MXPA00007148A - Method for treating inflammatory diseases using heat shock proteins - Google Patents

Abstract

This invention relates to a method to protect a mammal from a disease associated with an inflammatory response and in particular, from an inflammatory disease characterized by eosinophilia, airway hyperresponsiveness and/or a Th2-type immune response. The method includes administration of a heat shock protein to a mammal having such a disease. Formulations useful in the present method are also disclosed.

Description

METHOD FOR THE TREATMENT OF INFLAMMATORY DISEASES USING THERMAL SHOCK PROTEINS FIELD OF THE INVENTION The present invention relates to a method for protecting a mammal from inflammatory diseases, and particularly diseases characterized by eosinophilia associated with an inflammatory response.

BACKGROUND OF THE INVENTION Diseases comprising inflammation are characterized by the influx of certain cell types and mediators, in the presence of which they can lead to tissue damage and sometimes death. Diseases that comprise inflammation are particularly dangerous when they afflict the respiratory system, resulting in obstructed breathing, hypoxemia, hypercapnia and damage to lung tissue. Obstructive diseases of the airways are characterized by limited air flow (ie, obstruction or narrowing of air flow) due to airway smooth muscle constriction, edema and mucus hypersecretion leading to a job Increased breathing, dyspnea, hypoxemia and hypercapnia. While the mechanical properties of the lungs during obstructed breathing are shared between different types of obstructive airway disease, the pat of isiology may differ. A variety of inflammatory agents can cause limitation of airflow inclg allergens, cold air, exercise, infections and air pollution. In particular, allergens and other agents in allergic or sensitized animals (ie, antigens and haptens) cause the release of inflammatory mediators that attach cells comprised in inflammation. These cells include lymphocytes, eosinophils, mast cells, basophils, neutrophils, macrophages, monocytes, fibroblasts and platelets. Inflammation results in hypersensitivity. A variety of sts have linked the degree, severity and timing of the inflammatory process with the degree of hypersensitivity of the airways. In this way, a common consequence of inflammation is the limitation of airflow and / or hypersensitivity of the airways.

Asthma is a significant disease of the lung that affects about 16 million Americans. Asthma is typically characterized by periodic limitation of airflow and / or hypersensitivity to various stimuli which results in excessive narrowing of the airways. Other features may include airway inflammation and eosinophilia. More particularly, allergic asthma is frequently characterized by high levels of IgE, eosinophilic inflammation of the airways and hypersensitivity of the airways. The prevalence of asthma is increasing (that is, both incidence and duration). The current predominance reaches 10% of the population and has increased by 25% in the last 20 years. However, of more interest is the increase in the proportion of death. When coupled with illnesses in emergency room visits and hospitalizations, recent data suggest that the severity of asthma is increasing. While most cases asthma is easily controlled, for those with a more severe disease, the costs, the side effects and also too frequent, the inefficiency of the treatment, present serious problems. Fibroproliferative responses to chronic antigen exposure can explain both the severity of asthma and poor responses to therapy, especially if treatment is delayed. Most patients with asthma have very mild symptoms that are treated very easily, but a significant number of asthmatics have more severe symptoms. In addition, chronic asthma is associated with the development of the progressive and irreversible limitation of air flow due to some unknown mechanism. Currently, therapy for the treatment of inflammatory diseases such as moderate to severe asthma predominantly comprises the use of immunosuppressive glucocorticosteroids. Other anti-inflammatory agents that are used to treat inflammation of the airways include cromolyn and nedocromil. Symptomatic treatment with beta-agonists, anti-cholinergic agents and meth ixantins are clinically beneficial for the relief of discomfort, but fail to stop the underlying inflammatory processes that cause the disease. The frequently used systemic glucocorticosteroids have numerous side effects, including but not limited to, weight gain, diabetes, hypertension, osteoporosis, cataracts, atherosclerosis, increased susceptibility to infection, increased lipids and cholesterol, and simple contusion. Aerosolized icosteroid glucocort have less side effects but may be less potent and have significant side effects, such as milkweed. Other anti-inflammatory agents, such as cromolyn and nedocromil are much less potent and have less side effects than glucocorticosteroids. Anti-inflammatory agents that are used primarily as immunosuppressive agents of anti-cancer agents (ie, cytoxan, methotrexate and Immuran) have also been used to treat airway inflammation with mixed results. However, these agents have a serious potential for side effects, including but not limited to, increased susceptibility to infection, liver toxicity, drug-induced lung disease, and suppression of the bone marrow. In this way, these drugs have found limited clinical use for the treatment of most lung diseases of airways hypersensitivity. The use of symptomatic and anti-inflammatory relief reagents is a serious problem due to its side effects or its failure to attack the underlying or fundamental cause of an inflammatory response. There is a continuous requirement for less dangerous and more effective reagents for the treatment of inflammation. In this way, a need remains for processes that use reagents with lower side effect profiles and less toxicity than current anti-inflammatory therapies.

BRIEF DESCRIPTION OF THE INVENTION The present invention relates in general to a method for protecting a mammal from a disease associated with an inflammatory response, and in particular, from a disease characterized by eosinophilia, hypersensitivity of the airways? / or a Th2-type immune response, where the characteristic is associated with an inflammatory response. This method includes the step of administering to a mammal having this disease, a heat shock protein. In a preferred embodiment, this mammal is a human. One embodiment of the present invention relates to a method of protecting a mammal from a disease characterized by eosinophilia associated with an inflammatory response. The method includes the step of administering a heat shock protein to a mammal having this disease. Preferably, this method for treating a disease characterized by eosinophilia reduces eosinophilia in the mammal. In one embodiment, this method reduces the blood count of eosinophils in the mammal to be between about 0 and about 300 cells / mm 3, more preferably to be between about 0 and about 100 cells / mm 3. In another embodiment, this method reduces the blood counts of eosinophils in the mammal to be between about 0% and about 3% of total white blood cells in the mammal. Diseases for which a method of the present invention may be protective include, allergic airway diseases, hyper-eosinophilic syndrome, parasitic helminth infection, allergic rhinitis, allergic conjunctivitis, dermatitis, eczema, contact dermatitis, or allergy to the foods. In another embodiment, the disease is a respiratory disease characterized by eosinophilic inflammation of the airways and airway hypersensitivity, this disease including, but not limited to, allergic asthma, intrinsic asthma, allergic bronchopulmonary aspergillosis, eosinophilic pneumonia, bronchie t asis of allergic bronchitis, occupational asthma, reactive airway disease syndrome, interstitial lung disease, hyper-eosinophilic syndrome, or parasitic lung disease. In another embodiment, this disease is a disease that is associated with sensitivity to an allergen, and in a preferred embodiment, is allergic asthma. In one embodiment, a heat shock protein useful in a method of the present invention is selected from the group of a heat shock protein of the HSP-ßO family, a heat shock protein of the HSP-70 family, a protein of heat shock of the HSP-90 family, or a heat shock protein of the HSP-27 family. In alternative embodiments of the present method, the heat shock protein is selected from the group of a heat shock protein of the HSP-60 family, a heat shock protein of the HSP-70 family, or a heat shock protein of the HSP-70 family. the HSP-27 family; a heat shock protein of the HSP-90 family or a heat shock protein of the HSP-27 family; or from the group of a bacterial heat shock protein of a mammalian heat shock protein. In a preferred embodiment, the heat shock protein is a mycobacterial heat shock protein, and more preferably, a mycobacterial heat shock protein-65 (HSP-65). In some embodiments, a disease for which the present method is protective is characterized by hypersensitivity of the airways. In these embodiments, this method preferably decreases the methacholine sensitivity of the airways in the mammal. In other embodiments, the limitation of air flow in the mammal is reduced such that a mammalian FEVi / FVC value is at least about 80%. In another embodiment, the administration of a heat shock protein results in an improvement in a PC20metacoi? NaFEV? Value. of the mammal such that the value PC2ometacoi? naFEV? obtained before the administration of a heat shock protein when the mammal is motivated with a first concentration of methacholine is the same as the PC20metacoi? naFEV1 value obtained after the administration of the heat shock protein when the mammal is motivated with the double of the amount of the first methacholine concentration. In yet another modality, the administration of a heat shock protein improves the mammalian FEV_ to be between approximately 5% and approximately 100% of the predicted FEVi of the animal. In another embodiment, administration of a heat shock protein reduces the limitation of air flow in the mammal such that an RL value is reduced by at least about 20%. In one embodiment, a disease for which a method of the present invention is protective can be associated with the increased production of a cytosine selected from the group int erleucine-4 (IL-4), int er leucine-5.

(IL-5), interleukin-6 (IL-ß), int erleucin-9 (IL-9), interleukin-10 (IL-10), interleukin-13 (IL-13) or interleukin-15 (IL-15) ). Accordingly, it is an embodiment of the methods of the present invention that the administration of a heat shock protein induces the production of interferon-? (IFN-?) By T lymphocytes in the mammal. In another embodiment, the administration of a heat shock protein suppresses the production of int erleucine-4 (IL-4) and interleukin-5 (IL-5) by T lymphocytes in the mammal. According to the methods of the present invention, a heat shock protein can be administered in an amount between about 0.1 microgram x kilogram-1 and about 10 milligrams x kilogram-1 body weight of a mammal, and more preferably, in an amount between about 1 microgram x kilogram "1 and about 1 milligram x kilogram" 1 of a mammal's body weight. If the heat shock protein is aerosolized, a heat shock protein can be administered in an amount between about 0.1 milligram x kilogram "1 and about 5 milligrams x kilogram" 1 of a mammal's body weight. If the heat shock protein is parenterally distributed, a heat shock protein can be administered in an amount between about 0.1 milligram x kilogram "1 and about 10 micrograms x kilogram" 1 of a mammalian body weight. In one embodiment of the methods described so far in the present invention, a heat shock protein is administered in a pharmaceutically acceptable excipient. Preferred modes of administration include at least one route selected from the group of oral, nasal, topical, inhaled, transdermal, rectal or parenteral routes, and more preferably, include inhaled or nasal routes. Another embodiment of the present invention relates to a method of protecting a mammal from a disease characterized by airway hypersensitivity associated with an inflammatory response, the method comprising administering a heat shock protein to a mammal having this disease . The various particular modalities of this method have been described above with respect to a disease characterized by eosinophilia. Yet another embodiment of the present invention relates to a method of protecting a mammal from an inflammatory disease characterized by a Th2-type immune response, the method comprising administering a heat shock protein to a mammal having this disease. The various particular modalities of this method have been described above with respect to a disease characterized by eosinophilia. Another embodiment of the present invention relates to a method for prescribing treatment for airway hypersensitivity or airflow limitation associated with a disease comprising an inflammatory response. This method includes the steps of: (a) administering a heat shock protein to a mammal; (b) measuring "a change in lung function in response to a causative agent in the mammal to determine whether the heat shock protein modulates airway hypersensitivity or airflow limitation; and, (c) describe a Pharmacological therapy comprising administering the heat shock protein to the mammal effective to reduce inflammation based on changes in lung function In one embodiment the step of measuring comprises measuring a value selected from a group consisting of FEV_, FEVX / FVC, PC2ometacoi-.naFEV_, post-enhanced h (Penh), conductance, dynamic compliance, pulmonary resistance (LR), airway pressure time index (APTI), or peak flow.A causative agent may include a direct and an indirect stimulus, and preferably includes an agent selected from a group of an allergen, methacholine, a histamine, a leukotriene, saline, hyperventilation, exercise, sulfur dioxide, adenosine, propanolol, cold air, an antigen, bradykinin, acetylcholine, a prostaglandin, ozone, ambient air pollutants and mixtures thereof . In one embodiment of this method, the disease is characterized by eosinophilia of the airways. Yet another embodiment of the present invention relates to a formulation for protecting a mammal from the development of a disease characterized by eosinophilia associated with an inflammatory response, this formulation including a heat shock protein and an anti-inflammatory agent. This anti-inflammatory agent may include, but is not limited to, an antigen, an allergen, a hapten, pro-inflammatory cytokine agonists, pro-inflammatory cytokine receptor agonists, anti-CD23, anti-IgE, inhibitors of leukotriene synthesis, leukotriene receptor antagonists, glucocorticosteroids, chemical derivatives and steroids, anti-cyclooxygenase agents, antigene ant i -col agents, beta-adrenergic agonists, meth ixanthanes, antihistamines, chromones, zileuton, anti-CD4 reagents, anti-IL-5 reagents, surfactants, anti-romboxan reagents, anti-serotonin reagents, ketotifen, cytoxin, cyclosporin, methotrexate, macrolide antibiotics, heparin, low molecular weight heparin, or mixtures thereof. In one embodiment, a formulation of the present invention includes a pharmaceutically acceptable excipient and preferably a pharmaceutically acceptable excipient selected from the group of biocompatible polymers, other polymer matrices, capsules, microcapsules, microparticles, bolus preparations, osmotic pumps, diffusion devices, liposomes, lipospheres, or transdermal distribution systems. Yet another embodiment of the present invention relates to a method of protecting a mammal from a disease identified by a selected characteristic of eosinophilia, airway hypersensitivity and a Th2-type immune response, the characteristic that is associated with an inflammatory response. This method includes the method of administering a nucleic acid molecule encoding a heat shock protein to a mammal having the disease. In one embodiment, the nucleic acid molecule is operably linked to a transcription control sequence. In another embodiment, the nucleic acid molecule is administered with a pharmaceutically acceptable excipient selected from the group of a physiologically balanced, aqueous solution, a substrate containing artificial lipid, a substrate containing natural lipid, an oil, an ester, an glycol, a virus, a metal particle and a cationic molecule. In a preferred embodiment, the pharmaceutically acceptable excipient is selected from the group of liposomes, micelles, cells or cell membranes. The nucleic acid molecule can be administered by a mode selected from the group of intradermal injection, intramuscular injection, intravenous injection, subcutaneous injection, or ex vivo administration.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a bar graph demonstrating that treatment with mycobacterial HSP-65 of mice during a 7-day ovalbumin sensitization protocol upregulates the proliferation of antigen-specific and non-specific T cells, in mice Figure 2 is a line graph showing that treatment with HSP-65 my cobactin after sub-optimal sensitization with ovalbumin upregulates the proliferation of antigen-specific T cells in the spleen. Figure 2B is a line graph showing that treatment with HSP-65 mycobacteria of mice after sub-optimal sensitization with ovalbumin upregulates the proliferation of antigen-specific T cells in the lymph nodes per ibronchial ( PBLN). Figure 3 is a bar graph showing that treatment with mycobacterial HSP-65 of mice after sensitization with albumin and stimulation with it, upregulates both non-specific and antigen-specific T cell proliferative responses . Figure 4 is a bar graph showing the effect of mycobacterial HSP-65 treatment of mice after sensitization and ovalbumin stimulation of interferon-α production by splenocytes stimulated with ovalbumin m vitro. Figure 4B is a bar graph showing the effect of mycobacterial HSP-65 treatment of mice after sensitization and stimulation with ovalbumin in the production of IL-4 by splenocytes stimulated with ovalbumin in vitro. Figure 4C is a bar graph showing the effect of mycobacterial HSP-65 treatment of mice after sensitization and stimulation with ovalbumin in the production of IL-5 by splenocytes stimulated with ovalbumin in vitro. Figure 5A is a bar graph showing the effect of treatment with mycobacterial HSP-65 of mice after sensitization and stimulation with ovalbumin in the production of ovalbumin-specific IgG2a by splenocytes stimulated with ovalbumin in vitro. Figure 5B is a bar graph showing the effect of mycobacterial HSP-65 treatment of mice after sensitization and stimulation with ovalbumin in the production of ovalbumin-specific IgGl by splenocytes stimulated with ovalbumin in vitro. Figure 5C is a bar graph showing the effect of mycobacterial HSP-65 treatment of mice after sensitization and stimulation with ovalbumin in the production of ovalbumin-specific IgE by splenocytes stimulated with ovalbumin in vitro. Figure 6 is a bar graph demonstrating that mycobacterial HSP-65 treatment of mice suppresses eosinophilic airway inflammation induced by ovalbumin sensitization and stimulation in vivo. Figure 7 is a line graph showing that treatment with mycobacterial HSP-65 of mice suppresses hypersensitivity of airways to methacholine after sensitization and stimulation with ovalbumin in vivo.

