WO1994027634A1 - Cryptic peptides for use in inducing immunologic tolerance - Google Patents
Cryptic peptides for use in inducing immunologic tolerance Download PDFInfo
- Publication number
- WO1994027634A1 WO1994027634A1 PCT/AU1994/000292 AU9400292W WO9427634A1 WO 1994027634 A1 WO1994027634 A1 WO 1994027634A1 AU 9400292 W AU9400292 W AU 9400292W WO 9427634 A1 WO9427634 A1 WO 9427634A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- genus
- peptide
- allergen
- composition
- protein antigen
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/35—Allergens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0008—Antigens related to auto-immune diseases; Preparations to induce self-tolerance
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- Feeding antigens has been a classical method for inducing immunological unresponsiveness or oral tolerance (Asherson, G. L., et al., (1977), Cell Immunol, 33:145; Asherson, G. L., et al. (1979), Immunology, 36:449; Challacombe, S. J., and Tomasi, T.J., (1980), J. Exp. Med., 152:1459; Bruce, M. G., and Ferguson, A. (1986), Immunology, 57:627; Mowat, A.
- MBP myelin basic protein
- Ts suppresser cells through the release of a soluble cytokine were reported to be able to inhibit the in vitro response of a MBP-specif ⁇ c CD4 + T cell line and could also bring about a by-stander suppression of unrelated T cells (Miller, A., (1991), cited supra).
- Peptides derived from a variety of protein antigens including bacterial and viral pathogens, autoantigens, allergens and other experimental antigens such as hen egg lysozyme (HEL), ovalbumin (OVA) and lambda repressor (cl) have been examined for the ability to stimulate antigen-specific T cells.
- HEL hen egg lysozyme
- OVA ovalbumin
- cl lambda repressor
- a major mycobacterial antigen 65-kD protein has also been epitope-mapped (Lamb, J.R. et al, (1987) EMBO J., 6(5): 1245-1249). T cell epitopes have been identified in the peptides comprised of amino acid residues 112-132 and 437-459 of the 65-kD protein. MBP has also been epitope- mapped in both human (Ota, K. et al., (1990) Nature, 346:183-187 ) and rodent (Zamvil et al., (1986) Nature, 324:258-260) systems.
- T cell epitopes present in allergenic proteins have very recently been described (O ⁇ ehir, R. et al., (1991) Ann. Rev. Immunol, 9:67-95 ).
- Several peptides derived from the house dust mite allergen Der p I have been shown to be T cell-reactive (Thomas, W.R., et al. In Epitopes of Atopic Allergens Proceedings of Workshop from XIV Congress of the European Academy of Allergy and Clinical Immunology, Berlin (Sept. 1989) pp. 77-82;
- Perez el __ ⁇ _ demonstrated that T cell epitopes are contained within amino acid residues 191-210 of the protein allergen Lol p I (Perez, M. et al., (1990) J. Biol. Chem. 265(27): 16210- 16215 .
- compositions which include a cryptic peptide derived from a protein antigen, such as an allergen or autoantigen, can be administered to induce tolerance in a naive or pre-sensitized individual.
- the composition is administered orally to treat sensitivity in an individual to an allergen or autoantigen.
- Figure 7 is a graphic representation of the responses of T cells isolated from mice immunized with Der p I and analyzed for response to selected peptides derived from Der p I by tritiated thymidine incorporation.
- Figure 2a and 2b are graphic representations of the responses of T cells isolated from mice immunized with a selected peptide derived from Der p I and analyzed for response to either Der p I protein (panel a) or the appropriate peptide (panel b).
- Figure 5 is a schematic representation of the location of T cell epitopes recognized by mice in the Der p I protein sequence where immunodominant epitopes are represented with hatched squares, cryptic epitopes are represented by dotted squares and the absence of epitopes is represented by black squares.
- Figure 4 is a graphic representation of the responses of T cells isolated from mice fed with buffer (panel a), peptide GEX p57-130 (panel b), peptide GEX pi 01 -154 (panel c), or recombinant protein, GEX Der p I (1-222) (panel d) followed by immunization with Der I and analyzed for response to Der p I in vitro by IL-3/GM-CSF (panels a-d) or IL-2 production (panels e-h).
- Figure 5 is a graphic representation of the responses of T cells isolated from mice fed recombinant protein GEX Der p II (1-129), peptide GEX plOl-154, or peptide GEX pl88- 222 followed by immunization with Der p I and analyzed for response to Der p I (panel a and d), peptide p 110- 131 (panel b and e), or peptide ⁇ 78- 100 (panel c and f) by IL-3/GM-CSF (panels a-c) or IL-2 production (panels d-f).
- Figure 6 is a graphic representation of the responses of T cells isolated from mice fed with either buffer or recombinant fusion peptide (GEX pi 31-187) followed by immunization with Derp I and analyzed for response to Der p I by IL-2 production.
- GEX pi 31-187 buffer or recombinant fusion peptide
- This invention pertains to methods for inducing immunologic tolerance to a protein antigen in a subject by administering at least one cryptic peptide derived from the antigen.
- Protein antigens are known to contain certain determinants or epitopes which, upon presentation with a particular class II major histocompatibility (MHC) molecule will activate T cells of a subject upon exposure to the native protein antigen. Rather than the T cell response being limited by the presence of one or two determinants on an antigen, it appears that the T cell response preferentially utilizes a selected number of determinants. Thus, a hierarchy of T cell determinant usage exists for a multideterminant protein antigen.
- MHC major histocompatibility
- the T cell determinants or epitopes for a particular protein antigen can be divided into categories based on in vitro T cell proliferation assays in which protein antigen- primed T cells are cultured with a selected concentration of a peptide derived from the protein antigen and the amount of proliferation by the T cells in response to the peptide is determined by, for example, tritiated thymidine incorporation.
- a peptide is categorized as comprising an immunodominant T cell epitope if the peptide consistently induces one of the highest T cell proliferative responses in antigen-primed T cells in the subject tested.
- Relative to an immunodominant epitope a peptide which comprises a minor T cell epitope recalls in vitro T cell proliferation to a more variable and lesser extent.
- Those peptides which recall T cell proliferation of less than 2 fold the background level of media alone are categorized as either not comprising a T cell epitope or comprising a cryptic T cell epitope.
- Cryptic epitopes are those determinants in a protein antigen which, due to processing and presentation of the native protein antigen to the appropriate MHC molecule, are not normally revealed to the immune system.
- a peptide comprising a cryptic epitope is capable of tolerizing T cells, and when a subject is primed with the peptide, T cells obtained from the subject will proliferate in vitro in response to the peptide or the protein antigen from which the peptide is derived.
- Peptides which comprise at least one cryptic epitope derived from a protein antigen are referred to herein as cryptic peptides.
