MXPA98000190A - Preparations and methods for the treatment of medium diseases by cellula - Google Patents

Abstract

Emulsions of metabolizable lipids, such as Intralipid and LIpofundin, are excellent vehicles for peptide therapy of autoimmune diseases and other diseases or conditions mediated by T cells, TH1, since it promotes a change of TH1 or TH2 cytok ", said emulsions can be used together with an antigen recognized by inflammatory T cells associated with the pathogenesis of a disease or condition mediated by T cells for the therapeutic treatment of said condition.

Description

PREPARATIONS AND METHODS FOR THE TREATMENT OF MEDIUM DISEASES BY T CELLS FIELD OF THE INVENTION The present invention relates to vaccine therapy for T cell-mediated diseases, and in particular to therapeutic preparations comprising antigens recognized by T cells involved in the pathogenesis of T cell mediated diseases, such as autoimmune diseases, and a lipid emulsion metabolizable as a biologically active vehicle.

BACKGROUND OF THE INVENTION Autoimmune disorders, e.g., insulin-dependent diabetes mellitus (IDDM or type I diabetes), multiple sclerosis, rheumatoid arthritis and thyroiditis, are characterized by immunosuppressive activity of an endogenous antigen, with consequent damage to tissues. These immune responses to autoantigens are maintained by the persistent activation of autoreactive lymphocytes. T cells of the "auxiliary" CD type have been divided into two groups by the characteristic cytosines that they secrete when activated (Mosmann and Coffman, 1989). THl cells secrete IL-2, which include proliferation of T cells, and cytokines such coo IFN-r, which mediates tissue inflammation. In contrast, TH2 cells secrete IL-4 and IL-10. IL-4 helps T cells secrete antibodies from certain IgG isotypes and suppresses the production of THl inflammatory cytosines (Banchereau et al., 1994). IL-10 indirectly inhibits the activation of THl by affecting the presentation of antigen and the production of inflammatory cytosine by macrophages (Moore et al., 1993). It is THL cells that contribute to the pathogenesis of organ-specific autoimmune diseases. The TH1-type response also seem to be involved in other diseases or conditions mediated by T cells, such as contagious dermatitis (Romagnani, 1994). Peptides suitable for immunologically specific therapy of an autoimmune disease are peptides that are recognized by T cells involved in the pathogenesis of autoimmune diseases. Each autoimmune disease will have its ideal peptide to be used in therapy. A disease such as multiple sclerosis involving T cells reactive to autoantigens such as myelin basic protein (MBP) (Allegreta et al., 1990) will require a myelin basic protein peptide for its therapy, such as those described by Ota and others. , 1990. The inventors of the present have shown that autoimmune diseases such as diabetes mellitus of type I can be treated by administering a suitable peptide in an oil vehicle. NOD mice spontaneously develop type I diabetes caused by autoimmune T cells that attack cells & islet insulin producers. The autoimmune attack is associated with the reactivity of T cells to a variety of autoantigens including a 60kDa heat shock protein peptide (hsp 60) and glutamic acid desoxetase (GAD) peptides. For example, therefore spontaneous diabetes that develops in NOD / Lt of mice could be treated with a peptide designated p277 corresponding to positions 437-460 of the human hsp 60 sequence (PCT Patent Publication No. W090 / 10449 D. Elias and IR Cohen, Peptide therapy for diabetes in NOD mice, The Lancet 343: 704-06, 1994); with peptide variants p2777 in which one or both cysteine residues at positions 6 and / or 11 have been replaced by valine and / or the Thr residue at position 16 is replaced by Lys (see PCT Publication W096 / 19236) and with designated peptides pl2 and p32 corresponding to positions 166-185 and 466-485, respectively, of the human hsp60 sequence. See Israel Patent Application No. 114,407 of the same applicant of the present application filed on June 30, 1995. See also PCT Patent Application No.

PCT / US96 ..., filed the lo. July 1996, which claims priority from Israel Application No. 114,407, the entire contents of which are incorporated herein by reference. Peptide therapy for the treatment of IDDM using pl2, p32, p227 or variants thereof, was found by the present inventors as effective when the peptide was administered to mice subcutaneously (se) in an oily vehicle such as a mineral oil emulsion known as incomplete Freund's adjuvant (IFA). However, the IFA as well as the complete Freund's adjuvant (CFA; a mineral oil preparation containing several amounts of killed organisms of Mycobacterium) are not allowed for human use because the mineral oil is not metabolizable and can not be degraded in the body, therefore, it would be convenient to discover an effective vehicle for therapy peptides that were metabolizable. Several fat emulsions have been used for many years for the intravenous nutrition of human patients. Two of the commercially available fat emulsions, known as Intralipid ("Intralipid" is a registered trademark of Kabi Pharmacia, Sweden, for a fat emulsion for intravenous nutrition, described in US Patent No. 3,169,094) and Lipofundin (a registered trademark of B. Braun Melsungen, Germany) contain soybean oil as fat (100 or 200 g in 1,000 ml of distilled water: 10% or 20%, respectively). Egg yolk phospholipids are used as emulsifiers in Intralipid (12 g / l of distilled water) and egg yolk lecithin in Lipofundin (12 g / l of distilled water). Isotonicity results from the addition of glycerol (25g / l) in both Intralipid and Lipofundin. These fat emulsions are very stable and have been used for intravenous nutrition of patients suffering from gastrointestinal or neurological disorders, which prevent them from receiving nutrition orally, and therefore receive the necessary calories to sustain life. The usual daily doses are up to one liter a day. U.S. Patent No. 4,073 issued February 14, 1978 to Wretind and others and Re. 32,393 issued May 29, 1990 as U.S. Reissue Patent No. 4,168,308 issued September 18, 1979 to Wretlind et al. describe a vehicle system for use in increasing parenteral administration, particularly intravenously, of a pharmacologically active oil-soluble agent comprising a stable oil-in-water emulsion containing a pharmacologically inert lipoid as a hydrophobic phase dispersed in a hydrophilic phase, said lipid being dispersed in the emulsion as finely divided particles having an average particle size of less than 1 miera to achieve the rapid onset of an acceptable therapeutic effect, said vehicle system being used with an effective dose of said pharmacologically soluble oil agent active, predominantly dissolved in said lipoid at a fraction ratio to the same a in the hydrophobic phase, said therapeutic effect being attributable to said effective dose of the active agent. The vehicle system is said to be suitable for administration of a pharmacologically active agent soluble in oil, insoluble in water or soluble in water which is predominantly dissolved in the lipoid phase. Examples of such pharmacologically active agents are dispersants, anesthetics, analgesics, spasmolytic stimulants, muscle relaxants, vasopressors and diagnosticians, e.g., X-ray diagnostic agents. The vehicle system is said to increase the diagnostic effect or therapeutic agent with a rapid onset accompanied by a reduced incidence of damage to body tissues. Intralipid has been proposed as a non-irritating vehicle for various adjuvants to be used in vaccines such as, for example, 6-0- (2-tetradecylhexadecanoyl) - and 6-0- (3-hydroxy-2-docosylhexacosanoyl) -N-acetylmuramyl -L-alanyl-D-isoglutamine (Tsujimoto et al., 1986 and 1989), avridine (Woodard and Jasman, 1985), N, N-dioctadecyl-N ', N' -bis (2-hydroxyethyl) propanediamine (CP-20 , 961) (German Patent Application No. DE 2945788; Anderson and Reynolds, 1979; Niblack et al., 1979). Kristiansen and Sparrman, 1983, have reported that the immunogenicity of hemagglutinin and neuraminidase in mice is markedly increased after adsorption on lipid particles constituting Intralipid. None of the above publications describe the use of Intralipid as a vehicle for peptides in the treatment of autoimmune diseases, nor has there been any description that Intralipid could mediate a change in the immune response from a TH1-type response to a TH2 response .

