zinc have induced clinical regressions in a high percentage of patients with cancer in the head and neck of squamous cells (H &S) (Hadden J W. et al., Arch Otolaryngol Head Neck Surg. : 395-403, 1994; Meneses A, et al., Aech Pathol lab Med 122: 447-454, 1998; Barrera J, and collaborators Arch Otalaryngol Head Neck Surg 126: 354-351, 2000; Hadden, et al., 2003; Menesis, et al., 2003) with evidence of improved recurrence limit survival. In total, including the minor response of the contraction and reduction of the tumor (25% -50%) of the tumor in pathological specimens, over 90% responded and most had tumor reduction greater than 50%. These responses are speculated to be mediated by immune regression since both B and T lymphocytes were observed to infiltrate the rumors. Therapy was not associated with significant toxicity. The treatment of patients with lymphocytopenic cancer with the combination of NCM has resulted in the mobilization of the labeled lymphocyte; where they are analyzed, these patients showed increases in CD45RA positive T cells (ie, na? ve T cells (Table IV)). In addition, intratumoral or peritumoral injection of the NCM in patients with H & NSC resulted either in the reversal of tumor regression induced immunotherapy or in tumor progression. The tumor like this is not the immunization site. As a result, the analysis of the revealed data of the regional lymphoma nodes indicates that the regional lymphoma node is the immunization site for the postulated tumor antigens (Meneses, et al., 2003), (See, Figure 1- 5) . None of these patients treated with NCM developed metastasis expected in 15% clinically and up to 50% pathologically, which indicates the systematic immunity before only local immunity has been induced. Patients were pre-tested with a skin test at 0.1 ml of the NCM before treatment and more than 90% of those with a positive skin test (> 0.3 mm at 24 hours) had a robust clinical and pathological response. Patients with negative skin tests had weak or no response. Thus, the skin test selects good answers. The largest increases were observed in the T lymphocyte (CD2) 752- > 1020 in these T lymphocytopenic patients (T cell counts 752 vs. 1600 (normal)). Importantly, there was a corresponding increase in CD45RA positive T cells "nalve" (532-> 782). As mentioned previously, these increases in general were not thought to occur in adults particularly with pharmacological therapy similar to NCM. These cells are summarily resilient thymic migrants and could be considered a new major ability to respond to new antigens similar to the 'tumor antigens. The preexisting CD45RA positive cells did not respond to the tumor antigens and may be unable to do so due to the induced immune suppression of the tumor (energy). The literature (Hadden JW, Int'l J Immunopharmacol 11/12: 629-644, 1997, Hadden JW, Int'l J Immunophar acol 21: 79-101,1999) indicates that for both the SCC and the adenocarcinomas, the two For older types of cancer, regional lymphoma nodes reflect tumor-related abnormalities, including sinus histocytosis, lymphoid depletion, and often the presence of lymphocytes associated with tumors capable of reacting to tumor cells (with IL-2). With metastasis, lymphoid depletion and depressive function occur. An unpublished analysis of cervical lymphoma nodes not implicated 10 H & NSC and 10 controls showed reduction in average size and an increase in sinus histocytosis associated with the symbol H & NSC (Figures 1-4). Table III: Treatment of Lymphocytopenic Patients with H & NSC with Increments of NCM in T Cells Na? Ve in the Blood (#mm) CELL MARKER T NA? VE CELL MARKER T PAN PATIENT # PRE POST INCREASE PRE POST INCREASE 1 479 778 +299 704 1171 +467 2 938 130 +371 1364 1249 -115 3 98 139 +41 146 178 +32 4 341 438 +97 655 590 -65 5 5 56677 665522 +97 453 643 +190 6 658 1058 + 400 1118 1714 +569 7 642 1101 +459 822 1601 +779 MEDIUM 532 782 +250 752 1020 +269 After treatment with an NCM protocol cycle (Hadden JW, et al., Aech Otolaryngol Head Neck Surg. 120: 395-403 , 1994; Meneses A, et al., Arch Pathol Lab Med 122: 447-454, 1998; Barrera J, et al., Aech Otolayngol Hgeadn Neck Surg 126: 345-351, 2000). The cervical lymphoma nodes not involved showed the changes indicated in Figures 1-4. Compared to the regional lymphoma nodes of H & NSC patients not treated with NCM, these nodes showed an insignificant increase in size, area and density of T cells, and decreased the number of germinal centers and histocytosis and sinus congestion. The lymphoma nodes of the treated patients were all stimulated and larger than the control nodes with the area and density of the increased T cell. Not only were these nodes restored to normal but they showed evidence of T cell predominance, a known positive correlation with survival in H &S (NS Hadden JW Int'l J Immunophamacol 11/12: 629-644, 1997). Importantly, when lymphoma node changes related to the areas of B and T cells correlated with changes in their tumors that reflect infiltration of T and B cells, a high degree of correlation was obtained for the T cell ( p- < 0.01) and the cell B (< 0.01) and the presence of the complete lymphoid (p <0.001) (Figure 5). In turn, these changes were correlated with tumor reduction by clinical and pathological criteria. These findings indicate that tumor reductions correlate directly and positively with lymphoma node changes and that the tumor reaction reflects changes in the lymphoma node as the dependent variable. These findings, taken together with the knowledge about how the immune system works in general (Roitt 1, Brostoff J, Male D. Immunology, JB Lippincott Co, Phila, Pa., 1989), and after transfection of tumor with a gene of cytokine (Maass G, et al., Proc Nati Acad Sci USA, 1995, 92: 5540-5542), indicates that the NCM protocol immunizes these patients to still unidentified tumor antigens at the level of the lymphoma nodes. None have previously presented evidence for changes in the lymphoma node that reflects immunization with autologous tumor antigens. This confirms that the present invention can induce immunization with tumor antigens that are ineffective or ineffective in an effect to bring about the regression of distant metastases. EXAMPLE 2: Further analysis of the survival and pathological, clinical data from the aforementioned INCAN study offers further insights into the nature of the invention as it relates to the immunization of cancer patients to their own antigen tumor antigens and