WO2000014116A1

WO2000014116A1 - T cell peptides as therapeutics for immune-related disease

Info

Publication number: WO2000014116A1
Application number: PCT/US1999/020009
Authority: WO
Inventors: Jingwu Z. Zhang
Original assignee: Baylor College Of Medicine
Priority date: 1998-09-04
Filing date: 1999-09-01
Publication date: 2000-03-16
Also published as: AU5906099A

Abstract

The present invention relates generally to the fields of therapeutics for autoimmune diseases. More particularly, it concerns compositions comprising T cell peptides capable of eliciting immune responses when administered to a mammal. Specifically, the compositions comprise T cell receptor peptides and T cell activation marker peptides. The T cell receptor peptides are preferably derived from the second or third complementarity determining region (CDR2 or CDR3) of a T cell receptor characteristic of an autoimmune disease, and the T cell activation marker peptides are preferably derived from the extracellular domains of CD38, CD25, LFA-1, ICAM-1, and CD16. The compositions can be used in methods of treating T cell mediated autoimmune diseases.

Description

T CELL PEPTIDES AS THERAPEUTICS FOR IMMUNE-RELATED DISEASE

BACKGROUND OF THE INVENTION

s 1. Field of the Invention

The present invention relates generally to the fields of therapeutics for autoimmune diseases. More particularly, it concerns therapeutics comprising T cell peptides capable of eliciting immune responses when administered to a mammal. 2. Description of Related Art o The pathogenesis of a number of autoimmune diseases is believed to lie in autoimmune

T cell responses to antigens presented normally by the organism. An example of such a disease is multiple sclerosis (MS), which is generally held to arise in T cell responses to myelin antigens, in particular myelin basic protein (MBP). MBP-reactive T cells are found to undergo in vivo activation, and occur at a higher precursor frequency in blood and cerebrospinal fluid in s patients with MS as opposed to control individuals. These MBP-reactive T cells produce Thl cytokines, e.g. IL-2, TNF, and γ-interferon. These Thl cytokines facilitate migration of inflammatory cells into the central nervous system and exacerbate myelin-destructive inflammatory responses in MS.

In contrast to Thl cytokines, Th2 cytokines, e.g. IL-4 and IL-10, have anti-inflammatory 0 properties and downregulate Thl responses. A therapeutic effect has been seen for β-interferon in the treatment of MS, and is at least partially attributable to its ability to induce Th2 cytokines.

Other regulatory mechanisms can be made use of in the treatment of MS. One such is vaccination with irradiated autoreactive T cells (hereinafter "T cell vaccination"). T cell vaccination has been shown to treat experimental autoimmune models such as experimental ₅ autoimmune encephalomyelitis (EAE), an animal model for MS, as reported by Ben-Nun et al., Nature 292:60-61 (1981). T cell vaccination has also been advanced to clinical trials in MS patients, and lower expanded Kurzke disability scale scores (EDSS), lower rates of lesion formation, and fewer outbreaks of exacerbation have been seen (Medaer et al., Lancet 346:807- 808 (1995); Zhang et al., J Immunol. 155:5868-5877 (1995)). 0 Although effective, T cell vaccination requires a relatively high degree of skill, expense, and time to prepare a clonal population of MBP-reactive T cells. In view of present knowledge, it is likely that the therapeutic effect of T cell vaccination is due to a limited number of T cell membrane-associated peptides with extracellular domains. Therefore, an alternative to vaccination with irradiated or otherwise attenuated T cells is vaccination with one or more of the limited number of T cell membrane-associated peptides with extracellular domains.

Vandenbark, U.S. Patent 5,614,192, discloses treatment of autoimmune diseases by the use of immunogenic T cell receptor peptides of 15 to 30 amino acids comprising at least part of the second complementarity determining region (CDR2) of the T cell receptor. Levels of MBP- reactive T cells fell, and ambulation times and stamina improved; however, only two patients were tested.

Therefore, it is desirable to have compositions that are effective in the treatment of autoimmune diseases, particularly MS. It is desirable that such compositions be readily prepared and at relatively low cost.