BRIEF DESCRIPTION OF THE INVENTION It has now been found, in accordance with the present invention, that metabolizable lipid emulsions, such as Intralipid and Lipofundin, can act as vehicles for therapy with peptides of autoimmune diseases and other diseases or conditions mediated by T cells. Furthermore it has been found that this activity is associated with a change of cytosine from TH1 to HT. "The present invention relates to a therapeutic preparation for the treatment of an autoimmune disease or a disease or condition mediated by T cells, comprising a peptide or other antigen and a biologically active lipid vehicle, wherein the lipid or other antigen is one recognized by inflammatory T cells associated with the pathogenesis of said disease or condition, and because the biologically active lipid carrier is a fat emulsion comprising 10-20% of triglycerides of vegetable origin and / or of animal origin, 1.2-2.4% of phospholipids of origin ve getal and / or animal, 2.25-4.5% osmo regulator, 0-0.05% antioxidant and 100% sterile water. Triglycerides and phospholipids of vegetable or animal origin can be derived from any suitable vegetable oil, such as soybean oil, cottonseed oil, coconut oil or olive oil, or egg yolk or bovine serum. Preferably, the triglycerides are derived from soybean oil and the phospholipids are derived from soybean or egg yolk. Preferably, the weight ratio of triglycerides / phospholipids is about 8: 1. Any suitable osmoregulator can be added to the fat emulsion, preferably glycerol, jilitol or sorbitol. The fat emulsion may optionally comprise an antioxidant, for example 0.05% tocopherol. In one embodiment of the invention, the fat emulsion as defined above is processed by centrifugation, for example, at a higher 10,000g, thus forming a small layer rich in triglycerides (approximately 90% triglycerides) on top of a dispersion aqueous enriched phospholipids containing about 1: 1 triglycerides: phospholipids, and this latter aqueous dispersion is used as the lipid carrier in the preparations of the invention. In a preferred embodiment of the invention, the preparation is for the treatment of insulin-dependent eluted diabetes (IDDM) and copies a peptide derived from the human thermal punch protein 60 (hsp60) which is recognized by inflammatory T cells associated with pathogenesis. of IDDM, wherein said peptide is selected from the group of peptides that appear in the following Table I.

TABLE I Amino acid sequence Peptides Sequence ID (One letter code) P3 1 (31-59) KFGADARALMLQGVDLLADA plO 1 (136-155) NPVEIRRGVMLAVDAVIAEL pll 1 (151-170) VIAELKKQSKPVTTPEEIAQ pl2 1 (166-185) EEIAQVATISANGDKEIGNI pl4 1 (195-214) RKGVITVKDGKTLNDELEII ppll8S 11 ((225555--227744)) QSIVPALEIANAHRKPLVIIA p20 1 (286-305) LVLNRLKVGLQ VAVKAPGF p24 1 (346-365) GEVIVTKDDAMLLKGKGDKA p29 1 (421-440) VTDALNATRAAVEEGIVLGG p30 1 (436-455) IVLGGGCALLRCIPALDSLT pp3322 11 ((446666--448855)) EIIKRTLKIPAMTIAKNAGV p35 1 (511- 530) VNMVEKGIIDPTKVVRTALL p39 1 (343-366) GKVGEVIVTKDDAM p277 1 (437-460) VLGGGCALLRCIPALDSLTPANED p277 (Val *) * 2 VLGGGVALLRCIPALDSLTPANED pp227777 ((VVaallHH)) **** 33 VLGGGCALLRVIPALDSLTPANED p277 (ais- ain) 5le ** VLGGGVALLRVIPALDSLTPANED * 437-460 SEC ID. No. 1 with C-442 changed to V ** 437-460 of SEC ID. No. 1 with C-447 changed to V *** 437-460 of SEC ID. No. 1 with C-442 and C-447 changed to V The invention further relates to a method for therapy of a subject suffering from an autoimmune disease or another disease or condition mediated by TH1, which comprises administering to said subject a effective of a therapeutic preparation in accordance with the invention.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows production of anti-p277 antibodies in NOD mice treated with peptide p277 (Val6-Val11) in: (i) Intralipid or (ii) phosphate buffered saline (PBS), as described in Example 2. Figure 2 shows TH-2-dependent antibody isotypes induced in NOD mice by treatment with peptide p277 (Val6-Val11) in Intralipid, as described in Example 3. Figures 3A -B show that therapy with p277 (Val-Val11) / Intralipid induces in NOD mice a specific change in the profile of cytosines produced by the T cells reactive to the p277 peptide (Val6-Val11), as described in Example 4. Figure 3A shows that there is a reduction of TH! (IL-2, IFN-t) and elevation of TH2 cytokines (IL-4, IL-10) after treatment of mice with peptide p277 (Val6-Val11) in Intralipid and incubation of spleen cells with p277 ( Val-Val11); Figure 3B shows that there is no change in the cytosines after treatment of the mice with the peptide p277 (Val6-Val11) in Intralipid and incubation of the spleen cells with Con A. Figure 4 shows that the proliferative responses of the cells T a p277 (Val6 -Val1 ^) are reduced after the same treatment with peptide p277 (Val6-Val11) in Intralipid, as described in Example 5. Figure 5 shows that the treatment of rats with peptide p71-90 of Myelin basic protein in Intralipid reduces the severity of experimental autoimmune encephalomyelitis (EAE), as described in Example 6. Figure 6 shows that the treatment of rats with p71-90 peptide in IFA of myelin basic protein reduces the severity of experimental autoimmune encephalomyelitis (EAE), as described in Example 6.