immune regression. resulting from their tumors. Figure 6 shows that treatment with the NCM protocol (IRX-2) is associated with increased survival in 48 months (p < OR.01). Figure 7 shows that clinical responses determine survival in those patients with complete (CR) and partial (PR) responses (> 50% tumor reduction) have better survival than those with minor responses (MR) (< 50% , but> 25% tumor reduction) or no response (NR) (< 25%) (p < 0.01). Figure 8 shows that patients with stronger pathological responses (index of 6-9) have better than those with weaker pathological responses (<6) (p <0.02). Figure 9 shows that lymphoid filtration in the tumor as an individual variable predicts survival (p <0.01). Finally, the Chi Square analyzes of the relationship of the chemical response to the pathological response shows a highly insignificant relationship (p <0.01) which indicates that the two relate in coordination to each other as well as to survival and thus provide a statistical triangulation of the data of clinical interrelation responses, immune regression parameters, and survival. Such relationships have never been shown for immunotherapy of a human cancer. EXAMPLE 3 Two patients were treated with head and neck lymphoma. The patients included were those with cancer in the neck and head who agreed to participate in the protocol. The following scheme was followed. Before treatment, patients were tested on the skin with 0.1 ml of NCM subcutaneously on the forearm, the region was marked, 24 hours after the test was read. The test was considered positive if the induction and erythema was equal to or greater than 3 mm. Case 1: The patient was a 23-year-old man who presented with a previous history of three months with the presence of a tumor in the left sub-axillary region, without any other symptoms. In the emergency room, he was found to have adenopathy of lymphoma of the left submaxillary triangle of approximately 6.5 cm in diameter of a hard consistency, partially fixed in the deep levels. The rest of the physical examination was normal. The incisional biopsy showed Hodgkin's lymphoma. The lesion was classified ECHA. A treatment of one NCM cycle was given, obtaining a minor response, such as adenopathy reduced in size by 1 cm in diameter. The biopsy report obtained after NCM treatment showed 60% of the lesion showed normal lymphositic infiltration, and the rest of the neoplasm (40%) showed necrosis. No viable tumor cells were found. After this, the patient received neck radiation treatment of 3600 rads. The patient is currently free of disease. Case 2: The patient was an 82-year-old man who presented with a history of two months with a tumor mass in the middle of the painful neck, as well as a loss of 10 kg of weight. In the physical examination, the patient presented with a tumor in the right palatine tonsil, which enlarges approximately 4x3 cm, with an ulcer in the center of the tonsil. In the neck, a node of right submaxillary lymphoma measured approximately 2x2 cm and a node mass of lymphoma in level II and III of approximately 5x5 cm. The rest of the exam was normal. The incisional biopsy of the tonsil and one of the nodes of lymphoma of the neck did not show mixed defined Hodgkin lymphoma of intermediate grade. The patient underwent two NCM cycles at the end of which a centimeter of reduction in the diameter of the adenopathy of the tonsil to the neck was observed. The post-NCM treatment of pathological report showed 20% live tumor, fragmented and necrotic 30% and normal lymphocyte infiltration 50%. The patient was given chemotherapy (CHOP) for 6 cycles and then external radiotherapy (RT) in a total dose of 4600 rads. He resorted to eight months after the RT with adeno egalia at the occipital level. The patient died three months later with evidence of the disease in the neck. EXAMPLE 4 Ten patients with untreated early stage cervical cancer, clinically stage IB1, IB2 and HA were treated with perilymphatic, local NCM injections (10 daily injections) followed by radical hysterectomy on day 21. One day before the start of NCM treatment of the present invention, patients received an individual IV dose of CY at 300 mg / m. The oral INDO or zinc ibuprofen were administered from days 1 to 21. The pathological and clinical response, toxicity and disease-free survival were evaluated. All patients completed NCM treatment and were evaluated for response and toxicity. The clinical response was observed in 50% of patients (partial response 3 (PR), minor response 2 (MR) (> 25% &50% reduction)). Seven patients underwent surgery. Pathologically, tumor reduction was associated with tumor fragmentation that was found in five cases. Rather, there was a heterogeneous pattern of cell types that infiltrate the tumor, which included lymphocytes, plasma cells, neutrophils, macrophages, and eosinophils. Treatment was tolerated well except for mild pain and minor bleeding during injection and gastric intolerance to INDO. In 24 months of four complementary treatment, nine patients were disease free. This previously unpublished study shows that peritumoral NCM induces the immune mediated tumor response in untreated early stage cervical carcinoma. EXAMPLE 5 Two patients with liver metastases from primary hepatocellular carcinoma were treated with intraspinal NCM (1 or 3 injections). The protocol was otherwise as previously described by the cases of H &S, NSCC, cervical or lymphoma. A patient with advanced hepatocellular carcinoma had a partial response confirmed by tomography, no histology is available. The other tube a partial response confirmed by surgery. The histological examination showed the reduction, fragmentation of the tumor and infiltration of the lymphoid. EXAMPLE 6 Four patients with penile squamous cell carcinoma (associated human papilloma virus) were treated with the NCM protocol as described above; all four clinically had partial responses and the surgical specimen showed reduction and fragmentation of the tumor and characteristic of lymphoid infiltration among cancer patients H & NSC. EXAMPLE 7 Dose and Frequency of Natural Cytokine Mixture: A ten day injection protocol was compared to a twenty day injection protocol. Bilateral injections were compared to unilateral injections and a series of doses was compared. The significant activity in survival was observed from 74-1310 units of IL-2 eguivalence (as measured by R &D ELISA Systems) with a maximum of 100-233 units (Figure 