- antigen-primed T cells are cultured in vitro in the presence of each peptide separately to establish peptide-reactive T cell lines.
- a peptide is considered to comprise at least one cryptic epitope if a T cell line can be established with a given peptide and T cells are capable of proliferation upon challenge with the peptide and the protein antigen from which the peptide is derived.
- the presence of cryptic epitopes in a protein antigen is due to a lack of exposure of certain epitopes to the immune system which may result from normal processing of the protein antigen which fails to reveal the epitope to the appropriate class II MHC molecule.
- the end product of antigen processing may be a large fragment which hides the cryptic epitope and hinders access to the MHC molecule or the T cell receptor on T cells specific for the epitope.
- other epitopes on the same protein antigen may compete with the cryptic epitope for binding to the same restriction element or may have a higher affinity and availability for a different restriction element, thus preventing cryptic epitope interaction with MHC.
- Cryptic peptides of the invention comprise at least one cryptic epitope derived from a protein antigen (i.e., the peptide comprises at least approximately 7 amino acid residues).
- Such peptides can comprise as many amino acid residues as desired and preferably comprise at least about 7, more preferably at least about 15, even more preferably at least about 20 and most preferably at least about 25 amino acid residues of a protein antigen.
- a peptide length of about 20-40 amino acid residues is preferred as increases in length of a peptide may result in difficulty in peptide synthesis as well as retention of an undesirable property (e.g., immunoglobulin binding or enzymatic activity) due to maintenance of conformational similarity between the peptide and the protein antigen, such as an allergen from which it is derived.
- the amino acid sequences of one or more peptides can be produced and joined by a linker to increase sensitivity to processing by antigen-presenting cells.
- Such linker can be any non-epitope amino acid sequence or other appropriate linking or joining agent.
- two cryptic peptides can be joined or a cryptic peptide and a peptide comprising an immunodominant or minor epitope derived from the protein antigen can be linked.
- Cryptic peptides can be produced by recombinant DNA techniques in a host cell transformed with a nucleic acid vector directing expression of a nucleotide sequence coding for such peptide, or by chemical synthesis, or in certain limited situations by chemical cleavage of protein antigen such as an allergen.
- host cells transformed with nucleic acid vectors directing expression of a nucleotide sequence coding for a peptide are cultured in a medium suitable for the cells.
- the peptides may be secreted and harvested from a mixture of cells and cell culture medium. Alternatively, the peptide may be retained cytoplasmically and the cells harvested, lysed and the peptide isolated and purified.
- Peptides can be isolated using techniques known in the art for purifying peptides or proteins including ion-exchange chromatography, gel filtration chromatography, ultrafiltration, electrophoresis, and immunoaffinity purification with antibodies specific for the peptide or the protein antigen from which the peptide is derived, or a portion thereof.
- the cryptic peptides described herein are isolated such that the peptide is substantially free of cellular material or culture medium when produced by recombinant DNA techniques, or substantially free of chemical precursors or other chemicals when synthesized chemically, or obtained by chemical cleavage of a protein allergen or other protein antigen.
- the protein structure of the antigen can be reviewed and the sequence divided into at least two peptide fragments of desired lengths.
- the protein sequence of a protein antigen can be systematically divided into at least two non- overlapping fragments of desired length or overlapping fragments of desired length.
- Dermatophagoides pteronyssinus having an amino acid sequence of 229 residues can be divided into peptide fragments of about 20-35 amino acid residues in length, with each fragment overlapping with another by about 10 amino acids.
- areas of overlap and length of each fragment can be designed to maintain the presence of T cell epitopes predicted using algorithms (Rothbard, J. and Taylor, W.R. (1988) EMBOJ. 7:93-100; and Berzofsky, J.A. (1989) Philos. Trans. R. Soc. Lond. 323:535-544).
- human T cell epitopes within a protein antigen can be predicted using known HLA class II binding specific amino acid residues.
- the resulting peptide fragments can be produced by recombinant DNA techniques or chemical synthesis.
- the peptide fragments derived from a protein antigen are tested to determine those fragments having T cell stimulating activity (i.e., proliferation, lymphokine secretion and/or induction of T cell anergy/tolerization) and thus comprise at least one T cell epitope.
- T cell stimulating activity i.e., proliferation, lymphokine secretion and/or induction of T cell anergy/tolerization
- human T cell stimulating activity can be tested by culturing T cells obtained from a subject, such as a human, sensitive to a protein antigen (i.e., a subject which has an immune response to the protein antigen) with a peptide fragment derived from the protein antigen and determining the presence of proliferation by T cells in response to the peptide.
- the presence of proliferation by T cells can be determined by, for example, uptake of tritiated thymidine.
- Immunodominant T cell epitopes can be identified as described in Example 3.
- T cells are obtained from an individual sensitive to the protein antigen and cultured with each of the cryptic peptides separately to establish peptide-reactive T cell lines.
- the presence of T cell proliferation or induction of T cell tolerance in response to the peptide and the protein antigen from which the peptide is derived confirms the presence of at least one cryptic epitope in the peptide.
- Cryptic peptides of the invention can be derived from a protein antigen such as an allergen or autoantigen.
- the cryptic peptide can be derived from any known protein allergen, such as an allergen of the following genus: the genus Dermatophagoides: the genus Felis: the genus Ambrosia: the genus Lolium: the genus Cryptomeria: the genus Alternaria: the genus Alder: the genus Betula: the genus Quercus: the genus Olea: the genus Artemisia: the genus Plantago: the genus Parietaria: the genus Canine: the genus Blattella: the genus Apis: the genus Periplaneta: and the genus Sorghum.
- Cryptic peptides recognized by mice in Der p I a major allergen of the species Dermatophagoides pteronyssinus. have been determined in mice and comprise amino acid residues 120-143 of Der p I (SEQ ID NO:l), amino acid residues 144-169 of Der p I (SEQ ID NO:l) and amino acid residues 131-187 of Der p I (SEQ ID NO:l).
- Cryptic peptides can also be derived from protein antigens other than allergens where immunologic tolerance to an autoantigen is desired.
- Autoantigens from which cryptic peptides can be derived include insulin, glutamic acid decarboxylase (64K), PM-1 and carboxypeptidase for use in treating diabetes; myelin basic protein for use in treating multiple sclerosis; rh factor for use in treating erythroblastosis fetalis; acetylcholine receptors for use in treating myasthenia gravis; thyroid receptors for use in treating Graves Disease; basement membrane protein for use in treating Good Pasture's syndrome; and thyroid proteins for use in treating thyroiditis.