The present invention discloses that treatment of autoimmune diseases, particularly MS, by the use of immunogenic peptides can yield beneficial therapeutic effects. The present invention relates to compositions of immunogenic T cell receptor peptides, particularly peptides comprising from about 15 to about 30 amino acids from the second complementarity determining region (CDR2) or the third complementarity determining region (CDR3) of a T cell receptor characteristic of an autoimmune disease, and immunogenic T cell activation marker peptides, particularly peptides comprising from about 15 to about 30 amino acids from the extracellular domain of a T cell activation marker characteristic of an autoimmune disease, particularly T cell activation markers CD38, CD25, LFA-1, ICAM-1, and CD16. The present invention also relates to methods using the compositions to treat autoimmune diseases, particularly MS.

SUMMARY OF THE INVENTION

In one embodiment, the present invention is directed to a composition comprising at least one T cell receptor peptide and at least one T cell activation marker peptide. Preferably, the T cell receptor peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the second complementarity determining region (CDR2) or the third complementarity determining region (CDR3). For example, the T cell receptor peptide can be encoded by the nucleotide sequences given in SEQ ID NO: 1-32. The T cell receptor peptide may be naturally- occurring, a synthetic peptide with a wild-type sequence, or a functional derivative thereof. Preferably, the T cell activation marker peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the extracellular domain of a T cell activation marker. Preferably, the T cell activation marker peptide is derived from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 (SEQ ID NO:37-38), ICAM-1 (SEQ ID NO:36). or CD16 (SEQ ID NO:33). The T cell activation marker peptide may be naturally-occurring, a synthetic peptide with a wild-type sequence, or a functional derivative thereof.

In another embodiment, the present invention is directed to a method for treating a T cell mediated disease in a mammal, comprising administering to the mammal a composition comprising at least one T cell receptor peptide and at least one T cell activation marker peptide.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

To aid clear description of the present invention, the following terms have meanings herein as defined below:

"Immunogenic," when used to describe a peptide, means the peptide is able to induce an immune response, either T cell mediated, antibody, or both. "Antigenic" means the peptide can be recognized in a free form by antibodies and in the context of MHC molecules in the case of antigen-specific T cells. "Immune-related disease" means a disease in which the immune system is involved in the pathogenesis of the disease. A subset of immune-related diseases are autoimmune diseases. Autoimmune diseases contemplated by the present invention include, but are not limited to, rheumatoid arthritis, myasthenia gravis, multiple sclerosis, systemic lupus erythematosus, autoimmune thyroiditis (Hashimoto's thyroiditis), Graves' disease, inflammatory bowel disease, autoimmune uveoretinitis, polymyositis, and certain types of diabetes. In view of the present disclosure, one skilled in the art can readily perceive other autoimmune diseases treatable by the compositions and methods of the present invention. "T cell mediated disease" means a disease brought about in an organism as a result of T cells recognizing peptides normally found in the organism. "Derived from," when referring to the relationship of a peptide to a receptor or activation marker, means the peptide was selected based on its amino acid sequence identity or similarity to a portion of the receptor or the activation marker.

"Functional derivative" means a peptide that has at least one amino acid deletion, addition, substitution, or modification relative to a reference peptide, provided that the peptide retains a relevant function exhibited by the reference peptide. Unless otherwise indicated, in the present invention the reference peptide will be either a T cell receptor peptide of wild-type sequence or a T cell activation marker peptide of wild-type sequence, and the retained function will be immunogenecity. Functional derivatives may be described as variants, chemical derivatives, or both, as defined below.

In addition, it is understood that a peptide, either wild-type or functional derivative, of the present invention can be used alone or as part of a longer peptide. The longer peptide may comprise additional sequences from the T cell receptor or T cell activation marker of interest, or it may include sequences of an unrelated peptide, such as a carrier protein used to enhance the immunogenecity of the T cell receptor peptide. Such carriers are known in the art and include heterologous proteins such as keyhole limpet hemocyanin (KLH), bovine serum albumin, tetanus toxoid, and others known to the skilled artisan. Also included within the scope of this invention are conjugations of the peptide or functional derivative to an antibody or to a toxin. Toxins that can be used include, among others, ribosomal inhibitory proteins such as ricin A chain and Pseudomonas toxin.