9). Bilateral injections were effective and in a recurrent patient who had undergone dissection of the ipsilateral lymphoma node, cohtralateral injection was used and a good response was obtained. This observation means that the tumor antigen can also reside in contralateral nodes and thus bilateral injections are favored. The twenty-day injection protocol was effective in terms of response and clinical survival. Still, the surgical specimen showed less lymphoid infiltration (17% area) than the ten-day bilateral protocol (34% area) or the bilateral protocol of ten. days (33% area) (p < .05). Infiltration of the minor lymphoid with equivalent tumor reduction responses means that antibody-mediated immune responses were implicated, since these responses are considered less effective than lymphocyte-mediated responses (ie, cytotoxic T cells) and the protocol of Twenty-day injection involves more work and expense, a bilateral ten-day protocol with 100 units of eguivalence or IL-2 site is considered optimal. Figure 10 illustrates a series of doses of the NCM of the present invention at full survival in twenty-four months. An optimal impact on survival in approximately 100-233 international units of the IL-2 equivalence and no effect of approximately 16 units and less effect of approximately 1310 units. Function of Non-steroidal Anti-inflammatory Drug (NSAIDs): INDO is the most potent of the NSAIDs that act on both cyclooxygenase I & II but has greater gastrointestinal toxicity. Newer CoXII inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®) were thought to have less gastrointestinal toxicity. The use of these two agents in place of INDO in a series of patients gave lower responses as measured by the pathological and clinical criteria and by survival. In the case of Vioxx®All seven patients had clinical signs of gastritis after one week of therapy. In cervical cancer patients, ibuprofen was used as the NSAID and good responses were obtained. Based on these observations, INDO is preferred, but Celebrex or ibuprofen can be substituted if INDO is not tolerated. Vioxx® is not recommended. Prilosec or other proton pump inhibitors with or without a prostaglandin analogue are recommended as prophylaxis for gastritis, while H2 blockers of histamine are not considered indicative. Function of the NSAID in conjunction with the CY: In four patients a dose of NCM was given that was considered inactive (See, Figure 10, 15 column units) in conjunction with the INDO and the CY. No survivor was observed, yet two patients had minor response (<50%, if not > 25% narrowing of the tumor) and all four showed moderate pathological changes in the tumor specimen with tumor reduction and fragmentation as well as lymphoid infiltration (See, Table 4). INDO can increase lymphoid infiltration and tumor shrinkage in some patients (See, Panje, 1981, and Hirsch, et al., 1983), but this has not been clinically accepted as a useful therapy in the H &N SCC. Similarly, CY at this dose is not considered clinically active in the H &N SCC. The activity of INDO and CY alone can be considered surprising in the magnitude and type of tumor response. INDO and CY are considered as a synergistic combination for employment with other forms of immunotherapy. Recently the low dose of recombinant IL-2 was reported for delay recurrence in the metastasis and increase of the mean survival time in patients with the H &N SCC (See, DeStefani, et al., 2002, and Valente, et al., 1990 ). In the search of the previous technique, no clinical responses were observed and minor tumor changes (lymphoid infiltration without tumor regression) were observed. However, the rIL-2 can act with the CY & INDO to induce additional clinical responses and improved survival. Other natural or recombinant cytokines corresponding to those present in the NCM individually or combination are also potentially active. For example, cytokines such as IL-1, IFNd, TNFa, IL-6, IL-8, GM-CSF, G-CSF, IL-12, and combinations thereof can be used naturally or recombinantly. Table IV: CY & INDO (± NCM) EXAMPLE 8 Function of the Intradermal Skin Test in Prognosis: The researchers previously suggested that patients with an intradermal skin test negative to the NCM could show poor clinical responses based on an individual patient. A series of negative patient tests of skin tests are now accumulated and found to show similar changes to the combination CY & INDO without the significant NCM seen in Example 7. Ten patients had negative skin tests (including four from Hospital Juarez) with an NCM of the present invention (ie, no response to the NCM) were treated with the NCM plus the CY and the INDO. These patients had poor clinical responses, smaller tumor fragmentation and reduction, and poor survival (20%) (See, Table V). These observations corroborated the conclusion made in Example 7 that the INDO and the CY have marked activity without the NCM. Importantly they confirm that the skin test is critical to predict the clinical and pathological and emphatic responses that are related to improved survival. In addition, a negative skin test predicts the failure of patients to respond to surgery with or without radiation therapy. The skin test of the NCM can be usefully employed to predict the therapeutic effect in H &N SCC. Previously, the skin test with dinitrochlorobenzene (DNCB) showed significance of prognosis in H &N SCC, but due to the difficult to handle procedure that requires sensitization, it is never used again. The NCM skin test offers a convenient twenty-four hour test. Interestingly, these patients were divided into two groups. In one group, in Table VI b, the responses were especially poor without any survivors. In the other group, Table VI a, these patients converted the NCM test into a positive one after treatment and showed clinical and pathological responses and survival similar to patients in protocol (See, Table VI B). One of these patients had a considerable inoperable tumor and was shown to convert the negative test to positive and allowed a second treatment to clinically reduce the tumor and by the pathological criteria and allows prolonged survival after surgery (> 7 years). This pretreatment of negative skin test patients with the NCM can increase the response rates. The NCM plus thymosin.? it can also be predicted to work (See, published US application No. 20030124136). The negative NCM skin test reflects a defect and treatment of monocytes with active monocyte cytokines in a natural or recoiabinant form would be predicted to be useful individually or in combination thereof. These include but are not limited to, GM-CSF, M-CSF, IFNd, IL-1, IL-6, IL-8, IL-12 and others. Table V: Skin Test Patients NCM Negative Subj. Patient Patient Solid Tumor Frag. Stroma Limf. of the Tumor Resp. State No. Init. %%%%% Absolute Skin Test NCM Negative Changed to Positive 13 ANA 48 15 33 16 36 42 PR Vivo > 24 Month 15 ICV 70 63 7 6 24 5 MR Vivo > 24 Month 22 JMM 50 10 40 10 40 30 PR Dead Without Disease 9 Month. 