- 64K glutamic acid decarboxylase
- PM-1 carboxypeptidase for use in treating diabetes
- myelin basic protein for use in treating multiple sclerosis
- rh factor for use in treating erythroblastosis fetalis
- acetylcholine receptors for use in treating myasthenia gravis
- thyroid receptors for use in treating Graves Disease
- basement membrane protein for use in treating Good Pasture'
- cryptic peptides derived from a protein antigen are administered to a subject to induce immunologic tolerance in the subject to the protein antigen.
- subject includes living organisms capable of mounting an immune response to a protein antigen, e.g., mammals. Examples of subjects include humans, rats, mice, dogs, cats, horses, cows and transgenic species thereof.
- Immunologic tolerance refers to a condition in a subject where a block in the development, growth or differentiation of specific lymphocytes in the subject results upon administration of a tolerizing amount of a cryptic peptide of the invention. Tolerance results from the interaction of antigen with antigen receptors on lymphocytes under conditions in which the lymphocytes, instead of becoming activated, are deleted or rendered unresponsive.
- Tolerance may also be due to the action of specific T or B lymphocytes or other regulatory mechanisms that prevent lymphocyte activation.
- One mechanism for inhibiting an immune response is the stimulation of a class of lymphocytes, called suppresser T cells, whose principal function is to suppress the activation of specific T and B lymphocytes. In this situation, inhibition is mediated not by the antigen itself but by regulatory cells that are induced by the antigen.
- Another proposed mechanism for tolerance is a response by the immune system to antigen in which unique or idiotypic determinants of lymphocytes or antibodies specific for the antigen are targeted. This response results in a network of complementary idiotypes and antiidiotypes which block the stimulation of antigen-specific cells.
- the products of activation of B and T lymphocytes namely antibodies and cytokines, respectively, are themselves capable of regulating specific immunity to result in tolerance in addition to functioning as the principle effector molecules of lymphocytes.
- a tolerizing amount of a cryptic peptide derived from a protein antigen is administered to the subject.
- a tolerizing amount is defined as a dosage of cryptic peptide necessary to induce immunologic tolerance in a subject, such as a human to the antigen from which the cryptic peptide is derived.
- Immunologic tolerance in a subject is indicated by non-responsiveness or diminution in symptoms to the protein antigen, such an an allergen or autoantigen, as determined by standard clinical procedures (see e.g., Varney et aL, (1990) British Medical Journal 302:265- 269). When tolerance to an allergen is sought, such non-responsiveness includes diminution in allergen induced allergic symptoms.
- a diminution in symptoms to an allergen includes any reduction in the allergic response of a subject, such as a human, to the allergen following a treatment regimen with a cryptic peptide as described herein.
- This diminution in symptoms may be determined subjectively in a human (e.g., the patient feels more comfortable upon exposure to the allergen), or clinically, such as with a standard skin test.
- Cryptic peptides derived from a protein antigen are typically administered to a subject in the form of a composition which includes a pharmaceutically acceptable carrier or diluent.
- Administration of a composition of the present invention to induce immunologic tolerance in a subject to a protein antigen can be carried out using known procedures, at dosages and for periods of time effective to tolerize the subject to the protein antigen. Effective amounts of the composition will vary according to factors such as the degree of sensitivity of the subject to the antigen, the age, sex, and weight of the subject, and the ability of the cryptic peptide(s) to induce tolerance in the subject. Dosage periods may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
- Cryptic peptides may be administered to a subject in a convenient manner such as by injection (subcutaneous, intravenous, etc.), oral administration, inhalation, intranasal, transdermal application, or rectal administration.
- Preferred routes of administration to induce immunologic tolerance in a subject are oral and intranasal administration. See O ⁇ ehir, R.E. et al. (1993) Eur. J. Clin. Invest. 23(12): 763-772).
- the active compound i.e., the cryptic peptide
- the active compound i.e., the cryptic peptide
- the active compound may be coated with in a material to protect the compound from the action of enzymes, acids and other natural conditions which may inactivate the compound.
- a cryptic peptide or peptides by enteral administration, it may be necessary to coat the peptide with, or co-administer the peptide with, a material to prevent its inactivation.
- the cryptic peptide may be administered to a subject in an appropriate diluent, co-administered with enzyme inhibitors or in an appropriate carrier such as liposomes.
- Pharmaceutically acceptable diluents include saline and aqueous buffer solutions.
- Enzyme inhibitors include pancreatic trypsin inhibitor, diisopropylfluorophosphate (DEP) and trasylol.
- Liposomes include water-in-oil-in-water CGF emulsions as well as conventional liposomes (Strejan et al., (1984) J Neuroimmunol. 7:27).
- the composition is preferably administered in non-immunogenic form, e.g., one that does not contain adjuvant.
- the active compound may also be administered parenterally.
- Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
- compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
- the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
- the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
- the proper fluidity can be maintained, for example, by the use of a coating such as licithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
- Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, asorbic acid, thimerosal, and the like.
- isotonic agents for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition.
- Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
- Sterile injectable solutions can be prepared by incorporating active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
- dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above.
- the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient (i.e., a peptide of the invention) plus any additional desired ingredient from a previously sterile-filtered solution thereof.
- the peptide may be orally administered, for example, with an inert diluent or an assimilable edible carrier.
- the peptide and other ingredients may also be enclosed in a hard or soft shell gelatin capsule, compressed into tablets, or incorporated directly into the individual's diet.
- the active compound may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
- Such compositions and preparations should contain at least 1% by weight of active compound.
- compositions and preparations may, of course, be varied and may conveniently be between about 5 to about 80% of the weight of the unit.
- the amount of active compound in such compositions is such that a suitable dosage will be obtained.
- Preferred compositions or preparations according to the present invention are prepared so that an oral dosage unit contains between from about 10 mg to about 200 mg of active compound.
- pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the composition is contemplated. Supplementary active compounds can also be incorporated into the compositions.
- Dosage unit form refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired effect in association with the required pharmaceutical carrier.
- the specification for the dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of the subject.
- compositions of the invention can include one or more cryptic peptides or a cryptic peptide and a peptide comprising an immunodominant or minor epitope which are administered to subjects, such as humans, who are naive or pre-sensitized to the protein antigen from which the peptide is derived, at dosages and for lengths of time effective to induce tolerance in the subject to the antigen.
- subjects such as humans
- an amount of one or more of the same or of different compositions effective to induce tolerance in a subject can be administered simultaneously or sequentially.
- a composition comprising at least two peptides e.g., a physical mixture of at least two peptides
- a cryptic peptide and a peptide comprising an immunodominant epitope can be co-administered.
- a cryptic peptide of the invention i.e., a peptide which does not contain an epitope recognized during immunization when the entire protein antigen is presented to a subject
- Peptides effective in immunotherapy may therefore not simply be limited to those identified by T-cell clones or polyclonal responses of sensitized individuals.
- Administration of a cryptic peptide may avoid the potential limitations inherent in administering a peptide containing immunodominant epitopes to sensitized individuals.