DETAILED DESCRIPTION OF THE INVENTION The present invention relates in general to a method of formulation for protecting a mammal from a disease associated with an inflammatory response, and in particular, from a disease characterized by eosinophilia, hyper-sensitivity of the airways and / or an immunorere spues ta Th2 type, where this characteristic is associated by an inflammatory response. The present inventors have discovered that administration of a heat shock protein to a mammal results in significant inhibition of inflammation, and more specifically, of eosinophilia associated with inflammation. In addition, in respiratory diseases that include the limitation of air flow and / or hypersensitivity of the airways, the present inventors have discovered that administration of a heat shock protein also results in significant inhibition of airway hypersensitivity. Finally, the present inventors have shown that the administration of a heat shock protein to a mammal having an inflammatory disease characterized by a Th2-type response produces a change (ie, modulation) of the Th2-type immune response to an Thl-type immune response, for example, by modulating the production of cytokines and / or immunoglobulin isotypes. Heat shock proteins are highly immunogenic proteins and have been associated with the production of several inflammatory cytokines (including both the Thl associated cytokines and those associated with Th2, described in detail below) and with certain diseases, such as autoimmunity and of course, mycobacterial infections. Therefore, the discovery by the present inventors that it is surprising to administer a heat shock protein, immunostimulatory to a mammal is an effective therapeutic treatment for an inflammatory disease, particularly since the standard treatments for these diseases have emphasized immunosuppression Without being bound by theory, the present inventors believe that the present method of administering a heat shock protein to protect a mammal from an inflammatory disease provides an immunostimulatory effect that results in a modulation of an inflammatory immune response. to an immunoresponse that is beneficial or protective, or at least harmless According to the present invention, a heat shock protein (HSP) can be any protein that corresponds to a group of proteins originally identified by their increased expression in response to temperatures elevates and other stimuli related to stress, referred to collectively in the art as "heat shock proteins". It is now known that heat shock proteins are not only produced in response to cell stress, but can be constitutively presented in a cell and carry out various maintenance functions. Heat shock proteins are commonly divided into at least five major families based on the size of the protein. These five families are the HSP-100 family (that is, having a protein size of approximately 100 kD); the HSP-90 family (i.e., having a protein size of approximately 90 kD); the HSP-70 family (that is, it has a protein size of approximately 70 kD); the HSP-60 family (i.e. having a protein size of approximately 60 kD); and the HSP-27 family (ie, it has a protein size of approximately 27 kD). Heat shock proteins have several unique characteristics. For example, HSP-27, HSP-60 and HSP-70 are involved in protein processing and folding and may be important in the proper presentation of the antigen. HSP-27 and HSP-90 are known to participate in the binding of steroids to their receptor. Mycobacterial proteins, and particularly heat shock protein-65, mycobacterial (HSP-65), a member of the HSP-60 heat shock family, are known to be potent inducers of cellular immune responses, and in particular, he knows that they improve the functions of monocytes / macrophages and T cells. A heat shock protein useful in the present invention can be a heat shock protein from any of the known heat shock families, including the previously identified heat shock protein families. Preferably, a heat shock protein useful in the present invention is from a heat shock protein family including HSP-90, HSP-70, HSP-60 and HSP-27. In one embodiment, a heat shock protein useful in the present invention is from a family of HSP-90 or a family of HSP-27. In another embodiment, a heat shock protein useful in the present invention is of an HSP-60 family, an HSP-70 family and / or an HSP-27 family. In a preferred embodiment, a heat shock protein useful in the present invention is from a family of HSP-60. A heat shock protein useful in the present invention can be distributed or obtained from any organism, preferably from a mammal or a bacterium, and even more preferably from a member of the Mycobacterium genus. Particularly preferred species of Mycobacterium from which a heat shock protein can be derived include but are not limited to Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium leprae. In one embodiment, a heat shock protein useful in the present invention is a mycobacterial heat shock protein (HSP-65), a 65 kD mycobacterial member of the HSP-60 family. A heat shock protein useful in the method of the present invention can be obtained for example from its natural source, it can be produced using recombinant DNA technology, or it can be synthesized chemically. As used herein, a heat shock protein can be a full-length heat shock protein or any homologue of this protein, such as a heat shock protein in which the amino acids have been erased (e.g. truncated protein, such "as a peptide), inserted, inverted, substituted and / or derivatized (for example, by glycosylation, phosphorylation, acetylation, myristoylation, prenylation, palmitation, amidation and / or addition of licosyl phosphatidyl-inositol). A homologue of a heat shock protein is a protein having an amino acid sequence that is sufficiently similar to a heat shock protein amino acid sequence, natural that a nucleic acid sequence encoding the homologue is capable of hybridizing under conditions severe to (ie, with) a nucleic acid molecule that encodes the heat shock protein, natural (ie, to the strand of the nucleic acid strand encoding the natural amino acid sequence of the heat shock protein). A complement of the nucleic acid sequence of any nucleic acid sequence refers to the nucleic acid sequence of the nucleic acid strand is complementary to (ie, can form a double helix complete with) the strand for which it is cited sequence. Heat shock proteins useful in the method of the present invention include, but are not limited to, proteins encoded by nucleic acid molecules having heat shock protein coding regions, full length; proteins encoded by nucleic acid molecules having partial heat shock protein coding regions, wherein these proteins protect a mammal from a disease identified by a selected characteristic of eosinophilia, airway hypersensitivity, and / or a Th2 type immune response; fusion proteins; and chimeric proteins or chemically coupled proteins comprising combinations of different heat shock proteins, or combinations of heat shock proteins with other proteins, such as an antigen or allergen. In another embodiment, heat shock proteins useful in the method of the present invention include heat shock proteins having an amino acid sequence that is at least about 70% identical, and most preferably about 80% identical, so that more preferred, approximately 90% identical to the amino acid sequence of a heat shock protein that occurs naturally. The term, heat shock protein (HSP), can also refer to proteins encoded by allelic variants, including allelic variants that occur naturally of known nucleic acid molecules encoding heat shock proteins, which have similar nucleic acid sequences, but not identical to the nucleic acid sequences encoding the heat shock proteins, which occur naturally or wild type. An allelic variant is a gene that occurs essentially at the same site (or sites) in the genome as a heat shock protein gene, but which, due to natural variations caused for example by mutation or recombination, has a similar sequence but not identical. Typically, allelic variants encode proteins that have activity similar to that of the protein encoded by the gene to which they are being compared. Allelic variants may comprise alterations in the 5 'or 3' untranslated regions of the gene (eg, in regulatory control regions). According to the present invention, the phrase "administering a heat shock protein" can include administration of a protein directly to a mammal such as by any of the modes of administration of a protein described in detail below, alternatively "administering a "heat shock protein" can refer to the administration of a nucleic acid molecule encoding a heat shock protein to a mammal such that the heat shock protein is expressed in the mammal. One embodiment of the present invention in which a nucleic acid molecule encoding a heat shock protein is administered to a mammal is discussed in detail below. In accordance with the present invention, a heat shock protein can be administered to any member of the class of mammalian vertebrates, including without limitation, primates, rodents, livestock and domestic animals. Preferably, the method of the present invention is directed to the protection and / or treatment of a disease characterized by eosinophilia, airway hypersensitivity and / or a Th2-type response associated with an inflammatory response in mammals. A preferred animal to be protected using a heat shock protein includes a human, a rodent, a monkey, a sheep, a pig, a cat, a dog and a horse. An even more preferred mammal to protect is a human. As used herein, the phrase "to protect a mammal from a disease" comprising inflammation, refers to: reducing the potential for an inflammatory response (i.e., a response comprising inflammation) to an inflammatory agent (i.e. , an agent capable of eliciting an inflammatory response, for example, methacholine, histamine, an allergen, a leukotriene, saline, hyperventilation, exercise, sulfur dioxide, adenosine, propanolol, cold air, an antigen or bradykinin); reduce the occurrence of the disease or inflammatory response, and / or reduce the severity of the disease or inflammatory response. Preferably, the potential for an inflammatory response is optimally reduced to a degree that the mammal no longer suffers from discomfort and / or altered function of exposure to the inflammatory agent. For example, the protection of a mammal may relate to the ability of a compound, when administered to a mammal, to prevent a disease from occurring and / or to cure or mitigate the symptoms of the disease, signs or causes. In particular, protection to a mammal refers to the modulation of an inflammatory response to suppress (e.g., reduce, inhibit or block) an overactive or dangerous inflammatory response, which may include the induction of a beneficial, protective or harmless immune response. . Also, in particular, protection to a mammal refers to the regulation of cell-mediated immunity and / or humoral immunity (i.e., T cell activity and / or immunoglobulin activity, including humoral and / or cellular activity Th2 type and / or Thl type). The term, "disease" refers to any deviation from the normal health of a mammal and includes a state when symptoms of disease are present, as well as conditions in which there has been a deviation (eg, infection, gene mutation, defect genetic, etc.), but symptoms are not yet manifested. A disease for which a method of the present invention is protective can include any disease characterized by eosinophilia, airway hypersensitivity and / or a Th2-like immune response, where this feature is associated in an inflammatory response. This disease may include, but is not limited to, allergic airway diseases, hyper-eosinophilic syndrome, parasitic helminth infection, allergic rhinitis, allergic conjunctivitis, dermatitis, eczema, contact dermatitis or food allergy. In one embodiment, a disease for which the method of the present invention may be protective includes a respiratory disease characterized by eosinophilic inflammation of the airways and / or airway hypersensitivity. This respiratory disease includes airway diseases, allergic, mentioned above, which may include but are not limited to, allergic asthma, allergic bronchopulmonary aspergillosis, eosinophilic pneumonia, bronchial asthma of allergic bronchitis, occupational asthma (ie, asthma, panting, chest tightness and cough caused by a sensitizing agent, such as an allergen, irritant or hapten, in the workplace), reactive airway disease syndrome (ie, an individual exposure to an agent that leads to asthma), and lung disease, interstitial. Even more preferably, a respiratory disease for which the method of the present invention may be protective includes but is not limited to, allergic asthma, intrinsic asthma, allergic bronchopulmonary aspergillosis, eosinophilic pneumonia, allergic bronchitis bronchitis, occupational asthma , reactive disease of the airways, interstitial lung disease, hyper-eosinophilic syndrome, and parasitic pulmonary disease. In yet another embodiment, an infection for which the method of the present invention can be protective includes a disease that is associated with sensitization to an allergen. Examples of these diseases are described above. In a preferred embodiment, the method of the present invention protects a mammal from asthma, and particularly allergic asthma. As discussed above, the method of the present invention protects a mammal from a disease that is characterized by eosinophilia, hypersensitivity to the airways, and / or a Th2-like immune response associated with an inflammatory response. Although each of the characteristics of eosinophilia, airway hypersensitivity and a Th2 type immune response are discussed in detail separately subsequently, it is to be understood that a method of the present invention is useful in protecting a mammal from a disease that has any or a combination of these characteristics that are associated with an inflammatory response. Therefore, the particular results obtained with the present method and / or additional characterizations of a disease for which the method of the present invention is effective can be applied to a disease having any or a combination of the features referred to above. One embodiment of the present invention relates to a method of protecting a mammal from the development of a disease characterized by eosinophilia associated with an inflammatory response. This method includes the step of administering a heat shock protein to a mammal having this disease. As used in this, the term "eosinophilia" refers to the clinically recognized condition in which the number of eosinophils present in a mammal having eosinophilia increases or rises compared to the number of eosinophils present in a normal mammal (i.e., a mammal that does not has this condition). In a normal condition that does not have a disease characterized by eosinophils, eosinophils typically comprise from about 0% to about 3% of the total number of white blood cells in the mammal. The eosinophil blood counts of a mammal can be measured using methods known to those skilled in the art. In particular, blood eosinophil counts of a mammal can be measured by vital staining, such as floxin B or Diff Quick. According to the method of the present invention, the administration of a heat shock protein to a mammal having a disease characterized by eosinophilia results preferably in a reduction in eosinophilia in the mammal. Preferably, administration of a heat shock protein in the method of the present invention reduces eosinophil blood counts in a mammal to be between about 0 and about 470 cells / mm 3, more preferably to be between about 0 and 300 cells / mm 3, still more preferably to be between approximately 0 and 100 cells / mm 3. In a preferred embodiment, the administration of a heat shock protein of the method of the present invention reduces blood eosinophil counts in a mammal to be between about 0% and about 3% of the total number of white blood cells in a mammal. Another embodiment of the present invention relates to a method of protecting a mammal from a disease characterized by airway hypersensitivity associated with an inflammatory response. This method includes showing a heat shock protein to the mammal having this disease. The term "airway hypersensitivity" (AHR) refers to an abnormality of the airways that allows them to narrow too easily and / or too much in response to a stimulus capable of inducing limitation of air flow. The AHR can be a functional alteration of the respiratory system caused by the inflammation or remodeling of the airways (for example, such as by depositing collagen). The limitation of air flow refers to the narrowing of the airways that can be irreversible or reversible. The limitation of airflow due to airways hypersensitivity can be caused by the deposition of collagen, bronchospasm, smooth muscle hypertrophy of the airways, smooth muscle contraction of the airways, mucus secretion, cell deposition, destruction epithelial, altered epithelial permeability, smooth muscle function or sensitivity alterations, lung parenchymal normalities, abnormalities in the neural regulation of smooth muscle function (including adrenergic, cholinergic, and non-adrenergic-non-cholinergic regulation) , and infectious diseases in and around the airways. The AHR can be measured by an stress or strain test comprising measuring a function of the mammalian respiratory system in response to a causative agent (ie, stimulus). The AHR can be measured as a change in respiratory function from the baseline plotted against the dose of a causative agent (a method for this measurement and a useful mammalian model is therefore described in detail later in the examples). Respiratory function can be measured, for example, by spirometry, plethysmography, peak flows, symptom scores, physical signs (ie, respiration rate), panting, exercise tolerance, use of rescue medication (ie, bronchitis). lat adores) and blood gases. In humansSpirometry can be used to measure the change in respiratory function in conjunction with a causative agent, such as methothcolin or histamine. In humans, spirometry is done by asking a person to take a deep breath and blow, as long, as hard and as fast as possible in a gauge that measures airflow and volume. The volume of air expelled from the first second is known as the forced expiratory volume (FEVi) and the total amount of air expelled is known as the forced vital capacity (FVC). In humans, the expected, normal FEVi and FVC are available and standardized according to weight, height, sex and race. A disease free individual has a FEVi and a FVC of at least 80% of the expected values, normal for an abnormal person and a FEV_./FVC ratio of at least about 80%. The values are determined before (i.e., representing a resting state of the mammal) and after (i.e., representing a state of high pulmonary resistance of the mammal) of the inhalation of the causative agent. The placement of the resulting curve implies the sensitivity of the airways to the causative agent. The effect of increasing the doses or concentrations of the causative agent in lung function can be terminated by inhibiting the expired volume, forced in one second (FEV_) and the vital capacity enhanced FEV_ (ratio FEV_ / FVC) of the stimulated mammal with the agent causative. In humans, the dose or concentration of a causative agent (ie, methacholine or histamine) that causes a 20% drop in FEV_ (PC20FEV?) Is indicative of the degree of AHR. The values of FEV_ and FVC can be measured using methods known to those skilled in the art. Lung function measurements of airway resistance (LR) and dynamic compliance (CL) and hypersensitivity can be determined by measuring the air pressure as the pressure difference between the airway opening and the airway. body plethysmograph. The volume is the calibrated change of the pressure in the body plethysmograph and the flow is the digital differentiation of the volume signal. Resistance (RL) and compliance (CL) are obtained using methods known to those skilled in the art (for example, such as using a least squares recursive solution of the motion equation). The resistance (R_) and the dynamic compliance (C_) of the airways are described in detail in the examples.