"Variant" means a peptide that has at least one amino acid deletion, addition, or substitution relative to a reference peptide. Variants may be prepared by direct chemical synthesis or by expression of DNA encoding the variant, both of which techniques are known to those skilled in the art. The DNA encoding the variant may be either chemically synthesized or a mutated form of the DNA encoding the reference peptide. Again, both techniques are well- known in the art. It will be clear that the mutated form will preferably not have a shifted reading frame relative to the DNA encoding the reference peptide, nor will it create complementary regions that could bring about mRNA secondary structure.

Typically, variants will be sought which finely modulate the immunogenic characteristics of the reference peptide. Variants in this category will typically have one amino acid substitution, which substitution will be conservative as such term is known in the art. Alternatively, a second category of variants will have more substantial changes, featuring more than one amino acid substitution, at least one of which substitutions will be non-conservative as such term is known in the art.

One skilled in the art will appreciate that it cannot always be predicted what effect amino acid changes will have on the characteristics, particularly the immunogenic characteristics, of the reference peptide. Therefore it may be necessary to evaluate the characteristics of a variant by routine screening assays. For example, the immunological character of a variant, as exemplified by binding to a given antibody, can be measured by competitive type immunoassay. Changes in T cell recognition of the variant are measured by a delayed hypersensitivity (DH) assay in vivo or a T cell proliferation assay in vitro. Modifications of other peptide properties, such as redox or thermal stability, hydrophobicity, stability to proteolysis, and tendency to aggregate can be assayed by methods known in the art.

"Chemical derivative" means a peptide that has at least one amino acid modification relative to a reference peptide. Such amino acid modifications comprise the addition of chemical moieties not normally a part of the peptide. Modifications can be introduced into the peptide by reacting targeted amino acids of the peptide with an organic derivatizing agent or agents capable of reacting with the side chains or terminal residues of the targeted amino acids. Appropriate organic derivatizing agents and the side chains or terminal residues the agents can target are known to those skilled in the art. Exemplary chemical derivatives are described by Vandenbark, U. S. Patent No. 5,614,192.

Chemical derivatives can exhibit improved solubility, absorption, biological half life, or other properties, and/or can attenuate undesirable characteristics, of the reference peptide.

"Minimal epitope structure" of a peptide means an amino acid sequence sufficiently large that a T cell or antibody specific for the peptide will recognize and react with the related T cell receptor on an intact T cell.

"Overlaps," when referring to a first amino acid sequence relative to a second, means that part (>5 amino acids) of the first sequence is identical to either a part of the second sequence or all of the second sequence, wherein the part of the second sequence includes one terminus.

"Lies within," when referring to a first amino acid sequence relative to a second, means that the entire first sequence (>5 amino acids) is identical to a part of the second sequence. The part of the second sequence can include one terminus. "Corresponds," when referring to a first amino acid sequence relative to a second, means that the entire first sequence (>5 amino acids) is identical to the entire second sequence.

"Pharmaceutically-acceptable excipient" means a compound or compounds that can be used as a carrier, binder, or solvent for T cell receptor peptides and T cell activation markers of the present invention. To be pharmaceutically-acceptable, the compound or compounds are not toxic to the mammal to which the peptides and markers are to be administered, and the compound or compounds do not diminish the activity of the peptides and markers.

"Treatment" or "treating," when referring to protection of an animal from a disease, means preventing, suppressing, or repressing the disease. Preventing the disease involves administering a composition of the present invention to an animal prior to induction of the disease. Suppressing the disease involves administering a composition of the present invention to an animal after induction of the disease but before its clinical appearance. Repressing the disease involves administering a composition of the present invention to an animal after clinical appearance of the disease. It will be appreciated that in human medicine it cannot always be known when in the course of disease induction a composition of the present invention will be administered.

Description Compositions of the present invention comprise at least one T cell receptor peptide and at least one T cell activation marker peptide. To be useful in the present invention, each peptide should be immunogenic; otherwise, each peptide may independently be naturally-occurring, synthetic with a wild-type sequence, or a functional derivative as defined above.

T cell receptor peptides useful in the present invention can be derived from any T cell receptor specific for an autoimmune disease for which treatment is desired. T cell receptors are described by Vandenbark, U.S. Patent No. 5,614,192, herein in its entirety incorporated by reference, and by Lider et al., Science 239:181-184 (1988); Zhang et al., J. Immunol. 155:5868- 5877 (1995); and Zhang et al., Science 261 :1451-1454 (1993).

