MXPA00011938A - Peptide composition as immunogen for the treatment of allergy - Google Patents

Abstract

The invention provides peptides comprising a sequence homologous to a portion of the third constant domain of the epsilon heavy chain of IgE, covalently linked to either (1) a carrier protein, or (2) a helper T cell epitope, and optionally to other immunostimulatory sequences as well. The invention provides for the use of such peptides as immunogens to elicit the production in mammals of high titer polyclonal antibodies, which are specific to a target effector site on the epsilon heavy chain of IgE. The peptides are expected to be useful in pharmaceutical compositions, to provide an immunotherapy for IgE-mediated allergic diseases.

Description

PEPTIDE COMPOSITION AS IMMUNOGENOUS FOR THE TREATMENT OF ALLERGY 1 FIELD OF THE INVENTION The present invention relates to the use of peptide conjugated compositions as an immunogen, with each conjugated peptide contained therein comprising an objective antigenic site on the third constant domain (CH3) of the heavy chain (e) epsilon of IgE, with the target antigenic site covalently linked to (1) a carrier protein through chemical coupling, or (2) an attenuating T-cell epitope and other immunostimulatory sequences through chemical coupling or through direct synthesis, for the treatment of allergy. More particularly, the present invention relates to the use of such peptide conjugated compositions as an immunogen to produce the production in mammals, including humans, of high titer polyclonal antibodies specific for a target effector site on the CH3 domain of the heavy chain. of IgE, and the use of such a composition as a pharmaceutical to provide immunotherapy for IgE-mediated allergic diseases.