DETAILED DESCRIPTION OF THE INVENTION In accordance with the present invention, it was found that treatment with peptide p277 (Val6-Val11), in an appropriate vehicle, downregulated the proliferative responses of spontaneous T cells in epitopes of hsp60 and GAD and abolished the production of autoantibodies to hsp60 , to GAD and insulin. The arrest of the pathological procedure was associated, not with tolerance to T cells or anergy, but with a change in the cytosines produced by autoimmune T cells reactive to p277 (Val6-Val11) from a similar profile of TH1 (IL -2 IFNt) to a profile similar to TH2 (IL-4, IL-10). The modulation was immunologically specific; The spontaneous response of the T cells of the treated mice to a bacterial hsp60 peptide remained in the TH1 mode. In this way, the diabetogenic process characterized by autoimmunity to several antiantigens can be cured using one of the antigens, e.g., peptide? 277 (Val6-Val11). The therapy association of p277 (Val6-Val11) with a change in reactivity to p277 (Val6-Val11) from proliferation of T cells to antibodies indicates that the therapeutic effect results in a change in the predominant cytokines produced by the Autoimmune T cells in the treated mice. THl cells secrete IL-2, which induces T cell proliferation, and cytosines such as IFN-t, which mediate tissue inflammation, thus contributing to the pathogenesis of the disease; TH2 cells, on the other hand, secrete IL-4 and IL-10. IL-4 helps B cells secrete antibodies from certain IgG isotypes and suppresses the production of THl inflammatory cytokines. IL-10 indirectly inhibits the activation of THL, affecting the presentation of antigen and the production of inflammatory cytokines by macrophages. In this way, TH2 cells suppress TH1 activity (see Liblau et al., 1995). The behavior change similar to THl to behavior similar to TH2 was supported by analysis of the isotypes of the antibodies produced before and after therapy with p277 (Val6-Val11). The fact that the mechanism of the therapeutic effect of the peptide in a lipid vehicle treatment is shown to involve a change of TH1-TH2 cytokines provides the possibility of using the TH1-TH2 change as evidence that the treatment was effective and induced a response charitable In other words, the change of TH1-Th2 can serve as a substitute marker for the response to treatment. For example, the lack of change may indicate the need for a second treatment. See Israel Patent Application No. 114,459 filed July 5, 1995, and the corresponding PCT Application filed on a date coinciding therewith, the entire contents of which are incorporated herein by reference. The lipid emulsions of the present invention, when used as a vaccine adjuvant with the antigenic substance to which the T cells involved in the disease or condition being treated are active, serve to mediate a change in the response of the cells Tl of THl before treatment to the T cell response of TH2 after treatment. This finding establishes that said lipid emulsions are biologically active tolerogenic vehicles that can be used in vaccines for the treatment of any disease or condition mediated by THL. In such vaccines the antigen provides the immunological specificity for a therapeutic effect while the biologically active carrier of the present invention provides the biological result, ie, the change of TH1-TH2. Due to the change mediated by said biologically active vehicle of the present invention, diseases with a spectrum of auto-reactivities can be canceled with a single antigen / vehicle combination capable of inducing T-cell cytosine change. A preferred use according to the present invention invention is in the treatment of organ-specific autoimmune diseases that are mediated by THL cells. Such diseases include but are not limited to autoimmune diseases such as IDDM, rheumatoid arthritis, multiple sclerosis and thyroiditis. The peptide used in said treatment is an autoantigenic peptide. Thus, for example, for IDDM the peptide is the aforementioned peptide p277 or the analog substituted with valine p277 (Vald-Val11); for multiple sclerosis said peptide is derived from myelin basic protein; for thyroiditis, the peptide is thought to be derived from thyroglobulin, and for rheumatoid arthritis, the autoantigen may be derived from mycobacterial organisms, e.g., Mycobacterium tuberculosis. It is not critical that the antigen be a peptide. Thus, for example, THl-mediated allergic responses that result in skin sensitivity and inflammation, such as contact dermatitis, can be treated by vaccine containing the irritant antigen and a biologically active carrier according to the present invention. invention that produces a change in the response of the cytosine from a TH1 type to a TH2 type. In this way, while the patient continues to have elevated antibody levels against the antigen, the response of inflammatory T cells that produce skin irritation will be suppressed. Accordingly, the biologically active tolerogenic vehicle of the present invention can be used whenever it is desired to create tolerance for the antigen that T cells are attacking, ie, any time a vaccine is being used to restrict a situation. mediated by T cells, particularly a situation mediated by THL cells. It can be determined that the antigen is activating the response in rejection to grafts or in graft versus host disease, then administration of said antigen with a vehicle according to the present invention will be expected to facilitate the change of the inflammatory THL response. undesirable to a more desirable TH2 response, regardless of the overall complexity of the number of antigens to which the T cells are active in said condition. To determine the secretion of T cells from the cytosines after activation with peptides, peripheral blood lymphocytes from patients are tested in an in vitro activation test. Peripheral blood lymphocytes are isolated from whole heparinized blood on ficol-hypaque, and are cultured with the test peptide (s) at concentrations of 5-50 μg / ml. The supernatants of the cultured T cells are collected at different time points and tested for activity of several cytosines, by ELISA or bioassays.

Examples of fat emulsions that can be used in the preparations of the present invention include, but are not limited to, Intralipid and Lipofundin commercially available for intravenous nutrition, and the fat emulsions described in the aforementioned US Patent Nos. 3,169,096, 4,073,943. and 4,168,308, incorporated herein by reference in its entirety. However, the finding in accordance with the present invention that these metabolizable lipids, previously administered for intravenous nutrition, can be effectively used as vehicles for therapy of diseases mediated by T cells, is completely unexpected. Similarly, the discovery that these preparations are biologically active tolerogenic vehicles that mediate a TH1-TH2 change is also totally unexpected. The fat emulsions of the present invention are preferably used as freshly prepared or after storage in a container that does not open to atmospheric air. Prolonged storage of Intralipid, for example, while exposed to atmospheric air, results in a reduction in pH and a corresponding reduction in biological activity. In one embodiment, the biologically active carrier of the invention is a fat emulsion comprising 10% soybean oil, 1.2% egg yolk phospholipids, 2.5% glycerol, and sterile water to complete 100 ml (Intralipid 10%) .