Particularly, the T cell receptor peptides have an amino acid sequence comprising from about 5 to about 50, preferably from about 10 to about 40, and more preferably from about 15 to about 30 amino acids of a T cell receptor characteristic of a T cell mediated disease. Preferably the T cell receptor peptide is derived from a portion of the T cell receptor which is extracellular, exposed to antibody or other T cells, and is of biological importance in the activity of the T cell bearing it. Exemplary portions of the T cell receptor include the hypervariable regions, including the complementarity determining region 3 (CDR-3) and CDR 2. as well as the V region.

Preferably, the T cell receptor characteristic of the T cell mediated disease is of human origin. More preferably, the T cell mediated disease is rheumatoid arthritis, myasthenia gravis, encephalomyelitis, multiple sclerosis, thyroiditis, diabetes, inflammatory bowel disease, or systemic lupus erythematosus. Most preferably, the T cell mediated disease is multiple sclerosis.

Preferred portions of the T cell receptor are given as SEQ ID NO: 1-32. The identity of the T cell receptor characteristic of the T cell mediated disease will depend on the major autoantigen characteristic of the disease. For example, the T cell receptor characteristic of multiple sclerosis (MS) is capable of binding the MBP, whether alone or complexed with self MHC. The major autoantigen characteristic of myasthenia gravis (MG) is believed to be the nicotinic acetylcholine receptor (AChR), and therefore a T cell receptor which binds AChR is characteristic of myasthenia gravis. Determination of T cell receptors characteristic of a given autoimmune disease can be made by one skilled in the art.

Specifically, T cell receptors characteristic of a given autoimmune disease can be identified using a genetic approach, as has been done for MG and MS, or a genomic analysis using restriction fragment length polymorphism in families having a prevalence of the autoimmune disease. Alternatively, the T cell receptors can be identified using animal models of the autoimmune diseases. Examples of animal models include EAE, experimental MG, experimental autoimmune thyroiditis, adjuvant arthritis, and collagen-induced arthritis, among others. In another technique, T cells are isolated from humans susceptible for and/or suffering from the disease, and the T cells are then expanded in culture. All of the above techniques are well-known in the art.

For the purpose of the present invention, characterization of the autoantigen is not required, so long as T cells receptors for the autoantigen can be raised and/or identified.

T cells bearing receptors characteristic of a given autoimmune disease can be cloned, fused to immortalizing cells such as hybridomas, and grown in culture as is well-known. Such cultured cells can be used as a source of cDNA encoding the T cell receptor; cloning and expression of cDNA can be performed by techniques known to the skilled artisan. In a further embodiment, the present invention relates to T cell receptors that serve as tumor markers on malignant cells, such as T cell leukemia or T cell lymphoma cells.

Preferably, the T cell receptor peptide is derived from a portion of the V region of the T cell receptor. More preferably, the peptide or derivative thereof is derived from a segment of the VDJ region of the T cell receptor β chain or the VJ region of the T cell receptor α chain. Most preferably, the peptide or derivative thereof is derived from at least part of one of the three complementarity determining regions (CDR) of the T cell receptor heterodimer. The CDRs are defined by analogy to the immunoglobulin molecule wherein the CDRs comprise amino acid sequences of the heavy or light chain variable regions which contact antigen and form portions of the antigen binding site.

When the T cell mediated disease is multiple sclerosis, the T cell receptor peptide preferably comprises at least a part of the second complementarity determining region (CDR2) or at least a part of the third complementarity determining region (CDR3) of the T cell receptor. The part of the CDR2 or CDR3 can be from any portion of the T cell receptor. Preferably, the part of the CDR2 or CDR3 is from the β chain or the α chain of the T cell receptor.

The amino acid sequence of the T cell receptor peptide can have any of the following relationships to the amino acid sequence of the CDR2 or CDR3. In one embodiment, the amino acid sequence of the T cell receptor peptide overlaps that of the CDR2 or CDR3. In a second embodiment, the amino acid sequence of the T cell receptor peptide lies within that of the CDR2 or CDR3. In a third embodiment, the amino acid sequence of the T cell receptor peptide corresponds to that of the CDR2 or CDR3. Provided the T cell receptor peptide has therapeutic effect when used in methods of the present invention described below, the T cell receptor peptide can have any of the relationships listed above.