BACKGROUND OF THE INVENTION In the immune system of humans and other mammals, IgE mediated type I hypersensitivities. These are the allergic responses to certain foods, drugs, and environmental allergens which are manifested by such symptoms as allergic rhinitis. , asthma, allergic dermatitis and anaphylaxis. The strategies that exist to treat allergic diseases are of limited utility, consisting of attempts to desensitize the atopic individual to an identified allergen or to improve a continuous allergic reaction with therapeutic compounds. The limitations for allergen-based desensitization immunotherapy include difficulties in identifying the allergen involved and the adverse reactions frequently caused by the use of the identified allergen (World Health Organization and International Union of Immunological Societies Working Group, Lancet, 1989; i: 259- 261). Other treatments for allergy relief employ therapeutic compounds to block the acute inflammatory cascade that is responsible for allergic reactions. These compounds include anti-histamines, decongestants, β2 agonists, and corticosteroids. Decongestant antihistamines, and β2 agonists, act on events below IgE in the allergic cascade, making the palliative remedies which are aimed at allergic symptoms rather than preventive treatments which must act on events closer to the initiation of allergic reactions mediated by IgE. These palliative remedies provide relief that is short and partial, often accompanied by adverse side effects. Many patients with severe allergies are treated effectively with corticosteroids. Steroid therapy reduces inflammation but is largely immunosuppressive. To avoid the drawbacks of the known therapeutic drugs, it would be more desirable to prevent allergic responses by the selective intervention selected for IgE. In common with the other immunoglobulins, IgE has two heavy chains and two light chains. The heavy chain e has five domains, a variable VH domain and constant domains CH1 to CH4. The constant domains of both e chains of an IgE molecule are combined to comprise the constant region or Fc of IgE. IgE circulates and binds to effector cells such as basophils and mast cells through a site on the IgE Fc region, binding to a high affinity FceRI receptor on the surface of the cells. In an allergic response, allergens, (eg, pollen, tick dust proteins, flea antigens), bind to antigen binding sites on the variable region of mast cell or IgE linked to basophils. This action reticles the IgE molecules and the underlying FceRI receptors. Thus, allergic IgE complexes signal the degranulation of mast cell and basophil cells with the concomitant release of histamine and the other inflammatory mediators. These mediators produce the allergy symptom, regulate the production of IgE at high and result in increased sensitivity to the allergen (Davis et al., Springer Semin Immunopathol, 1993; 15: 51-73). It has been suggested that allergic diseases can be treated by interventions which inhibit the binding of IgE to mast cell and basophil cells. For example, synthetic peptides corresponding to various sites on the IgE Fc have been studied as competitive inhibitors for the binding of IgE to the FceRI receptor. the assumption of the investigators has been that such peptides act as antagonists for the sites on IgE that participate in the I g E bond to the FceRI receptor, they serve to map portions of the binding site. The amino acid residues of competitively inhibiting IgE peptides, and of all IgE peptides that follow, including non-human IgE peptide homologs, are indexed according to the numbering for human IgE given by Dorrington and Bennich (Immunol Rev. , 1978; 41: 3-25, also accessible at the internet location http: www.pdb.bnl.gov/pdb.bin/pdbids). That human sequence is listed here as SEC. FROM IDENT. NO: 1 and is listed as shown in Table 1. The homologous sequences of dog, rat and mouse for IgE (Patel et al., Immunogenetics, 1995; 41: 282-286; Steen et al., J Mol Biol, 1984; 177: 19-32; and Ishida et al., EMBO, ---- "" '1982; 1: 1117-1123) are also shown in Table 1 and are listed as SEQ ID NOS: 2, 3 and 4. The animal sequences are shown in the register with IgE. The individual amino acid positions in human IgE, and in homologues of other species, are identified herein according to the numbering system for the amino acid sequences shown in Table 1, unless otherwise specified. al. (Nature, 1988; 331: 180-183) has shown that a long recombinant polypeptide of 76 amino acids, spanning the C-terminal region CH2 and N-Terminal CH3 of human IgE, of amino acids 301-376, reduces the IgE binding to mast cell by competitive inhibition Other studies report that only the CH3 domain is involved with the FceRI linkage, for example, a rat peptide sequence corresponding to amino acids 401-415 of the human sequence (Table 1) inhibits rat IgE binding to rat mast cells (Burt and Stanworth, Eur J Immunol, 1987; 17: 437-440). A residue peptide 419 to 463 of human IgE prevents sensitization of rat mast cells (Nio et al., FEBS Lett, 1992; 314: 229-231). Jardieu and Presta WO 93/04173) reported on peptide homologs of the CH3 and CH4 regions which may include amino acids 373-390, 420-428, 446-453, and adjacent regions, which bind differentially to the FceRI receptor. However, high concentrations of all of The peptides were required to achieve effective inhibition of the IgE linkage. These high concentrations predict excessively large doses for significant physiological effect, and are not therapeutically practical. Anti-IgE antibodies have also been applied as a method to map sites on IgE that participate in binding to the FceRI receptor. Studies with mouse monoclonal antibodies directed against various IgE Fc domains revealed that anti-IgE monoclonal antibodies with specificities for the CH3 domain inhibit the binding of IgE to its high affinity receptor (Baniyash et al., Molec Immunol, 1988; 25: 705-711; and, Stadler et al., Immunol Cell Biol, 1996; 74: 195-200). These studies of monoclonal antibodies are in agreement with early studies that used polyclonal antypeptide antibodies to map sites that are apparently involved in receptor binding. For example, rabbit antibodies with specificities for positions 401-415 of amino acid IgE (Burt et al., Molec Immunol, 1987; 24: 379-389), and 355-368 (Robertson and Liu, Molec Immunol, 1988; 25: 103-113) showed specificity for the IgE binding but reacted poorly with the IgE binding to the receptor. A canine IgE peptide fragment containing at least five continuous amino acids of dog IgE 356-479 amino acids is useful for the preparation of antibodies for the diagnosis of allergy in dogs (Jp 9179795, 1997). These results suggest surface-exposed effector sites in the CH3 domain of the dog e chain, but no such effector site is taught nor is a therapeutic application for anti-IgE antibodies described. These epitope mapping studies demonstrate more consistently that the CH3 domain of the heavy chain can be selected for interventions directed to inhibit the binding of IgE to basophils and mast cells. However, the various studies are quite inconsistent on the precise indications for the sites on CH3 that are most useful. Also, the results of cross-inhibition studies on IgE, with site-specific antibodies (eg, Burt et al., 1987) have often been over-interpreted to mean that they have defined an accurate location for the FceRI binding site on the chain e. The interpretation of such cross-inhibition studies is limited because it can not be assumed that an antibody recognition site is equivalent to a ligand binding site. The antibodies can inhibit by binding directly to the desired target site, but they can also occupy non-continuous effector sites and inhibit ligand binding through steric hindrance or conformational change induction. Therefore, epitope mapping studies have provided empirical observations but have not resolved the : £ Afeaát. binding site for the high affinity receptor within the CH3 domain. In the urgency of a defined binding site, no means are available for the reliable prediction of potentially therapeutic synthetic immunogens with immunological cross-reactivities for the effector sites that participate directly or indirectly in the binding to FceRI. In addition, in the absence of X-ray crystallography data for IgE, the structural models available for IgE are not sufficient for the prediction of sites on IgE that are suitable for anti-IgE interventions. Incompatible structures have been modeled based on the disclosed three-dimensional structure of IgG for IgE and for the CH2 / CH3 region of IgE that is associated with the interaction between IgE and its high affinity receptor. These models propose several conformationally dependent structures for the site, which involve contact with linearly non-adjacent residues of the IgE molecule. Some models for the site suggest interactions between non-contiguous sites on the same chain and mediated by intramolecular undulations linked by disulfide (Helm et al., Eur J Immunol, 1991; 21: 1543-1548) or intramolecular undulations maintained by electrostatic interactions (Presta et al., J Biol Chem, 1994; 269: 26368-26373). Other models propose intermolecular interactions between segments of the dimerized chains of an IgE molecule (McDonnell et al., Biochem Soc Trans, 1997; 25; 387-392). In fact, the experimental observations show that the potential contact points comprise various disseminated and discontinuous sites on the CH3 domain of the e chain and make it clear that the three-dimensional structure of the FceRI binding site can not be easily resolved by modeling (Helm et al., 1988; Baniyash et al., 1988; and, Presta el al., 1994). Therefore, the identification of useful synthetic peptide antagonists and immunogens that mimic the effector sites on IgE has not been described by theoretical modeling. In the absence of a structure for IgE resolved by X-ray crystallography, only useful peptide sites can be reached by empirical experimentation. The concept of treating allergic diseases with anti-IgE antibodies, of specificities that inhibit the binding of IgE to the high affinity receptor on basophils and mast cells (Stadtler et al., 1996, Davis et al., 1993) has also been known. . Such antilgE antibodies are anaphylactogenic (crosslinkers) or non-anaphylactogenic (non-crosslinkers). The majority of such anti-IgE antibodies are anaphylactogenic. They will bind and crosslink IgE on the surface of basophils and mast cells and trigger the release of pharmacological mediators of allergy. This cross-linking can lead to anaphylaxis and death. It is therefore crucial that anti-IgE antibodies for treatment are not anaphylactogenic. Certain non-anaphylactogenic antibodies retain specificity for the CH3 domain of the e chain and do not cross-link I g E bound to cell, while they show inhibitory activity for the release of histamine mediated by IgE (Davis et al., 1993; Stadler et al., nineteen ninety six). Rup and Kahn (U.S. 4,940,782) report such a non-anaphylactogenic monoclonal antibody that reacts with free rat IgE and rat IgE bound to B cells, but not IgE binding to the rat mast cell FceRI receptor. More significantly, it inhibits sensitization of rat mast cells. Non-anaphylactogenic antibodies with homologous specificities for human IgE also inhibit sensitization by the same mode of action. These anti-human IgE antibodies bind free IgE in serum, binding to bound IgE to B cells, fail to bind basophil bound IgE and high affinity mast cell receptor and prevent sensitization of human cells. It is assumed that these antibodies act by specificity for the site on IgE that binds the FceRI receptor (Rup and Kahn, U.S. 4,940,782; Davis et al., 1993, Chang, U.S. 5,420,251; Presta ei al., J Immunol, 1993; 151: 2623-2632). In addition, a non-anaphylactogenic anti-human IgE monoclonal antibody with a different specificity has been found to also neutralize free I g E (Rudolf et al., J Immunol, 1996; 157: 5646-5652). This anti-IgE does not bind directly to the receptor binding site because it also recognizes I g E bound to FceRI. Apparently, it works to reduce the * ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Thus, the anti-lgE antibodies that bind directly to the site of FceRI link and anti-IgE antibodies that interfere with the binding to FceRI other effector sites, serve to block the sensitization of mastocíticas cells and basophils by free IgE. These potentially immunotherapeutic antibodies identify CH3 as the IgE domain that interacts with the high affinity IgE Fc receptor, according to previous mapping studies. However, a more precise identification of the link site and useful alternative effector sites such as those described by Rudolf et al. a phage display library has also been used to identify mimotope peptides which bind to their anti-IgE monoclonal antibody; however, the mimotopes peptides do not show homology to the primary amino acid sequence of human IgE (Rudolf et al., J. Immunol., 1998; 160: 3315-3321). An anti-IgE humanized monoclonal antibody with apparent specificity for FceRI receptor site is under clinical study in humans for treatment of allergy by passive immunotherapy (MacGlashan ef al, J Immunol, 1997; 158:. 1438-1445). It has been found that infusion with that antibody, rhuMAb-E25, reduces the concentration in serum of IgE in patients, downregulates the expression of the IgE receptor on ^^ ^ ¿gj ^ j L to ^^^ ^^ ^ jg ^^^^^^^^^^^^^ effector cells, reduces allergic to the allergen challenge sensitivities, and improves the symptoms of asthma and allergic rhinitis. The antibody has a good safety profile. The results of the clinical tests establish the possibility of an anti-IgE proposal for the treatment of allergic diseases. But this mode of treatment is problematic: immunotherapy by the anti-IgE invention is achieved by passive immunization, that is, by infusion of the antibody. The antibody should be delivered in sufficiently high doses and at sufficiently frequent frequencies, via intravenous or subcutaneous routes inconvenient, to achieve a continuous pharmacologically effective concentration of antibodies. The effective dose is determined by the patient's body weight, the free IgE base line in circulation, and by the route of administration. In recent clinical trials, the continuous concentration required for therapeutic efficiency was achieved by two weekly doses and was subsequently maintained by biweekly doses. A full course of treatment for a typical allergy patient would consume a total of 2000-3000 mg of humanized antibody and requires seven to 10 inconvenient intravenous administrations (MacGlashan ef al., 1997; Boulet ef al, Am J Respir Crit Care Med. 1997; 155: 1835-1840). The cost for this amount of antibodies and the consumption and inconvenience of multiple infusions in a hospital facility suggests that this treatment is too expensive for all but a small proportion of the patient population. The clinical effectiveness of the rhuMAb-E25 monoclonal antibody establishes the possibility of immunotherapy by the passive administration of anti-IgE. It also provides the rationale for an alternative anti-IgE proposal by active immunization, as long as such immunogens can be designed. An anti-IgE treatment affected by active immunization with IgE immunogen, ie by "vaccination" against endogenous IgE, would be preferable on the basis of cost and convenience. "Vaccination" against IgE offers advantages over passive immunization: small amounts of cheap immunogen, intramuscular injections administered conveniently and infrequently, and without the need to construct murine antibodies for compatibility with the target species, that is, to "humanize" antibodies for use in humans, since the procedure uses the patient's own immune system to produce antibodies. Nevertheless, while the uncoupling of the monoclonal antibodies cited above may suggest the desire for IgE immunogens, they do not describe the identity of safe and effective immunogens. Such immunogens must mimic relevant IgE effector sites with sufficient fidelity to evoke cross inhibitory antibodies, while retaining sufficient site specificity to avoid the injection of anaphylactogenic antibodies. In addition, effective I g E immunogens must be highly immunostimulatory. There remains a need for such immunogens, of relevant and safe specificity to the site, and of sufficient immunopotency. IgE immunogens for allergy immunotherapy should be immunostimulatory to evoke sufficient anti-IgE levels to reduce IgE-mediated sensitization. Such immunogens must be designed to overcome the strong tolerance presented to automoléculas. Haba and Nisonoff (Proc Nati Acad Sci USA, 1990; 87: 3363-3367) induced an effective anti-IgE response in mice only by immunizations with I g E during a short neonatal window of development, from birth to day 10. Vaccinations initiated beyond this time failed to induce desired autoimmune response unless the I g E used to immunize the mice had been covalently coupled to a foreign carrier protein, lamellated hemocyanin (KLH). Similarly, a desensitizing anti-IgE response in rats was evoked by a recombinant protein comprising the CH2-CH3 chain domains fused to the glutathione-S-transferase protein of Schistosoma japonicum (Hellman, Eur J Immunol, 1994; 24: 415- 420). Other investigators have been interested in minimizing the risk of evoking anaphylactogenic anti-IgE antibodies that cross-link IgE already bound to the surfaces of mast cell and basophil cells by looking for peptide IgE immunogens of specificity at the finest site. For example, a peptide corresponding to a site in the CH4 domain of IgE (residues 497-506 of SEQ ID NO: 1) was coupled to KLH and used to induce polyclonal antibodies that were effective in suppressing transduction of the IgE-mediated signal in rat mast cell cells. However, the peptide KLH conjugate exhibited poor immunostimulatory abilities which required demonstration of efficiency by passive immunization of rats with a peak of immune rabbit antiserum (Stanworth et al., Lancet, 1990; 336: 1279-1281). The Stanworth CH4 immunogen was al. it was later produced, by the work of the present inventor, as a series of completely synthetic immunogens by synthesis which provide the covalent link to the promiscuous human T-helper epitopes. The immunogenicity of these peptides was improved over that of the original KLH-peptide conjugate, but no evidence of the efficiency of the resulting anti-IgE CH4 antibodies was provided (Wang, WO 95/26365). In addition, as shown herein in Example 1 (Table 2, entry 34), anti-peptide antibodies with specificity for the CH4 effector site previously described (Stanworth et al., 1990) had no cross-reactivity to human IgE. The early antipeptide studies of Burt and Stanworth (1987) selected for the peptide IgE-CH3 401-415 also provides evidence of evoking cross-reaction desensitization, but this also requires peak rabbit antiserum selected from a peptide-protein carrier conjugate more defined to observe the effects of passive immunization in a rat model. The synthetic peptides have never been shown to be effective in producing production in immunized hosts of polyclonal antiserum capable of inhibiting the release of histamine. The improvement of the immunogens of the prior art discussed above is necessary before a synthetic peptide immunogen of sufficient immunogenicity and specificity of efficiency and safety can be obtained. The present invention achieves these improvements through the incorporation of a collection of additional methods for the identification and design of synthetic peptide immunogens. These methods include: (1) an effective procedure for the identification of an effective target epitope; (2) means for enhancing the immunogenicity of a B-cell target epitope by combining it with a peptide comprising promiscuous T-cell epitope broadly reactive (Th); (3) the means to enlarge the repertoire of T-cell epitopes by applying combinatorial peptide chemistry and additionally thereby accommodating the immunovapable responsiveness of a population multiplied by race mixture; and (4) the stabilization of conformational characteristics by the introduction of repressors ^^^ * ^^^^ fog ^^^ A¿? faith? ^^^^^^^^^^ i ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ native molecule. Synthetic peptides have been used for "epitope mapping" to identify immunodominant determinants or epitopes on the surface of proteins, for the development of new vaccines and diagnosis. The epitope mapping employs a series of overlapping peptides corresponding to regions on the protein of interest to identify sites that participate in the immunogenic-antibody determinant interaction. Most commonly, epitope mapping employs peptides of relatively short length to detect precisely linear determinants. A rapid method of epitope mapping known under the trade name "PEPSCAN" is based on the simultaneous synthesis of hundreds of overlapping peptides, of lengths of 8 to 14 amino acids, coupled to solid supports. Coupled peptides are tested for their ability to bind antibodies. The PEPSCAN proposal is effective for locating linear determinants, but not for the identification of the epitopes required for the imitation of discontinuous effector sites such as the FceRI binding site (Meloen et al., Ann Biol Clin, 1991; 4 ^: 231- 242). An alternative method lies in a set of nested and overlapping peptides of multiple lengths ranging from 15 to 60 residues. These longer peptides can be synthesized by a laborious series of independent solid phase peptide synthesis, rather than by rapid syntheses and Simultaneous PEPSCAN. The resulting set of nested and overlapping long peptides can then be used for antibody binding analyzes in systems such as experimental immunizations and natural infections to identify long peptides that best exhibit immunodominant determinants, including simple discontinuous epitopes. This method is exemplified by the Wang studies for the mapping of immunodominant sites of HTLV l / ll (US 5,476,765) and HCV (US 5,106,726); and was used for the selection of an accurate position on the gp120 sequence for optimal presentation of an epitope that neutralizes HIV (Wang et al., Science, 1991; 254: 285-288). Peptide immunogens are generally more flexible than proteins and do not tend to retain any preferred structure. Therefore, it is useful to stabilize a peptide immunogen by the introduction of cyclic repressors. A properly cyclized peptide immunogen can mimic and preserve the conformation of a selected epitope and thereby evoke antibodies with cross reactivities for the site on the authentic molecule (Moore, Chapter 3 in synthetic Peptides: A User's Guide, ed Grant, WH Freeman and Company : New York, 1992, pp. 63-67). Another important factor that affects the immunogenicity of an IgE-derived peptide for a pharmaceutical allergy is its presentation to the immune system by attendant cell epitope T «- ^ A > * ^ ^ «-,. ^ ^ .. *. ^ 4 ^ - ^^ -, which react with receptors of host T helper cells and MHC Class II molecules (Babbitt et al., Nature, 1985; 317: 359-361). These are frequently provided by carrier proteins with concomitant disadvantages due to the difficulties for the manufacture of peptide / well-defined carrier conjugates, and more the direction of most of the antibody response to the carrier, and their carrier-induced epitope pressure (Cease, Intern Rev Immunol., 1990; 7: 85-107; Schutze et al., J Immunol., 1985; 135: 2319-2322). Alternatively, T (Th) attendant cell epitopes can also be delivered by synthetic peptides comprising Th sites. Thus, Th epitopes termed promiscuous Th evoke sufficient assistance of T cells and can be combined with synthetic epitopes of B cells which themselves are poorly immunogenic to generate powerful peptide immunogens (US ,759,551). Well-designed promiscuous Th / cell epitope chimeric B peptides are capable of producing Th responses and resulting antibody responses in most members of a genetically diverse population expressing various MHC haplotypes. Th promiscuous can be provided by specific sequences derived from potent foreign antigens, such as for example, measles virus F protein, hepatitis B virus surface antigen, and major outer membrane protein (MOMP) of Chlamydia trachomatis. Many known promiscuous Th, taken from viral and bacterial pathogens, have been shown to be effective in enhancing a poorly immunogenic peptide corresponding to the hormonal decapeptide LHRH (US 5,759,551). Promiscuous Th epitopes derived from foreign pathogens may include, but are not limited to, epitopes of T helper nuclei antigen and hepatitis B surface (HBS Th and HBC Th) pertussis toxin attendant T cell epitopes (PT Th) Tetanus toxin attendant T cell epitopes (TT Th), T cell epitopes Assisting Measles Virus F protein (MVF Th), T cell epitopes Assisting the major outer membrane protein of Chlamydia trachomatis (CT Th ), diphtheria toxin attendant T-cell epitopes (DT Th), attendant T-cell epitopes of Plasmodium falciparum circumesporozoite (PF Th), epitopes of Triosa phosphate isomerase-isomerase helical T cell epitopes (SM Th), and epitopes of T helper cells TraT Escherichia coli (TraT Th). The Th derivatives of pathogens were listed as SEC. FROM IDENT. NOS: 2-9 and 42-52 in US 5,759,551; as an assistant site of Chlamydia P11 in Stagg et al., Immunology, 1993; 79; 1-9; and as peptide of HBc 50-69 in Ferrari et al., J Clin Invest, 1991; 88; 214-222. Promiscuous Th epitopes vary in size from about 15 to about 50 amino acid residues in length (US 5,759,551) and frequently share common structural characteristics and may contain specific tag sequences. For example, a common characteristic are antipathetic helices, which are alpha-helix structures with hydrophobic amino acid residues that dominate one side of the helix and with charged and polar residues dominating the surrounding faces (Cease et al., Proc Nati Acad Sci USA, 1987; 84: 4249-4253). Th epitopes frequently contain additional primary amino acid patterns such as a Gly or a charged residue followed by two to three hydrophobic residues, followed in turn by a charged or polar residue. This pattern defines what is called Rothbard sequences. Also, Th epitopes frequently obey rule 1, 4, 5, 8, where a positively charged residue is followed by hydrophobic residues at the fourth, fifth and eighth positions after the charged residue, consistent with an amphipathic helix having positions 1 , 4, 5 and 8 located on the same face. Since all these structures are composed of hydrophobic, charged and polar common amino acids, each structure can exist simultaneously within a single Th epitope (Partidos et al., J Gen Virol, 1991; 72: 1293-1299). Most if not all, the promiscuous T-cell epitopes fit into at least one of the periodicities described above. These characteristics can be incorporated into the "idealized artificial Th-site" designs. Useful Th sites can also include combinatorial Ths that incorporate selected degenerate sites within the design of the idealized Th sites. In Wang ei al. (WO 95/11998), a particular class of a combinatorial epitope was designed as a "library of Structured Synthetic Antigens" or SSAL. A Th constructed as an SSAL epitope is composed of positional substitutions organized around a waste frame structure without variation. The sequence of the SSAL is determined by aligning the primary amino acid sequence of a promiscuous Th, retaining residues relatively unchanged at the positions responsible for the unique structure of the Th peptide and providing degeneracy at the positions associated with the recognition of the various elements of MHC restriction. Lists of variable and preferred amino acids are available for MHC binding motifs (Meister et al., Vaccine, 1995; 13: 581-591; Alexander et al., Immunity, 1994, 1: 751-761). All members of the SSAL are produced simultaneously in a simple solid phase peptide synthesis in tandem with the selected B cell epitope and other sequences. The Th library sequence maintains the binding motifs of a promiscuous Th and accommodates the reactivity in a wider range of haplotypes. For example, the degenerate Th epitope described in WO 95/11998 as "SSAL1TH1" was modeled after a promiscuous epitope taken from the measles virus F protein (Partidos et al., 1991). SSAL1TH1 was designed to be used in tandem with an LHRH target peptide. As the epitope of iJ i "j measles, SSAL1TH1 follows the sequence of Rothbard and rule 1, 4, 5, 8: 1 5 10 15 Asp-Leu-Ser-Asp-Leu-Lys-Gly-Leu-Leu-Leu-His-Lys-Leu-Asp-Gly-Leu Glu lle Glu lle Arg lle lle lle Arg lle Glu lle Val Val Val Val Val Val Val Phe Phe Phe Phe Phe Phe Phe Glu or Asp-laden residues are added in position 1 to increase the charge surrounding the hydrophobic face of Th. The hydrophobic face of the amphipathic helix is then maintained by hydrophobic residues in 2, 5, 8, 9, 10, 13 and 16, with variability in 2, 5, 8, 9, 10, 13, and 16, to provide a front with the ability to link to a wide range of MHC constraint elements. The net effect of the SSAL feature is to enlarge the range of immunoresponsibility to an artificial Th (WO 95/11998). Peptide immunogens that have been designed with the peptide technologies and peptide design elements discussed above, ie, precise epitope mapping, cyclic restriction, and the incorporation of promiscuous or promiscuous Th epitopic Th epitopics, and idealized Th SSAL epitopes are the basis for the effective synthetic peptide IgE immunogens of the present invention. Such peptides are preferred for proper selection and safety due to the effective presentation of the effector site I g E by optimized position and cyclization, and for immunopotency due to widely reactive Th responsivity.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides new synthetic peptide conjugate compositions for the treatment of IgE-mediated allergic diseases by active immunization. The immunization induces the production of high titre non-anaphylactogenic polyclonal antibodies specific for an IgE effector site in an immunized host. This in turn prevents triggering and activation of mast cell / basophil cells and down-regulates IgE synthesis. The treatment is effected by immunization of the host with the peptide composition, each peptide comprising therein an objective antigenic peptide sequence (referred to herein as an "IgE-CH3 domain antigen" or "IgG-CH3 domain antigen peptide"). ") modified from a segment of the CH3 domain of the epsilon (e) heavy chain of human IgE (eg, amino acids 413-435 of SEC.