27 MVR 70 28 42 12 18 10 PR Lost for follow-up Medium 60 29 31 11 30 22 1 SD 12 24 16 4 10 17 1 Skin Test NCM Negative 29 JISM 80 10 10 NR Dead by Illness < 1 Year 30 AGM 48 32 10 10 NR Dead for Illness < 1 Year 35 NGS * 70 70 30 NR Died for Illness < 1 Year 36 GCS * 50 15 35 10 40 40 NR Death due to Illness < 1 Year 37 MJBV * 80 16 64 16 NR Killed by Illness < 1 Year 39 FHV * 70 28 42 25 NR Killed by Illness < 1 Year Medium 72 43 29 12 17 7 SD 12 28 25 15 16 * Extracted from an Experience of the JUÁREZ Hospital. Table VI: IgG serum to ALF peptide Dilution NCM 0VA-PSMA NCM KLH-PSMA-NCM OVA-PSMA-CpG 1/200 0.929 0.692 0.241 1/400 0.989 0.518 0.208 1/800 0.695 0.351 0.144 1/1600 0.309 0.191 0.120 Table VI B: Peptide IgG Serum LLH 1/200 0.950 0.720 0.277 1/400 1.013 0.502 0.200 1/800 0.607 0.327 0.157 1/1600 0.316 0.201 0.125 Table VI C: IgG to Ovalbumin Serum 1/500 0.920 0.629 1.050 1/1500 0.632 0.185 0.955 1/3000 0.457 0.146 0.813 1/6000 0.259 0.104 0.537 EXAMPLE 9 Other Predictors of Prognosis for Use of Present Invention: Historically there have been few predictors for the effect on H & N SCC, lymphocyte counts, IgE and IgA levels or nutrition were suggested and as mentioned, the DNCB skin tests have been used. For chemotherapy (5 FU &CISPLATINUM) clinical responses occur before surgery in most patients, still the time of average survival and complete survival are essentially not affected. Thus, the immune response manifests to relate the ability of surgery with or without radiotherapy to cure or prolong survival in patients with cancer H & NSCC. The data presented in the examples shows that the use of the invention retards the recurrence of metastases in those who have residual tumor after surgery and increases survival in a manner that is related to the magnitude of the clinical response and the intensity of the immune in the tumor as estimated by the quantification of tumor reduction, fragmentation and lymphoid infiltration. These observations point to important modifications of the invention for additional improved survival. In patients with severe immunodeficiency In patients with low, weak or absent lymphocyte counts of NCM skin tests, sinus histocytosis, poor pathological responses, retreatment and monitoring of immune responses would be indicated. In Patients with Minor or No Clinical Responses These patients have a high risk of recurrence of metastases and thus would logically benefit from surgical post-treatment with the NCM of the present invention. In the absence of currently available tests for specific reactivity to the tumor rejection response observed in patients, the complementary treatment test with the triad of tests described in US Pat. No. 6,482,389 would help determine the frequency of treatment with the NCM of the present invention. In Patients Who Recur: Significant responses were observed including two complete responses in patients who were portrayed with the NCM of the present invention. This is in contrast to previous results with natural and recombinant IL-2 who failed to respond to treatment. Thus, the present invention is useful for treating recurrence of metastases in patients. EXAMPLE 10 Use of the Invention with Other Treatments Similar to Radiotherapy or Chemotherapy: Patients with Cancers H & N stage IV SCCs have markedly reduced survival compared to patients with stage III disease (10-20% versus 30-50%) despite the radiotherapy 's vision. Radiotherapy is well known to reduce T cell counts in these patients over a prolonged period. Despite the negative impact of radiotherapy on the number and function of the T cell, the NCM-treated patients of the present invention who have the Stage IV disease as well as the patients with the Stage III disease. Thus, the therapeutic impact was relatively greater in Stage IV patients, which contradicts the current dogma that immunotherapy and cytokine therapy work best with minimal tumor. It also suggests that the invention increases the potency of the effect of radiotherapy. Similarly in four patients with SCC cancer, related to the penis, the NCM with the present invention was used and followed by chemotherapy with cisplatin 5FU and a second NCM cycle of the present invention. The reduction of the clinical tumor was observed with the immunotherapy and the examination of the tumor from the surgery showed persistence of the immune regression. Another patient with H & amp; amp;; N SCC treated with the NCM of the present invention followed by chemotherapy with 5FU and cisplatin showed the same result. These observations indicate that the NCM of the present invention can be used with chemotherapy. EXAMPLE 11 Additional Characterization of NCM Correction of Dendritic Cell Defect in Cancer: Lymphoma nodes of five, patients treated with NCM and five, control patients of H & N SCC untreated were isolated and cellular constituents were analyzed by flow cytometry using a panel of cell surface markers for dendritic cells (ie, CD83, CD86, and CD68). Ceno histiocytosis (SH +) is associated with an accumulation of CD68 +, CD83 +, but CD86-DC while those without perceptible SH have few CD83 + cells (Figure 11). The NCM treatment of the present invention results in a five-fold increase in the number of CD86 + cells (concomitant with CD68 + CD83 +) compared to untreated cancer controls, which indicate a conversion to an "activated" DC phenotype. . The controls are H & N SCC not treated compared to cancer patients treated with NCM. Sine histiocytosis is characterized by an accumulated intrasinusoidal myeloid dendritic cells of CD68 + CD83 + CD86 ~ (DC) and the effective use of the NCM of the present invention was associated with a 5-fold increase in CD86 + cells indicative of mature DC activated. Breast histiocytosis was confirmed to represent an "accumulation of partially mature DC that is carrying endogenous tumor peptides." Complete maturation and activation with the expression of the