Regions of relevant T cell receptor sequences are identified for synthesis on the basis of their predicted immunogenic or antigenic properties. Such regions can be identified, for example, by using approaches and algorithms described by Margaht et al. (J Immunol. 138:2213-2229 (1987)) or Rothbard et al. (EMBO J. 7:93-100 (1988)). In Margaht et al., antigenic sites are postulated to be helices with one predominantly polar and one predominantly apolar face. Rothbard et al. recognize motifs similar to epitopes recognized preferentially by T helper or T cytotoxic cell clones. Use of one such approach predicted that immunogenic regions of the T cell receptor fall within CDR1, CDR2, CDR3, or in T cell receptor hypervariable regions not strictly part of a CDR, such as residues 39-49 of the Vβ segment. Other approaches will be known to the skilled artisan.

Once immunogenic regions of a T cell receptor characteristic of a T cell mediated disease are identified, a T cell receptor peptide as described above can be derived therefrom. Immunogenicity of the peptide can be screened by well-known methods, such as the DH response in an animal. In this method, an animal is sensitized by injection, typically subcutaneously, of an appropriate dose of an antigen, typically with an adjuvant such as complete Freund's adjuvant (CFA). Typically 5-15 days later, the response is elicited by challenging the animal with the antigen, typically by intradermal injection. The response is assessed 24-48 h later, by assay methods such as size of redness and swelling at the site of injection, ear swelling, footpad swelling, tail swelling, accumulation of systemically-injected radiolabeled iododeoxyuridine, radiolabeled serum protein, or labeled inflammatory cells, at the challenge site. A peptide is typically considered immunogenic if a dose of about 10-200 μg per animal, and preferably about 25-100 μg per animal, is able to sensitize the animal. In addition, in a sensitized animal, a challenge dose of about 1-100 μg, and preferably about 5-50 μg, would be expected to elicit a response. It is to be understood that a peptide derived from a human T cell receptor may not be immunogenic in an animal model; it is preferable that the peptide used in animal testing be derived from the analogous animal T cell receptor.

Typical animal disease model systems for human autoimmune diseases include systemic lupus erythematosus in susceptible mice; myasthenia gravis in SJL/J female mice induced with soluble AChR protein from another species; arthritis in a susceptible mouse strain induced by injection of Type II collagen; adjuvant arthritis in susceptible rats induced by injection of mycobacterial heat shock protein; thyroiditis in mice induced by administration of thyroglobulin; insulin-dependent diabetes mellitus, either naturally occurring or induced in certain mouse strains; MS in mice and rats by inducing EAE by administration of myelin basic protein, proteolipid protein, or Theiler's virus. The above list is not exclusive.

In any embodiment, it is preferred the T cell receptor peptide comprises a minimal epitope structure. A minimal epitope structure will not only make the peptide immunogenic, but also will raise antibodies specific to the T cell receptor. T cell activation marker peptides useful in the present invention can be derived from any

T cell activation marker involved in an autoimmune disease for which treatment is desired. T cell activation markers are described by Sakaguchi et al., J Autoimmun. 9, 211-220 (1996);

Lund et al., Immunol. Rev. 161, 79-93 (1998); van de Stolpe et al., J. Mol. Med. 74, 13-33

(1996); and Van Gool et al., Immunol. Rev. 153, 47-83 (1996).

The at least one T cell activation marker peptide has an amino acid sequence comprising from about 5 to about 50 amino acids, more preferably from about 10 to about 40, and most preferably from about 15 to about 30 amino acids, of the amino acid sequence of a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 α subunit (SEQ ID NO:37), LFA-1 β subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), CD16

(SEQ ID NO:33), or other T cell activation marker. Preferably the T cell activation marker peptide is derived from the extracellular domain of the T cell activation marker, and more preferably from the active site. Specifically, for CD38, the activation marker peptide preferably overlaps, lies within, or corresponds to amino acids

123-151 of the wild-type CD38 sequence. For ICAM-1, the activation marker peptide preferably overlaps, lies within, or corresponds to at least one of the five Ig-like extracellular domains.

Alternatively, instead of a T cell activation marker peptide, a functional derivative thereof may be used, in ways similar to those described for functional derivatives of T cell receptor peptides above.