IDENT. NO: 1 or SEC. FROM IDENT. NO: 5) or the homologous sequence of another species (for example SEQ ID NOS: 6-8 and 84). In general, the IgE-CH3 domain antigen is a peptide sequence between about 25 and about 29 amino acids in length, is substantially homologous to ^^ ¡^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ previous of the CH3 domain of the epsilon heavy chain of a mammalian IgE antibody, and contains two cysteine residues separated by approximately 23 amino acid residues. In the present context, substantially homologous means that in addition to the two cysteine residues, which can be introduced by insertion or substitution, up to about four other amino acid substitutions (preferably conservative substitutions) can also be made. Preferably, the target site is modified from that of the naturally occurring IgE sequences as follows: (1) by the insertion of a cysteine residue next to the N-terminus of position 413 or homologous position, unless the Cystine is already present at positions 413 or 414 in the natural sequence; (2) by conservative substitution (preferably serine) for any native cysteines at positions 415 to 434 of the natural target sequence; (3) by the insertion of cysteine on the C-terminal side of position 435 or homologous position unless the cysteine is already present at positions 435 or 436 in the natural sequence; and (4) by the formation of a disulfide bond between the retained cysteines to produce a cyclic structure. The structures can also comprise from 1 to 5 additional amino acids taken from any term of segment 413-415 and IgE, with the proviso that the wavy structure of the simple disulfide is retained. An IgE-CH3 domain antigen peptide optimized for human IgE, having the sequence Cys-Gly-Glu-Thr-Tyr-Gln-Ser-Arg-Val-Thr-His-Pro-His-Leu-Pro-Arg-Ala -Leu-Met-Arg-Ser-Thr-Thr-Lys-Cys (SEQ ID NO: 5) is provided by the present invention. The human IgE target site is cyclized through non-natural terminal cysteines and a serine residue replaces the cysteine residue of the natural sequence. The antibody evoked by the peptide immunogens comprising this IgE-CH3 domain antigen is cross-reactive with human IgE and inhibits the sensitization of human basophils by human IgE. In the same way, the corresponding target sites for I g E of other species can be derived from the homologous chain segment of the relevant species, for example, such target sequences can be taken from the dog, mouse and rat sequences and shown in FIG. Table 1 (SEQ ID NOS: 2, 3 and 4), or the horse IgE-CH3 sequence provided by Navarro et al., Molec. Immunol., 1995, 32: 1-8.

The additional IgE-CH3 domain antigen peptides (SEQ.

IDENT. NOS: 6, 7, 8 and 84) can be derived from these sequences. Preferably, the IgE-CH3 domain antigens of the invention become more immunogenic by means of the covalent linkage to a carrier protein through chemical coupling, or more preferably by means of covalent linkage to synthetic immunostimulatory elements, such as Th epitopes. promiscuous, through direct synthesis. Specific examples of carrier protein and immunostimulatory elements are provided, for example, a Carrier Slime Hemocyanin (KLH) carrier, an artificial Th (SEQ ID NO: 9), Artificial SSAL Th (SEQ ID NOS. : 10 and 11), a Th derived from a pathogen (SEQ ID NO: 12), and an immunostimulatory invasin peptide (Inv) taken from Yersinia (SEQ ID NO: 13). The fully synthetic peptide conjugates of the invention can be represented by the formulas: (A) n- (IgE-CH3 antigen domain) - (B) 0- (Th) mX or (A) n- (Th) m- (B) o- (IgE-CH3 domain antigen) -X or (A) n- (B) 0- (Th) m- (B) o- (IgE-CH3 domain antigen) -X or (IgE-CH3 domain antigen) - (B) 0- (Th) m- (A) nX or (Th) m- (B) 0- (IgE-CH3 antigen domain) - (A) nX where each A is independently an amino acid or a general immunosuppressant sequence; each B is selected from the group consisting of amino acids, -NHCH (X) CH2SCH2CO-, -NHCH (X) CH2SCH2CO (e-N) Lys-, -NHCH (X) CH2S-succinimidyl (eN) Lys-, and -NHCH (X) CH2S- (succinimidyl) -, each Th is independently an amino acid sequence constituting an attendant T-cell epitope, or an immutable analog or segment of the same; The IgE-CH3 domain antigen is a peptide between about 25 and about 29 amino acids in length, it is substantially homologous to one of the segments represented by SEC. FROM IDENT. US: 5-8 and 84 of the CH3 domain of the epsilon heavy chain of a mammalian I g E antibody, and contains two cysteine residues separated by approximately 23 amino acid residues; X is an amino acid a-COOH or a-CONH2, n is from 0 to about 10; m is from 1 to about 4; and o is from 0 to about 10. More specifically, the IgE-CH3 domain antigen is selected from the group consisting of SEC. FROM IDENT. NO: 5, SEC. FROM IDENT. NO: 6, SEC. FROM IDENT. NO: 7, SEC. FROM IDENT. NO: 8, homologous sequences of the epsilon heavy chain of mammalian IgE-CH3 antibodies, and cross-reactive and immunologically functional analogues thereof.