A variety of causative agents are useful for measuring AHR values. The appropriate causative agent includes direct and indirect stimuli. Preferred causative agents include, for example, methacholine (Mch), histamine, an allergen, a leukotriene, saline, hyperventilation, exercise, sulfur dioxide, adenosine, propranolol, cold air, an antigen, bradykinin, acetylcholine, a contaminant. carried in the air, environmental (for example, particles, O, N02), prostaglandins, ozone and mixtures thereof. Preferably, methacholine is used as a causative agent. The preferred concentrations of methacholine to use in a curve the response to concentration are between about 0.001 and about 100 milligrams per milliliter (mg / ml). The most preferred concentrations of methacholine to use a concentration-response curve are between about 0.01 and about 50 mg / ml. Even more preferred concentrations of methacholine for use in a concentration response curve are between about 0.02 and about 25 mg / ml. When methacholine is used as a causative agent, the degree of HAR is defined by the provocative methacholine concentration necessary to cause a 20% drop in mammalian FEVi (PC20metacoyin FEV_). For example, in humans and using standard protocols in the art, a normal person typically has a PC20metacoi? NAFEV1 > 8 mg / ml methacholine. In this way, in humans, the AHR is defined as PC_ometacoi? NaFEV? < 8 mg / ml methacholine. The effectiveness of a drug to protect a mammal from AHR in a mammal that has or is susceptible to AHR is typically measured in double amounts. For example, the effectiveness of a drug to protect a mammal from AHR is significant if PC2ometacoyin FEV? of mammal is at 1 mg / ml before administration of the drug and is at 2 mg / ml methacholine after administration of the drug. Similarly, the drug is considered effective if PC2o etacoimaFEV1 of the mammal is at 2 mg / ml before administration of the drug and is at 4 mg / ml methacholine after administration of the drug. In one embodiment of the present invention, a heat shock protein decreases methacholine sensitivity in a mammal. Preferably, the administration of a heat shock protein increases the PC2ometacoyin FEV? of a mammal treated with the heat shock protein for about a double concentration towards the PC2metacoyin FEV_ of a normal mammal. A "normal mammal" refers to a mammal that is known not to suffer from or is susceptible to abnormal AHR. A test mammal refers to a mammal suspected of suffering from or being susceptible to abnormal AHR. In another embodiment, administration of a heat shock protein to a mammal results in an improvement in a value of PC_.metacoi? NAFEV? of a mammal such that the value of PC2_raeta_oi_naFEV? obtained before the administration of the heat shock protein when the mammal is motivated with a first concentration of methacholine is the same as the PC2ometacoylifene FE1 value obtained after administration of the heat shock protein when the mammal is motivated with the double the amount of the first methacholine concentration. A preferred amount of a heat shock protein to be administered comprises an amount that results in an enhancement in a PC2 value of the mammalian pseudocoke-strain FEV_ such that the PC2metacoyin FEV_ value obtained after administration of the heat shock protein when the mammal is motivated to a concentration of methacholine that is between about 0.01 mg / ml to about 8 mg / ml, is the same as the value of PC.ometacoycin FEV_ obtained after administration of the heat shock protein is when the mammal is motivated with a double concentration of methacholine of between about 0.02 mg / ml to about 16 mg / ml. In accordance with the present invention, respiratory function can be evaluated with a variety of static tests that comprise measuring the function of the respiratory system of a mammal in the absence of a causative agent. Examples of static tests include, for example, spirometry, islet imaging, peak flows, symptom scores, physical signals (ie, respiratory rate), panting, exercise tolerance, use of rescue medication (ie, bronchodilators). ) and blood vessels. The assessment of lung function in static tests can be performed by measuring, for example, total lung capacity (TLC), thoracic gas volume (TgV), functional residual capacity (FRC), residual volume (RV) and specific conductance ( SGL) for lung volumes, lung diffusion capacities for carbon monoxide (DLCO), bacterial blood gases, including pH, P02 and Pco2 for gas exchange. Both FEVi and FEV_ / FVC can be used to measure the air flow limit. If spirometry is used in humans, the FEV_ of an individual can be compared to the FEV_ of predicted values. The predicted FEVi values are available for normal normograms based on the age, sex, weight, height and breed of the mammal. A normal mammal typically has an FEV_ of at least about 80%. FEVi intended for the mammal. The airflow limitation results in an FEVx or FVC of less than 80% of the predicted values. An alternative method for measuring airflow limitation is based on the ratio of FEV_ and FVC (FEVi / FVC), disease-free individuals are defined as having a FEVi / FVC ratio of at least about 80%. The obstruction of the air flow causes the FEV_ / FVC ratio to fall below 80% of the predicted values. In this way, a mammal having an air flow limitation is defined to have a FEV3./FVC of less than about 80%. The effectiveness of a drug to protect a mammal that has or is susceptible to airflow feeding is determined by measuring the percent improvement in FEVX and / or the ratio of FEV_ / FVC before and after drug administration. . In one embodiment the administration of a suitable heat shock protein by the present method reduces the airflow limitation of a mammal such that the FEV_ / FVC value of the mammal is at least about 80%. In another embodiment, the administration of a heat shock protein improves the FEV_ of a mammal, preferably by between about 5% and about 100%, more preferably between about 6% and about 100%, more preferably between about 7% and about 100%, and even more preferably between about 8% and about 100% (or about 200 ml) of the predicted FEVi of the mammal. It should be noted that the measurement of the airway resistance (RL) value in a non-human mammal (e.g., a mouse) can be used to diagnose airflow obstruction similar to the measurement of FEV_ and / or ratio of FEVx / FVC in a human. In one embodiment of the present invention, the administration of a heat shock protein reduces the air flow feed in a mammal such that an RL value of the mammal is reduced by at least about 10% and more preferably, by less about 20%, still more preferably, by at least about 30%, and even more preferably, by at least about 40%. It is within the scope of the present invention that a static test can be performed before or after the administration of a provocative agent used in a stress test. In another embodiment, the administration of a heat shock protein in the method of the present invention reduces the airflow limitation of a mammal such that the variation of FEVi or PEF values of the mammal when measured at dusk before the bed and on sunrise upon awakening is less than about 75%, preferably less than about 45%, more preferably less than about 15% and even more preferably less than about 8%. Yet another embodiment of the present invention relates to a method for protecting a mammal from an inflammatory disease characterized by a Th2-type immune response. This method includes administering a heat shock protein to a mammal having this disease. According to the present invention, a disease characterized by a Th2-like immune response (alternatively referred to as a Th2 immune response), can be characterized as a disease that is associated with the predominant activation of a subset of Th2-type T lymphocytes (or lymphocytes). Th2), in comparison to the activation of Thl type T lymphocytes (or Thl lymphocytes). In accordance with the present invention, Th2 type T lymphocytes can be characterized by their production of one or more cytokines, collectively known as Th2 type cytokines. As used herein, Th2-type cytokines include interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-9 (IL-9), int er leucine-10 (IL-10), int leucine-13 (IL-13) and interleukin-15 (IL-15).