costimulatory receptor (B7.1 or CD86) reflects the use of the NCM of the present invention to correct this defect in maturation and allow the presentation of the effective antigen to the T cells. The NCM of the present invention inverts the histiocytosis of the breast and leads to the effective immunization of the "na? ve" T cells. that the histiocytosis of the breast is characterized by a DC defect of the myeloid origin and the negative NCM skin test predicts a defect in another myeloid cell, the monocyte suggests that these are linked observations (ie, there is a class defect myeloid which is crucial for the immune response of the host to cancer.) EXAMPLE 12 Administration of exogenous tumor antigens that are used by the mice of the i nvention: The procedure was to immunize mice with prostate-specific membrane antigens (PSMA) such as T Cell Peptides (ALF & LLH) (lOOμg) were conjugated to both the OVALBUMIN and the Hemocyanin of Lapa de Punta (KLH). Previous attempts with the isolated unconjugated peptides were not successful in the mice. The NCM (0.1 mL) was given as an individual immunization with both conjugated antigens preceded by the low dose CY (400 ug / mouse) followed by nine daily NCM injections (0.1 mL) without antigens, while the adjuvants CpG, Alu or Ribi-Corixa were a unique primary immunization with the OVA conjugate. Two booster immunizations (conjugate plus adjuvant) were given on day 21 and 28 in each group of mice. The DTH for the T cell peptides was measured 9 days later for the first boost and the serum was taken at slaughter on days 15-21. Figure 12 shows the DTH results on the skin test with the individual ALF and LLH peptides (10ug @) without the conjugate. The NCM induces negligible DTH responses after immunization with both conjugates and with Alum for the OVA conjugate. Alum, Ribi-Cosixa, and CpG showed negligible activity. Results of the Serum Antibody: The serum was diluted as indicated and added to the cavities of a microplate coated with either peptide (LAF or LLH) or ovalbumin. The results are expressed as the average OD in 405 for 5 groups of mice. The data are presented in Table VII. Mice immunized with the KLH + NCm conjugate were negative for ovalbumin antibodies, but positive for the peptides. Mice immunized with the OVA + NCM conjugates were positive for the antibodies for both the OVA and the peptides, while those immunized OVA + CpG conjugate were positive for OVA only. These results indicate that the NCM makes conjugated PMSA peptides effective in stimulating both TH and IgG responses specific for the peptides, while other adjuvants similar to alum, Ribi-Corixa and CpG were inactive or poorly active. Humans: Three patients with advanced prostate cancer received peptides LAF & amp; amp;; Non-conjugated LLH (100 μg with NCM (lml equivalent of IL-2 100 units) preceded by low dose CY (300 mg / m2 and INDO daily (tid 25 mg plus 9 additional NCM injections (1 ml). On day 15, an NCM plus peptide booster was given.A patient (# 4) received OVA conjugated peptides in this regimen.The delayed hypersensitivity reactions (DTH) were measured with NCM (0.1 ml), ALF, LLH (10 μg) by reading the intradermal skin test in 24 hours in centimeters of erythema and in duration the results are presented in Table VII Table VII: DTH to peptides PSMA &NCM Time 0 1 month NCM 1) 0 0.5 2) 1.0 1.0 3) 0.5 1.0 4) 0.3 0.3 Peptide ALF 1) 0 0.5 2) 0 0.1 3) 1.0 1.0 4) 0 0.4 Peptide LHH 1) 0 0.5 2) 0 0.3 3) 1.5 2.0 4) 0 0.5 These data indicate that the CM is effective in inducing DTH reactions for unconjugated and conjugated PMSA peptides in humans with advanced prostate cancer. This result is different from the results of more previous attempts that have failed with the isolated peptides. For all this application, several publications, including American patents, are referred by author and year and patents by number. Full citations for publications are listed below. The descriptions of these publications and patents in their entireties are hereby incorporated by reference in this application in order to more fully describe the state of the art to which this invention pertains. The invention has been described in an illustrative manner, and it is to be understood that the terminology, which has been used is proposed to be in the nature of the words of the description rather than limitation. Obviously, many modifications and variations of the present invention are possible in view of the above teachings. Therefore, it is to be understood that within the scope of the invention described, the invention may be practiced otherwise than as specifically described. REFERENCES US Patent Nos. 4,116,951 4,353,821 4,390,623 4,439,196 4,447,224 4,447,233 4,464,355 4,466,918 4,470,926 4,475,196 4,486,194 4,487,603 4,612,365 4,910,296 4,925,678 4,959,217 5,100,664 5,167,616 5,169,383 5,225,182 5,503,841 5, 632.983 5 643.565 5,698,194 5,800,810 6,060,068 Publications Albert et al, Nature, Vol. 392, pp. 86-89 (1998). Banchereau et al., Annual Reviews of Immunology, Vol. 18, pp. 767-811 (2000). Barrier J, Verastegui E, Meneses A, de la Garza J, Zinser J & Hadden JW. Neoadjuvant immunological treatment with IRX-2 in patients with advanced oral cavity sguamous cell carcinoma of the head and neck induces clinical and histological responses. In First World Congress on Head and Neck Oncology. JJ Alvarez Vicent, Ed. Monduzzi, Bologna, Italy; 1998; pp 1017-1019 Barrera J, Verastegui E, Meneses A, Zinser J, de la Garza J, Hadden JW. Combination i munotherapy of squamous cell head and neck cancer: A phase 11 trial. Arch Otolaryngol Head Neck Surg 126: 345-351 (2000). Bellone, et al., Immunology Today, Vol 20, No. 10, p 457-462 (1998). Berd D, Mastrangelo M J. Effect of low dose cyclophosphamide on the immune system of cancer patients: reduction of suppressor function without depletion of the CD8 + subset. Cancer Research 47: 3317-3321 (1987). Berd D. Low doses of chemotherapy to inhibit suppressor T cells. Progress in Clin Biol Res 288: 449-458 (1989). Borysiewickz L K, Fiander A. Nimako M. A recombinant vaccine virus encoding human papillomavirus type 16 and 18, E6 and E7 proteins as immunotherapy for cervical cancer. Lancet 347: 1524-1527 (1996). Burke and Olson, "Preparation of Clone Librarles in Yeast Artificial-Chromosome Vectors" in Methods in Enzymology, Vol. 194, "Guide to Yeast 'Genetics and Molecular Biology", eds. C. Guthrie and G. Fink, Academic Press, Inc., Chap. 17, pp. 251-270 (1991).