The peptide compositions of the present invention comprise peptide immunogens from about 25 to about 100 amino acid residues, preferably from about 25 to about 80 amino acid residues and more preferably from about 45 to about 65 amino acid residues. Adjuvants and / or delivery vehicles and other ingredients routinely incorporated with vaccine formulations, and instructions for dosing are also provided in such a way that immunotherapeutic antibodies directed against the selected IgE effector site are generated. This in turn inhibits sensitization by the IgE circulation of basophils and mast cells, thus preventing the triggering and activation of mast cell / basophil cells by allergen-IgE complexes. The inhibitory mechanism, mediated by the antibodies and induced by the peptide composition of the present invention, will specifically reduce or eliminate the IgE-mediated pathology while leaving the defensive components of the immune system, for example IgG unaffected. DETAILED DESCRIPTION OF THE INVENTION This invention is directed to a novel peptide and peptide conjugated compositions for the generation of high titer polyclonal antibodies with specificity for an objective effector site in the third domain of the Fc portion of IgE, i.e., the CH3 domain of the chain e. For convenience, the term "peptide conjugate" as used herein refers to molecules comprising Th epitopes covalently linked to IgE-CH3 domain antigen peptide., either through conventional peptide bonds to form a larger single peptide, or through other forms of covalent bonding. The high site specificity of the compositions of this invention minimizes the generation of anti-IgE antibodies that can cross-link the bivalent IgE binding to FCeRI on the basophilic surface / mast cell cells, and thus evoke the production of anti-IgE antibodies. Anaphylactogenic Therefore, the invention is further directed to a safe method for the treatment of IgE-mediated allergic diseases in mammals, including humans. The target antigenic sequence was determined by a complete selection of candidate sites on the CH2 and CH3 domains of human IgE for useful immunoreactivities. The CH2 and CH3 sites were selected for synthesis as peptide immunogens based on the descriptions of Helm et al. (1988) and Presta ei al. (1994) that a long region which starts in the carboxyl terminus region of the CH2 domain of IgE and proceeds through the CH3 domain contains potential effector sites. The potential ripple structures in the conformation of IgE were deduced from a theoretical model for the three-dimensional structure of human IgE made available by Brookhaven National Laboratory at the internet address http://www.pdb.bnl.gov/pdb.bin/ pdbids and reported in Helm e al. (Eur J Immunol, 1991; 21: 1543-1548). The undifferentiated undulations were incorporated into the design of selected peptide immunogens to mimic the predicted ripple positions, to maximize the possibility of cross reaction between the designed target antigenic peptides and the native IgE molecule. Potential target antigenic sites were synthesized and made immunogenic either by chemical conjugation to KLH after solid phase peptide synthesis, or by covalent attachment to promiscuous Th epitopes and other immunostimulatory sequences by continuous synthesis (Table 2). Various sites were synthesized as cyclic peptides, with the incorporation of specific disulfide bonds, to stabilize the mobile peptides within conformations that resemble predicted IgE ripple structures. Potentially useful target effector sites were then identified by the preparation of hyperimmune sera and the antiserum test for cross-reactivity in human IgE. Antibodies from sera with high cross-reactivity for human IgE were purified and evaluated for their ability to inhibit IgE-mediated sensitization of human basophils in an in vitro test for histamine release. The antipeptide antibodies evoked by the peptides, SEC. FROM IDENT. NOS: 14 and 15 comprising SEC. FROM IDENT. NO: 5, showed strong cross-reactivity for IgE (Table 2), and most consistently exhibited high inhibitory activity in the histamine release test (Table 3). The objective epitope common to the peptides of SEC. FROM IDENT. NOS: 14 and 15 correspond to a segment of the IgE CH3 domain shown in Table 1. Table 1 shows the amino acid sequence of the heavy chain CH2, CH3 and CH4 domains of human IgE aligned with the homologous sequences taken from dog, rat and mouse. The target site on the human chain sequence which was determined useful for representation as the IgE-CH3 domain antigens of the invention is highlighted in Table 1 and includes human e chain residues 413-435. The homologous target sequences in the dog, rat and mouse proteins are also underlined in Table 1. The homologous sequences in the horse are residues 296-318 in the amino acid sequence of Navarro et al., Molec Immunol., 1995, 32: 1-8. The target IgE CH3 effector sites underlined] and the IgE-CH3 domain antigen peptides derived from this invention, are short peptide sequences which, when synthesized by themselves, are usually weak or non-immunogenic, more because they are self-antigens: These peptides Shocks can be immunopotentiated by chemical coupling to a carrier protein, for example, slotted limpet hemocyanin (KLH). A disadvantage of such immunogens based on "carrier-antigen protein of IgE-CH3 domain" is the weak immunogenicity of the antigen compared to the large carrier protein, an inherent problem associated with the carrier protein-peptide conjugates. The majority of the antibodies generated by such a conjugate are non-functional antibodies directed against the carrier protein. The preferred immunogens of the present invention are completely synthetic peptides which minimize the generation of irrelevant antibodies, and thereby produce more targeted immune responses to the target IgE-CH3 domain antigens, for example, SEC. FROM IDENT. NOS: 5-8 and 84. However, because the short IgE-CH3 domain antigen peptides of the present invention (eg, SEQ ID NOS: 5-8 and 84) are cell-dependent epitopes Non-immunogenic T, are dependent for immunogenicity of extrinsic Th epitopes. These are provided by the preferred peptides of the invention as promiscuously bonded th epitopes covalently. The immunogens of the invention produce site-specific immunoreactivity to provide accurate selection of the effector site and thus produce non-cross-linked anti-IgE antibodies. Antibodies specific for the resulting site inhibit sensitization and allergic response but do not induce spontaneous degranulation.

Specific examples are provided in the present invention as embodiments of the immunogenic peptide conjugates of the invention. These examples provide for the binding of synthetic immunostimulatory elements to IgE-CH3 domain antigen peptides (eg, SEQ ID NOS: 5-8 and 84) in such a way that potent cross-reactive antibodies are widely generated in a population genetically diverse host, against the target site in the IgE CH3 domain. These anti-IgE antibodies are non-anaphylactogenic and are directed specifically against I g E (examples 2 and 3). These antibodies, in turn, lead to the inhibition of histamine release and decreased IgE-mediated responses, resulting in effective treatment and / or prevention of IgE-mediated allergic diseases. By active immunization, the term "immunogen" referred to herein refers to a peptide conjugate composition which is capable of inducing antibodies against an effector site present in the third domain of the IgE heavy chain e (e.g., SEC). DE IDENT NOS: 5-8 and 84), leading to the inhibition or suppression of basophilic mediated by Ige and degranulation of mast cell. The peptide compositions of the present invention include IgE-CH3 domain antigenic peptides, preferably linked to carrier proteins by chemical coupling, more preferably the IgE-CH3 domain antigen peptides bound to promiscuous Assay T cell epitopes (Th epitopes). ) by means of chemical coupling, and more preferably completely synthetic peptides which contain IgG-CH3 domain antigen sequences promiscuous assistant T cell epitope sequences (Th epitope). The carrier proteins are covalently bound to the IgE-CH3 domain antigen peptides, preferably with a spacer (eg, Lys-Lys-Lys), by chemical coupling. The Th peptides (e.g., SEQ ID NO: 9-12) are covalently linked to the IgE-CH3 domain antigen peptides (e.g., SEQ ID NOS: 5-8 and 84) either by chemical coupling medium or preferably by means of direct synthesis, preferably with spacer (eg, Gly-Gly), to be adjacent either to the N- or C-terminal sequences of the IgE-CH3 domain antigen to evoke responses efficient antibody. The immunogen may also optionally comprise an immunostimulatory amino acid sequence, for example one corresponding to a domain of an invasive protein of the bacterium Yersinia ssp (Bret et al., Eur J Immunol, 1993, 23: 1608-1614) (SEQ. FROM ID: NO: 13). The general immunostimulatory sequence may comprise an optional spacer through which it binds to a Th peptide. The fully synthetic peptides of this invention may be represented by the formulas: (A) p- (IgE-CH3 antigen domain) - (B) 0 - (Th) mX -. Safa »i« rl.rt¿A¡í "" - "~ - (A) n- (Th) m- (B) 0- (IgE-CH3 antigen domain) -X or (A) n- (B) o- (Th) m- (B) o- (lgE-CH3 antigen domain) -X or (lgE-CH3 domain antigen) - (B) 0- (Th) m- (A) nX or (Th) m- (B) 0- (IgE-CH3 antigen domain) - (A) nX wherein each A is independently an amino acid or a general immunostimulatory sequence; each B is selected from the group consisting of amino acids, -NHCH (X) CH2SCH2CO-, -NHCH (X) CH2SCH2CO (eN) Lys-, -NHCH (X) CH2S-succinimidyl (eN) Lys-, and -NHCH (X) CH2S- (succinimidyl) -; each Th is independently an amino acid sequence constituting an attendant T cell epitope, or an enhancer analog or segment thereof; the IgE-CH3 domain antigen represents the sequence of an IgE CH3 domain antigen peptide as defined herein (or a cross-reactive and immunologically functional analogue thereof); n is from 0 to about 10; m is from 1 to about 4; and o is from O to about 10. The peptide immunogen of the present invention comprises from about 25 to about 100 amino acid residues., preferably from about 25 to about 80 amino acid residues and more preferably from about 25 to about 65 amino acid residues. When A is an amino acid, it can be any amino acid that does not occur naturally or any that occurs naturally. Amino acids that do not occur naturally include, but are not limited to, D-a-amino acids, β-alanine, ornithine, norleucine, norvaline, hydroxyproline, trioxin, α-aminobutyric acid, homoserine, citrulline, and the like. Naturally occurring amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, trionine, tryptophan, tyrosine and valine. In addition, when n is larger than one, and two or more of groups A are amino acids, then each amino acid can be independently the same or different. When A is an invasin domain, it is an immunostimulatory epitope of the invasin protein of a Yersinia species. This immunostimulatory property results from the ability of this invasin domain to interact with the β1 integrin molecules present in T cells, particularly memory T cells or immunoactivated cells. The specific sequence for an invasin domain that was found to interact with the β1-mtegrins has been described by Brett et al. (Eur J Immunol, 1993). A preferred embodiment of the invasin domain (Inv) for binding to a promiscuous Th epitope has been previously described in US 5,759,551 which is incorporated herein by reference. The Inv domain has the sequence Thr-Ala-Lys-Ser-Lys-Lys-Phe-Pro-Ser-Tyr-Thr-Ala-Thr-Tyr-Gln-Phe (SEQ.

IDENT. NO: 13) or is an immunostimulatory homologue thereof of the corresponding region in another species of invasive protein of Yersinia species. Such homologs can thus contain substitutions, deletions or insertions of amino acid residues to accommodate the variation of the bacterial strain, with the proviso that the homologs retain the immunostimulatory properties. An immunostimulatory homolog can also comprise an optional spacer through which it binds to a Th epitope. In one embodiment, n is 3 and (A) 3 is an invasin domain (Inv), glycine and glycine, in that order. (B) 0 is an optionally spaced and comprises naturally occurring amino acids or amino acids that do not occur naturally as described above. Each B is independently the same or different. The carrier proteins are covalently linked to the peptides with a spacer (eg, Lys-Lys-Lys) by chemical coupling. The amino acids of B can also provide a spacer, for example Gly-Gly or (eN) Lys, between the promiscuous Th epitope (e.g., SEQ ID NO: 9) and the IgE-CH3 peptide (e.g. DE IDENT NO.5), and cross-reactive analogues and immunological functionalities thereof. In addition to the physical separation of the Th epitope from the B-cell epitope, ie, the IgE-CH3 peptide (e.g., SEQ ID NO: 5) and immunological analogues thereof, the spacer may break any secondary artifact structures created by the binding of the Th epitope to the IgE-CH3 peptide (eg, SEQ ID NO: 5) and cross-reactive analogues and immunological functionalities thereof and thereby eliminate interference between them. Th and / or B cell responses. B amino acids can also form a spacer which acts as a flexible hinge that improves the separation of Th and IgE domains. Examples of the sequences encoding flexible hinges are found in the hinge region of the immunoglobulin heavy chain. Flexible hinge sequences are often rich in the future. A particularly useful flexible hinge is provided by the sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO: 16), wherein Xaa is any amino acid, and preferably aspartic acid. The conformational separation provided by the amino acids of B allows more efficient interactions between the presented peptide immunogens and the appropriate Th and B cells and thus improves the immune responses to the Th epitope and the epitope that produces antibody and its cross-reactive and functional immunological analogues. thereof. Th is a sequence of amino acids (natural or non-natural amino acids) comprising a Th epitope. A Th epitope can consist of a continuous or discontinuous epitope. Consequently, not all Th amino acids are necessarily part of the epitope. Accordingly, Th epitopes, which include analogs and segments of Th epitopes, are capable of enhancing or stimulating an immune response of the IgE-CH3 antigen peptides (e.g., SEQ ID NOS: 5-8 and 84). , and immunological analogues thereof). Th epitopes that are immunodominant and promiscuous are highly and widely reactive in animal and human populations with widely divergent MHC types (Partidos et al., 1991; US Patent No. 5,759,551). The Th domain of the subject peptides has from about 10 to about 50 amino acids and preferably from about 10 to about 30 amino acids. When multiple Th epitopes are present (ie, m > 2), then each Th epitope is independently the same or different. The Th segments are contiguous portions of a Th epitope that are sufficient to enhance or stimulate an immune response to the IgE-CH3 peptide (eg, SEQ ID NO: 5) and immunological analogues thereof.