In contrast, Thl lymphocytes produce cytokines that include IL-2 and IFN-α. Alternatively, a Th2-like immune response can sometimes be characterized by the predominant production of antibody isotypes that include IgGl (the approximate human equivalent of which is IgG4) and IgE, whereas a Thl-like immune response can sometimes be characterized by the production of an IgG2 a or IgG3 antibody isotype (the approximate human equivalent of which is IgG1, IgG2 or IgG3). According to the method of the present invention, the administration of a heat shock protein to a mammal having a disease characterized by a Th2-type response preferentially results in a modulation of the immune response in the mammal from a Th2 type response to a more predominant Thl type response. Preferably, administration of a heat shock protein in the method of the present invention results in a decrease (or deletion) in the production of Th2-type cytokines by T lymphocytes, such as IL-4 and IL-5. In addition, or alternatively, the administration of a heat shock protein in a method of the present invention results in an increase (or induction) in the production of Thl-like cytokines by T-lymphocytes, such as IFN-α. Additionally, administration of a heat shock protein in the present method can sometimes result in a decrease in the production of Th2-type antibody isotypes, such as IgG1 and IgE, and / or an increase in isotype production. of Thl-type antibodies, such as IgG2a and IgG3. In one embodiment, the administration of a heat shock protein to a mammal having a disease as described herein may preferably reduce the level of IgG1 (the human equivalent isotype approximately of which is IgG4) in the serum of a mammal to be between about 0 to about 100 international units / ml, preferably to be between about 0 to about 50 international units / ml, more preferably to be between about 0 to about 25 international units / ml, and even more preferably to be between about 0 to about 20 international units / ml. The concentration of IgGl in the serum of a mammal can be measured using methods known to those skilled in the art. In particular, the concentration of IgGl in the serum of an animal at the concentration of IgGl produced by B cells of a mammal in vitro can be measured for example, antibodies that specifically bind to IgGl in an immunized enzyme-linked or a radioimmunoassay. In yet another embodiment, administration of the heat shock protein to a mammal having a disease as described herein may preferentially increase the level of IgG2a (the approximate human isotype equivalent of which is IgG1, IgG2 or IgG3). ) in the serum of a mammal to be between about 0 to about 100 international units / ml, preferably to be between about 10 to about 50 international units / ml, more preferably to be between about 15 to about 25 international units / ml, and even more preferably to be approximately 20 international units / ml. As discussed above, one embodiment of the present invention is that a Th2-like immune response may be associated with other characteristics described hitherto of a disease for which the method of the present invention is protective (e.g., eosinophilia and / or hypersensitivity of the airways). Eosinophilia, for example, is associated with the production of the cytokine IL-5, and hypersensitivity of the airways can be associated with the production of the cytokine, IL-4. One embodiment of the method for protecting a mammal having a disease characterized by eosinophilia, airway hypersensitivity and / or a Th2-like immune response associated with an inflammatory disease, this disease can be further associated with increased production of a selected cytokine from the group of int erleucine-4 (IL-4), interleukin-5 (IL-5, interleukin-6 (IL-6), interleukin-9 (IL-9), interleukin-10 <; IL-10), interleukin-13 (IL-13), interleukin-15 (IL-15). According to the present invention, acceptable protocols for administering a heat shock protein include both the mode of administration and the amount of a heat shock protein to be administered to a mammal, including the size of the individual dose, number of dose and frequency of dose administration. The determination is these protocols can be achieved by those skilled in the art. Suitable modes of administration may include, but are not limited to, oral, nasal, topical, inhaled, transdermal, rectal and parenteral routes. Preferred parenteral routes may include, but are not limited to, subcutaneous, intradermal, intravenous, intramuscular, and intraperitoneal routes. Preferred topical routes include inhalation by aerosol (i.e., spraying), nasal administration, or topical surface administration to the skin of a mammal. In a preferred embodiment, a heat shock protein used in the method of the present invention is administered by a route selected from the nasal and inhaled routes. Particularly preferred routes of administration of a nucleic acid molecule encoding a heat shock protein are discussed in detail below.

As discussed above, administration of a heat shock protein to a mammal in the method of the present invention may result in one or more effects in the mammal, including, but not limited to, reduction and eosinophilia (including but not limited to, eosinophilic inflammation of the airways), reduction of airway hypersensitivity, reduction of IFN-α production. by T cells, and / or suppression of IL-4 and / or IL-5 production by T cells. In accordance with the method of the present invention, an effective amount of a heat shock protein to be administered to a mammal comprises an amount that is capable of reducing airway hypersensitivity (AHR), eosinophilia, reducing airflow limitation and / or symptoms (eg, shortness of inhalation, panting, dyspnea, limitation to exercise or night awakenings), Induction of INF-production? by T cells, and / or suppression of the production of IL-4 and / or IL-5 by T cells without being toxic to the mammal. An amount that is toxic to a mammal comprises any amount that causes damage to the structure or function of a mammal (i.e., poisonous).

An individual, adequate dose of a heat shock protein to be administered to a mammal is a dose that is capable of protecting a mammal from a disease characterized by eosinophilia, airway hypersensitivity, and / or a Th2-like immune response associated with an inflammatory response when administered once or several times for an adequate period of time. In particular, an individual, adequate dose of a heat shock protein comprises a dose that improves the AHR by a double dose of a causative agent or improves the static respiratory function of a mammal. Alternatively, an individual, adequate dose of a heat shock protein comprises a dose that reduces the eosinophil counts in a mammal at the levels described thus far, increases the production of Thl-like cytokines (e.g., IFN-γ) and / or inhibits the production of Th2-type cytokines (for example IL-4 and IL-5). A single, preferred dose of a heat shock protein comprises between about 0.1 microgram x kilogram "1 and about 10 milligrams x kilogram" 1 of a mammal's body weight. A more preferred single dose of a heat shock protein comprises between about 1 microgram x kilogram "1 and about 10 milligrams x kilogram" 1 of a mammalian body weight. An even more preferred individual dose of a heat shock protein comprises between about 1 microgram x kilogram "1 and about 5 milligrams x kilogram" 1 of a mammalian body weight. A particularly preferred individual dose of a heat shock protein comprises between about 1 microgram x kilogram "1 and about 1 milligram x kilogram" 1 of a mammal's body weight. In yet another embodiment, a single, particularly preferred dose of a heat shock protein comprises between about 0.1 milligram x kilogram "1 and about 5 milligrams x kilogram" 1 of a mammalian body weight, if the heat shock protein is distributed by thickness. Another individual, particularly preferred, dose of a heat shock protein comprises between about 0.1 micrograms x kilogram "1 and about 10 micrograms x kilogram" 1 of a mammalian body weight, if the heat shock protein is parenterally distributed. In another embodiment, a heat shock protein of the present invention can be administered simultaneously or sequentially with a compound capable of enhancing the ability of the heat shock protein to protect a mammal from a disease characterized by eosinophilia, hypersensitivity of the airways and / or a Th2-type immune response associated with an inflammatory response. The present invention also includes a formulation that contains a heat shock protein and at least one compound to protect a mammal from a disease comprising inflammation. Of suitable compound to be administered simultaneously or sequentially with a heat shock protein includes a compound that is capable of regulating the production of IgG1 or IgE (i.e., suppression of the synthesis of IgE induced by interleukin-4), up-regulation of the production of int erferón-gamma, regulation of the proliferation of activation of NK cells, regulation of the annihilating cells, activated by lymphokines (LA-K), regulation of the activity of T-helper cells, regulation of mast cell degranulation, protection of sensory nerve endings, regulation of mast cells and / or eosinophil activation, prevention or relaxation of smooth muscle contraction, reduction of microvascular permeability or modulation of Thl and / or Th2 cell subsets. A preferred compound to be administered simultaneously or sequentially with a heat shock protein includes, but is not limited to, any anti-inflammatory agent. In accordance with the present invention, the anti-inflammatory agent may be any compound that is known in the art to have anti-inflammatory properties, and may also include any compound that, under certain circumstances and / or when administered in conjunction with a heat shock protein, may provide an anti-inflammatory effect. A preferred anti-inflammatory agent that can be administered simultaneously or sequentially with a heat shock protein includes, but is not limited to, an antigen, an allergen, a hapten, pro-inflammatory cytokine antagonists (e.g. anti-cytokine, soluble cytokine receptors), pro-inflammatory cytokine receptor antagonists (eg anti-cyccine receptor antibodies) anti-anti-CD23 IgE, inhibitors of leukotriene synthesis, receptor antagonists leukotriene, glucocorticosteroids, chemical derivatives of steroids, anti-cyclooxygenase agents, anti-cholinergic agents, beta-adrenergic agonists, methylxanthenes, antihistamines, chromones, zyleuton, anti-CD4 reagents, anti-IL-5 reagents, surfactants , ant i -tromboxan reagents, anti-serotonin reagents, ketotifen, cytoxin, cyclosporine, methotrexate, macrolide antibiotics, heparin, low molecular weight flour and mixture s of them. The choice of the compound to be delivered in conjunction with a heat shock protein can be made by one skilled in the art based on various characteristics of a mammal. In particular, a genetic background of the mammal, history of occurrence of inflammation, dyspnea, panting on physical examination, symptom scores, physical cues (ie, respiratory rate), exercise tolerance, use of rescue medication (ie, bronchodilators) and blood gases.