Preferably, the T cell activation marker is characteristic of the same T cell mediated disease as of which the T cell receptor peptide is characteristic. Therefore, the T cell activation marker is preferably of human origin, and the preferred T cell mediated diseases are as described above.

The T cell activation marker peptide or functional derivative thereof can be produced, and its efficacy tested, by techniques described above with regard to T cell receptor peptides above.

A pharmaceutical composition of the present invention comprises a T cell receptor peptide or functional derivative thereof and a T cell activation marker peptide or functional derivative thereof, as described above, and also at least one pharmaceutically acceptable carrier.

Carriers include excipients and auxiliaries which facilitate processing of the active compounds into preparations for pharmaceutical use. The compositions can also include adjuvants, such as alum or other adjuvants known in the art. To enhance delivery or bioactivity, the peptides can be incorporated into liposomes, using techniques and compounds known in the art. The immunogenicity of one or more of the peptides can be enhanced by including it in a longer peptide or chain or by conjugating it to an immunological carrier such as KLH, serum albumin, tetanus toxoid, and the like. If intended for oral administration as a tablet or capsule, rectal administration as a suppository, or injection or oral consumption as a solution, the composition can contain from about 0.001 to about 99 percent, and preferably from about 0.01 to about 95 percent, of the active compounds. Excipients that can be used will depend on the intended route of administration, and their selection will be routine to the skilled artisan. In a subject afflicted with, or susceptible to, an immune-related disease, introduction of a composition comprising a T cell receptor peptide or functional derivative thereof and a T cell activation marker peptide or functional derivative thereof results in generation of an immune response directed to the T cell receptor peptide and T cell activation marker and protection from the immune-related disease. Preferably, the T cell mediated disease is of human origin. More preferably, the T cell mediated disease is rheumatoid arthritis, myasthenia gravis, encephalomyelitis, multiple sclerosis, thyroiditis, diabetes, inflammatory bowel disease, or systemic lupus erythematosus. Most preferably, the T cell mediated disease is multiple sclerosis (MS).

For measuring the protective benefit of administration of compositions of the present invention to humans, certain well-known clinical outcome measures can be used. For example, in MS, such measures include (a) clinical disability, (b) on-study exacerbation rate, (c) MRI of brain plaque load, and (d) neuropsychological measures of cognitive impairment. Clinical disability is typically measured by the McAlpine scale, the Kurtzke score, or the modified Kurtzke score termed the expanded disability status score (EDSS). The above clinical outcome measures are known to the skilled artisan.

The pharmaceutical compositions of the present invention can be administered to any animal which may benefit therefrom. Preferably, the animal is a human.

The pharmaceutical compositions can be administered through any appropriate route. Administration may be parenteral, subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, transdermal, buccal, or oral. The compositions can be administered parenterally by bolus injection or by gradual perfusion over time. The dose administered will depend on the age, sex, health, and weight of the recipient, the kind of concurrent treatment if any, the frequency of treatment, and the desired effect of administration. The doses should be large enough to produce an immune response to the peptides, but not so large as to cause adverse side effects, such as generalized immunosuppression and anaphylactic reactions, among others. For humans, preferred doses are in the range of about 0.001-25 mg/kg body weight.

In addition to the use of T cell peptides for active immunization, in a further embodiment the present invention uses the T cell peptides to effect passive immunity. In one technique, T cells can be extracted from an actively immunized animal and transferred to an unimmunized animal, either directly or after growth in culture. Antibodies, either chimeric, polyclonal or monoclonal, against T cell receptor peptides and T cell activation marker peptides can be generated using techniques known in the art. The antibodies can be conjugated to cytotoxic proteins, such as ricin A, Pseudomonas toxin, Diphtheria toxin, tumor necrosis factor; to radionuclides such as ²¹²Bi, ¹³¹1, ¹⁸⁶Re, and ⁹⁰Y; or to cytotoxic drugs such as daunorubicin, doxorubicin, methotrexate, and mitomycin C. None of the lists of conjugates are to be construed as limiting. Details of treatment, such as doses and routes of administration, will be known to one skilled in the art.