,? The Th epitopes of the present invention include as examples, but are not limited to, antigen core epitopes and hepatitis B surface derived from pathogens.

(HBs Th and HBc Th), pertussis toxin assistant T cell epitopes (PT Th), tetanus toxin assistant T cell epitopes (TT Th), T cell epitopes F protein attendant of measles virus (MVF) Th), epitopes of T-cell assistant protein of the outer main membrane of Chlamydia trachomatis (CT Th), diphtheria toxin assistant T-cell epitopes (DT Th), attendant T-cell epitopes of Plasmodium falciparum circumesporozoite (PF Th ), epitopes of Triosaphosphateisomerase attendant T cell Schistosoma mansoni (SM Th), and T cell epitopes attendant Trat of Escherichia coli (TraT Th). The pathogen-derived Th was enlisted as SEC. FROM IDENT. NOS.2-9 and SEC. FROM IDENT. US: 42-52 in U.S. Patent No. 5,759,551; as an assistant site of Chlamydia P11 in Stagg et al., Immunology, 1993; 79: 1-9 (also listed here as SEQ ID NO: 12); and as peptide HBc 50-69 in Ferrari et al., J Clin Invest, 1991; 88: 214-222, and is incorporated herein by reference. Exemplary Th sites of the invention also include the artificial Th site termed "Syn Th (1,2,4)" (SEQ ID.

NO: 9), artificial SSAL sites "(1,4,9 PALINDROMIC) Th", "IS (1.4.9 PALINDROMIC) LF Th "y" IS (1,4,9 PALINDROMIC) LF and simplified Th "(SEQ ID NOS: 10, 11 and 60), and the like immunologically functional proteins of the same. Functional Th analogues include immunomodulatory analogs, cross-reactive analogs and segments of any of these Th epitopes. Functional Th analogues further include conservative substitutions, additions, terminations and insertions from one to about 10 amino acid residues in the Th epitope which do not essentially modifies the Th stimulator function of the Th epitope. The synthetic peptides of this invention are generally from about 50 to about 90 amino acids, and comprise (a) an immunostimulatory invasin domain, (b) an attendant T cell epitope (Th) , and (c) an IgG-CH3 domain antigen peptide. More specifically, the synthetic peptides of this invention are described by the formulas (A) n- (Th) m- (B) 0- (IgE-CH3 antigen domain) -X, (A) n- (B) 0- ( Th) m- (B) 0- (IgE-CH3 domain antigen) -X, (A) n- (IgE-CH3 domain antigen) - (B) 0- (Th) mX, (IgE-CH3 domain antigen) - (B) 0- (Th) m- (A) nX, and (Th) m- (B) 0- (IgE-CH3 domain antigen) - (A) nX. The Th epitope is attached, optionally through a spacer B, to either the N-terminal or C-terminus of the IgE-CH3 peptide and cross-reactive and functional immunological analogs thereof. Preferred peptide immunogens of this invention are peptides that contain the IgE-CH3 domain antigen peptides (eg, SEQ ID NO: 5) (or immunological analogues thereof) and Th peptides, and optionally Inv ( ID SECTION NO: 13). In a more preferred embodiment, the Th epitope is HBs Th, HBc Th, MVp Th, PT Th, TT Th, CT Th (eg, SEQ ID NO: 12) or artificial Th (SEQ ID. NOS: 9-11 and 60), or functional immunogenic analog thereof, and optionally, A is Inv (SEQ ID NO: 13) linked through a spacer (B) 0, such as Gly-Gly or (eN) Lys. The structure of the IgE-CH3 domain antigen comprises a peptide sequence taken from the CH3 domain of human IgE (amino acids 413-435 of SEQ ID NO: 1) or the homologous sequences of other species (eg SEQ ID NO. NOS: 6-8 and 84) and subjected to the following modifications: The target site is modified from naturally occurring IgE sequences by the insertion of a cysteine residue on the N-terminal side of position 413 or homologous position unless that the cysteine is already present at positions 413 or 414 in the natural sequence, the substitution of the native cysteine at position 418 or the position corresponding to a non-human homologous sequence, or any other cysteine of the native target sequence by serine (unless the native cysteines are present in positions 413 or 414 and 435 or 436), the insertion of cysteine on the C-terminal side of position 435 or homologous position unless the cysteine is already present at positions 435 or 436 in the natural sequence ', and the formation of a disulfide bond between the cisterns retained for produce a cyclical structure. The cyclic structures also comprise from 1 to 5 additional amino acids taken from either the segment terminus of 413-435 IgE with the proviso that the wavy structure of single disulfide is retained. A target antigen optimized for human I g E sequence Cys-Gly-Glu-Thr-Tyr-Gln-Ser-Arg-Val-Thr-His-Pro-His-Leu-Pro-Arg-Ala-Leu-Met-Arg - Ser-Thr-Thr-Lys-Cys (SEQ ID NO: 5) is provided by the present invention. The IgE target antigen is cyclized through unnatural terminal cysteines and the first serine residue replaces the cysteine residue of the natural sequence. The antibody evoked by the peptide immunogens comprises this IgE-CH3 domain antigen cross-reactive with human IgE and inhibits the sensitization of human basophils by human IgE. In the same way, the IgE-CH3 domain antigen sequences corresponding to IgE from other species can be derived from the homologous chain segment of the relevant species. For example, such target sequences can be taken ii ^ yy ^.? ^^ A ^ A ^^^^^^^^^^^^^^^^^^^. of the e chain sequences of dog, rat and mouse, shown in Table 1 as SEC. FROM IDENT. NOS: 2, 3 and 4, and the equine sequence published by Navarro et al., And the IgE-CH3 domain antigen sequences such as SEC. FROM IDENT. NOS: 6, 7, 8 and 84 can be derived. The cross-reactive and immunologically functional analogs of the IgE-CH3 domain antigen peptides (eg, SEQ ID NOS: 5-8 and 84) according to the invention, may additionally comprise conservative substitutions, additions, deletions, or insertions of one to about four amino acid residues, with the proviso that the resulting peptide analogs are capable of producing cross-reactive immune responses with the IgE-CH3 peptides (eg, SEQ ID NOS: 5-8). and 84). Conservative substitutions, additions, and insertions can be achieved with natural or unnatural amino acids as designated herein. Peptide compositions containing mixtures of the subject peptide immunogens with two or more of the Th epitopes can improve immunoefficiency in a larger population and thus provide an improved immune response to the IgE-CH3 domain antigen (eg, SEQ ID. NOS: 5-8 and 84). The peptide immunogens of this invention can be manufactured by chemical synthesis methods which are well known to the ordinarily skilled artisan. See, for example, Fields et al., Chapter 3 in Synthetic Peptides: A User's Guide, ed. Grant, W. H. Freeman & Co., New York, NY, 1992, p. 77. When a peptide immunogen includes an SSAL Th, the coupling of the alternative amino acids at a given variable position is achieved by providing a mixture of the amino acids specified for that position. Accordingly, the peptides can be synthesized using the automated Merrifield solid phase synthesis techniques with the α-NH2 protected either by i-Boc or Fmoc chemistry using side-chain protected amino acids, for example, an Applied Biosystems Peptide Synthesizer Model 430A or 431. After the complete assembly of the desired peptide immunogen, the resin is treated according to standard procedures to unfold the peptide from the resin and unblock the functional groups on the amino acid side chains. The free peptide is purified, for example by HPLC, and characterized biochemically, for example, by amino acid analysis, mass spectrometry, and / or by sequencing. The methods of purification and characterization for the peptides are well known to those of ordinary skill in the art. Other chemical means for generating the synthetic peptide constructs of the invention containing IgE and Th sites include the ligation of haloacetylated and cysteinated peptides to ^^? M ?? ?? through the formation of a "thioether" bond. For example, a cysteine can be added to the C-terminus of a Th-containing peptide and the thiol group of the cysteine can be used to form a covalent bond for an electrophilic group, such as an a-e-NH2N-chloroacetyl-modified group. or a maleimide derived from a lysine residue bound to the N-terminus of an IgE-CH3 peptide (eg, SEQ ID NO: 5) or functionally cross-reactive immunological analogues thereof. In this way, a construct with Th- (IgE-CH3 domain antigen) or its inverse, (Ig E-CH3 domain antigen) -Th, can be obtained. The subject immunogen can also be polymerized. Polymerization can be achieved for example by the reaction of the immunogen with a cross-linking agent, for example, by reaction between the glutaraldehyde and -NH2 groups of lysine residues, using routine methodology. By another method, a synthetic immunogen, such as for example "A-Thm- (IgE-CH3 antigen domain)", can be polymerized or copolymerized with another immunogen by the use of an additional cysteine added to the N-terminus of the synthetic immunogen. The thiol group of the N-terminal cysteine can be used for the formation of a "thioether" bond with an α-NH2 or e-NH2 group of modified haloacetyl amino acid or maleimide derived from a lysine residue that binds to the N-terminus of a branched poly-lysyl core molecule (e.g., K2K, K4K2K or K8K4K2K). The subject immunogen can also be prepared as a branched polymer through ^. Y-? tSyy ^^^^^ of the synthesis of the desired peptide construct directly on a branched poly-lysyl core resin (Wang et al., Science, 1991; 254: 285-288). Alternatively, the longer synthetic peptide immunogens can be synthesized by well-known recombinant DNA techniques. Many standard manuals on molecular cloning technology provide detailed protocols for producing the peptides of the invention by the expression of recombinant DNA and RNA. To construct a gene encoding a peptide of this invention (eg, immunogenic peptides comprising SEQ ID NOS: 5-8 and 84, and other species-specific homologs) the amino acid sequence is translated inversely within a sequence of nucleic acid, preferably using the codon optimized for the organism in which the gene will be expressed. Next, a gene encoding the peptide is typically manufactured by synthesizing overlapping oligonucleotides which encode the peptide and the necessary regulatory elements. The synthetic gene is assembled and inserted into the desired expression vector. The synthetic nucleic acid sequences encompassed in this invention include those encoding the peptides of the invention, immunologically functional homologs, and nucleic acid constructs characterized by changes in the non-coding sequences that do not alter the immunogenic properties of the peptide encoded therein. . The acids "" "" "" '"" Nucleics comprising sequences encoding the peptides of this invention are also provided. The synthetic gene is inserted into a suitable cloning vector and the recombinants are obtained and characterized. The peptide is then expressed under conditions appropriate for the host and selected expression system. The peptide is purified and characterized by standard methods. The nucleic acids of this invention may be useful in themselves as components of the so-called "DNA vaccines". In this embodiment of the invention, the expression of the immunogenic peptides of the invention is induced in the patient's own cells, by the introduction into these cells of nucleic acids encoding the peptides. Methods for manufacturing and using DNA vaccines are described in U.S. Patent Nos. 5,580,859, 5,589,466, and 5,703,055; see also WO 97/02840 and W. McDonnell and F. Askari, New Engl. J. Med., 1996, 334: 2-45, all of which are incorporated herein by reference. Such methods for making and using the peptides and peptide conjugates of this invention are contemplated to be within the scope of this invention. The efficiency of any peptide composition of the present invention can be established by an in vitro test in which a host animal is immunized with a peptide composition of the invention and the resulting antibodies show inhibit sensitization of basophils and mast cells by I g E, as shown in Examples 2 and 6. The efficiency can be established in vivo by injecting a host with an appropriate peptide composition for species (eg, immunizing mice with a formulation of immunogens comprising SEQ ID NOS: 24 and / or 25) followed by observation of the humoral immune response to the IgE-CH3 peptide and functional cross-reactive immunological homologs thereof, as detailed in Example 5. Another aspect of this invention provides a peptide composition comprising an amount immunologically effective of one or more of the peptide immunogens of this invention in a a pharmaceutically acceptable supply. Accordingly, the subject peptides can be formulated as a pharmaceutical composition using adjuvants, pharmaceutically acceptable carriers or other ingredients routinely provided in vaccine compositions. Among the ingredients that can be used in this invention, are adjuvants or emulsifiers including alum, incomplete Freund's adjuvant, liposine, saponin, squalene, L121, emulsifier, monophosphoryl lipid A (MPL), QS21, ISA51, ISA35, ISA 206, and ISA 720, as well as other adjuvants and known effective emulsifiers. The formulations include formulations for immediate release and / or sustained release, and for the induction of systemic immunity and / or induction of localized mucosal immunity, which can be achieved by, for example, capture of immunogen by, or co-administration with microparticles. . Such formulations are readily determined by one of ordinary skill in the art, and methods for the preparation, preservation, and sterilization of such formulations are known to those skilled in the art. The present drugs can be administered by any convenient route, which includes subcutaneous, oral, intramuscular route, or other parenteral or enteral route. Similarly, the drugs can be administered as a single dose or multiple doses. Immunization plans are easily determined by the commonly skilled artisan. The pharmaceutical composition of the present invention contains an effective amount of one or more of the peptide immunogens of the present invention and a pharmaceutically acceptable carrier. Such a composition in a suitable unit dosage form generally contains about 0.5 μg to about 1 mg of the immunogen per kg of body weight. When multiple doses are supplied, it can conveniently be divided into an appropriate amount per unit dosage form. For example, an initial dose, for example 0.2-2.5 mg; preferably 1 mg, of immunogen represented as a peptide composition of the present invention, can be administered by injection, preferably intramuscularly, followed by repeated doses (impeller). The dosage will depend on the age, weight and general health of the patient as is well known in the vaccine and therapeutic techniques. The immune response for peptide immunogens synthetic IgE-CH3 can be improved by delivery through capture in, or on biodegradable microparticles of the type described by O'Hagan et. al., (Vaccine, 1991; 9: 768-771). The immunogens can be encapsulated with, or without an adjuvant in biodegradable microparticles, to enhance immune responses, which include immunity of the localized mucosa, which can be especially applicable to allergic reactions localized in the mucosa, and to provide controlled time release during sustained or periodic responses, for oral administration, and for local administration (O'Hagan et al., 1991; Eldridge et al., Molec. Immunol., 1991; 28: 287-294). The pharmaceutical compositions of this invention are used in a manner similar to that of vaccines, for the prevention of atopic allergic reactions including allergic rhinitis, those of food allergies, asthma, anaphylaxis, lice allergy dermatitis and other mediated hypersensitivities. for IgE. All patents and literature references referenced above are incorporated herein by reference. Peptide immunogens and specific peptide conjugated immunogens are provided in the following examples , ia ¡.A.í! l¿ßUnly? ? ^^ and to illustrate the invention. These examples are for illustration purposes only, and should not be construed as limiting the scope of the invention in any way.