A heat shock protein and / or formulation of the present invention to be administered to a mammal may also include other components such as a pharmaceutically acceptable excipient. For example, the formulations of the present invention can be formulated in an excipient such that the mammal to be protected can tolerate. Examples of these excipients include water, saline, phosphate buffered saline solutions, Ringer's solution, dextrose solutions, Hank's solution, physiologically balanced salt solutions containing polyethylene glycol, and other physiologically balanced salt solutions, aqueous. "Non-aqueous vehicles, such as fixed oils, sesame oil, ethyl oleate, triglycerides may also be used Other useful formulations include suspensions containing viscosity improving agents, such as sodium carboxymethylcellulose, sorbitol or dextran.Excipients may also contain minor amounts of additives , such as substances that improve isotonicity and chemical stability or buffers Examples of buffers include phosphate buffers, bicarbonate buffer and Tris buffer, while examples of preservatives include thimerosal, m- or o-cresol, formalin and benzyl alcohol Normal formulations can be either injectable liquids or solids that can be taken in a suitable liquid such as a suspension or solution for injection. Thus, in a non-liquid formulation, the excipient may comprise dextrose, human serum albumin, preservatives, etc., to which sterile water or saline may be added before administration. Examples of pharmaceutically acceptable excipients that are particularly useful for the administration of nucleic acid molecules encoding heat shock proteins are described in detail below. In one embodiment of the present invention, a heat shock protein or a formulation of the present invention may include a controlled release composition that is capable of slowly releasing the heat shock protein or formulation of the present invention in a mammal. As used herein, a controlled release composition comprises a heat shock protein or a formulation of the present invention in a controlled release vehicle. Suitable controlled release vehicles include, but are not limited to, biocompatible polymers, other polymer matrices, capsules, microcapsules, microparticles, bolus preparations, osmotic pumps, diffusion devices, liposomes, lipospheres, dry powders, and distribution systems transdermal Other controlled release compositions of the present invention include liquids which, when administered to a mammal, form a solid or gel in situ. Preferred controlled release compositions are biodegradable (i.e., bioactive). A preferred controlled release composition of the present invention is capable of delivering a heat shock protein to a formulation of the present invention in the blood of a mammal at a constant enough rate to achieve therapeutic dose levels of a heat shock protein or the formulation for preventing inflammation for a period of time ranging from days to months based on the toxicity parameters of the heat shock protein. A controlled release formulation of the present invention is capable of effecting protection for at least about 6 hours, more preferably at least about 24 hours, more preferably for at least about 7 days. Another embodiment of the present invention comprises a method for prescribing treatment for airway hypersensitivity and / or airflow limitation associated with a disease comprising an inflammatory response, the method comprising: (1) administering to a mammal a heat shock protein; (2) measuring a change in "lung function in response to a causative agent in the mammal to determine if the heat shock protein is capable of modulating airway hypersensitivity and / or airflow limitation, and (3) ) describe an effective pharmacological therapy to reduce inflammation based on changes in lung function.In an additional modality, this disease is characterized by eosinophilia of the airways.A change in pulmonary function includes the measurement of static respiratory function before or after administration of the heat shock protein In accordance with the present invention, the mammal receiving the heat shock protein is known to have a respiratory disease comprising inflammation.Measurement of a change in lung function in response a causative agent can be made using a variety of techniques known to those skilled in the art. Causing people may include direct and indirect stimuli, and may encompass any of the causative agents mentioned so far. In particular, a change in lung function can be measured by determining the FEV_, FEV? / FVC, PCbometacoimaFEVi, h po st -mejor da (Penh), conductance, dynamic conformation, lung resistance (RL), time index of airway performance (APTI), and / or maximum group for the receptor of the causative agent. Other methods for measuring a change in lung function include, for example, airway resistance, dynamic conformation, lung volumes, peak flows, point scores, physical signs (ie, respiratory velocity), panting, exercise tolerance, use of rescue medicine (ie, bronchodilators, and blood vessels). A suitable pharmacological therapy effective to reduce inflammation in a mammal can be evaluated by determining whether and to what degree the administration of a heat shock protein has an effect on the lung function of the mammal. If a change in lung function results from the administration of a heat shock protein, then that mammal can be treated with a heat shock protein. Depending on the degree of change in lung function, additional compounds can be administered to the mammal to improve the treatment of the mammal. If there is no change or there is a sufficiently small change in lung function that results from the administration of a heat shock protein, then that mammal must be treated with an alternative compound to the heat shock protein. The present method for prescribing treatment for a respiratory disease may also include evaluating other characteristics of the patient, such as the history of the patient's respiratory disease, in the presence of infectious agents, the patient's habits (eg, smoking), work and living environment of the patient, allergies, a history of life-threatening respiratory cases, severity of disease, duration of illness (ie, acute or chronic) and previous response to other drugs and / or therapy . Another embodiment of the present invention relates to a method for protecting a mammal from a disease infected by one or more characteristics selected from eosinophilia, airway hypersensitivity and a Th2-type immune response, wherein the characteristic is associated with an inflammatory response. . This method includes the step of administering a nucleic acid molecule encoding a heat shock protein to a mammal having this disease. This nucleic acid molecule encoding a heat shock protein can then be expressed by a host cell in the mammal to which the isolated nucleic acid molecule is distributed. The expressed heat shock protein can function at the site to which it is distributed in the manner as previously described herein for heat shock proteins useful in the present method (i.e., to protect a mammal from a disease characterized by eosinophilia, hypersensitivity of the airways, and / or a Th2 immune response associated with an inflammatory response). In accordance with the present invention, a nucleic acid molecule can include DNA, RNA, or derivatives of either DNA or RNA. A nucleic acid molecule encoded for a heat shock protein can be obtained from its natural source, either as a complete (ie, complete) gene or a portion thereof that is capable of encoding a shock protein protein that protects a mammal from a disease identified by a selected characteristic of eosinophilia, airway hypersensitivity, and / or a Th2-like immune response, when this protein and / or nucleic acid molecule encoding this protein is administered to the mammal. A nucleic acid molecule can also be produced using recombinant DNA technology (e.g., polymerase chain reaction (PCR) amplification, cloning) or chemical synthesis. Nucleic acid molecules include natural nucleic acid molecules and homologs thereof, including but not limited to natural allelic variants and modified nucleic acid molecules into which nucleotides have been inserted, deleted, substituted and / or inverted in a manner such that the modifications do not substantially interfere with the stability of the nucleic acid molecule to encode a heat shock protein that is useful in the method of the present invention. In one embodiment, a nucleic acid molecule encoding a heat shock protein is useful in the present invention has a nucleic acid sequence that is at least about 70% identical, and preferably at least about 80% identical, and even more preferably at least about 90% identical to the nucleic acid sequence a heat shock protein that occurs naturally. A nucleic acid molecule, isolated biologically pure, is a nucleic acid molecule that has been removed from its natural environment. As such, "isolated" and "biologically pure" does not necessarily reflect the degree to which the nucleic acid molecule has been purified. A homolog of a nucleic acid molecule can be produced using a number of methods known to those skilled in the art (see, for example, Sambrookk et al., Molecul ar Cloning: A Labora t ory Manual, Cold Spring Harbor Labs Press , 1989). For example, nucleic acid molecules can be modified using a variety of techniques including, but not limited to, classical mutagenesis techniques and recombinant DNA techniques, such as sequence directed mutagenesis, chemical treatment of a nucleic acid molecule for inducing mutations, cleavage with restriction enzymes of a nucleic acid fragment, ligation of the nucleic acid fragments, amplification by polymerase chain reaction (PCR) and / or mutagenesis of selected regions of a nucleic acid sequence, synthesis of mixtures of oligonucleotides and ligation of mixture groups to "constitute" a mixture of nucleic acid molecules and combinations thereof. Homologs of nucleic acid molecules can be selected from a mixture of modified nucleic acids upon detection of the function of the protein encoded by the nucleic acid (e.g., activity of the heat shock protein, as appropriate). Techniques for detecting the activity of the heat shock protein are known to those skilled in the art. Although the phrase "nucleic acid molecule" refers primarily to physical nucleic acid molecule and the phrase "nucleic acid sequence" refers primarily to the nucleotide sequence in the nucleic acid molecule, the two phrases can be used interchangeably , especially with respect to a nucleic acid molecule, or a nucleic acid sequence, which is capable of coding for a heat shock protein. In addition, the phrase "recombinant molecule" refers primarily to a nucleic acid molecule operably linked to a transcription control sequence, but can be used interchangeably with the phrase "nucleic acid molecule" that is administered to a mammal. As described above, a nucleic acid molecule encoding a heat shock protein that is useful in the method of the present invention can be operably launched to one or more transcription control sequences to form a recombinant molecule. The phrase "operably linked" refers to the linkage of a nucleic acid molecule to a transcription control sequence such that the molecule is capable of being expressed when transfected (i.e., transforms, transduces or transfects) into a cell host Transcription control sequences are sequences that control the initiation, elongation and termination of transcription. Particularly important transcription control sequences are those that control the initiation of transcription, such as the promoter, int ensi f icor, operator and repressor sequences. Suitable transcriptional control sequences include any transcriptional control sequence that can function in a recombinant cell useful for the expression of a heat shock protein and / or useful for administration to the mammal in the method of the present invention. A variety of these transcription control sequences are known to those skilled in the art. Preferred transcriptional control sequences include those that function in mammalian, bacterial or insect cells, and preferably in mammalian cells. More preferred transcriptional control sequences include but are not limited to simian virus 40 (SV-40), β-actin, retroviral long terminal repeat (LTR), Rous sarcoma virus (RSV), cytomegalovirus (CMV) , ta c, lac, trp, t re, oxy-pro, omp / lpp, rmb, bacteriophage lambda (?) such as? pL and? pR and fusions that include these promoters), bacteriophage T7, Tulac, bacteriophage T3, bacteriophage SP6, bacteriophage SPO1, metalotin, alpha coupling factor, Pi-ia alcohol-oxidase, subgenomic alphavirus promoters (such as Sindbis virus subgenomic promoters), baculovirus, He lio th isz ea insect virus, vaccinia and other poxviruses, herpes viruses, and adenovirus transcription control sequences, as well as other sequences capable of controlling gene expression in eukaryotic cells. Additional, suitable description control sequences include tissue-specific enhancer promoters (for example, specific promoter enhancers in T cells). The transcription control sequences of the present invention may also include naturally occurring transcriptional control sequences naturally associated with a gene encoding a heat shock protein useful in a method of the present invention. The recombinant molecules of the present invention, which may be either DNA or RNA, may also contain additional regulatory sequences, such as translation regulatory sequences, origins of replication, and other regulatory sequences that are compatible with the recombinant cell. In one embodiment, a recombinant molecule of the present invention also contains secretory signals (ie, nucleic acid sequences of the signal segment) to allow an expressed heat shock protein to secrete from a cell that produces the protein. Suitable signal segments include: (1) a segment of bacterial signal, in particular a signal segment of heat shock protein; or (2) any heterologous signal segment capable of directing the secretion of a heat shock protein from a cell. Preferred signal effects include, but are not limited to, signal segments naturally associated with any of the heat shock proteins mentioned hitherto. One or more recombinant molecules of the present invention can be used to produce a coded product (i.e., a heat shock protein). In one embodiment, an encoded product is produced by expressing a nucleic acid molecule of the present invention under conditions effective to produce the protein. A preferred method for producing an encoded protein is by transfecting a host cell with one or more recombinant molecules having a nucleic acid sequence encoding a heat shock protein to form a recombinant cell. Host cells for transfecting include any cell that can be transfected. The host cells can be either non-transfected cells or cells that are already transformed with at least one nucleic acid molecule. Host cells useful in the present invention can be any cell capable of producing a heat shock protein, including bacterial, fungal, mammalian and insect cells. A preferred host cell includes a mammalian cell. A more preferred host cell includes mammalian lymphocytes, muscle cells, hematopoietic precursor cells, mast cells, natural killer cells, macrophages, monocytes, epithelial cells, endothelial cells, dendritic cells, mesenchymal cells, eosinophils, lung cells and keratinocytes. According to the present invention, a host cell can be transfected in vivo (i.e., by distribution of the nucleic acid molecule in a mammal), ex vivo (i.e., out of a mammal for reintroduction into the mammal, such such as introducing a nucleic acid molecule into a cell that has been removed in a mammal in a tissue culture, followed by reintroduction of the cell into the mammal); or in vitro (ie, outside a mammal, such as in tissue culture for the production of a heat shock protein, recombinant). The transfection of a nucleic acid molecule into a host cell can be achieved by any method by which a nucleic acid molecule can be inserted into the cell. Transfection techniques include, but are not limited to, transfection, electroporation, microinjection, lipofection, adsorption, and protoplast fusion. Preferred methods for transfecting host cells in vivo include lipofection and adsorption. A recombinant cell of the present invention comprises a host cell transfected with a nucleic acid molecule encoding a heat shock protein. It can be appreciated by one skilled in the art that the use of recombinant DNA technologies can improve the expression of transfected nucleic acid molecules by manipulating, for example, the number of copies of the nucleic acid molecules within a host cell, the efficiency with which these nucleic acid molecules are transcribed, the efficiency with which the resulting transcripts are transduced, and the efficiency of post-translational modifications. Recombinant techniques useful for increasing the expression of nucleic acid molecules encoding a heat shock protein include, but are not limited to, operably linked nucleic acid molecules, to plasmids with a high copy number, integration of the acid molecules nucleic acid in one or more chromosomes of host cells, adhesion of vector stability sequences to plasmids, substitutions or modifications of transcription control signals (eg, promoters, operators, substitution or modification of transduction control signals is (eg, ribosome binding sites, Shine-Delgarno sequences), modification of nucleic acid molecules to correspond to cell codon usage host, and the acceptance of sequences that destabilize the transcripts. The stability of a recombinant heat shock protein, expressed can be improved by "fragmenting, modifying or derivatizing nucleic acid molecules that code for a protein." According to the present invention, a nucleic acid molecule encoding a shock protein can be administered, in one embodiment, with a pharmaceutically acceptable excipient A pharmaceutically acceptable excipient can include, but is not limited to, a physiologically balanced, aqueous solution, a substrate containing artificial lipid, a substrate containing natural lipid, a oil, an ester, a glycol, a virus, a metal particle or a cationic molecule The pharmaceutically acceptable excipients, particularly preferred for administering a nucleic acid molecule encoding a heat shock protein include liposomes, micelles, cells and cell membranes The recombinant molecules of acid nucleic acids to be administered in a method of the present invention include: (a) recombinant molecules useful in the method of the present invention in a non-targeting carr(eg, as "naked" DNA molecules, as is shown , for example in Wolff et al., 1990, Sci in ce 24 7, 1465-1468); and (b) recombinant molecules of the present invention complex turns to a delivery vehicle of the present invention. Delivery vehicles suitable for local administration comprise liposomes. Delivery vehicles for local administration may additionally comprise ligands for targeting the vehicle to a particular site (as described in detail herein). Preferably, a nucleic acid molecule encoding a heat shock protein is administered by a method that includes intradermal injection, intramuscular injection, intravenous injection, subcutaneous injection, or ex vivo administration. In one embodiment, a recombinant nucleic acid molecule useful in a method of the present invention is injected directly into muscle cells in a patient, which results in prolonged expression (e.g., weeks or months) of this recombinant molecule. Preferably, this recombinant molecule is in the form of "naked DNA" and is administered by direct injection into muscle cells in a patient. A pharmaceutically acceptable excipient which is capable of targeting is referred to herein as a "distribution vehicle". The delivery vehicles of the present invention are capable of delivering a formulation, including a heat shock protein and / or a nucleic acid molecule encoding a heat shock protein, to a target site of a mammal. A "target site" refers to a site in a mammal to which it is desired to distribute a therapeutic formulation. For example, a target site can be a lung cell, a cell representing the antigen, or a lymphocyte, which is targeted by direct injection or distribution using liposomes or other delivery vehicles. Examples of distribution vehicles include, but are not limited to, distribution vehicles containing artificial and natural lipids. Delivery vehicles containing natural lipids include cells and cell membranes. Distribution vehicles containing artificial lipids include liposomes and micelles. A delivery vehicle of the present invention can be modified to target a particular site in a mammal, targeting and thus making use of a nucleic acid molecule at that site. Suitable modifications include manipulation of the chemical formula of the lipid portion of the delivery vehicle and / or introduction of the vehicle into a compound capable of specifically targeting a delivery vehicle to a preferred site, eg, a preferred type. of cell. Specifically, targeting refers to causing a delivery vehicle to bind to a particular cell by the interaction of the compound in the vehicle with a molecule on the surface of the cell. Suitable targeting compounds include ligands capable of selective binding to (ie, specific) targeting of another molecule at a particular site. Examples of these ligands include antibodies, antigens, receptors and receptor ligands. For example, an antibody specific for an antigen found on the surface of a pulmonary cell can be translated to the outer surface of a liposome delivery vehicle to direct the delivery vehicle to the pulmonary cell. The manipulation of the chemical formula of the lipid portion of the delivery vehicle can modulate the selection or direction to the extracellular or intracellular target of the delivery vehicle. For example, a chemical can be added to the lipid form of a liposome that alters the charge of the lipid bilayer of the liposome so that the liposome phases of particular cells have particular charge characteristics. A preferred delivery vehicle of the present invention is a liposome. A liposome is capable of remaining stable in a mammal for a sufficient amount of time to deliver a nucleic acid molecule described in the present invention to a preferred site in the mammal. A liposome of the present invention is preferably stable in a mammal in which it has been administered for at least about 30 minutes, more preferably for at least about 1 hour, even more preferably for at least about 24 hours. A liposome of the present invention comprises a composition of a lipid that is capable of targeting a nucleic acid molecule described herein to a particular site or selected in a mammal. Preferably, the lipid composition in the liposome is capable of being directed to any organ of a mammal, more preferably to the lung, spleen, lymph nodes and skin of a mammal, even more preferably to the lung of a mammal. A liposome of the present invention comprises a lipid composition that is capable of being approximated with the plasma membrane in the cell sought to distribute a nucleic acid molecule in a cell. Preferably, the transfection efficiency of a liposome of the present invention is about 0.5 micrograms (μg) of nonamol DNA (nmol) of liposome distributed at about 10β cells, more preferably about 1.0 μg of DNA per 16 nmol of liposome distributed to approximately 106 cells, still more preferably about 2.0 μg of DNA per 16 nmol of the liposome distributed to approximately 10 6 cells. A preferred liposome of the present invention is between about 100 and 500 nanometers (nm), more preferably between about 150 and 450 nm and even more preferably between about 200 and 400 nm in diameter. Liposomes suitable for use with the present invention include any liposome. Preferred liposomes of the present invention include those liposomes used in a standard manner for example in gene delivery methods known to those skilled in the art. The most preferred liposomes contain liposomes having a composition of polycationic lipids and / or liposomes having a cholesterol structure conjugated to polyethylene glycol. The complex formation of a liposome with a nucleic acid molecule of the present invention can be raised using standard methods in the art (see for example methods described in Example 2). A suitable concentration of a nucleic acid molecule of the present invention to be added to a liposome includes an effective concentration for the distribution of an effective amount of the nucleic acid molecule to a cell such that the cell can produce sufficient superantigen and / or protein of cytokine to regulate the immunity of effector cells in a desired manner. Preferably, from about 0.1 μg to about 10 μg of the nucleic acid molecule of the present invention is combined with about 8 nmol of liposomes, more preferably from about 0.5 μg to about 5 μg of the nucleic acid molecule. it combines with about 8 nmol of liposomes, and even more preferably about 1.0 μg of the nucleic acid molecule is combined with about 8 nmol of liposomes.