- the use of cryptic peptides also offers the potential for modifying immune responses without having to redirect the development of T-cell clones which have already progressed along T ⁇ l and T ⁇ 2 or equivalent pathways.
- a modified peptide can be produced in which the amino acid sequence has been altered, such as by amino acid substitution, deletion, or addition, to modify the ability of the peptide to induce tolerance, or to which a component has been added for the same purpose.
- a peptide can be modified so that it maintains the ability to induce T cell anergy or tolerance and bind MHC proteins.
- critical binding residues for the T cell receptor i.e., the amino acid residues which comprise the cryptic epitope
- those residues shown to be essential can be modified by replacing the essential amino acid with another, preferably similar amino acid residue (a conservative substitution) whose presence is shown to enhance, diminish, but not eliminate or affect T cell reactivity.
- amino acid residues which are not essential for T cell interaction can be modified by being replaced by another amino acid whose incorporation may enhance, diminish or not affect T cell reactivity, but not eliminate binding to relevant MHC.
- Preferred amino acid substitutions for non-essential amino acids include, but are not limited to substitutions with alanine, glutamic acid or a methyl amino acid.
- Another example of a modification of peptides is substitution of cysteine residues preferably with alanine, or glutamic acid, or alternatively with serine or threonine to minimize dimerization via disulfide linkages.
- peptides can also be modified to incorporate one or more polymorphisms in the amino acid sequence of a protein antigen resulting from natural allelic variation.
- D-amino acids, non-natural amino acids or non-amino acid analogues can be substituted or added to produce a modified peptide within the scope of this invention.
- peptides can be modified using the polyethylene glycol (PEG) method of A. Sehon and co-workers (Wie et al. supra) to produce a peptide conjugated with PEG. Modifications of peptides can also include reduction/alkylation (Tarr in: Methods of Protein Microcharacterization, J.E. Silver ed.
- poly-histidine can be added to a peptide to purify the peptide on immobilized metal ion affinity chromatography (Hochuli, E. et al., (1988) Bio/Technology, 6:1321-1235).
- specific endoprotease cleavage sites can be introduced, if desired, between a reporter group and amino acid sequences of a peptide to facilitate isolation of peptides free of irrelevant sequences.
- canonical protease sensitive sites can be recombinantly or synthetically engineered within the peptide.
- charged amino acid pairs such as KK or RR
- the resulting peptide can be rendered sensitive to cathepsin and/or other trypsin-like enzymes cleavage to generate portions of the peptide containing one or more T cell epitopes.
- such charged amino acid residues can result in an increase in solubility of a peptide.
- Site-directed mutagenesis of DNA encoding a peptide can be used to modify the structure of the peptide. Such methods may involve PCR (Ho et al, (1989) Gene 77:51-59) or total synthesis of mutated genes (Hostomsky, Z., et al., (1989) Biochem. Biophys. Res. Comm. 161:1056-1063). To enhance bacterial expression, the aforementioned methods can be used in conjunction with other procedures to change the eucaryotic codons in DNA constructs encoding peptides to ones preferentially used in Jj . coli. yeast, mammalian cells or other eucaryotic cells.
- mice Female BIO and BALB congenic mice, and inbred C57BL/6J were purchased from the Animal Resource Centre, Murdoch, Western Australia at 6-8 weeks of age.
- the house dust mite allergen Der p I was affinity purified from spent mite medium (SMM) using previously described techniques (Hoyne, G.F. et al. (1993) cited supra; Lombardo et al. J. Immunol. 144:1353-1360 and Chapman (1989) Advances in Biosciences 74:281-295).
- Ovalbumin (OVA) crystalline Grade V was purchased from the Sigma Chemical Company, St. Louis, MO. Overlapping synthetic peptides derived from the published Der p I sequence (Chua et al. (1988) J. Exp. Med.
- peptides used in this study comprised the following amino acid residues derived from the Der p I sequence (Chua et al. (1988) cited supra): 1-20, 13- 39, 21-49, 40-60, 50-71, 61-84, 78-100, 85-109, 101-119, 110-131, 120-143, 132-157, 144- 169, 158-180, 170-191, 181-204, 197-222.
- Bacterial pellets were resuspended in tris buffered saline with 1 mM EDTA and transferred to a homogenizing bottle containing 0.1 mm glass beads and were homogenized using a Braun MSK Homogenizer for five minutes. The lysate was removed after ultracentrifugation at 10,000 g for 10 minutes at 4°C. The pellet was washed twice with 1.75 M guanidine HCL containing 1 M NaCl and 1% triton-X 100 (BDH Chemicals) by thoroughly aspirating in a pipette and then centrifugation.
- BDH Chemicals triton-X 100
- the pellet was then dissolved by incubating it in 8 M urea with 50 mM NaCl and 1 mM ethylene diamine tetraacetic acid (EDTA) for 2 hours at 37°C.
- the sample was dialyzed in 3- (cyclohexylamino)-propanesulfonic acid (CAPS) buffer pH 10.7 and the pH was slowly adjusted to pH 9.6.
- CAPS 3- (cyclohexylamino)-propanesulfonic acid
- the recombinant material was then clarified by centrifugation at 10 000 g and the concentration of the soluble material was estimated against standard quantities of bovine serum albumin (BSA) using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and staining with Coomassie blue.
- BSA bovine serum albumin
- SDS-PAGE SDS-polyacrylamide gel electrophoresis
- Recombinant peptides used in this study included GEX Der p I (amino acid residues 1-222), GEX Der p II (amino acid residues 1-129), GEX pi -14 (amino acid residues 1-14 of Der p I), GEX p60-l 11 (amino acid residues 60-111 of Der p I), GEX p98-140 (amino acid residues 98-140 of Der p I), GEX pi 01 -154 (amino acid residues 101-154 of Der p I), GEX p57-130 (amino acid residues 57-130 of Der p I), GEX pl88-222 (amino acid residues 188-222 of Der p I).
- mice were lightly anesthetized under ether and fed intragastrically by a tube with 3 mg of protein or peptide on 3 consecutive days. Antigens were dissolved in CAPS buffer and administered in a volume of 0.2 ml. Mice were immunized subcutaneously at the base of tail 7 days after the last feed with 100 mg of native protein emulsified in complete Freund's Adjuvant (CFA) in a volume of 0.2 ml.
- CFA complete Freund's Adjuvant
- Lymph node cells were cultured in Dulbecco's Modified Eagles medium (DME) supplemented with 2% fetal calf serum (FCS), 50 mM, 2-ME, 2 mM L-Glutamine and 20 mg/ml gentamycin.