EXAMPLE 1 IDENTIFICATION OF SITES POTENTIAL EFFECTORS IN THE HUMAN IQE MOLECULE Peptide Design Sites within the CH2 and CH3 domains of the heavy chain of human IgE were selected for imitation by peptides, according to the descriptions of Helm et al. (1988) and Presta ei al. (1994) that a long segment of the e chain, which overlaps both of these domains, participates in the IgE binding to the FceRI receptor. The sequences from such sites were synthesized as target site peptides and became antigens by (1) linking them through chemical coupling to long carrier proteins, such as KLH or (2) construction peptides where promiscuous Th and Inv (SEQ. DE IDENT NO: 13) were linked to the amino terminal of the target sites. Specific sites within these domains were selected as peptides for cyclization based on the predictions of Brookhaven's three-dimensional model for human I g E (http: www.pdb.bnl.gov/pdb.bin/pdbids) of loops exposed to surface. The specified cyclic constraints were installed within the design of these peptides to maximize cross-reactions between the antigens and the native IgE molecule. Consequently, several of the synthetic constructs were synthesized with introduced cysteines not found in the native sequence to produce disulfide bond loops of specified position, and imitation of the loop structures predicted by the Brookhaven model. In some cases, the naturally occurring cysteines were replaced with serines to prevent the formation of conformations not favored by the model. The constructions are listed in Table 2. Peptides labeled by * in the description column of Table 2 are cyclized by cysteine disulfide bonds. The cysteine residues that have been inserted into the native sequence for cyclization are denoted in the amino acid sequences of Table 2 by parentheses, other residues that have been inserted, substituted by a native residue, or are natural cysteines, which participate in Disulfide linkages are indicated in the amino acid sequences of Table 2 underlined. Other peptides are linear. The peptides labeled by "a" in the third column represent the peptide antigen IgE-CH2 / 3 or -CH3, chemically linked to a carrier protein KLH for conventional coupling reasons of glutaraldehyde or MBS (m-Maleimidobenzoyl-N-hydrosuccinimide ester, Pérce Chemical Co., Catalog No. 22510). Peptides labeled "b" in the third column were synthesized as IgE antigen peptides in tandem with the Th sites shown. The Th sites used include 3 *, *. *, The HBs19.32 taken from the hepatitis B virus, the MVf Th taken from the measles virus and the PT? 49-? 6, taken from pertussis toxin as referenced in U.S. Patent No. 5,759,551, CT Th termed P11 (Stagg et al., 1993) and novel artificial Th sites termed "1,4,9 PALINDROMIC Th" (SEQ. DE IDENT NO: 10), "IS (1, 4, 9 PALINDROMIC) LF Th" (SECTION ID NO: 11), "IS (, 4, 9 PALINDROMIC) LF simplified Th" (SEC. NO: 60), and "Syn Th (1,2,4)" (SEQ ID NO: 9). Peptides marked by "c" are variants of the "b" constructs synthesized in tandem with the immunostimulatory peptide of the Inv domain (SEQ ID NO: 13). Constructs "b" and "c" were also synthesized with Gly-Gly spacers for the separation of the target antigen IgE-CH2 / 3 or CH3 site from the h site, and the Th separation from the Immunostimulatory site Inv. Constructs "b" and "c" of Table 2 have the Th and / or Inv domains attached to the amino terminal of the IgE target site. The peptide immunogens of Table 2 were selected as candidate antigenic peptides for the treatment of allergy, by the hyperimmunization of animals followed by the testing of the hyperimmune sera for cross-reactivity to human IgE. * - "• * --- ----« • - ^ mtiataj ...

Specific Procedures for the Selection of Peptides Anti-Calenic Candidates Objective: 1. Synthesis of Peptides antigen domain lqE-CH3 and Conjugates. The peptides listed in Table 2 were synthesized by the Merrifield solid phase synthesis technique on Applied Biosystems automated peptide synthesizers using Fmoc chemistry. When a peptide immunogen includes an SSAL Th, the coupling of one of the alternating amino acids in a given variable position is achieved by providing a mixture of amino acids at equivalent molar ratios. After the complete assembly of the desired peptide, the resin is treated according to the standard procedure using trifluoroacetic acid to unfold the peptide from the resin and unblock the groups protectors in the side chains of amino acids. For the cyclic peptides, the split peptides were dissolved in 15% DMSO in water, for 48 hours, to facilitate the formation of the intradisulfide bond between cysteines. 2. Experimental Immunizations 20 Rats or guinea pigs were immunized intramuscularly with experimental peptide immunogens. The dose was 100 μg of peptide suspended in a volume of 0.5 ml. The first dose was administered with Freunds Complete adjuvant. Subsequent doses were administered in Incomplete Adjuvant of Freunds. The animals received injections in weeks 0, 3, 6 and 10 or 0, 2, 4 and 8. The test bleedings were taken at biweekly intervals and the reactivities were determined by IgE peptide ELISA and cross reactivities by human IgE ELISA. 3. ELISA tests Peptide ELISAs for the determination of anti-IgE peptide reactivity were run in 96 well microtitre plates coated with peptide by incubation for 1 hour at 37 ° C with an appropriate target antigen site "a" peptide without carrier to 0.5 μg / ml using 100 μg per well in 10 mM of NaHCO3 buffer, pH 9.5. For the determination of the cross-reactivity of anti-human IgE, the human IgE ELISAs were run on microtitre plates of 96 wells coated with IgE coated in a similar manner with a human IgE myeloma protein (American Biosystems, Inc. cat.No.A113) at 5 μg / ml. The wells coated with human IgE or peptide were incubated with 250 μL of 3% by weight of gelatin in PBS, at 37 ° C for 1 hour to block nonspecific protein binding sites, washed three times with PBS containing 0.05% in volume of TWEEN 20, and then dried. The test samples were serially diluted with PBS containing 20% by volume of normal goat serum, 1% by weight of gelatin and 0.05% by volume of TWEEN 20. 100 μL of the diluted sample was added to each of the wells and allowed to react for 1 hour at 37 ° C. The wells were then washed six times with 0.05 volume% TWEEN 20 in PBS to remove unbound labeled antibodies. Added »-», - n mulli r • y -irtuui rl, i "t látl¡rtí ^ li ^ ¿j ^ 100 μL of horseradish peroxidase labeled goat anti-goat IgG antibody or anti-guinea pig IgG antibody of goat at a predetermined optimal dilution in 1% by volume of its normal goat roast, 0.05% by volume of TWEEN 20 in PBS to each well and incubated at 37 ° C for 30 minutes. The wells were washed six times with 0.05% by volume of TWEEN 20 in PBS to remove unbound labeled antibody conjugate and reacted with 100 μL of the substrate mixture containing 0.04% by weight of orthophenylenediamine (OPD) and 0.12% by weight. volume of hydrogen peroxide in sodium citrate buffer pH 5.0, for 15 minutes. The reactions were stopped by viewing 100 μL of 1.0 M H2SO4 and the absorbance was measured at A 92. The ELISA titers, expressed as the log-io of the reciprocal dilution, were calculated based on the linear regression analysis of the Absorbances, with cut A492 set to 0.5. This cut-off value was rigorous since the values for the normal guinea pig control samples diluted in each test was less than 0.15.