Capecchi, "Altering the genome by homologous recombination" Science 244: 1288-1292 (1989). Cortesin G, DeStefani A, Galcazzi E. Temporary regression of recurrent squamous cell carcinoma of the head and neck is achieved with a low dose but not a high dose of recombinant interleukin 2 injected perilymphatically. Br J Cancer 69: 572-577 (1994). Cortesina G, DeStefani A, Giovarelli M, et al., Treatment of recurrent squamous cell carcinoma of the head and neck with low doses of interleukin-2 injected perilymphatically. Cancer 62: 2482-2485 (1988). Cortesina G, Destefani A & Galeazzi E. Temporary regression of recurrent squamous cell carcinoma of the head and neck is achieved with a low but not high dose of recombinant interleukin-2 injected perilymphatically. Br. J. Cancer; 1994; 69: 572-577. Cregg J M, Vedvick T S, Raschke W C: Recent Advances in the Expression of Foreign Genes in Pichia pastoris, Bio / Technology 11: 905-910 (1993). Cross DS, Platt JL, Juhn SK, et al., Administration of a prostaglandin synthesis inhibitor associated with an increased immune cell infill in squamous cell carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg 1992; 118: 526-8.

Culver, Site-Directed recbination for repair of mutations in the human ADA gene. (Abstract) Antisense DNA & PNA based therapeutics (1998). Davies et al., "Targeted alterations in yeast artificial chromosomes for inter-species gene transfer", ? ucleic Acids Research, Vol. 20,? o. 11, pp. 2693-2698 (1992). DeLaugh and Lotts, Current Opinion In Immunology, Vol. 12, pp. 583-588 (2000). DeStefani A, Forni G, Ragona R, and collaborators, I proved survival with perilymphatic interleukin 2 in patients with resectable squamous cell cancer of the inner cavity and oropharynx. Cancer 2002; 95: 90-97 Dickinson et al., "High frequency gene targeting using insertional vectors", Human Molecular Genetics, Vol. 2, ?or. 8, pp. 1299-1302 (1993). Duff and Lincoln, "Insertion of a pathogenic mutation into a yeast artificial chromosome containing the human APP gene and expression in ES cells", Research Advances in Alzheimer's Disease and Related Disorders (1995). Gilboa, E, Eglitis, M A, Kantoff, P W, Anderson, W F: Transfer and expression of cloned genes using retroviral vectors. BioTechniques 4 (6): 504-512 (1986). Gillis et al., (1978) Hadden JW, Endicott J, Baekey P, Skipper P, Hadden E M. Interleukins and contrasuppression induce immune regression of head and neck cancer. Arch Otolaryngol Head Neck Surg. 120: 395-403 (1994). Hadden JW, Saha R, Sosa M, Hadden E M. Immunotherapy with natural interleukins and / or Thymosin to potently augments T lymphocyte responses of hydrocortisone-treated aged mice. Int'l J Immunopharmacol 17: 821-828 (1995). Hadden JW. Immunology and immunotherapy of breast cancer: An update: vint'l J Im unopharmacol 21: 79-101 (1999). Hadden JW. The immunopharmacology of head and neck cancer: An update. Int'l J Immunopharmacol 11/12: 629-644 (1997). Hadden JW. The treatment of zinc deficiency is an immunotherapy. ' Int'l J Im unopharmacol 17: 696-701 (1995). Hadden, J., E. Verastegui, J. L. Barrera, M. Kur an, A.