- DME Dulbecco's Modified Eagles medium
- FCS fetal calf serum
- FDC-P1 cells were maintained in DME + 5% FCS
- CTLL-2 cells Krillis, S. (1978) J. Immunol. 120:20
- RPMI Rosewall Park Memorial Institute
- the periaortic and inguinal lymph nodes were collected from immunized mice and single cell suspensions were prepared by expressing the nodes through a stainless steel wire mesh. Cells were washed and cultured at 4 x 10-* * cells in a volume of 0.2 ml in DME culture medium in a 96 well flat bottom tissue culture plate. Protein or peptide antigens were added at various concentrations and the cells were incubated at 37°C for 24 hours. Supematants were collected and stored at -20°C until required. The Der p I used for all in vitro assays was the allergen isolated from spent mite medium (SMM).
- SMM spent mite medium
- FDC-P1 cells proliferate maximally in response to IL-3 and GM-CSF and submaximally to IFN- ⁇ or IL-4 (Kelso, A. (1990) cited supra). 2x10*-* cells were added in 50 ⁇ l DME + 5% FCS to 50 ⁇ l of culture supernatant in 96 well flat bottom tissue culture plates.
- the cells were incubated for 40 hours at 37°C and then pulsed with 1 ⁇ Ci - ⁇ H-Thymidine for another 4-6 hours at 37°C.
- the cells were then harvested onto glass fiber filter mats and samples counted for ⁇ H-Thymidine incorporation using liquid scintillation spectrometry or for latter experiments due to its acquisition, on a Packard matrix 9600 direct beta counter
- CTLL-2 cell line will proliferate maximally with IL-2 but only poorly in the presence of IL-4 (Kelso, A. (1990) J. Immunol. 145:2167). Supematants were cultured with 5000 CTLL-2 cells per well for 24 hours at 37°C and pulsed with 1 ⁇ Ci of 3 H-thymidine
- H2 D mice are high responders to Der p I while H2- ⁇ , _ ⁇ and H2*l mice are low responders (Hoyne, G. (1992) Ph.D. Thesis, T cell Recognition During Mucosal and Systemic Responses, University of Western Australia).
- B10 mice were immunized subcutaneously with 100 ⁇ g of Der p I in CFA and after 8 days the periaortic and inguinal lymph nodes were examined for antigen specific lymphokine release (IL-3/GM-CSF) using a panel of overlapping peptides.
- IL-3/GM-CSF antigen specific lymphokine release
- mice were immunized with all the peptides and responses to Der p I and the immunizing peptide were measured in the presence of spleen adherent cells.
- Peptide pl20-143 amino acid residues 120-143 of Der p I
- peptide pl44-169 amino acid residues 144-169 of Der p I
- Figure 2a The results of Figure 2 show the mean IL-3/GM-CSF response of triplicate samples.
- the following peptides are shown in the Figure: peptide pi 20- 143 (D); peptide pi 44- 169 ( ⁇ ); peptide pl32-157 (O); and peptide pl58-180 (X).
- Peptides A number of recombinant peptides were generated by restriction enzyme digestion of Der p I cDNA. These fragments were cloned into the pGEX expression vector as described above and transformed into I eoji. The recombinant peptides chosen for use in this study were expressed as fusions attached to the glutathione-S-transferase protein of Schistosoma japonicum. The fusion proteins and peptides were solubilized from bacterial cell pellets and dialyzed into CAPS buffer pH 9.6. The recombinant peptides listed in Figure 3 were chosen on the basis of the known T-cell epitope data described above.
- Recombinant peptides were selected for the presence of immunodominant (hatched squares) or cryptic epitopes (dotted squares) or the absence of T cell epitopes (black squares) within the sequence.
- control peptides GEX pl-23 (amino acid residues 1-23 of Der p I) and GEX pi 88-222 (amino acid residues 197-222 of Der p I) did not contain any T cell epitopes.
- GEX p57-130 (amino acid residues 57-130 of Der p I) contained two epitopes while GEX pi 01 -154 (amino acid residues 101-154 of Der p I) and GEX p98-140 (amino acid residues 57-130 of Der p I) contains the single immunodominant epitope (amino acid residues 110-131), while GEX pl31-187 (amino acid residues 131-187 of Der p I) contains the cryptic epitopes.
- mice were fed 3 mg of fusion peptide on 3 consecutive days and after a further 7 days were immunized subcutaneously with native protein in CFA.
- In vitro lymphokine assays were then performed 7 days later using the periaortic and inguinal lymph nodes stimulated with either protein or synthetic peptides.
- Experiments were performed to show that feeding mice CAPS buffer or the recombinant GEX Der p I (1-222) fiision protein did not effect the IL-2 or IL-3/GM-CSF responses of mice to subcutaneous injection of OVA in CFA.
- mice were fed CAPS buffer ( Figure 4, panels a and e), while test animals received 3 mg on three consecutive days of either GEX Der p I (1-222) ( Figure 4 panels d and h) or the fusion peptides GEX p57-130 ( Figure 4, panels b and f) or GEX pl01-154 ( Figure 4, panels c and g).
- GEX Der p I (1-222)
- Figure 4 panels d and h the fusion peptides
- GEX p57-130 Figure 4, panels b and f
- GEX pl01-154 Figure 4, panels c and g
- mice were immunized with Der p I and the responses of draining lymph node cells were measured to the protein and peptides in vitro.
- the data shows the response for individual mice in each group at the following antigen concentrations: Der p I, 20 ⁇ g/ml; and peptide pi 10-131 and peptide p78-100, 10 ⁇ M.
- feeding mice either GEX pl88-222 or GEX Der p II (1-129) did not affect the capacity of their lymph node cells to secrete either IL-3/GM-CSF or IL-2 upon in vitro challenge with either protein or with the immunogenic peptides pi 10-131 or p78-100.
- mice were fed 3mg on three consecutive days of GEX ⁇ l31-187 (Figure 6 (•)) which contains the cryptic epitope found on peptide 144-169 while control mice were fed with CAPS buffer ( Figure 6 (D)).
- One week later mice were immunized with Der p I in CFA.
- Lymph node cells were cultured in vitro with Der p I and supematants assayed for IL- 2.
- Each data point in Figure 6 represents the mean response of 5 animals per group ⁇ standard deviation.
- the responses of cryptic peptide fed mice were statistically different (p ⁇ 0.05 t-test).
- lymph node cells from control mice showed strong responses to Der p I in vitro by secreting IL-2, but mice fed the cryptic epitope displayed much weaker lymphokine response in vitro.
- This effect may therefor be mediated by a soluble factor.
- the inhibition was otherwise specific because it could not be induced by the Der p II fusion protein. Similar data has been obtained by Miller, A., et al., (1991), J. Exp. Med, 174:791; Whitacre, C. C, et al, (1991), J. Immunol, 147:2155; and Miller, A., et al., (1992), Proc. Natl. Acad. Sci. USA, 89:421, who found that oral tolerance to MBP was mediated by TGF-B1 and could be shown to suppress bystander responses in an in vitro model.