Results The candidate antigen target sites are described in Table 2. They are shown either as "a" peptides attached to the KLH carrier or as "b" peptides bound to synthetic Th sites or as "c" peptides attached to synthetic Th and Inv. The rats or guinea pigs were inumunited as described in the above Specific Procedures and the hyperimmune antisera were collected in week 8 were analyzed by anti ELISA peptide and anti ELISA human IgE as described in the Specific Procedures. Many of the peptide immunogens CH2 / 3 and CH3 were immunogenic, since they evoked antypeptide antibodies with titers in the range of log 10 2-5. The target antigenic sites CH2 / 3 comprising long segments of the human chain of 301-376 (numbering scheme of Table 1) were all strongly cured reaction with human I g E, as shown by the log 1 or on anti human ELISA I g E of more than 3. The crude reactivity was lost for some CH3 peptides which started at position 342 and beyond (eg, entries 21 and 22). However, for CH3 peptides that included a relatively short region comprising 354-372, the crude reactivity was greatly restored (eg, entries 27, 28 and 29) with the exception of entry 31 (354-368). Another short region of crude reactivity is seen at entry 10 (positions encompassing cyclic peptide 374-3865). As evidenced by the lack of cross-reactivity of the inputs 14, 17, 23, 24, 25 and 26, a sequence expansion extending from 365 to 413 is devoid of cross-reactivity, despite the overlap with the region 354-372 of cross-reactivity and a region of cross-reactivity represented by entry 20 (374-385). It is interesting that the short cross reactivity exemplified by entries 27, 28, S? r-i *. ÍAA * £ .ly. .. »*» ...., .. - «-. ^,.,",.!, Tk.¿.y ^ MÍK > . *. and > yh.?í(tfñn,ñ.._ _JJJ_L_1 _ ^ _ mm m_J ____ J | m 29 (354-372) and 20 (374-385) are losses in the conformation of 17 long cyclic peptide entry (365-396), despite their overlap in those cross-reaction regions Cross-reaction sites that overlap sites without cross-reaction are found again beyond a region that starts around position 399 and extends to position 445, as shown by the reactivities crossings of entries 15 and 30 and the weak cross reactivities of entries 19 (432- ^ 45) and 23 (404-413) .It is significant that of the two similarly cyclized peptides which include the position 418, 15 (413- 435 and 18 (404-434), only entry 15 (SEQ ID NO: 5), in which the cysteine at position 418 has been replaced by serine, is cross-reactive with human IgE. corresponding to a IgE defect site described by Stanworth (Stanworth et al., Lancet, 1990; 336: 1279-12 81) failed to show cross-reactivity (entry 34). These results demonstrate that the cross-reactivity for peptides I g E is a complex phenomenon influenced by conformational characteristics, and can not be predicted from a direct analysis of overlapping linear peptides. The candidate IgE-CH3 domain antigens were selected from the conjugates shown to cross-react with human I g E in Table 2 and used for further analysis.

EXAMPLE 2 IDENTIFICATION OF THE EFECTOR SITE IN THE HUMAN IQE MOLECULE The IgE-CH3 domain antigen peptides were selected for further analysis from among those peptide conjugates of Table 2 that exhibited high affinity cross-reactivities for human IgE, as evidenced by the titers anti-IgE for their respective antisera of more than log = 3. The guinea pig mammalian sera were produced as described above. The guinea pig IgG antibodies were purified from the hyperimmune sera by protein A affinity chromatography and analyzed by a functional test to determine the ability of anti-IgE to inhibit the sensitization of human basophils by allergen-specific I g E. The endpoint of the test is expressed as the percent inhibition of histamine release mediated by IgE. The guinea pig IgG antibodies were purified from the serum by Protein A Affinity chromatography (ImmunoPure® Immobilized Recomb® Protein A, Pierce) and the eluted anties were prepared at a standard concentration of 8 mg / ml in 25 mM PIPES buffer, 0.15 M NaCl, pH 7.2. A control anty preparation was prepared from the pooled serum of guinea pigs immunized with a relevant peptide immunogen. These anties were used in tests that measured the reduction in IgE-mediated sensitization of human basophils. The ** «f • - T-fi" 1 ^ - "r t - '.- *** - *. The human basophils were prepared from the venous blood of volunteers using centrifugation through Percoll density gradients (MacGlashan J Allergy Clinic). Immunol, 1993; 91: 605-615.) The band leukocytes were harvested, washed and resuspended in 0.1 ml of PAGCM regulator as described (MacGlashan, 1993) except that the PAGCM regulator used to suspend the cells was manufactured with water that contains 44% D2O The lg E used for the test was specific for allergen, either for specific IgE for human BPO or chimeric human IgE specific for HIV gp120 glycoprotein The specific IgE for allergen used for sensitization at 0.25 μg / ml was pre-incubated with a volume of a purified guinea pig anty at 8 mg / ml, total volume 0.1 ml, for 15 minutes at 37 ° C, before being added to the basophils.The anty mixture was added to the cells and incubated for 20 minutes to allow a sensitization of the cells by IgE without complex. The sensitized cells were then stimulated by the addition of the allergen, either BPO2? -HSA or a gp120 polypeptide as described (MacGlashan, 1993). After an appropriate incubation period (usually 45 minutes), the cells were separated from the supernatant and the supernatant was tested for histamine content by an automated fluorimetric technique (Siraganian, Anal Biochem, 1974; 57: 383-394). All reactions were performed in duplicate. The percentage of histamine release it was calculated from the sample's relation to total histamine after the spontaneous release was subtracted from both. The results are expressed as percent inhibition of histamine release, as determined from the release ratio of histamine by the experimental anty to the release of histamine by the control anty of specific specificity. The histamine release tests on human basophils were performed kindly under conditions coded by Dr. Donald W. MacGlashan, The Johns Hopkins University School of Medicine, Johns Hopkins Asthma and Allergy Center, Baltimore. Results The results for the inhibition of the histamine release tests are shown in Table 3 for antipeptide anties of guinea pigs that showed cured reactivities for human IgE of log10 3. The determinations were made of anties purified from bleeds of 8 weeks, except for anties against peptide entries 15b and 15c, which were also characterized from serum collected at week 12. The inhibition results shown for the anti-15b and anti-15c anties, 61% and 71%, were made on the Purified anties from indentations taken at week 8 and 12, respectively. The separated animals had been immunized with 15b and 15c, but anties from both sets of animals had been pooled for the week 8 results ?? eleven ?? i l.ii n 11 - írigtfair ,,, ,, MM ,, t, M ^ aMi ^ iMM¿jtM ¿BÍa Mittt ^ M. ^^ fa ^^ M ^^^^^^^^^^^^ ja ^^^^ MMl ^ y 12 shown in Table 3. (The guinea pigs of these groups received an additional dose of peptide conjugate at week 10 and thus retained high levels of anty for week 12 bleeding. The inhibitory reactivity Significance of the anti-15 anties was unexpected in comparison to the reactivities of the IgE cross-reactive anties evoked by the remaining of the peptides shown in Table 3.

These other IgE-CH3 domain antigenic peptides failed to provide inhibition, or exhibited histamine release inhibition levels that were negligible and non-reproducible. The results of inhibition of histamine release and cured IgE reactivities for antibodies produced by antigen peptides of IgE-CH3 domains that overlap with the antigenic site (SEQ ID NO: 5) of peptide entries 15b (SEC IDENTIFICATION NO: 14) and 15c (SEQ ID NO: 15) can be compared. The IgE antigens represented by the peptide entries 19, 23, 24 and 33 comprise short overlaps with the input antigen sequence 15 (SEQ ID NO: 5). Their reactivity cured for I g E is unfavorably compared to the inlet 15 and they are devoid of inhibitory activity. The sequence of IgE antigen (SEQ ID NO: 44) of entry 18 comprises the complete antigen sequence of entry 15, except that (1) the carboxyl terminal lysine is deleted (2) the cysteine that ui »? utt? ÍiA *« ü¡? -dbtedaí -? t? ? ,.,? mm ^^? ^ Utf ^ ¡^ occurs naturally at position 418 is retained, and (3) there are nine additional N-terminal amino acids. It is not cross-reactive with IgE and fails to inhibit the release of histamine. In contrast, the immunogens of entry 15, which have SEC. FROM IDENT. NO: 5, provides unexpected reactivities. The IgG-CH3 domain antigen sequence of entry 1 with a specified cyclic structure by introducing terminal cysteines, and without contribution of the cysteine at position 418 (which has been replaced), provides an antigen that is reacted with cured IgE and produces antibodies that inhibit IgE sensitization. The antibodies produced by the entry 15b (SEQ ID NO: 14) and 15c (SEQ ID NO: 15) were prepared from the indentations of 13 weeks and tested individually. At week 13, both reactivities cured for IgE, as determined by I g E ELISA, and percent inhibition of histamine release, had decreased from week 12 values. However, antibodies of both preparations were found be individually effective to reduce the release of histamine: anti-15b inhibited 28% and anti-15c inhibited 20%. The degree to which the histamine release was inhibited by any of these antibodies was dose dependent, as evidenced by the effect of antibody dilution. When an anti-15b preparation of week 13 was tested at full concentration (8 mg / ml), dilutions 1: 3 and 1: 9, the Histamine release inhibition percent was 28%, 21% and 14% respectively. A guinea pig anti-15b preparation was tested by direct challenge of basophils sensitized to IgE, in the absence of allergen, as an assessment of their ability to cross-link IgE to the receptor and induce degranulation. The release of histamine by anti-15b was equivalent to the level of spontaneous histamine release by the donor cells. This indicates that the antibody of specificity for the IgE antigen SEC. FROM IDENT. NO: 5 is not anaphylactogenic. Thus, active immunization with peptide conjugated immunogens comprising the IgE domain antigen-CH3 SEC. FROM IDENT. NO: 5 (SEQ ID NOS: 14 and 15) produces non-anaphylactogenic anti-IgE antibodies that inhibit IgE-mediated sensitization without themselves causing the release of histamine. These actively evoked pohclonal antibodies have specificity for a site I g E effects that has not been described by previous studies, which includes previous studies of non-anaphylactogenic therapeutic anti-IgE antibodies tried for the treatment of passive immunization allergy US Patent No. 4,940,782 , North American Patent. 5,420,251, and Presta ei al. , 1993).

- * - * * *. «L ^ jaaA EXAMPLE 3 SPECIFICITY OF ISOTIPO AND POTENTIAL FOR IMMUNOSUPPRESSION Polyclonal antibodies produced by active immune response to SEC. FROM IDENT. NOS: 14 and 15 were examined for specificity for IgE compared to IgG. The anti-15b guinea pig antibodies described in Example 2 which were prepared from week 12 bleeding were subjected to a parallel comparison of cross-reactivities to IgE and IgG, by the IgE ELISA described in Example 1 and by a similar IgG ELISA. For the IgE ELISA, the plates were coated with human IgE myeloma at 5 μg / ml. For the IgG ELISA, the plates were coated with purified human IgG (Sigma reagent human IgG grade), also at 5 μg / ml. The purified guinea pig anti-15b was tested by reactivities in both ELISA at concentrations of 0.5 and 0.1 μg / ml. The results were compared with the antibodies purified from control guinea pig serum and with a "non-antibody" control. The values of A 90 for the anti-15b antibody in IgE were 1.126 at 0.5 μg / ml and 0.344 at 0.1 μg / ml. The A490 values for anti-15b antibody in IgG were equal to the control antibody and base values. There was no cross-reactivity of guinea pig anti-15b with human IgG. The peptide composition of the invention did not evolve antibodies that recognize IgG antibodies, and therefore are isotype specific for IgE. They will suppress allergic reactions mediated by IgE and will not result in ^^^^ &J ^ y ^ ¿^^^^^ &Z ^^^ Z & ^ r undesirable immunosuppression of protective IgG antibody responses.

EXAMPLE 4 REPRESENTATIVE PEPTIDE CONJUGATES OF THE INVENTION The immunogenic peptide conjugates of the invention shown in Table 4A, which are fully synthetic peptides, were synthesized by the solid phase method delineated in Example 1. Each peptide in the Table can be represented in formula (A) n- (Th) m- (B) 0- (IgE-CH3 antigen domain) -X, but the peptides of other formulas described above are understood to be comprised within the peptides of this invention. The antigen sequence of IgE-CH3 domain is SEC. FROM IDENT. NO: 5, 6, or 8 in the peptides of Table 4A, but it is understood that the homologous IgE-CH3 domain antigen sequences of other mammalian species are comprised within the peptides of this invention. The immunogenic peptides comprise Th sites derived from foreign pathogens (eg, SEQ ID NO: 87) and also artificial Th (eg, SEQ ID NO: 14, 18, 21 and 90). In addition to the examples shown in Table 4A, other pathogen-related Th may be selected from the promiscuous Th sites exemplified in Table 5, and artificial Th may be selected from the exemplified Th sites.