In another embodiment, the carrier is a fat emulsion comprising 20% soybean oil, 2.4% egg yolk phospholipid, 2.5% glycerol, and sterile water to complete 100 ml. In still another embodiment, the vehicle is a fat emulsion which comprises 5% soybean oil and another 5% triglycerides of animal origin, eg, 5% medium chain butter triglycerides, 1.2% lecithin egg yolk, 2.5% glyceron and distilled water to complete 100ml (Lipofundin 10%). In one embodiment of the invention, the vehicle is a processed lipid emulsion obtained by centrifugation, e.g., at a higher 10,000g, of the original fat emulsion defined herein, whereby a small, high triglyceride-rich dispersion is formed. (about 90% triglycerides) on top of an aqueous dispersion enriched with phospholipids containing about 1: 1 triglyceride: phospholipids. The two phases are separated and the aqueous dispersion rich in phospholipids is used as the carrier. The preparations of the invention may comprise one or more peptides. Thus, for example, the IDDM treatment, the preparation may comprise one or more of the peptides p2, p32, p277, p277 (Val6), p277 (Val11), p277 (Val6-Val11), or any other of the Peptides of Table 1. In a preferred embodiment, the preparation for the treatment of IDDM comprises a peptide p277 or p277 (Val6-Val11) and a fat emulsion comprising 10% soybean oil, 1.2% egg yolk phospholipids , 2.5% glycerol and sterile water to complete 100 ml (Intralipid 10%). The invention further relates to the use of a fat emulsion as defined herein or an aqueous dispersion enriched with phospholipids prepared therefrom by centrifugation for the preparation of a therapeutic preparation comprising one or more peptides or other antigens and said fat emulsion or aqueous dispersion processed as a vehicle in the therapy of autoimmune diseases or other diseases or conditions mediated by THl. The invention will now be illustrated by the following non-limiting examples.

EXAMPLES PEPTIDE THERAPY OF TYPE I DIABETES USING P277 (Val ^ - Val * 1) IN OILS The efficacy of various lipid preparations as vehicles for diabetes lipid therapy of NOD mice was tested. In this model, the autoimmune destruction of the insulin-inducing β cells in the pancreas is mediated by T lymphocytes. An inflammatory infiltrate develops around the pancreatic islets at 5-8 weeks of age and the destruction of cells &; which leads to insulin deficiency and obvious diabetes manifests at 14-20 weeks of age affecting almost 100% of female NOD mice at 35-40 weeks of age. NOD female mice were treated with 100 μg of peptide p277 (Val'S-Val11) per mouse in 0.1 ml of: (i) phosphate buffered saline (PBS), or (ii) a 10% lipid emulsion composed of 10% soybean oil, 1.2% egg phospholipids and 2.25% glycerol (Intralipid, Kabi Pharmacia AB, Sweden). The incidence of diabetes was followed at 6 months of age and the production of anti-p277 antibodies (Val6-Val11). Diabetes was diagnosed as persistent hyperglycemia, blood glucose levels above 11 mmol / L and measured at least twice weekly with a Beckman Glucose Analyzer II glucose analyzer. Successful treatment with peptides was tested by maintaining a normal concentration of blood glucose (less than 11 mmol / L), remission of pancreatic intraislotes inflammation (insulitis) and the induction of antibodies to the therapeutic peptide as an indicator of an immune response of type TH2. The results are shown in Table II.

TABLE II INCIDENCE OF DIABETES AT 6 MONTHS Treatment Diabetes Incidence of Death (%) P277 (Val6 -Val 1) / PBS 90 80 p277 (Val6-Val? I) / Intralipid 45 # 20 # none 100 90 # p < 0.01 compared to untreated NOD mice. As can be seen in Table II, treatment with peptide administered in Intralipid was effective in reducing the incidence of diabetes and death. On the other hand, the treatment administered with PBS was ineffective.

EXAMPLE 2 PRODUCTION OF ANTIBODY ANTI-P277 (al -ValH) The protection against diabetes by treatment with the peptide p277 (Val6-Val11) depends on the immunological reactivity of TH2 to the peptides. Therefore, antibody production was measured in mice immunized with p277 (Val6-Val11) ELISA. Maxisorp microtiter plates (Nunc) were coated with peptide p277 (Val6-Val11), 10 μg / ml for 18 hours and non-specific binding blocked with 7% milk powder for 2 hours. Mouse serum, diluted 1:50, was allowed to bind for 2 hours and specific binding was detected by adding anti-mouse IgG (Serotec) with alkaline phosphatase for 2 hours and p-nitrophenyl phosphate substrate (Sigma) for 30 hours. minutes The intensity of the color was measured by means of an ELISA reader (Anthos) at D0 = 405 nm. As can be seen in Figure 1, NOD mice immunized to p277 (Val6-Val11) for Intralipid developed peptide-specific antibodies, while mice immunized to p277 (Val6-Val11 for PBS showed no antibody response at all.

EXAMPLE 3 TYPES OF ANTIBODIES INDUCED BY THERAPY WITH P277 (Va * 6- ValH) The therapy association of Intralipid p277 (Val6-Val11) with antibodies p277 (Val6-Val11) shown in Example 2, suggested that the therapeutic effect could be the result of a change in the predominant cytokines produced by autoimmune T cells. T cells of the "auxiliary" type of CD4 have been divided into two groups by the characteristic cytosines that they secrete when activated (Mosmann and Coffman, 1989): THl cells secrete IL-2, which induces T cell proliferation, and cytosines such as IFNt, which mediate tissue inflammation; TH2 cells, on the other hand, secrete IL-4, which "helps" B cells to produce certain isotypes of antibodies, and IL-10 and other cytokines, which can "depress" tissue inflammation. The possibility of a behavior of type TH1 to behavior of type TH2 was supported by the analysis of the isotypes of the antibodies produced after p277 therapy (Val6-Val11). Groups of three-month-old NOD mice were treated with p277 (Val6-Val11) or with PBS in oil as described in Example 2. Sera from individual mice were tested for their isotypes of their antibodies to p277 (Val6 -Val11) after treatment (12-15 mice per group). Antibody isotypes were detected using an ELISA test with isotype-specific developing antibody reagents (Southern Biotechnology Associates, Birmingham, AL). The results are shown in Figure 2, wherein: the antibodies for p277 (Val6-Val11) in open circles to NOD mice treated as controls; in closed circles in mice treated with p277 (Val6-Val11). The columns in each experiment show the results of the same number of mice; An obvious reduction in numbers of circles is caused by over imposition. Analysis of antibody titers of anti-p277 antibodies (Val6-Val11) that develop after treatment showed to be exclusively of the IgG1 and IgG2b classes, dependent on TH2 T cells that produce IL-4 (Snapper et al., 1993a ) and possibly TGFβ (Snapper et al., 1993b). There were no THL IgG2a antibodies induced by p277 therapy (Val6-Val11). The development of antibodies to the specific peptide used in the treatment is a sign that autoimmune T cell responses have changed from a damaging inflammatory mode called THL to a TH2 T cell response that produces innocuous antibodies and suppresses inflammation. and tissue damage.