Another preferred distribution vehicle comprises a vaccine of recombinant virus particles. In the recombinant virus particle vaccine of the present invention includes a recombinant nucleic acid molecule useful in the method of the present invention, in which the recombinant molecules are packaged in a viral coating that allows the entry of DNA into a cell so that the DNA is expressed in the cell. After using various particles of recombinant viruses, including but not limited to, those based on alphaviruses, poxviruses, adenoviruses, herpesviruses, sand viruses and ret roviruses. The following examples are provided for the purposes of illustration and are not intended to limit the scope of the present invention.

EXAMPLES EXAMPLE 1 The following example demonstrates that mycobacterial heat shock protein-65 (HSP-65) upregulated T-cell proliferative responses in a mouse model of airway hypersensitivity after short-term sensitization with albumin in alum Disease models in animals are invaluable in providing evidence to support hypotheses or justify human experiments. Mice have many proteins that share more than 90% homology with the corresponding human proteins. For the following experiments, the present inventors have used a murine system fractionated with antigen and characterized by an immune response (IgE), a dependence on the Th2 type response, and an eosinophil response. The model is characterized by both a marked hypersensitivity and a development of the airways. The development of a versatile murine system of chronic exposure to aeroantigen, which is associated with deep eosinophilia and airway tenderness, marked, consistent, progressive, provides an unparalleled opportunity to investigate potential therapeutic compositions (ie, formulations). therapeutic) to prevent or treat respiratory inflammation and / or respiration associated with eosinophilia and a Th2-type immune response. The mouse system described herein is characterized by significant eosinophilia, followed by fibroses of the airways and collagen deposition. The present inventors have used this mouse system to illustrate that administration of the mycobacterial heat shock protein-65 (HSP-65) effectively suppresses airways hypersensitivity and eosinophilia in a sensitized mouse. Female BALB / c mice between the age of 8-12 weeks were obtained from Jackson Laboratories (Bar Harbor, ME). The mice were housed under pathogen-free conditions and kept on a diet free of ovalbumin (OA). The experiments described in the following examples were performed in groups coupled by age and sex between the age of 8-12 weeks. To determine whether mycobacterial HSP-65 facilitates immune responses to antigenic sensitization, the effects of mycobacterial HSP-65 on the T cell response were studied in vitro from mice sensitized with OA. In this experiment, mice were sensitized by intraperitoneal (ip) injection of 20 μg of ovalbumin (OA) (Grade V, Sigma Chemical Co., St. Louis, MO) together with 20 mg of alum (Al (OH) "1 ( Alum injected; Perce, Rockford, IL) in 100 μL of PBS (phosphate buffered saline), or with PBS alone Immediately after the OA injection, the mice received 100 μL intravenously (iv) of either 100 μg of heat shock protein-65 of M. l epra e (mycobacterial HSP-65) in PBS (provided by Dr Kathleen Lukacs, National Heart &Lung Institute, London) or PBS alone 7 days later, mice The spleens were sacrificed and removed and placed in sterile PBS Single cell suspensions were prepared from the spleens, the mononuclear cells were purified by gradient density centrifugation Cells were cultured in 2 × 106 / ml in 96-cavity round bottom tissue culture plants, incuband or cells in triplicate with the medium alone (medium: RPMI 1640 containing heat inactivated fetal calf serum (10%); L-glutamine (2 M); 2-mercapt oet anol (5 mM); HEPES buffer (1 5 m? M); penicillin (100 U / ml); and streptomycin (100 μg / ml); all components of GIBCO / BRL), with 100 μg / ml ovalbumin (OA), or with the combination of phorbol 12,13-dibutyrate (10 nM) and ionomycin (0.5 μM) (Pl) for 48 hours. Cell proliferation was assessed by measuring the cellular admission of (3H) -thymidine. Cell-free supernatants were collected and stored at 20 ° C pending the ELISA assays with cytokine. The cytokine levels secreted in the supernatants of the mononuclear cell cultures were determined by ELISA. Briefly, 96-well plates were coated (immulon) overnight (401C) with the primary anti-cytokine capture antibody (1 μg / ml). IL-4, IL-5 and INF-? Rat anti-mouse, purified from Pharmingen (San Diego, CA) The plates were then washed three times with PBS / Tween 20 (Fischer) and blocked overnight with PBS / 10% FCS. After washing, 100 μL of the cell culture supernatant samples were added to the cavities. The serial dilution of the standards was prepared with a dilution factor of 0.33. After incubation overnight at 4 ° C, the plates were washed and added to 1 μg / ml biotin-conjugated anti-cyclin antibodies (Pharmigen). The plates were incubated overnight and after six washes, the avidin-peroxidase complex (sigma St. Louis, MO) and the substrate were added and incubated at room temperature. A green color was revealed and read at a wavelength of 410 nm in a spectrophotometer (Biorad 2550, Japan). The amounts of cytokine were calculated by using the normal curve in each plate. The detection limits were 5 pg / ml for IL-4 and IL-5 and 3 pg / ml for IFN- ?. As standards, recombinant mouse IL-4 (Pharmigen), IL-5 (Pharmigen) and IFN-γ were used. mupno, recombinant (Genentech, San Francisco, CA). In order to determine antibody levels, ELISA plates (Dynatech, Chantilly, VA) were coated with OA (20 μg / ml (NaHCO3 buffer, pH 9.6) or with 3 μg / ml polyclonal goat anti-mouse IgE (The Binding Site Ltd., San Diego, CA) and incubated overnight at 4 ° C. Plates were blocked with 0.2% gelatin buffer (pH 8.2) for 2 hours at 37 ° C. Standards containing IgE and OA-specific IgGs were generated in the laboratory of the present inventors using a method described by Oshiba et al., 1996, J. Cl in. In ves t 97: 1398-1408, which is incorporated herein by reference in its The ELISA data were analyzed with a Macintosh microplate management computational program (Bio-Rad Labs, Richmond, Va.) The data in all the figures presented here are expressed as averages ± SEM. parametric variance (Krus kal-allis method) was used to determine the variance active among the groups. If a significant variance was found, the Mann-Whitney U test was used to analyze the differences between individual groups. In the case of multiple comparisons, the Bonferroni correction was applied. A value of p of < 0.05 was considered significant. The regression analysis was analyzed in order to establish the correlation between the variables. The data were analyzed with a normal statistical package MINI (Minitab Inc., State College, PA, USA). Figure 1 shows that immunization of mice sensitized with mycobacterial HSP-65 significantly up-regulated the splenocyte proliferative responses in cultures containing only OA (p <0.05; n = 6). Both non-specific and ovalbumin-specific proliferative responses were up-regulated in mice treated with mycobacterial HSP-65. The levels of IL-4, IL-5 and INF-? as the immunoglobulin levels were also up-regulated in the culture supernatants from the mice treated with mycobacterial HSP-65 but not in cultures of the mice treated with PBS (not shown) in summary, these data indicate that 7 days later of sensitization with OA, mice that have been immunized with mycobacterial HSP-65 but not with PBS alone, OA-dependent immune processes have been improved.