- Feeding two fusion proteins that did not contain T-cell epitopes did not inhibit the immune responses. However, feeding the fusion peptide GEX p 131 - 187 which contained the cryptic epitope found in peptide pi 44- 169 did significantly inhibit. The degree of inhibition was not as marked as for the fusions containing dominant epitopes but presumably could be increased by extending the feeding regime or increasing the dose. Feeding the fusion peptides was also found to sensitize T cells in the MLN so they release GM-CSF on stimulation in vitro with Der p I or synthetic peptides including the cryptic peptide pi 44- 169 after feeding peptide GEX pi 31-187. The presence of these sensitized cells in oral tolerance has recently been described for OVA (Hoyne, G. F., et al., (1993), Immunology 78:534-540).
- a T cell line can be established by culturing mite-allergic patient peripheral blood white cells in complete medium at 2 x 10-Vml in the presence of 20 ⁇ g purified native Derp I/ml. After 7 days of culture at 37°C in a humidified CO2 incubator the viable cells can be isolated by centrifugation with lymphocyte separation medium (LSM, Organon Technica, Durham, NC) and cultured in complete medium containing recombinant IL-2 and recombinant IL-4 for 2-3 additional weeks.
- lymphocyte separation medium LSM, Organon Technica, Durham, NC
- a secondary proliferation assay can be performed by culturing 2 x 10 ⁇ T cells in 200 ⁇ l complete medium with 5 x 10 ⁇ gamma-irradiated (3500 Rads) peripheral white blood cells as antigen presenting cells in the presence of various concentrations of peptides derived from the intact protein. The cultures can then pulsed with tritiated thymidine (1 ⁇ Ci/well) on day 3 and harvested onto glass fiber filters on day 4.
- Peptides stimulating tritium incorporation at least 2-fold over the medium control are defined as containing T cell epitopes naturally exposed to the T cells when presented with the entire protein (i.e., the peptides comprise at least one minor or immunodominant epitope). Those peptides stimulating tritium incorporation of less than 2-fold above the medium control either do not contain a T cell epitope or contain a cryptic epitope (i.e., an epitope not normally exposed to T cells when the entire protein is presented). To confirm the presence of a cryptic epitope in these peptides, T cell lines can be established by culturing peripheral blood white cells from the same individual in the presence of each peptide separately to establish peptide- reactive T cell lines.
- the "rested" T cells can then be challenged with each peptide and the Der p I protein.
- a peptide which comprises at least one cryptic epitope is capable of stimulating the proliferation of the T cell line in the presence of the peptide or the entire protein at a level at least 2-fold above the medium control or is capable of tolerizing T cells.
- Trp He Val Arg Asn Ser Trp Asp Thr Asn Trp Gly Asp Asn Gly Tyr 210 215 220
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pulmonology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Rheumatology (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP7500034A JPH09509135A (en) | 1993-06-02 | 1994-06-01 | Cryptic peptides for use in inducing immune tolerance |
EP94916845A EP0705108A4 (en) | 1993-06-02 | 1994-06-01 | Cryptic peptides for use in inducing immunologic tolerance |
AU68385/94A AU674584B2 (en) | 1993-06-02 | 1994-06-01 | Cryptic peptides for use in inducing immunologic tolerance |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7283293A | 1993-06-02 | 1993-06-02 | |
US08/072,832 | 1993-06-02 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1994027634A1 true WO1994027634A1 (en) | 1994-12-08 |
Family
ID=22110024
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/AU1994/000292 WO1994027634A1 (en) | 1993-06-02 | 1994-06-01 | Cryptic peptides for use in inducing immunologic tolerance |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP0705108A4 (en) |
JP (1) | JPH09509135A (en) |
AU (1) | AU674584B2 (en) |
CA (1) | CA2164326A1 (en) |
NZ (1) | NZ266739A (en) |
WO (1) | WO1994027634A1 (en) |
Cited By (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996007428A1 (en) * | 1994-09-02 | 1996-03-14 | Immulogic Pharmaceutical Corporation | Peptide compositions capable of down regulating an antigen specific immune response |
EP0788800A1 (en) * | 1996-01-22 | 1997-08-13 | Meiji Milk Products Company Limited | Method and kit for inducing immunological tolerance |
WO1997035193A1 (en) * | 1996-03-21 | 1997-09-25 | Imperial College Of Science, Technology And Medicine | Cryptic peptides and method for their identification |
US5853723A (en) * | 1995-09-21 | 1998-12-29 | University Of Utah Research Foundation | Targeting of peg antibody conjugates to islet cells |
WO1999025387A1 (en) * | 1997-11-14 | 1999-05-27 | Washington University | Recombinant vaccines comprising immunogenic attenuated bacteria having rpos positive phenotype |
WO1999054349A2 (en) * | 1998-04-17 | 1999-10-28 | Heska Corporation | Dermatophagoides nucleic acid molecules, proteins and uses thereof |
US6759234B1 (en) | 1994-09-02 | 2004-07-06 | Immulogic Pharmaceutical Corporation | Compositions and methods for administering to humans, peptides capable of down regulating an antigen specific immune response |
EP1473041A2 (en) * | 2001-03-30 | 2004-11-03 | Antialis | Anti-allergic pharmaceutical composition |
WO2005077410A1 (en) | 2004-02-06 | 2005-08-25 | Greer Laboratories, Inc. | Methods and compositions for dosing of allergens |
US7097845B2 (en) | 1997-04-23 | 2006-08-29 | Jacob Sten Petersen | Combinations of antigen and mucosal binding component for inducing specific immunological tolerance |
US7128921B1 (en) | 1998-04-17 | 2006-10-31 | Heska Corporation | Dermatophagoides proteins and fragments thereof |
US8551493B2 (en) | 2007-08-15 | 2013-10-08 | Circassia Limited | Peptide with reduced dimer formation |
US8551492B2 (en) | 2007-06-01 | 2013-10-08 | Circassia Limited | Vaccine peptide combinations against cat allergy |
WO2013192532A2 (en) | 2012-06-21 | 2013-12-27 | Northwestern University | Peptide conjugated particles |
WO2015023796A2 (en) | 2013-08-13 | 2015-02-19 | Shea Lonnie D | Peptide conjugated particles |