EXAMPLE 4 PEPTIDE THERAPY p277 (Val6 -Val "? / INTRALIPID INDUCES A SPECIFIC CHANGE IN THE CITOSINE PROFILE To confirm the idea of a cytosine change, the cytosines produced by p277-reactive T cells (Val6-Val11) in mice treated with p277 (Val6-Val11) / Intralipid and control were tested. Concanavalin A (ConA), a mitogen of T cells, was used to activate the total T cells of the spleen as a control. Groups of 10 NOD mice, 3 months old, were treated with p277 (Val6-Val11) in Intralipid (closed bars) or with PBS in Intralipid (open bars, see Example 2). Five weeks later the spleens of the mice were excised and the spleen cells were deposited. Spleen cells were incubated with Con A or p277 (Val6-Val11) for 24 hours (for secretion of IL-2 and IL-4) or for 48 hours (for secretion of IL-10 and IFNx). The presence of the cytosines in the culture supernatants were quantified by ELISA, using paired Pharmingen antibodies according to the Cytosine ELISA Protocol of Pharmingen. Pharmingen recombinant mouse cytosines were used as standards for calibration curve. Briefly, 96-well flat bottom microliter plates were coated with an anti-mouse cytosine of rat mAbs for 18 hours at 4 ° C, and the supernatants of the recombinant mouse culture or cytosine were added for 18 hours at 4 ° C. The plates were washed and the biotinylated rat anti-mouse cytosine mAbs were added for 45 minutes at room temperature, then washed extensively and alkaline phosphatase-avidin was added. The plates were washed, a chromogen substrate (p-nitrophenyl phosphate) was added and the samples were read at 405nm in an ELISA reader. The results are shown in the Figure. Cytosine concentrations are shown as DO * P < 0.01. Figure 3A shows that spleen cells from control mice secreted IL-2 and IFNt under incubation with p277 (Val6-Val11). In contrast, mice treated with p277 (Vald-Val11) produced significantly less (P <0.01 IL-2 and IFNt in response to incubation with p277 peptide (Val6-Val11), this reduction in THl cytokines was specific; mice treated with p277 (Val6-Val11) maintained their cytosine responses IL-2 and IFNt to ConA (Fig. 3B) Figures 3A and 3B show the amounts of IL-10 and IL-4 produced by spleen cells The control mice produced very little IL-4 or IL-10 in response to p277 (Val6 -Val1 i) or Con A. Conversely, there was a significant increase in IL-10 and IL-4 in response only at p277 (Val6-Val1: 1) and only in mice treated with p277 (Val6-Val11) / Intralipid (P <0.01) .A decrease in IL-2 and IFNt coupled with an increase in IL-10 and IL -4 confirms the change of behavior from type TH1 to type TH2 behavior, which may help explain a decrease in the proliferation of T cells to p2 77 previously shown by the inventors (Elias et al., 1991) and the appearance of IgG1 and IgG2b antibodies to p277 (Val6-Val11) according to the present invention.

EXAMPLE 5 PROLIFER RESPONSES OF SPONTANEOUS T CELLS TO D277 (Val * -ValíM ARE REDUCED BY THERAPY WITH p277 (Vais -Val * M Groups of 5 female mice of the NOD / Lt strain were treated at the age of 3 months with 100 μg of p277 peptide (Val6-Val11) in Intralipid or with PBS mixed with INtralipid in the posterior part. Five weeks later, the spleens of the mice were excised and the proliferative response of T cells was tested in vitro to the T cell mitogen Con A (1.25 μg / ml) or to p277 (Val6-Val11)) (10 μg / ml). ml) using a normal test. The results are shown in Figure 4, where: With A-bars of black strips; p277 (Val6-Val11) - gray bars. T cell responses were detected by the incorporation of [3 H] thymidine added to the wells in quadrupled cultures during the last 18 hours of a three-day culture. The stimulation index (SI) was calculated as the ratio of the average cpm of crops to the average cpm of wells containing antigen to control wells grown without antigens or with A. The normal deviations of the average cpm were always less than 10%. As shown in Figure 4h. , the control mice tested with PBS / Intralipid showed proliferative responses of T cells to both p277 (Val6-Val11) and mitogen T-cell Con A. In contrast, mice treated with p277 (Val6-Val11) in Intralipid showed a decrease in proliferative reactivity of T cells to p277 (Val6 -Val11) but did not decrease to Con A. Thus, the beneficial effect of peptide therapy p277 (Val6-Val1: 1) is caused not by the inactivation of the autoimmune response, but by the activation of autoimmunity in a mode of behavior different from the cytosine (Cohen, 1995). The regulation of destructive autoimmunity is programmed within the immune system (Cohen, 1992); it only needs to be activated by a suitable signal that requires the peptide together with the lipid vehicle; neither the peptide alone nor the lipid without the peptide are effective, as shown in Table 1. These results indicate that metabolizable lipid emulsions can be used defectively as vehicles for the therapy of autoimmune diseases. Each disease will require its own specific peptide, but the metabolizable lipid emulsion can be used for the various therapies.