Meneses, J. W. Zinser, J. de la Garza, and E. Hadden, "A trial of IRX-2 in patients with squamous cell carcinomas of the head and neck," International Immunopharmacology 3; - 1073-1081 (2003). Hank A J, Albertini M R, Sondel P M. Monoclonal antibodies, cytokines and fusion proteins in the treatment of malignant disease. Cancer Chemother & Bíol Resp Mod 18: 210-222 (1999). Hirsch B, Johnson JT, Rabin BD, et al. Immunostimulation of patients with head and neck cancer. Arch Otolaryngol 1983; 109: 298-301 Huston et al, "Protein engineering of single-chain Fv analogs and fusion proteins" in Methods in Enzymology (J J Langone, ed., Academic Press, New York, N. Y.) 203: 46-88 (1991). Huxley et al., "The human HPRT gene on a yeast artificial chromosome is functional when transferred to mouse cells by cell fusion", Genomics, 9: 742-750 < BR > < BR > < BR > (1991). Jakobovits et al., "Germ-line transmission and expression of a human-derived yeast artificial chromosome", Nature, Vol. 362, pp. 255-261 (1993). Johnson and Bird, 1991"Construction of single-chain Fvb derivatives of monoclonal antibodies and their productisn in Escherichia coli in Methods in Enzymology (JJ Langone, ed .; Academic Press,? Ew York,?., Y.) 203: 88-99 (1989) Kavanaugh DY, Carbone D P. Immunologic dysfunction in cancer, Hemato- Oncol Clinics of? Orth Amer 10 (4): 927-951 (1996), by collaborators, "Introduction and expression of the 400 kilobase amyloid precursor. protein gene in transgenic mice ", Nature Genetics, Vol 5, pp. 22-29 (1993) Maass G, Schmidt W, Berger M, et al., Priming of tumor-specific T-cells in the draining lymph nodes after immunization with interleukin 2-secreting tumor cells: three consecutive stages may be required for successful tumor vaccination Proc Nati Acad Sci USA, 92: 5540-5542 (1995) Mackall (Stem Cells 2000, Vol. 18. pp. 10-18) Mackall et al., (New England Journal of Medicine, Vol. 332, pp. 143-149 (1995) Maclean GD, Miles DW, Rubens RD, Reddish M A, Longenecker bone marrow. Enhancing the effect of Theratope STn-KLH cancer vaccine in patients with metastatic breast cancer by pretreatment with low-dose intravenous cyclophosphamide. J Immunother Emphasis Tumor I munol 19 (4): 309-316 (1996). Marshak et al, "Strategies for Protein Purification and Characterization, A laboratory course manual." CSHL Press (1996). Mastrangelo M J, Maguire H C Jr., Sato T, Nathan F E, Berd D. Active specific immunization in the treatment of patients with melanoma. (Review) Seminars in Oncology 23 (6): 773-781 (1996). Meneses A, Verastegui E, Barrera JL, Zinser J, de la Garza J, Hadden JW. Histological findings in patients with head and neck squamous cell carcinoma receiving perilympatic natural cytokine mixture prior to surgery. Arch Pathol Lab Med 122: 447-454 (1998). Meneses A, Verastegui E, Barrera JL, de la Garza J, and Hadden JW, "Lymph node histology in head and neck cancer: impact of immunotherapy with IRX-2," International Immunopharmacology, 3; 1083-1091 (2003). Mernaugh and Mernaugh, "An overview of phage-displayed recombinant antibodies" in Molecular Methods in Plant Pathology (R P Singh and U S Singh, eds., CRC Press Inc., Boca Ratón, Fia.) Pp. 359-365 (1995). Mishell and Shiigi (Selected Methods in Cellular Immunology (1981) . Montovani A, Sozzani S, Locati M, and collaborators, Macrophage polarization: tumor associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends in Immunol 2002; 23: 549-555 Murphy G P, Tjoa B A, Simiaons S J. The prostate. 38: 43-78 (1999). Panje WR. Regression of head and neck carcinoma with a prostaglandin-synthesis inhibitor. Arch Otolaryngol 1981; 107: 658-63 Pearson and Choi, Expression of the human b-amyloid precursor protein gene from a yeast artificial chromosome in transgenic mice. Proc. Nati Acad. Sci. USA, 90: 10578-82 (1993). Roitt I, Brostoff J, Male D. Immunology, JB Lippincott Cs, Phila, Pa., (1989). Rothstein, "Targeting, disruption, replacement, and allie rescue: integrative DNA transformation in yeast" in Methods in Enzymology, Vol. 194, "Guide to Yeast Genetics and Molecular Biology", eds. C. Guthrie and G. Fink, Academic Press, Inc., Chap. 19, pp. 281-301 (1991). Saha A, Hadden E M, Hadden J W. Zinc induces thymulin secretion from human thy ic epithelial cells in vitro and augments splenocytes and thymocyte response in vivo. Int'l J I munopharmacol 17: 729-734 (1995). Sahin U, Tureci 0, Pfreundschuh. Serological identification of human tumor antigens. Curr Opin Immunol 9: 709-715 (1997).

Sanda M G, Smith D C, Charles L G. Recombinant vaccinia-PSA (Prostvac) can include a prostate-specific immune response in androgen-odulated human prostate cancer. TJrology 52: 2 (1999) . Schedl et al., "A yeast artificial chromosome covering the tyrosinase gene confers copy number-dependent expression in transgenic mice", Nature, Vol. 362, pp. 258-261 (1993). Sprent, et al., Science, Vol. 293, 245-248 (2001).

Strauss et al., "Germ line transmission of a yeast artificial chromosome spanning the murine to (I) collagen locus", Science, Vol. 259, pp. 1904-1907 (1993). Tagawa M. Cytokine therapy for cancer. Current Pharmaceut Design 6 (6): 681-699 (2000). Valente G, DeStefani A, Jemma C, Giovarelli M, Geuna N, Cortesin G, Forni G, Palestro G. Infiltrating leukocyte populations and T-lymphocyte subsets in head and neck squamous cell carcinomas from patients receiving perilymphatic injections of recombinant interleukin-2. A pathologic and immunophenotypic study. Modern Pathol 3 (6): 702-708 (1990). Valente G, DeStefani A, Jemma C. Infiltrating leukocyte populations and T lymphocyte subsets in H &N SCC from patients receiving perilymphatic injections of rIL-2. Mod Pathol 1990; 3: 702-708 Van der Eynde B, Van der Bruggen, T cell defined tumor antigens. Curr Opin Immunol 9: 684-693 (1997). Verastegui E, Barrera J L, Zinzer J, R River, Meneses A, Garza J, Hadden J W. A natural cytokine mixture (IRX-2) and interference with immune suppression induces immune mobilization and regression of head and neck cancer. Int'l J Immunopharmacol 11/12: 619-627 (1997). Verastegui E, Hadden EM, Barrier J, Meneses A, Hadden JW. A natural cytokine mixture (IRX-2) and interference with immune suppression induces immune regression and improved survival in head and neck cancer. Int'l. Journal of Immunorehabilitation; 1999; 12: 5-11. Wang R F, Rosenberg S A. Human tumor antigens for cancer vaccine development. Immunologic Reviews 170: 85-100 (1999). Weber J. Tumor vaccines. Medscape Anthology 3: 2 (2000). Whiteside, et al., Cancer Res. 53: 5654-5662, (1993).