US11510996B2 (en) | 2015-12-23 | 2022-11-29 | Cour Pharmaceuticals Development Company Inc. | Covalent polymer-antigen conjugated particles |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU1959888A (en) * | 1987-06-17 | 1989-01-19 | Institute For Child Health Research | Cloning of mite allergens |
AU2079788A (en) * | 1987-06-24 | 1989-01-19 | Autoimmune, Inc. | Treatment of autoimmune diseases by oral administration of autoantigens |
AU6979191A (en) * | 1989-12-20 | 1991-07-18 | Autoimmune, Inc. | Improved treatment of autoimmune diseases by aerosol administration of auto antigens |
AU7578991A (en) * | 1990-03-02 | 1991-09-18 | Autoimmune, Inc. | Enhancement of the down-regulation of autoimmune diseases by oral administration of autoantigens |
AU8721991A (en) * | 1990-09-07 | 1992-03-30 | Regents Of The University Of California, The | Methods for the diagnosis and treatment of diabetes |
AU8755991A (en) * | 1990-09-06 | 1992-03-30 | De Staat Der Nederlanden Vertegenwoordigd Door De Minister Van Welzijn, Volksgezonheid En Cultuur | Inhibitor of lymphocyte response and immune-related disease |
AU9023791A (en) * | 1990-10-15 | 1992-05-20 | Brigham And Women's Hospital | Treatment of autoimmune diseases by oral administration of autoantigens |
AU3778593A (en) * | 1992-02-28 | 1993-09-13 | Autoimmune, Inc. | Bystander suppression of autoimmune diseases |
AU3922693A (en) * | 1992-03-25 | 1993-10-21 | Merck Patent Gmbh | Peptides useful for inducing tolerance |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07502890A (en) * | 1991-10-16 | 1995-03-30 | イミユロジク・フアーマシユーチカル・コーポレーシヨン | T-cell epitopes of major allergens from Dermatophagoides (house dust mite) |
-
1994
- 1994-06-01 CA CA002164326A patent/CA2164326A1/en not_active Abandoned
- 1994-06-01 AU AU68385/94A patent/AU674584B2/en not_active Ceased
- 1994-06-01 JP JP7500034A patent/JPH09509135A/en active Pending
- 1994-06-01 NZ NZ266739A patent/NZ266739A/en unknown
- 1994-06-01 WO PCT/AU1994/000292 patent/WO1994027634A1/en not_active Application Discontinuation
- 1994-06-01 EP EP94916845A patent/EP0705108A4/en not_active Withdrawn
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU1959888A (en) * | 1987-06-17 | 1989-01-19 | Institute For Child Health Research | Cloning of mite allergens |
AU2079788A (en) * | 1987-06-24 | 1989-01-19 | Autoimmune, Inc. | Treatment of autoimmune diseases by oral administration of autoantigens |
AU6979191A (en) * | 1989-12-20 | 1991-07-18 | Autoimmune, Inc. | Improved treatment of autoimmune diseases by aerosol administration of auto antigens |
AU7578991A (en) * | 1990-03-02 | 1991-09-18 | Autoimmune, Inc. | Enhancement of the down-regulation of autoimmune diseases by oral administration of autoantigens |
AU8755991A (en) * | 1990-09-06 | 1992-03-30 | De Staat Der Nederlanden Vertegenwoordigd Door De Minister Van Welzijn, Volksgezonheid En Cultuur | Inhibitor of lymphocyte response and immune-related disease |
AU8721991A (en) * | 1990-09-07 | 1992-03-30 | Regents Of The University Of California, The | Methods for the diagnosis and treatment of diabetes |
AU9023791A (en) * | 1990-10-15 | 1992-05-20 | Brigham And Women's Hospital | Treatment of autoimmune diseases by oral administration of autoantigens |
AU3778593A (en) * | 1992-02-28 | 1993-09-13 | Autoimmune, Inc. | Bystander suppression of autoimmune diseases |
AU3922693A (en) * | 1992-03-25 | 1993-10-21 | Merck Patent Gmbh | Peptides useful for inducing tolerance |
Non-Patent Citations (8)
Cited By (45)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996007428A1 (en) * | 1994-09-02 | 1996-03-14 | Immulogic Pharmaceutical Corporation | Peptide compositions capable of down regulating an antigen specific immune response |
US6759234B1 (en) | 1994-09-02 | 2004-07-06 | Immulogic Pharmaceutical Corporation | Compositions and methods for administering to humans, peptides capable of down regulating an antigen specific immune response |
US5853723A (en) * | 1995-09-21 | 1998-12-29 | University Of Utah Research Foundation | Targeting of peg antibody conjugates to islet cells |
US5951984A (en) * | 1996-01-22 | 1999-09-14 | Meiji Milk Products Company, Limited | Method for inducing immunological tolerance, immunological tolerance inducing food kit, and immunological tolerance inducer kit |
EP0788800A1 (en) * | 1996-01-22 | 1997-08-13 | Meiji Milk Products Company Limited | Method and kit for inducing immunological tolerance |
US6221354B1 (en) | 1996-01-22 | 2001-04-24 | Meiji Milk Products Company, Limited | Immunological tolerance inducer kit |
GB2352518A (en) * | 1996-03-21 | 2001-01-31 | Circassia Ltd | Screening for therapeutic activity |
US6737406B1 (en) | 1996-03-21 | 2004-05-18 | Circassia, Ltd. | Cryptic peptides and method for their identification |
EP1612555A1 (en) * | 1996-03-21 | 2006-01-04 | Circassia Limited | Cryptic peptides for use in inducing immunologic tolerance |
GB2349463A (en) * | 1996-03-21 | 2000-11-01 | Circassia Ltd | Method for identification of cryptic peptides |
GB2349463B (en) * | 1996-03-21 | 2001-01-10 | Circassia Ltd | Cryptic peptides and method for their identification |
WO1997035193A1 (en) * | 1996-03-21 | 1997-09-25 | Imperial College Of Science, Technology And Medicine | Cryptic peptides and method for their identification |
GB2326642B (en) * | 1996-03-21 | 2001-02-07 | Imperial College | Cryptic peptides and method for their identification |
GB2352518B (en) * | 1996-03-21 | 2001-04-04 | Circassia Ltd | Screening of compunds for therapeutic activity |
GB2326642A (en) * | 1996-03-21 | 1998-12-30 | Imperial College | Cryptic peptides and method for their identification |
US7097845B2 (en) | 1997-04-23 | 2006-08-29 | Jacob Sten Petersen | Combinations of antigen and mucosal binding component for inducing specific immunological tolerance |
US6383496B1 (en) | 1997-11-14 | 2002-05-07 | Washington University | Recombinant vaccines comprising immunogenic attenuated bacteria having RPOS positive phenotype |
AU736242B2 (en) * | 1997-11-14 | 2001-07-26 | Washington University | Recombinant vaccines comprising immunogenic attenuated bacteria having RPOS positive phenotype |
WO1999025387A1 (en) * | 1997-11-14 | 1999-05-27 | Washington University | Recombinant vaccines comprising immunogenic attenuated bacteria having rpos positive phenotype |