EXAMPLE 6 ADMINISTRATION OF PEPTIDE IN INTRALIPI AFFECTS THE DEVELOPMENT OF EXPERIMENTAL AUTOINMUNE ENCEPHALOMYELITIS Experimental autoimmune encephalomyelitis (EAE) is an experimental autoimmune disease of animals that are thought to model aspects of multiple sclerosis (Zamvil and Stein an, 1990). EAE can be tested on susceptible rats, such as the Lewis rat, by immunization to myelin basic protein (MBP) in complete reunding adjuvant (CFA), a mineral oil emulsion containing dead mycobacteria. The disease developed approximately 12 days after immunization and is characterized by paralysis of various degrees due to inflammation of the central nervous system. The paralysis can last up to 6 or 7 days and the rats generally recover unless they die during the peak of their acute paralysis. EAE is caused by T cells that recognize defined determinants of the MBP molecule. The determinant of major MBP in the Lewis rat is composed of sequence 71-90 (Zamvil and Steinman, 1990). therefore, the present inventors performed an experiment to test whether p71-90 of the encephalitogenic MBP peptide in IFA could also inhibit the development of EAE. Figure 5 shows that the administration of p71-90 in IFA 14 days before the induction of EAE led to a significant decrease in the maximum degree of paralysis compared to the control treatment with PBS emulsified in IFA, which had no effect on the severity of the disease. In this way, p71-90 given in IFA affects the EAE. However, IFA can not be administered, as previously indicated, to humans because it is not metabolizable in the body and produces local inflammation. Therefore, the inventors of the present treated Lewis rats with p71-90 in Intralipid. Figure 6 shows the results. Rats that received p71-90 in Intralipid developed significantly less paralysis than the control rats treated with PBS / Intralipid. Therefore, it can be concluded that the relevant peptide such as p71-90 administered in Intralipid is capable of modulating EAE in rats. In this way, the effects of the peptide / Intralipid treatment are not limited to only one peptide, in one species or to one single autoimmune disease.

EXAMPLE 7 NEW 10% INTRALIPID EMULSION EFFECTIVENESS COMPARED TO YEARS % Intralipid emulsion was used to treat female 12-week-old NOD mice with p277 (Val6-Val11).

The emulsion was used the day the sealed bottle was opened, or four months later, after exposure to atmospheric air. The pH of the emulsion was tested at the time of preparation of the peptide + emulsion for treatment. Aging was marked by a drop in pH from 8.2 to 6.7. In each experiment 10 mice were treated with the peptide + emulsion preparation, 10 mice received the emulsion alone and 10 mice were not treated. The results are shown in Table 3.

TABLE 3 Group Treatment Emulsion Diabetes Incidence of pH (%) Death (%) 1 peptide + emulsion 8.2 20 * 10 * 2 emulsion 90 70 3 peptide + emulsion 6.7 60 40 4 emulsion 80 60 5 untreated - 90 80 * p < 0.01 It can be seen that mice treated with placebo (emulsion only, groups 2 and 4) and untreated mice (group 5) developed a similar incidence of diabetes, 80-90% at 6 months of age. In contrast, treatment of the mice with peptide in the freshly opened emulsion protected 80% of the mice against diabetes. However, using the "aged" emulsion only 40% were protected. Therefore, the emulsion was chemically unstable after exposure to air, as shown by the marked decrease in the pH value. this change is relevant to its biological activity. Therefore, Intralipid is a biologically active vehicle and the functional properties depend on the pH and not only on the presence of inert lipid. Now this invention has been described, complete, those skilled in the art will appreciate that it can be performed within a broad range of parameters, concentrations and equivalent conditions without departing from the spirit and scope of the invention and without inadequate experimentation. Although this invention has been described in connection with specific embodiments thereof, it will be understood that it may undergo modifications. The application is intended to cover any variations, uses or adaptations of the following inventions, in general, the principles of the invention and including such deviations from the present disclosure as are included within common or customary practice within the art to which it is applied. The invention relates to and may be applied to the essential features set forth above as set forth in the scope of the appended claims. All references cited herein, including journal articles or abstracts, patent applications of E.U.A. or foreign published or unpublished, patents of E.U.A. or foreign issued, or any other references, are incorporated in their entirety here by reference, including all data, tables, figures and text presented in the references cited. In addition, the complete contents of the references cited within the references cited herein are also incorporated in their entirety by reference. Reference is made to steps of known methods, steps of conventional methods, known methods or conventional methods is in no way an admission that any aspect, description or embodiment of the present invention is described, taught or suggested in the relevant art. The above description of the specific modalities will fully reveal the general nature of the invention which, applying the knowledge within the experience in the art, (including the content of the references cited here), others can easily modify and / or adapting said specific modalities for several applications, without inadequate experimentation, without departing from the general concept of the present application. Therefore, said adaptations and modifications are intended to be within the meaning and scope of equivalents of the described modalities, based on the teachings and guidance presented herein. It should be understood that the phraseology of the present specification should be interpreted by those skilled in the art in light of the teachings and guidance presented herein, in combination with the knowledge of one skilled in the art.

REFERENCES 1. Allegretta, M., Nicklas, J.A., Sriram, S and Albertini, R.J., Science 247: 718-721 (1990). 2. Banchereau, J. and Rybak, M.E., in: The Cytokine Handbook, 2nd. Ed., A, Thompson Ed. Academic Press New York, pp 99 (1994). 3. Cohen, I.R., Immunology Today 13: 490-494 (1992). 4. Cohen, I.R., in: Selective Immunosuppression: Basic Concepts and Clinical Applications (Eds. Adorini, L., Capra, D.J., Waldmann, H.) Chem. I munol Karger, Basel, 60: 150-60 (1995). 5. Kristiansen, T., Sparrman, M. and Heller, L., J.

Biosci 5 (suppl.1: 149-155 (1983) 6. Lib RS, Singer, SM and McDevitt, HO, Immunology Today 16:34 38 (1995) 7. Moore, KV, 0 'Claw, A. , by Waal Malefyt, R., Vieira, P. and Mosmann, TR, Ann. Rev Immunol., 11: 165-190 (1993). 8. Mosmann, T.R. and Coffman R.L., Ann. Rev Immunol. 7: 145-173 (1989). 9. Ota, K., Matsui, M., Milford, E.L., Mackin, G.A., Winer, H.I. and Hafler, D.A., Nature 346: 183-187 (1990). 10. Rabinovitch, A., Sorensen, 0., Sua-Pinzon, W.K., Rajotta, R.V. and Bleakley, R.C., Diabetologia 37: 833-837. 11. Romagnani, S., Ann, Rev. I munol. 12: 227-257 (1994). 12. Snapper, C.M. and Mond, J.J., Immunology Todav 14: 15-17 (1993). 13. Snapper, C.M., Waegell, W., Beernink, H. and Dasch, J.R., J. Immunol. 151: 4625-36 (1993). 14. Zarnvil, S.S. and Steinman, L., Ann. Rev. Immunol. 8: 579-621 (1990).

LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: YEDA RESEARCH & DEVELOPMENT CO. LTD at the Weizmann Institute of Science (B) STREET: P.O. Box 95 (C) CITY: Rehovot (E) COUNTRY: Israel (F) ZIP CODE: 76100 (G) TELEPHONE: 972-8-470739 (H) TELEFAX: 972-8-4706178 (A) NAME: Irun R. COHEN (B) STREET: 11 Hankin Street (C) CITY: Rehovot (E) COUNTRY: Israel (F) ZIP CODE: 76344 (A) NAME: Dana ELIAS (B) STREET: 57 Derech Yavne (C) CITY: Rehovot (E) COUNTRY: Israel (F) POSTAL CODE: 76344 (A) NAME: Meir SHINITZKY (B) STREET: 20 Derech Haganim (C) CITY: Kfar Shmaryahu (E) COUNTRY: Israel (F) POSTAL CODE: 46910 (ii) TITLE OF THE INVENTION: PREPARATIONS AND METHODS FOR THE TREATMENT OF T CELL MEDIATED DISEASES (iii) SEQUENCE NUMBER: 4 (iv) COMPUTER LEADABLE FORM: (A): TYPE OF MEDIUM: Flexible disk (B) COMPUTER: IBM COMPATIBLE PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patentln Relay # 1.0, Version # 1.30 (EPO) (vi) PREVIOUS APPLICATION DATA (A) APPLICATION NUMBER: IL 114458 (B) SUBMISSION DATE: 05-JUL-1995 (2) INFORMATION FOR SEO ID NO. 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 573 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: individual (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: Met Leu Arg Leu Pro Thr Val Phe Arg Gln Met Arg Pro Val Ser Arg 1 5 10 15 Val Leu Ala Pro His Leu Thr Arg Ala Tyr Ala Lys Asp Val Lys Phe 20 25 30 Gly Ala Asp Ala Arg Ala Leu Met Leu Gln Gly Val Asp Leu Leu Ala 35 40 45 Asp Ala Val Ala Val Thr Met Gly Pro Lys Gly Arg Thr Val He He SO 55 60 Glu Gln Gly Trp Gly Ser Pro Lys Val Thr Lys Asp Gly Val Thr Val 65 70 75 80 Wing Lys Ser He Asp Leu Lys Asp Lys Tyr Lys Asn He Gly Wing Lyß 85 90 95 Leu Val Gln Asp Val Wing Asn Asn Thr Asn Glu Glu Wing Gly? Sp Gly 100 105 110 Thr Thr Thr Wing Thr Val Leu Wing Arg Ser He Wing Lys Glu Gly Phe 115 120 125 Glu Lys He Ser Lys Gly Wing Asn Pro Val Glu He Arg Arg Gly Val 130 135 140 Met Leu Ala Val Asp Ala Val He Ala Glu Leu Lys Lys Gln Ser Lys 145 150 155 160 Pro Val Thr Thr Pro Glu Glu He Wing Gln Val Wing Thr He Ser Wing 165 170 175 Asn Gly Asp Lys Glu He Gly Asn He He Ser Asp Wing Met Lys Lys 180 185 190 Val Gly Arg Lys Gly Val He Thr Val Lys Asp Gly Lys Thr Leu Asn 195 200 205 Asp Glu Leu Glu He He Glu Gly Met Lys Phe Asp Arg Gly Tyr He 210 215 220 Ser Pro Tyr Phe He Asn Thr Ser Lys Gly Gln Lys Cys Glu Phe Gln 225 230 235 240 Asp Ala Tyr Val Leu Leu Ser Glu Lys Lys He Ser Ser He Gln Ser 245 250 255 He Val Wing Ala Leu Glu He Wing Asn Wing His Arg Lys Pro Leu Val 260 265 270 He He Wing Wing Glu Asp Val Asp Gly Glu Wing Leu Ser Thr Leu Val Leu 275 280 285 Asn Arg Leu Lys Val Gly Leu Gln Val Val Wing Val Lys Wing Pro Gly 290 295 300 Phe Gly Asp Asn Arg Lys Asn Gln Leu Lys Asp Met Wing He? Thr 305 310 315 320 Gly Gly? The Val Phe Gly Glu Glu Gly Leu Thr Leu Asn Leu Glu? Sp 325 330 335 Val Gln Pro His? Sp Leu Gly Lys Val Gly Val Val He Val Thr Lys 340 345 350 sp? Sp? The Met Leu Leu Lys Gly Lys Gly? Sp Lys? Gln He Glu 355 360 365 Lys? Rg He Gln Glu He He Glu Gln Leu? Sp Val Thr Thr Ser Glu 370 375 380 Tyr Glu Lys Glu Lys Leu? Sn Glu? Rg Leu? La Lys Leu Ser? Sp Gly 385 390 395 400 Val Ala Val Leu Lys Val Gly Gly Thr Ser Asp Val Glu Val Asn Glu 405 410 415 Lys Lys Asp Arg Val Thr Asp Wing Leu Asn Wing Thr Arg Wing Wing Val 420 425 430 Glu Glu Gly He Val Leu Gly Gly Gly Cys Wing Leu Leu Arg Cys He 435 440 445 Pro Wing Leu Asp Being Leu Thr Pro Wing? Sn Glu Asp Gln Lys He Gly 450 455 460 He Glu He He Lys Arg Thr Leu Lys He Pro Wing Met Thr He Wing 465 470 475 480 Lys? Sn Wing Gly Val Glu Gly Ser Leu He Val Glu Lys He Met Gln 465 490 495 Being Ser Glu Val Gly Tyr? S Wing Met Wing Gly Asp Phe Val Asn 500 505 510 Met Val Glu Lys Gly He He Asp Pro Thr Lys Val Val Arg Thr? La 515 520 525 Leu Leu Asp Ala Ala Gly Val Ala Ser Leu Leu Thr Thr Ala Glu Val 530 535 540 Val Val Thr Glu He Pro Lys Glu Glu Lys Asp Pro Gly Met Gly Ala 545 550 555 560 Met Gly Gly Met Gly Gly Gly Met Gly Gly Gly Met Phe 565 570 (2) INFORMATION FOR SEQ ID NO. 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: individual (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Val Lßu Gly Gly Gly Val Ala Leu Leu Arg Cys He Pro Ala Leu Asp 1 5 10 15 Ser Leu Thr Pro Ala? Sn Glu? Sp 20 (2) INFORMATION FOR SEQ ID NO. 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: individual (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: Val Leu Gly Gly Gly Cys? Leu Leu? Rg Val He Pro? La Leu? Sp 1 5 10 15 Ser Leu Thr Pro? La? Sn Glu? Sp 20 (2) I NFORMATION FOR SEQ ID NO. 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: individual (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: Val Leu Gly Gly Val Val Leu Leu Val Val Pro Pro Leu Asp 1 5 o 15 Pro Ser Leu Thr Pro Ala Asn Glu Asp 20