The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. Example 1

A composition effective for the treatment of MS is produced as follows. The gene encoding the T cell receptor specific for MS is cloned from a population of T cells, e.g. by isolation of mRNA and production of cDNA therefrom using reverse transcriptase. The gene is then ligated into a plasmid or other cloning vehicle, a portion of the gene encoding either the CDR2 or CDR3 is subcloned therefrom and ligated into a second cloning vehicle and into an operable linkage with control sequences effective for expression in a given microorganism. The resulting vehicle is then transformed into a suitable microorganism, e.g. Escherichia coli. Likewise, genes encoding the T cell activation markers CD38, CD25, LFA-1, ICAM-1, and CD 16 are cloned from a population of T cells, and regions thereof encoding the extracellular domains are subcloned and transformed into a suitable microorganism. The T cell receptor peptides and T cell activation marker peptides are then expressed following fermentation methodologies known in the art.

The expressed peptides are then isolated and purified, and pharmaceutical compositions are made. For a first group of compositions, each composition comprises one T cell receptor peptide (derived from either CDR2 or CDR3 of a T cell receptor specific for MS) and one T cell activation marker peptide (derived from extracellular domain of CD38, CD25, LFA-1, ICAM-1, or CD 16).

A second group of compositions consists of compositions comprising one T cell receptor peptide and two or more T cell activation marker peptides, derived as given above. A third group of compositions consists of compositions comprising one T cell receptor peptide and five T cell activation marker peptides, one derived from each T cell activation marker given above.

In all cases, each composition further comprises an isotonic aqueous solution suitable for solubilizing the peptides and administration to humans.

Patients suffering from MS then receive an injection of at least one composition described above in order to elicit an immunogenic response. Booster injections are made if deemed necessary by the clinician. If the most efficacious composition is known from clinical trials, that composition is used.

A result of using a composition, or the most efficacious composition if known, is a reduction in the EDSS for the majority of patients, and a significant reduction (greater than 0.5 on the Kurzke scale) for at least one-third of patients, at about 12 months after treatment. An additional result is a reduction of at least 50% in the rate of clinical relapse in the 12 months after treatment relative to the 12 months before treatment. These results are comparable to those reported for vaccination with irradiated T cells (Zhang et al.. manuscript in preparation).

All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

CLAIMS;

1. A composition, comprising: at least one T cell receptor peptide; and, at least one T cell activation marker peptide.

2. The composition of claim 1, wherein the T cell receptor peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the second complementarity determining region (CDR2) or the third complementarity determining region (CDR3), or a functional derivative thereof.

3. The composition of claim 1, wherein the T cell receptor is of human origin.

4. The composition of claim 1, wherein the T cell receptor peptide is of a T cell receptor characteristic of a T cell mediated disease.

5. The composition of claim 4, wherein the T cell mediated disease is rheumatoid arthritis, myasthenia gravis, encephalomyelitis, multiple sclerosis, thyroiditis, diabetes, inflammatory bowel disease, or systemic lupus erythematosus.

6. The composition of claim 5, wherein the T cell mediated disease is multiple sclerosis.

7. The composition of claim 2, wherein CDR2 or CDR3 is from a T cell receptor ╬▓ chain or ╬▒ chain.

8. The composition of claim 2, wherein the amino acid sequence of the T cell receptor peptide overlaps that of the CDR2 or CDR3.

9. The composition of claim 2, wherein the amino acid sequence of the T cell receptor peptide lies within that of the CDR2 or CDR3.

10. The composition of claim 2, wherein the amino acid sequence of the T cell receptor peptide corresponds to that of the CDR2 or CDR3.

11. The composition of claim 1 , wherein the T cell receptor peptide is a variant.

12. The composition of claim 1, wherein the T cell receptor peptide is a chemical derivative.

13. The composition of claim 1, wherein the T cell receptor peptide comprises a minimal epitope structure.

14. The composition of claim 1, wherein the T cell receptor peptide is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 1-32.

15. The composition of claim 1, wherein the T cell activation marker peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), CD 16 (SEQ ID NO:33), or other T cell activation markers.

16. The composition of claim 15, wherein the T cell activation marker peptide is derived from CD38.

17. The composition of claim 15, wherein the T cell activation marker peptide is derived from CD25.

18. The composition of claim 15, wherein the T cell activation marker peptide is derived from LFA-1 ╬▒ subunit or ╬▓ subunit.