US6024961A (en) * | 1997-11-14 | 2000-02-15 | Washington University | Recombinant avirulent immunogenic S typhi having rpos positive phenotype |
US7083794B2 (en) | 1997-11-14 | 2006-08-01 | Washington University | Recombinant vaccines comprising immunogenic attenuated bacteria having RpoS positive phenotype |
WO1999054349A2 (en) * | 1998-04-17 | 1999-10-28 | Heska Corporation | Dermatophagoides nucleic acid molecules, proteins and uses thereof |
US6455686B1 (en) | 1998-04-17 | 2002-09-24 | Heska Corporation | Dermatophagoides nucleic acid molecules, proteins and uses thereof |
US7589187B2 (en) | 1998-04-17 | 2009-09-15 | Heska Corporation | Dermatophagoides nucleic acid molecules |
US7128921B1 (en) | 1998-04-17 | 2006-10-31 | Heska Corporation | Dermatophagoides proteins and fragments thereof |
US7256263B2 (en) | 1998-04-17 | 2007-08-14 | Heska Corporation | Dermatophagoides proteins and uses thereof |
WO1999054349A3 (en) * | 1998-04-17 | 2000-02-03 | Heska Corp | Dermatophagoides nucleic acid molecules, proteins and uses thereof |
EP1473041A3 (en) * | 2001-03-30 | 2004-11-24 | Antialis | Anti-allergic pharmaceutical composition |
EP1473041A2 (en) * | 2001-03-30 | 2004-11-03 | Antialis | Anti-allergic pharmaceutical composition |
US9345761B2 (en) | 2004-02-06 | 2016-05-24 | Greer Laboratories, Inc. | Methods and compositions for dosing of allergens |
EP1718331A1 (en) * | 2004-02-06 | 2006-11-08 | Greer Laboratories, Inc. | Methods and compositions for dosing of allergens |
EP1718331A4 (en) * | 2004-02-06 | 2009-11-11 | Greer Lab Inc | Methods and compositions for dosing of allergens |
US8491909B2 (en) | 2004-02-06 | 2013-07-23 | Greer Laboratories, Inc. | Methods and compositions for dosing of allergens |
WO2005077410A1 (en) | 2004-02-06 | 2005-08-25 | Greer Laboratories, Inc. | Methods and compositions for dosing of allergens |
US8551492B2 (en) | 2007-06-01 | 2013-10-08 | Circassia Limited | Vaccine peptide combinations against cat allergy |
US9168295B2 (en) | 2007-06-01 | 2015-10-27 | Circassia Limited | Vaccine peptide combinations |
US9744222B2 (en) | 2007-08-15 | 2017-08-29 | Circassia Limited | Peptide for vaccine |
US8652485B2 (en) | 2007-08-15 | 2014-02-18 | Circassia Limited | Peptide for vaccine |
US8551493B2 (en) | 2007-08-15 | 2013-10-08 | Circassia Limited | Peptide with reduced dimer formation |
US9340580B2 (en) | 2007-08-15 | 2016-05-17 | Circassia Limited | Peptide with multiple epitopes |
WO2013192532A2 (en) | 2012-06-21 | 2013-12-27 | Northwestern University | Peptide conjugated particles |
EP3607973A1 (en) | 2012-06-21 | 2020-02-12 | Northwestern University | Peptide conjugated particles |
WO2015023796A2 (en) | 2013-08-13 | 2015-02-19 | Shea Lonnie D | Peptide conjugated particles |
EP3650047A1 (en) | 2013-08-13 | 2020-05-13 | Northwestern University | Peptide conjugated particles |
US11510996B2 (en) | 2015-12-23 | 2022-11-29 | Cour Pharmaceuticals Development Company Inc. | Covalent polymer-antigen conjugated particles |
Also Published As
Publication number | Publication date |
---|---|
JPH09509135A (en) | 1997-09-16 |
EP0705108A4 (en) | 1997-08-06 |
AU6838594A (en) | 1994-12-20 |
NZ266739A (en) | 1997-07-27 |
AU674584B2 (en) | 1997-01-02 |
CA2164326A1 (en) | 1994-12-08 |
EP0705108A1 (en) | 1996-04-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU674584B2 (en) | Cryptic peptides for use in inducing immunologic tolerance | |
US6036957A (en) | Suppression of T-cell proliferation using peptide fragments of myelin basic protein | |
JP2635444B2 (en) | Treatment of autoimmune diseases by oral administration of autoantibodies | |
US6759234B1 (en) | Compositions and methods for administering to humans, peptides capable of down regulating an antigen specific immune response | |
US6884785B2 (en) | Compositions and methods for the treatment or prevention of autoimmune diabetes | |
PT2397154E (en) | Peptides for desensibilization against allergens | |
JP3554319B2 (en) | Peptide p277 analog and pharmaceutical composition for treating or diagnosing diabetes containing the same | |
Hoyne et al. | Inhibition of T-cell responses by feeding peptides containing major and cryptic epitopes: studies with the Der p I allergen. | |
JP3434510B2 (en) | Inhibition of T-cell proliferation using peptide fragments of myelin basic protein | |
WO1996012737A9 (en) | Compositions and treatment for multiple sclerosis | |
HUT77047A (en) | Compositions and treatment for multiple sclerosis | |
Karachunski et al. | Interleukin-4 deficiency facilitates development of experimental myasthenia gravis and precludes its prevention by nasal administration of CD4+ epitope sequences of the acetylcholine receptor | |
CA2132873C (en) | Peptides useful for inducing tolerance | |
EP1536826B1 (en) | Igf-2 peptide for use in tolerogenic approaches for type i diabetes. | |
US20030220229A1 (en) | Proinsulin peptide compounds for detecting and treating type I diabetes | |
EP0783322A1 (en) | Peptide compositions capable of down regulating an antigen specific immune response | |
EP1037663A2 (en) | Methods to treat undesirable immune responses | |
AU748104B2 (en) | Peptide compositions capable of down regulating an antigen specific immune response | |
AU738564B2 (en) | Proinsulin peptide compounds for detecting and treating type I diabetes | |
AU2005202898A1 (en) | Compositions and methods for the treatment or prevention of autoimmune disorders | |
HUT77806A (en) | Peptide compositions capable of down regulating an antigen specific immune response | |
AU9724801A (en) | Proinsulin peptide compounds for detecting and treating type I diabetes |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AT AU BB BG BR BY CA CH CN CZ DE DK ES FI GB GE HU JP KG KP KR KZ LK LU LV MD MG MN MW NL NO NZ PL PT RO RU SD SE SI SK TJ TT UA UZ VN |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 266739 Country of ref document: NZ |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2164326 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1994916845 Country of ref document: EP |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
WWP | Wipo information: published in national office |
Ref document number: 1994916845 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1994916845 Country of ref document: EP |