Claims (31)

NOVELTY OF THE INVENTION CLAIMS

1. - A therapeutic preparation for the treatment of disease or condition mediated by T cells comprising an antigen and a biologically active vehicle, characterized in that the antigen is an antigen recognized by inflammatory T cells associated with the pathogenesis of said disease or condition, and because said vehicle is a fat emulsion comprising 10-20% of triglycerides of vegetable and / or animal origin, 1.2-2.4% of phospholipids of vegetable and / or animal origin, 2.25-4.5% of osmo-regulator, 0-0.05% of antioxidant, and sterile water to complete 100 ml.

2. A preparation according to claim 1, further characterized in that the triglycerides are of vegetable origin.

3. A preparation according to claim 2, further characterized in that the triglycerides are derived from soybean oil.

4. A preparation according to claim 1, further characterized in that the triglycerides are of animal origin.

5. A preparation according to claim 4, further characterized in that the triglycerides are derived from egg yolk or bovine serum.

6. - A preparation according to any of claims 1 to 5, further characterized in that the phospholipids are of vegetable origin.

7. A preparation according to claim 6, further characterized in that the phospholipids are soy derivatives.

8. A preparation according to any of claims 1 to 5, further characterized in that the phospholipids are of animal origin.

9. A preparation according to claim 8, further characterized in that the phospholipids are derivatives of egg yolk or bovine serum.

10. A preparation according to any of claims 1 to 5, further characterized in that the osmo-regulator is selected from the group consisting of glycerol, sorbitol and xylitol.

11. A preparation according to any of claims 1 to 5, further characterized in that it comprises 0.05% tocopherol as an antioxidant.

12. A preparation according to claim 1, further characterized in that the biologically active vehicle is a fat emulsion comprising 10% soybean oil, 1.2% egg yolk phospholipids, 2.5% glycerol and sterile water for complete 100 mi.

13. A preparation according to claim 1, further characterized in that the biologically active carrier is a fat emulsion comprising 20% soybean oil, 2.4% egg yolk phospholipids, 2.5% glycerol and sterile water for complete 100 mi.

14. A preparation according to claim 1, further characterized in that the biologically active vehicle is a fat emulsion comprising 5% soy oil oil, 5% medium chain triglycerides, 1.2% lecithin egg yolk, 2.5% glycerol and sterile water to complete 100 ml.

15. A preparation according to any of claims 1 to 5 or 12 to 14, which produces the change of a response to the T cell cytosine of the individual from TH1 to TH2.

16. A preparation according to any of claims 1 to 5 or 12 to 14 that produces a decrease in the response to the cytosine of T cells of IL-2 or IFN-t and an increase in response to cytosine in cells T of IL-4 or IL-10.

17. A preparation according to any of claims 5 or 12 to 14 for the treatment of insulin dependent elitus diabetes (IDDM) comprising a peptide derived from the human toxic shock protein 60 (hsp60) which is recognized by inflammatory T cells associated with the pathogenesis of IDDM wherein said peptide is selected from the group consisting of peptides listed in Table 1.

18. A preparation according to claim 17 for the treatment of IDDM comprising the p227 peptide and a fat emulsion comprising 10% soy oil, 1.2% egg yolk phospholipids, 2.5% glycerol and sterile water to complete 100 ml.

19. A preparation according to claim 17 for the treatment of IDDM comprising the peptide p227 (Val6-Val1l) and a vehicle consisting of an emulsion comprising 10% soybean oil, 1.2% phospholipid yolk egg, 2.5% glycerol and sterile water to complete 100 ml.

20. A preparation according to any of claims 5 or 12 to 14 for the treatment of multiple sclerosis comprising a peptide derived from myelin basic protein (MBP) that is recognized by T cells involved in the pathogenesis of multiple sclerosis .

21.- The use of a fat emulsion comprising 10-20% of triglycerides of vegetable and / or animal origin, 1.2-2.4% of phospholipids of vegetable and / or animal origin, 2.25-4.5% of osmo-regulator, 0 -0.05% antioxidant and sterile water to complete 100 ml, for the manufacture of a therapeutic preparation according to claim 1.

22.- The use of a fat emulsion comprising 10% soybean oil, 1.2% phospholipids of egg yolk, 2.5% glycerol and sterile water to complete 100 ml, for the manufacture of a therapeutic preparation according to claim 12.

23. A method for the treatment of a patient suffering from a T cell-mediated disease or condition comprising administering to said patient a preparation comprising an antigen recognized by inflammatory T cells associated with the pathogenesis of said disease or condition in a biologically active carrier that It consists of a fat emulsion comprising 10-20% of triglycerides of vegetable and / or animal origin, 1.2-2.4% of phospholipids of vegetable and / or animal origin, 2.25-4.5% of osmo-regulator, 0-0.05% of antioxidant and sterile water to complete 100 ml.

24. A method according to claim 23, characterized in that the vehicle consists of fat emulsion comprising 10% soybean oil, 1.2% egg yolk phospholipids, 2.5% glycerol and sterile water to complete 100%. my.

25. A method according to claim 23 or claim 24, further characterized in that said T cell-mediated disease is an autoimmune disease and said antigen is a peptide.

26.- A method in accordance with the claim 23 or claim 24, further characterized in that said T cell-mediated disease is a disease mediated by TH1.

27. A method according to claim 26, further characterized in that said autoimmune disease is an organ-specific autoimmune disease.

28. - A method of treating a patient suffering from IDDM comprising administering to said patient a preparation comprising one or more peptides listed in Cusdro 1 and a biologically active vehicle consisting of a fat emulsion comprising 10% oil soy, 1.2% egg yolk phospholipids, 2.5% glycerol and sterile water to complete 100 ml.

29. A method according to claim 28, further characterized in that the preparation comprises the p227 peptide.

30. A method according to claim 28, further characterized in that the preparation comprises the peptide p227 (Val6-ValH).

31. A method according to claim 23 or claim 24, further characterized in that said T cell-mediated disease is an allergic condition mediated by T cells and said antigen is the allergen that triggered said condition.