Wolf et al., Arch. Oto. Laryngol. 111: 716-725 (1985).

Claims (32)

1. CLAIMS 1. A method of immunotherapy to treat cancer, characterized by se. administering an effective amount of a natural cytokine mixture (NCM) including cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d TNF-a, GM-CSF, G-CSF, recombinants thereof and combinations thereof. The method according to claim 1, characterized in that the step of administering is defined as administering from 75 to 500 units of the equivalence of IL-2. 3. The method according to claim 1, characterized in that the administration step is defined as bilaterally administering the NCM in lymphatics draining at lymphoma nodes. 4. The method according to claim 1, characterized in that the administration step is defined as unilaterally administering the NCM. 5. The method according to claim 1, characterized in that the administration step is defined as administering the NCM for at least 1 to 10 days. 6. The method according to claim 5, characterized in that the administration step is further defined as administering the NCM up to about 20 days. The method according to claim 6, characterized in that the administration step is further defined as administering the NCM bilaterally and for approximately 10 days. The method according to claim 1, characterized in that the administration step is defined as administering the NCM before surgery or radiotherapy. The method according to claim 1, characterized in that the administration step is defined as administering the NCM during the recurrence of tumors. 10. The method of compliance with the claim 1, characterized in that it also includes the step of administering an effective amount of cyclophosphamide (CY). The method according to claim 1, characterized in that it further includes the step of administering an effective amount of a non-steroidal anti-inflammatory drug (NSAID) selected from the group consisting of indomethacin (INDO), ibuprofen, celecoxib (Celebrex®), rofecoxib (Vioxx®), CoxII inhibitors and combinations thereof. 12. A method of immunotherapy for treating cancer, characterized in that an effective amount of CY and an effective amount of INDO are administered. 13. A method of synergistic anti-cancer treatment, characterized in that an effective amount of CY and an effective amount of NSAID selected from the group consisting essentially of indomethacin (INDO), ibuprofen, celecoxib (Celebrex®), rofecoxib (Vioxx®) is administered. ), CoxII inhibitors and combinations thereof. 14. A method of immunotherapy for treating cancer, characterized in that an effective amount of CY is administered in combination with an effective amount of INDO and an effective amount of IFN-d, IL-2, IL-1 and TNFa. 15. A method of immunotherapy for treating cancer, characterized in that an effective amount of CY in combination with an effective amount of INDO and an effective amount of recombinant IL-2, recombinant IFN-d, recombinant TNF-a and recombinant IL-1. 16. A synergistic anti-cancer treatment, characterized in that it comprises the steps of administering an effective amount of CY and INDO in combination with a NCM that includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof, and combinations thereof. 17. A synergistic anti-cancer composition, characterized in that it comprises an effective amount of CY; an effective amount of INDO; and an effective amount of an NCM that includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM-CSF, G -CSF, recombinants thereof and combinations thereof. 18. An anti-metastatic treatment method, characterized in that it comprises the steps of promoting the differentiation and maturation of immature dendritic cells in a lymphoma node; allow the presentation by mature dendritic cells resulting from the antigen to the T cells to gain immunization of the T cells to the antigen; and prevent the development of metastasis. 19. An anti-metastatic method, characterized in that it comprises unblocking the immunization in a lymphoma node; and generate systematic immunity. 20. The anti-metastatic method according to claim 19, characterized in that it also includes the step to prevent the development of metastasis. 21. A method for using a mixture of natural cytokine as a diagnostic skin test to predict the effect of the treatment, characterized in that a NCM is administered intracutaneously and a response to the NCM is determined within 24 hours, wherein a test of negative skin indicates insensitivity to NCM and predicts the failure of patients to respond to surgery with or without radiation therapy. 22. A dendritic cell (DC) pretreatment method, characterized in that an effective amount of CY and INDO is applied in combination with a NCM that includes cytokines selected from the group consisting of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof and combinations thereof. 23. A method for treating monocyte defects that exhibit sinus histiocytosis or a negative NCM skin test, characterized in that an effective amount of CY and INDO is applied in combination with a NCM that includes cytokines selected from the group consisting essentially of IL -1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof and combinations thereof. 24. A method for inducing an immune response to tumor antigens, characterized in that an effective amount of an NCM is administered which includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof and combinations thereof. 25. The method of compliance with the claim 24, characterized in that the tumor antigens are selected from the group consisting essentially of endogenous and exogenous tumor antigens. 26. A method for inducing an immune response to tumor antigens, characterized in that an effective amount of an NCM is administered; and an effective amount of CY, wherein the NCM includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM- CSF, G-CSF, recombinants thereof and combinations thereof. 27. The method according to claim 26, characterized in that the tumor antigens are selected from the group consisting essentially of endogenous and exogenous tumor antigens. 28. A method for inducing an immune response to tumor antigens, characterized in that an effective amount of an NCM is administered; an effective amount of CY; and an effective amount of INDO, wherein the NCM includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL-8, IL-12, IFN-d, TNF-a, GM- CSF, G-CSF, recombinants thereof and combinations thereof. 29. The method according to claim 28, characterized in that the tumor antigens are selected from the group consisting essentially of endogenous and exogenous tumor antigens. 30. A composition for eliciting an immune response to endogenous or exogenous tumor antigens, characterized in that it comprises an effective amount of an NCM that includes cytokines selected from the group consisting essentially of IL-1, IL-2, IL-6, IL- 8, IL-12, IFN-d, TNF-α, GM-CSF, G-CSF, recombinants thereof and combinations thereof. 31. The composition according to claim 30, characterized in that the composition further comprises an effective amount of CY. 32. The composition according to claim 31, characterized in that the composition includes an effective amount of INDO.