EXAMPLE 2 The following example demonstrates that mycobacterial HSP-65 upregulated T-cell proliferative responses in a mouse model of allergic sensitization after sub-optimal sensitization with ovalbumin with aerosol stimulations. Since immunization of mice with HSP-65 mycobacteriana improved T cell responses to OA after i.p. of mice (Example 1), the question arises as to whether mycobacterial HSP-65 will up-regulate the response under conditions in which the response of antigen-specific T cells will not normally be detected (i.e., sub-optimal sensitization with ovalbumin). In addition, the following experiment was designed to test how short-term treatment with mycobacterial HSP-65 will affect airway responses (bronchial alveolar lavage (AL), cellularity, and airway response to methacholine stimulation). Mice were exposed to OA aerosol (1%) on days 1, 2, 3, and 6 (sub-optimal protocols) and injected with 100 μg of mycobacterial HSP-65 or PBS, iv, on day 1 and 6 It should be noted that both immunization and subsequent stimulation with the antigen (OA) are required to observe a response in mice in the optimal protocol of the mouse model. On day 7, airway responses to methacholine (MCh), bronchial alveolar lavage (BAL) were measured, samples were analyzed for their cellular content and spleens and peribronchial lymph nodes (PBLN) were removed to study the responses proliferative Bronchial sensitivity was assessed as a change in airway function after stimulation with aerosolized methacholine via the airways using a modification of the methods described in rats and mice (see, Haezku et al., 1995, Imm un olgy 85: 598-603; and Martin et al., 1988, J. Appl. Phys i ol. 64: 2318-2323; both publications are incorporated herein as a reference in their totalities). Briefly, the mice were anesthetized with an intraperitoneal injection of sodium pentobarbital (70 to 90 mg / kg). An 18G stainless steel tube was inserted as a tracheotomy chamber and passed through an omen in the Plexiglass chamber containing the mouse. A four-way connector was attached to the tracheostomy tube, with two holes connected to the inspiratory and expiratory sides of a ventilator (Model 683, Harvard Apparatus, South Natwick, MA). Ventilation is achieved at 160 inhalations per minute and a flood volume of 0.15 ml with a positive final expiratory pressure of 2-4 cm H20. The Plexiglass chamber was continued with a 1.0-liter glass bottle and one with a copper gauge to stabilize the volumetric signal for the thermal direction. T rans-pulmonary pressure was estimated as the PAO required at the pressure within the plethysmograph using a differential pressure transducer (Validyne Model MP-45-1-871, Validyne Engineering Corp. Northridge, CA). Changes in lung volume were measured by detecting changes in the pressure in the plethysmographic chamber referred to the pressure in a reference frame using a second differential pressure transducer. The two translators and amplifiers were phase-connected electronically to less than 5 degrees from 1 to 30 Hz and then converted from a digital analog signal using a NB-MIO-6X-18 (Natonal Instruments Corporation, Austin, TX) from 6-bit analog to digital board) to 600 bits per channel. The digitalized signals were fed into a Macintosh Quadra 800 computer (Model M1206, Apple Computer, Inc., Cupertino, CA) and analyzed using a LabVIE real-time computer program (National Instruments Corporation, Austin, TX). The flow was determined by differentiation of the volumetric signal and the compliance was calculated as the volume change divided by the change in pressure at zero flow points for the inspiratory phase and the expiratory phase. Average compliance was calculated as the watertight averaging of inspiratory and expiratory compliance for each inhalation. The LabVIEW computer program used the average pressure, flow, volume and compliance to continuously calculate lung resistance (LR) and dynamic compliance (Cdyn) according to the method of Amdur et al., (Pp. 364-368, 1958). , Am. J. Physiol., Vol 192). The breath expiration results for RL, compliance, conductance and specific compliance were tabulated and the reported values are on average of at least 10-20 puffs at the maximum of responses for each dose. It should be noted that the measurement of the RL value in a mouse can be used to diagnose obtaining airflow similar to the measurement of FEVi and / or ratio of FEVi / FVC in a human. Bronchopulmonary aerosol agents were administered through a bypass line via an ultrasonic nebulizer placed between the expiratory orifice of the pipe fan via an ultrasonic nebulizer placed between the expiratory orifice of the ventilator and the four-way connector. The aerosolized agents were administered for 10 seconds with a volume of 0.5 ml. After a dose of inhaled PBS was given, the subsequent values of RL were used with a baseline. Starting 3 minutes after the saline expression, increasing concentrations of methacholine were given by inhalation (10 inhalations), with the initial concentration adjusted to 0.4 mg / ml. Increasing concentrations were given at intervals of 5-7 minutes. Violations of twice the volume of flood were applied between each concentration of methacolma and were performed by manually blocking the overflow of the ventilator in order to reverse any residual atelectasis and ensure a constant volume history before stimulation. From twenty seconds to three minutes after each aerosol stimulation, the data of RL and Cdyn were collected continuously and the maximum values of RL and Cdyn were taken to express the changes in the murine expression of the airways. After measurement of cell function parameters, the lungs were washed with 1 ml aliquots of sterile NaCl at 0.9% (w / v) (room temperature) through a polyethylene syringe attached to the trachial cannula. The washing fluid was centrifuged (500 x g for 10 minutes at 4 ° C), the cell pellet was redispersed in 0.5 ml of the RPMI tissue culture medium. The cell-free supernatant of each BAL sample was stored at -20 ° C for subsequent cytokine analysis by ELISA (described in Example 1). PBLN and splenocytes were analyzed by proliferation assay as described in Example 1. Figure 2 shows that treatment with mycobacterial HSP-65, even after suboptimal sensitization with OA, significantly regulated the proliferative responses of T za OA cells both in splenocytes (Figure 2A) and in cells of peribronchial lymph nodes (PBLN) (Figure 2B), and particularly in cells from local drainage PBLNs (p <; 0.05; ANOVA). No cellular changes were found in the BAL, although there was an increase in pulmonary resistance (RL) to methacholine in the group that was treated with mycobacterial HSP-65 (not shown). These data indicate that mycobacterial HSP-65 upregulates antigen-specific immune responses even after sub-optimal sensitization with OA. In addition, mycobacterial HSP-65 also has an influence on the methacholine sensitivity of the airways if it occurs 24 hours before measurements of lung function.

EXAMPLE 3 The following example demonstrates that mycobacterial HSP-65 upregulates prolifferent T-cell responses in a mouse model of hyper-sensibility of the airways after optimal sensitization and stimulation with ovalbumin in alum. In the mouse model of airway hypersensitivity and allergic sensitization used herein, it has been established that systematic sensitization and local stimulations of the airways results in hypersensitivity of the airways (AHR) associated with eosinophilic inflammation of the airways, cardinal features of human asthma (see for example, Bentley et al., 1992, Am. Re v. Respi. Di. 146: 500-506L; Houston et al. , 1953, Thoras 8: 207-213, or Dunhill, 1960, J. Cl in Pa th ol., 13: 27-33, these publications which are incorporated herein by reference in their totalities. treatment with mycobacterial HSP-65 in the pathological changes of the airways, mice were sensitized intraperitoneally with 20 μg of OA (Grade V, Sigma Chemical Co., St. Louis, MO) together with 20 mg of lum (Al ( OH) 3) (Alum injected; Perce, Rockford, IL) in 100 μL of PBS (phosphate buffered saline), or with PBS alone, on days 1 and 14. The mice received a subsequent stimulation with OA aerosol during 20 minutes with a solution of 1% OA / PBS on days 24, 25 and 26. The mice were sacrificed and investigated 48 hours later. It was assumed that the maximum eosinophil infiltration and airway responses are presented. Splenic mononuclear cells from mice sensitized and stimulated to OA were purified, cultured and proliferative responses to OA were assessed as described in Example 1. Figure 3 shows that mononuclear cells from mice sensitized and stimulated with OA (immunized with PBS only) showed a significant proliferative response to OA (see Figure 3, PBS group). In addition, the proliferation of mononuclear cells from mice treated with mycobacterial HSP-65, sensitized and stimulated with OA (see Figure 3, HSP group) was significantly improved in the presence of OA as well as in the medium alone. These results indicate that mononuclear cells from mice treated with mycobacterial HSP-65 were activated in vivo and will exhibit both antigen-specific and non-specific proliferation in vitro.

EXAMPLE 4 The following example demonstrates that mycobacterial HSP-65 regulates the production of Thl-associated cytokines and antibody isotypes, and down-regulates the production of clots associated with Th2 in a mouse model of airway hypersensitivity after sensitization. Optimum and stimulation with ovalbumin in alum. Allergic asthma is characterized by high levels of IgE, eosinophilic inflammation of the airways and hypersensitivity of the airways. T cells play a cardinal role in this disease, since in the recognition of an allergen, they are capable of producing large quantities of a subset of cytokines, collectively known in the art as Th2-type cytokines. Among Th2-type cytokines, IL-4 has a unique role in the choice of IgE production, and IL-5 is essential in the development of tissue eosinophilia. While the production of Thl-type cytokines will normally be the consequence of T-cell activation, the synthesis of Th2 cytokines requires special conditions, the nature and meaning of which is not known. Without being bound by theory, the present inventors believe that the allergic inflammation may reflect a pathological imbalance of the production of Thl against Th2 cytokines, and in addition these responses to common environmental antigens possibly due to the insufficiency of regulatory mechanisms that normally operate to suppress them. The murine model currently described for airway hypersensitivity provided an ideal system in which it is terminated if the administration of the heat shock protein can modulate the predominant Th2 immune response observed in this model. Splenic mononuclear cells from mice treated with PBS and HSP-65 mycobact dwarf described in Example 3 were cultured for 48 hours. Culture supernatants were harvested and analyzed for cytokine release by ELISA as described in Example 1. Figure 4 illustrates that splenocytes from mice treated with mycobacterial HSP-65 produced a significantly increased amount of IFN-α. (Figure 4A) in cultures stimulated with phorbol ester / ionomycin (Pl) but not in cultures stimulated with OA, when compared to cells from mice treated with PBS (p <; 0.05; n = 6). Meanwhile, the production of IL-4 (Figure 4B) and IL-5 (Figure 4C) in both cultures stimulated with Pl and OA was down-regulated in the splenocytes isolated from mice treated with HSP-65 cobacterial I compared to treated mice with PBS, suggesting that treatment with mycobacterial HSP-65 may have a modulated effect on the production of T cell cytokines in vitro.

In order to assess the production of immunoglobulin, the splenic mononuclear cells which were isolated from mice were treated as described in Example 3, were cultured for 14 days in the presence of varying concentrations of OA as set forth in the axis of the X of Figure 5. The supernatants were harvested and analyzed for the release of OA-specific immunoglobulin by ELISA as described in Example 1. Figure 5 shows that the production of specific IgG2a (Figure 5) from cells of mice treated with Mycobacterial HSP-65 was significantly increased compared to cells from mice treated with PBS (p <0.05, n = 6). The in vitro production of OA-specific IgGl (Figure 5) and IgE (Figure 5) in mice treated with mycobacterial HSP-65 appears to be slightly diminished in comparison to mice treated with PBS, although these results are not conclusive. These data indicate that immunization of mice with mycobacterial HSP-65 modulates the function of T cells and B cells, and further that mycobacterial HSP-65 can modulate inflammatory immune function from a Th2 immune response towards a type Thl EXAMPLE 5 The following example demonstrates that mycobacterial HSP-65 suppresses eosinophilic airway inflammation induced by sensitization and stimulation with ovalbumin in the current modulator of airway hypersensitivity. Allergic sensitization of the airways is associated with a massive inflammation predominated with eosinophils. In order to determine the effects of mycobacterial HSP-65 on eosinophilic inflammation of the airways after allergic sensitization, the cellular content of BAL was assessed in each group of mice treated as described in Example 3. The aveolar wash, bronchial was analyzed 48 hours after the last aerosol stimulation of OA as described above in Example 2. BAL cells were redispersed in RPM I and counted in a hematocrit. Differential counts of cells were made from preparations with cytospin as described (see Haczku et al., Supra). The cells were identified as macrophages, eosinophils, neutrophils and lymphocytes by normal morphology and at least 300 cells were found under an increase of 400 x. The absolute percentage percentage of each cell type was then calculated. Figure ß shows that mice sensitized and exposed to OA and treated with PBS (normal control for hypersensitivity of the airways) developed significant inflammation of the airways (black bars, n = 8). Approximately 60% of all cells in BAL consisted of eosinophils but also significantly increased neutrophil numbers. Single mice (white bars, n = 8) that received a 3-day OA aerosol exposure alone had no eosinophils in their BAL samples. Surprisingly, no eosinophilia was detected in the animals treated with mycobacterial HSP-65 (plume bars, n = 8), and these mice have a cellular content that was virtually identical to the simple control mice. The difference in cellular content of BAL between PBS and animals treated with mycobacterial HSP-65 was significant both in the numbers of eosinophils (P <0.001) and neutrophils (P <0.001). These results indicate that mycobacterial HSP-65 suppresses eosinophilic inflammation of the airways after sensitization to OA exposure.