19. The composition of claim 15, wherein the T cell activation marker peptide is derived from ICAM-1.

20. The composition of claim 15, wherein the T cell activation marker peptide is derived from CD16.

21. The composition of claim 15, further comprising a T cell activation marker peptide derived from a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 a subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), CD16 (SEQ ID NO:33), or other T cell activation markers.

22. The composition of claim 1 , comprising T cell activation marker peptides derived from CD38 (SEQ ID NO:35). CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38). ICAM-1 (SEQ ID NO:36), and CD16 (SEQ ID NO:33).

23. The composition of claim 1, wherein the T cell receptor peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the second complementarity determining region (CDR2) or the third complementarity determining region (CDR3) of a T cell receptor of human origin and characteristic of a T cell mediated disease, and wherein the T cell activation marker peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), or CD16 (SEQ ID NO:33).

24. The composition of claim 1 , wherein the T cell receptor peptide and the T cell activation marker peptide are present in a therapeutically effective amount, and further comprising a pharmaceutically-acceptable excipient.

25. A method for treating a T cell mediated disease in a mammal, comprising: administering to the mammal a composition comprising at least one T cell receptor peptide, and at least one T cell activation marker peptide.

26. The method of claim 25, wherein the T cell receptor peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the CDR2 or CDR3.

27. The method of claim 25, wherein the T cell receptor is characteristic of the T cell mediated disease, and the mammal is a human.

28. The method of claim 25, wherein the T cell mediated disease is rheumatoid arthritis, myasthenia gravis, encephalomyelitis, multiple sclerosis, thyroiditis, diabetes, inflammatory bowel disease, or systemic lupus erythematosus.

29. The method of claim 25, wherein the T cell mediated disease is multiple sclerosis.

30. The method of claim 26, wherein the CDR2 or CDR3 is from a T cell receptor ╬▓ chain or ╬▒ chain.

31. The method of claim 26, wherein the amino acid sequence of the T cell receptor peptide overlaps that of the CDR2 or CDR3.

32. The method of claim 26, wherein the amino acid sequence of the T cell receptor peptide lies within that of the CDR2 or CDR3.

33. The method of claim 26, wherein the amino acid sequence of the T cell receptor peptide corresponds to that of the CDR2 or CDR3.

34. The method of claim 25, wherein the T cell receptor peptide comprises a minimal epitope structure.

35. The method of claim 25, wherein the T cell receptor peptide is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 1-32.

36. The method of claim 25, wherein the T cell receptor peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of the second complementarity determining region (CDR2) or the third complementarity determining region (CDR3) of a T cell receptor of human origin and characteristic of a T cell mediated disease, and wherein the T cell activation marker peptide is derived from a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37). LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), or CD16 (SEQ ID NO:33).

37. The method of claim 25, wherein the T cell activation marker peptide has an amino acid sequence comprising from about 15 to about 30 amino acids of a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), CD16 (SEQ ID NO:33), or other T cell activation markers.

38. The method of claim 37, wherein the T cell activation marker peptide is derived from CD38.

39. The method of claim 37, wherein the T cell activation marker peptide is derived from CD25.

40. The method of claim 37, wherein the T cell activation marker peptide is derived from LFA-1 ╬▒ subunit or ╬▓ subunit.

41. The method of claim 37, wherein the T cell activation marker peptide is derived from ICAM-1.

42. The method of claim 37, wherein the T cell activation marker peptide is derived from CD16.

43. The method of claim 37, further comprising a T cell activation marker peptide derived from a T cell activation marker selected from CD38 (SEQ ID NO:35), CD25 (SEQ ID NO:34), LFA-1 ╬▒ subunit (SEQ ID NO:37), LFA-1 ╬▓ subunit (SEQ ID NO:38), ICAM-1 (SEQ ID NO:36), CD16 (SEQ ID NO:33), or other T cell activation markers.

44. The composition according to any of claims 1-24 for use in treating a T cell mediated disease in a mammal.

45. The composition according to any of claims 1-24 for use in treating rheumatoid arthritis, myasthenia gravis, encephalomyelitis, multiple sclerosis, thyroiditis, diabetes, inflammatory bowel disease, or systemic lupus erythematosus.

46. The composition according to any of claims 1-24 for use in treating multiple sclerosis.