EXAMPLE 6 The following example demonstrates that mycobacterial HSP-65 suppresses hypersensitivity of airways to methacholine after sensitization and stimulation of ovalbumin bacteria in a mouse model of airway hypersensitivity. In this experiment, bronchial sensitivity was assessed "as a change in airway function after aerosolized methacholine stimulation via the airways." Mice that were treated with mycobacterial HSP-65 to PBS as described in Example 3 were anesthetized 48 hours after the final stimulation of the antigen, were subjected to the cannula and ventilated as described in Example 2. The single mice received immunization for 3 days 48 hours before their measurements were taken. Lung volume of trans-respiratory pressure and flow were measured and lung resistance (LR) was continuously computed, as described in Example 2. Figure 7 illustrates that mice that were sensitized and stimulated with OA and treated with PBS ip (normal control of airway hypersensitivity) showed a significant increase in lung resistance (LR) in response to methacholine stimulation (triangles) as compared to single mice (circles). Mice that were sensitized and stimulated with OA and treated with mycobacterial HSP-65 showed sensitivity to normal (square) methachome (i.e., almost identical to single mice) and significantly less than mice treated with PBS (P <; 0.001), indicating that treatment with mycobacterial HSP-65 suppressed airway hypersensitivity in mice sensitized with and exposed to OA. In summary, in the experiments described above, the specific immune responses to OA were studied after the in vitro culture of mononuclear cells from sensitized mice that were treated with mycobacterial HSP-65. The sensitivity of the airways in vivo was measured by studying lung resistance to methacholine (MCh). Inflammation of the airways and eosinophilia of lung tissue was also assessed. In mice treated with mycobacterial HSP-65, the proliferation of OA-specific T cells was significantly regulated, and the supernatants of the spleen cell cultures contained IFN-α. and IgG2a significantly increased. Surprisingly, the significant eosinophilia of the airways, the increased sensitivity to methacholine, which developed in mice sensitized and stimulated with OA, was suppressed in mice that also received live administration of mycobacterial HSP-65. While various embodiments of the present invention have been described in detail, it is apparent that modifications and adaptations of these embodiments will be presented by those skilled in the art. However, it will be expressly understood that these modifications and adaptations are within the scope of the present invention, as set forth in the following claims.

Claims (56)

1. CLAIMS 1. Use of a heat shock protein for the manufacture of a medicament to protect a mammal from a disease characterized by eosinophilia associated with an inflammatory response.
2. 2. A method for protecting a mammal from a disease characterized by eosinophilia associated with an inflammatory response, the method comprising administering a heat shock protein to a mammal having this disease.
3. 3. The use according to the rei indication 1 or the method according to claim 2, wherein the disease is associated with the increased production of int er leucine-4 (IL-), int erleucine-5 (IL-5), interleukin - 6 (IL-6), int erleucine-9 (IL-9), interleukin-10 (IL-10), interleukin-13 (IL-13) or interleukin-15 (IL-15).
4. 4. The use according to claim 1 or the method according to claim 2, wherein the disease is allergic diseases of the airways, hyper-eosinophilic syndrome, parasitic helminth infection, allergic rhinitis, allergic conjunctivitis, dermatitis, eczema, contact dermatitis or food allergy.
5. 5. The use according to claim 1 or the method according to claim 2, wherein the disease is a respiratory disease characterized by eosinophilic inflammation of the airways and hypersensitivity of the airways.
6. The use or method according to claim 5, wherein the respiratory disease is allergic asthma, intrinsic asthma, bronchopulmonary allergic aspergillosis, eosinophilic pneumonia, bronchial bronchitis, allergic bronchitis, occupational asthma, reactive pathway disease syndrome airway, interstitial lung disease, hyper-eos inophilic syndrome or parasitic lung disease.
7. The use according to claim 1 or the method according to claim 2, wherein the disease is associated with sensitization to an allergen 8.
8. The use according to claim 1 or the method according to claim 2, wherein the disease is Allergic asthma 9.
9. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein is a heat shock protein of the HSP-60 family, a heat shock protein of the HSP- family. 70, a heat shock protein of the HSP-90 family or a heat shock protein of the HSP-27 family 10.
10. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein is a heat shock protein of the HSP-60 family, a heat shock protein of the HSP-70 family or a heat shock protein of the HSP-27 family 11.
11. The use according to claim 1 or the method according to the claim 2, wherein the heat shock protein is a heat shock protein of the HSP-90 family or a heat shock protein of the HSP-27 family.
12. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein is a bacterial heat shock protein or a mammalian heat shock protein.
13. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein is a mycobacterial heat shock protein.
14. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein is a mycobacterial heat shock protein-65 (HSP-65).
15. 15. The method according to claim 2, wherein the heat shock protein is administered by at least one route selected from the oral, nasal, topical, inhaled, transdermal, rectal and parenteral routes.
16. The method according to claim 2, wherein the heat shock protein is administered by a route selected from the inhaled and nasal routes.
17. 17. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein reduces eosinophilia in the mammal.
18. 18. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein reduces the eosinophilic blood counts in the mammal to be between about 0 and about 300 cells / mm 3.
19. 19. The use according to claim 1 or the method according to rei indication 2, wherein the heat shock protein reduces eosinophil blood counts in the mammal to be between about 0 and about 100 cells / mm3.
20. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein reduces the eosinophil blood counts in the mammal to be between about 0 and about 3% of the total white blood cells in the mammal .
21. 21. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein induces the production of internal feron-? (I FN-?) By T lymphocytes in the mammal.
22. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein suppresses the production of interleukin-4 (IL-4) and interleukin-5 (IL-5) by T lymphocytes in the mammal.
23. 23. The use according to claim 1 or the method according to claim 2, wherein the thermal type protein decreases methacholine sensitivity of the airways in the mammal.
24. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein reduces the limitation of air flow in the mammal such that a value of FEVJVFVC of the mammal is at least about 80%.
25. 25. The method according to claim 2, wherein the heat shock protein results in an improvement in a value of PC_ometacoixnaFEV? of mammal such that the value of PC20metacoi? naFEV? obtained before the administration of the heat shock protein when the mammal is motivated with a first concentration of methacholine is the same as the PC2ometacoynefeV_ value obtained after administration of the heat shock protein when the mammal is motivated with the double the amount of the first methacholine concentration.
26. 26. The method according to claim 25, wherein the first methacholine concentration is between about 0.01 mg / ml and about 8 mg / ml.
27. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein improves the F EV_ of the mammal by between about 5% and about 100% of the predicted FEV of the mammal.
28. 28. The use according to claim 1 or the method according to claim 2, wherein the heat shock protein reduces the limitation of air flow in the mammal such that a RL value of the mammal is reduced by at least about 20%.
29. 29. The method according to claim 2, wherein the heat shock protein is administered in an amount of between about 0.1 micrograms x kilogram "1 and about 10 milligrams x kilograms-1 of a mammalian body weight. according to claim 2, wherein the heat shock protein is administered in an amount between about 1 microgram x kilogram-1 and about 1 milligram x kilogram-1 body weight of a mammal 31. The method according to claim 2, in where the heat shock protein is administered in an amount between about 0.1 milligram x kilogram "1 and about 5 milligrams x kilogram" 1 of a mammal's body weight, when the heat shock protein is distributed by aerosol 32. The method according to Claim 2, wherein the heat shock protein is administered in an amount between about 0.1 microgram x kilogram "1 and about 10 micron ogram x kilogram "1 of body weight of a mammal, when the heat shock protein is distributed parenterally. 33. The method according to claim 2, wherein the heat shock protein is administered in a pharmaceutically acceptable excipient. 34. The use according to claim 1 or the method according to claim 2, wherein the mammal is a human. 35. A formulation for protecting a mammal from developing a disease characterized by eosinophilia associated with an inflammatory response, comprising a thermal type protein and an anti-inflammatory agent. 36. The formulation according to claim 35, wherein the anti-inflammatory agent is an antigen, an allergen, a hapten, pro-inflammatory cytosine antagonists, pro-inflammatory cytosine preceptor antagonists, anti-CD23, anti-IgE. , inhibitors of leukotriene synthesis, leukotriene receptor antagonists, glucocorticosteroids, steroid chemical derivatives, anti-cyclooxygenase agents, anti-cholinergic agents, beta-adrenergic agonists, methylsanthes, antihistamines, chromones, cilcuton, anti-reagents CD4, anti-IL5 reagents, surfactants, anti-romboxan reagents, anti-serotonin reagents, ketotifen, cytosine, cyclosporin, methotrexate, macrolide antibiotics, heparin, low molecular weight heparin, or mixtures thereof. 37. The formulation according to the claim 35, wherein the formulation includes a pharmaceutically acceptable excipient. 38. The formulation according to claim 35, wherein the formulation includes a pharmaceutically acceptable excipient selected from biocompatible polymers, other polymer matrices, capsules, microcapsules, microparticles, bolus preparations, osmotic pumps, diffusion devices, liposomes, lipospheres and transdermal distribution systems. 39. The method according to claim 35, wherein the heat shock protein is a heat shock protein of the HSP-60 family, a heat shock protein of the famine HSP-70, a heat shock protein of the HSP family. -90 or a heat shock protein of the HSP-27 family. 40. The method according to claim 35, wherein the heat shock protein is a mycobacterial heat shock protein. 41. The method according to claim 35, wherein the heat shock protein is a heat shock protein, bacterial mycobacterium (HSP-65). 42. The use of a heat shock protein for the manufacture of medicament to protect a mammal by a disease characterized by airway hypersensitivity associated with an inflammatory response. 43. A method to protect a mammal from a disease characterized by airway hypersensitivity associated with an inflammatory response, and a method comprising administering a heat shock protein to a mammal having the disease. 44. The use of a heat shock protein for the manufacture of a medicament to protect a mammal from an inflammatory disease characterized by a Th2-type immune response. 45. A method for protecting a mammal from an inflammatory disease characterized by a Th2-type immune response, the method comprising administering a heat shock protein to a mammal having the disease. 46. The use of a nucleic acid molecule encoding a heat shock protein to protect a mammal from a disease characterized by eosinophilia, airway hypersensitivity or a Th2-like immune response, which is associated with an inflammatory response. 47. A method for protecting a mammal from a disease identified by a characteristic selected from eosinophilia, airway hypersensitivity or a Th2-type immune response, or characteristic that is associated with an inflammatory response, and a method comprising administering a nucleic acid molecule that encodes a heat shock protein to a mammal having the disease. 48. The use according to claim 46 or the method according to claim 47, wherein the nucleic acid molecule is operatively linked to a transcription control sequence. 49. The method according to claim 47, wherein the nucleic acid molecule is administered with a pharmaceutically acceptable excipient selected from a physiologically balanced aqueous solution, a substrate containing artificial lipid, a substrate containing natural lipid, an oil, an ester, a glycol, a virus, a metal particle or a cationic molecule. 50. The method according to claim 47, wherein the pharmaceutically acceptable excipient is liposomes, micelles, cells or cell membranes. 51. The method according to claim 47, wherein the nucleic acid molecule is administered by a mode selected from intradermal injection, intramuscular injection, intravenous injection, subcutaneous injection, or ex vir ve administration. 52. A method for prescribing the treatment of airway hypersensitivity or limitation of airflow associated with a disease comprising an inflammatory response, comprising: (a) administering a heat shock protein to a mammal; (b) a change in lung function in response to a causative agent in a mammal to determine whether the heat shock protein modulates airway hypersensitivity or airflow limitation; and (c) prescribing a pharmacological therapy comprising administering a heat shock protein to the mammal effective to reduce inflammation based on changes in lung function. 53. The method according to claim 52, wherein the disease is characterized by eosinophilia in the airways. 54. The method according to claim 52, wherein the causative agent is a direct stimulus or an indirect stimulus. 55. The method according to claim 52, wherein the causative agent is an allergen, methacholine, a histamine, a leukotriene, saline, hyperventilation, exercise, sulfur dioxide, adenosine, propranolol, cold air, an antigen, bradykinin, acetylcholine, a prostaglandin, ozone, ambient air pollutants, or mixtures thereof. 56. The method according to claim 52, wherein the step to measure comprises a value selected from FEVi, FEV_ / FVC, PC20metacoi? NaFEV1, h po st -me j orada, (Penh), conductance, dynamic compliance, resistance pulmonary, (RL), airway pressure time index (APTI), or peak flow.