MAXIMUM * F? m? t¡t &? in Table 6. Each peptide in this example has Gly-Gly or (eN) Lys spacers between immunogenic elements, but the peptides of the invention may have other spacers (e.g., SEQ. DE IDENT NO: 16) or no spacer. The peptides of these examples also comprise an optional Inv immunostimulatory site (eg, SEQ ID NOS: 15-19 and 22). It is understood, however, that the invention is not limited to Inv as an additional immunostimulatory element.

As shown by the KLH conjugate, peptide conjugates of the invention also include an IgE-CH3 domain antigen coupled to a carrier protein.

Materials and methods The representative peptide constructs of the invention listed in Table 4A (SEQ ID NOS: 18, 85, 87, 88, 90 and 91) were synthesized, split, cyclized and purified as described in Example 1. Peptide constructs were formulated for immunization within small animals such as guinea pigs, or within larger animals such as pigs or baduinos for the evaluation of their immunogenicities. The peptides were suspended in a volume of 0.5 ml containing emulsifiers or representative adjuvants such as ISA51, ISA720, DDA or monophosphyl lipid A (MPL). The dose was 100 μg of peptide for guinea pigs or 300 μg of i? 'iiirtmi. tt ^^^ H ^ átiF ^ a ^^ f? peptide for pig or baduino and the animals were immunized intramuscularly. The animals received injection at week 0, 3 and 6 or 0, 2 and 4 weeks as specified in Table 4B. Test bleeds 8 weeks after the initial immunization were evaluated by cross-reactivities for I g E by either human IgE or dog ELISA I g E as described in Example 1, except that a dog IgE ELISA was used for the dog IgE ELISA. dog IgE myeloma protein (Bethyl Laboratories Inc., Montgomery TX for plaque coating at 1 μg / ml, and reagent A / G labeled horseradish peroxidase protein (Pierce Chemical Co., Rockford IL) at an optimal dilution The peptide-induced cross reactivities were also evaluated for the ability to inhibit the release of IgE-mediated histamine.I g E of guinea pig, pig or baduin from immune sera was purified as the marker for the detection of dog IgE. samples were analyzed by affinity chromatography of protein / ... and were analyzed by functional test for the determination of the ability to inhibit the sensitization of human basophils by specific IgE for allergen, as described in detail in Example 2. The endpoint of the test was expressed as percent inhibition of histamine release mediated by I g E compared to the control antibody of the same species that arose with the specificity for an irrelevant antigen, as shown in Table 4B.

• MÉUlUIMIMMiM WMi -. ^. ^ .-. ^ .... U ^ .... ^. , i .and. **** .8 Results The representative peptide constructs were of relevant immunogenicity, since all tested peptides produced cross-reactivities directed to the strong site to the corresponding human IgE or dog IgE, as shown by the titles Log-io of the I g E antihuman or anti-dog IgE ELISA of more than 3 (Table 4B). The inhibition of IgE-mediated sensitization was observed for antibodies from guinea pigs, pigs, and baduin as assessed by the ability of peptide anti-IgE antibodies to inhibit histamine release by basophils. This functional cured reactivity of baduin antibodies is worthy of mention since the neutralization of human IgE by baduin IgG is almost a human system. Thus, the efficiency of a peptide construct of the invention, as an agent for the immunotherapy of allergy by active immunization, is indicated in a model that is almost homologous to peptide species and target species.

EXAMPLE 5 IMMUNIZATION OF MICE AND EVALUATION OF EFFECTIVENESS IN VIVO The efficiency of SEC peptides. FROM IDENT. NOS: 24 and 25 (37b and 38b) is evaluated with five groups of 16 mice per the sensitization and immunization protocol delineated later.

Groups of 16 mice (Balb / c), females, 8-10 weeks old, are immunized subcutaneously with the indicated peptide composition of the invention. Mice are given, doses of 20 μg / 0.2 ml at weeks 0, 3, 6, and 11. The first dose is prepared with Freunds Complete Adjuvant. Subsequent doses with Incomplete Freund's Adjuvant. Mice are sensitized to a hapten conjugate, diphenylated KLH (DNP-HLJ), at weeks 7 and 10. Sensitization achieved by intraperitoneal administration of DNP-KLH in 0.4% alum, with 5 μg / 0.2 ml / dose. Imitation immunizations and sensitizations are achieved in control groups by the administration of adjuvants with saline with phosphate buffer. The groups are as follows: 1. Sensitize imitation with peptide 37b and 0.4% alum. 2: Immunize / sensitize, with peptide 37b and DNP-KLH 3: immunize imitation / sensitize, with Freunds and DNP-KLH 4: Immunize / sensitize imitation, with peptide 38b and 0.4% alum 5: Immunize / sensitize, with peptide 38b and DNP-KLH. Serum is harvested at weeks 0, 5, 7, 9, 10, 11, 13, 16 and 20. Splenocytes are prepared from pairs of mice from each group at weeks 10 and 11. i ^ JiiMjtSid i JF, ^ ^ ÉaíFá? aááFM The IgG response to peptide antigens and DNP is observed by standard ELISA tests using an anti-mouse IgG mouse horseradish peroxidase conjugate, and microtiter plates whose wells were coated with peptide 37 unconjugated 5 (peptide antigen of mouse IgE-CH3 domain, SEQ ID NO: 8) for peptide ELISA, and plates coated with DNP-BSA conjugate for DNP ELISA. Cross-reactivity of anti-37b antibodies with mouse IgG was observed by a standard IgG Elisa on plates coated with mouse monoclonal IgE SPE 7 10 (Sigma). The IgG response to the peptide immunogens is compared to the cross-reactivity of mouse I g E between the groups over the course of 20 weeks, to determine 1) primary and secondary responses, 2) the presence of undesirable immunosuppression of IgG responsiveness and , 3) the occurrence of a desirable reduction in anti-IgE reactivity during weeks 10-20 as evidence of reversibility and safety of the antibody response to the peptide composition of the invention. In weeks 7, 9, 10, 11, 13 and 16, the IgE response by IgE ELISA complete and by ELISA specific for DNP. At weeks 10 and 11 the splenocifer B cells that secreted IgE with specificity for DNP are numbered by ELISPOT test specific for DNP. Also, because the levels of I g E in serum may not be completely predictive of anaphylaxis, that is, IgE determinations may be lost J ^ - > i > * - '-. . , Y *,,. - < Y * ,. ^, .. ^^, 1 ..,, -t..¿ > ^ Fcf, .A ^^ significant effects on sensitivity in vivo, sensitization of mice is measured by passive percutaneous anaphylaxis test of mouse serum in rats (heterologous PCA). Heterologous PCA is preferred to the autologous PCA test in mice because rat skin mastocytic cells are selectively cross-sensitized by mouse IgE as opposed to mouse IgG. Therefore, the heterologous mouse / rat PCA reaction is IgE-specific and is not confused with IgG mediated anaphylaxis which can occur in the autologous mouse PCA test (Maekawa and Ovary, J Immunol Methods, 1984; 71: 229-239). The results of ELISA, ELISPOT, and PCA are compared between groups for immunosuppression of IgE responsiveness and for the sotypic specificity of immunosuppression. The experimental methods are described later. Complete ELISA IqE For an ELISA to measure total mouse IgE in serum, the microtiter plates are coated with I g E anti monoclonal rat mouse, R35-72 (Pharmingen) at 1 μg / ml. The plates are coated, washed and blocked as described. Mouse sera diluted in series are added to the plates and incubated. The captured IgE is detected by the reaction by biotinylated monoclonal rat anti-mouse IgE, R35-118 (Pharmingen), followed by sequential additions of streptavidin-horseradish peroxidase (Pierce) and OPD. The values of A492 are determined.

ELISA IqE Specific for DNP For an ELISA to determine mouse IgE specific for DNP hapten in mouse serum that has been sensitized with DNP-KLH, microtitre wells are coated with DNP-BSA conjugate (Molecular Probes, Inc.) at 5 μg / ml. The IgE captured with DNP hapten specificity is detected as described above. ELISPOT specific for DNP For an ELISPOT test to determine B cells that secrete mouse IgE specific for DNP hapten, 5 μg / ml DNP-BSA conjugate was used to coat the sterile plate wells, whose wells are lined with nitrocellulose filters 0.45 μm, for example, a MULTISCREÉN HA Píate (Millipore Inc., catalog No. MAHAS4510). Serum diluted splenocytes prepared from sensitized control mice are added to the wells and incubated overnight at 37 ° C under 5% C02. The cells are washed from the plates and the cells secreting IgE with DNP hapten specificity are counted as spots located on the filters after staining by alkaline phosphatase conjugated to rat monoclonal antibody R35-118 with 5-bromo-4-chloro -3-indolyl phosphate (Sigma) as a colored substrate. PCA Heteroloqa Serial dilutions of sera from immunized / sensitized mice are injected and intradermally controlled within the shaved backs of adult male Sprague Dawley rats. The anesthetized animals received 10-12 injections of diluted serum into each of three parallel rows in on the dorsal skin (50 μl / site). Each pattern of injections is replicated in duplicates of animals. After a latent period of 24 hours, for effective sensitization of skin mastocytic cells, the rats were rotated by intravenous injection of 1 mg DNP-BSA in 1% Evans blue dye in PBS. In 30 minutes to 1 hour, the rats suffocate and peel so that the blue reactions can be observed inside the dorsal skin. A PCA titre is determined from the highest serum dilution which results in an easily definable spot.

EXAMPLE 6 IMMUNIZATION OF MICE AND INHIBITION OF ANAFILAXIS PASSIVE SKIN To study the effect of immunization by an immunogenic peptide of the invention on an IgE-mediated inflammatory reaction, an antibody response was produced to the mouse IgE-CH3 target antigenic site, SEC. FROM IDENT. NO: 8, by immunizing the mice with a peptide of the invention. The resulting mouse antiserum was then used to suppress passive cutaneous anaphylaxis (PCA) triggered by the cross-linking of mouse IgE bound to rat mast cells.

Materials and methods Balb / c mice were immunized with a peptide composition of the invention, SEQ. FROM IDENT. NO: 25, as described in ple 5, except that subcutaneous injections were given at weeks 0, 3, and 6 only and the mice were not sensitized. At week 8, mouse sera were harvested and evaluated for cross-reactivity to I g E by mouse IgE elisa. The mouse IgE ELISA was as described for the human IgE ELISA in ple 1, except that the microtiter wells were coated with 1 μg / ml of monoclonal anti-DNP IgE mouse IgG antibody (Sigma Chemical Co., St. Louis MO ), and goat anti-mouse IgG-labeled horseradish peroxidase (HRP) (Kirkegaard and Perry Laboratories, Gaithersburg MD) was used for the detection of captured mouse IgG. Thirteen of 20 immunized mice had cross-reacting antibodies to mouse IgE. The sera were pooled from seven mice showing ELISA titers against IgE = og-io 2.3 for use as site-specific anti-IgE. Another group of 10 balb / c mice was used to produce Murine IgE. This group was sensitized by a simple intraperitoneal administration of ovalbumin (Oa) in 0.4% alum, 1.0 μg / 0.2 ml. The IgE content of the mouse sera was measured on day 20 by the elisa I g E complete described in ple 5, except that the captured IgE was detected by anti-I g E mouse • '*' - * Ft Í ?? M? T '* t * at't * t ~ lfr "- sheep marked HRP supplied by The Binding Site Inc. (San Diego, CA). Of the 10 mice, 7 had appreciable IgE responses of the Hogio 1.6 titre.These sera were pooled for use as the IgE anti-Oa work pattern.The serum I g E meeting was serially diluted 1:62, 1: 124 and 1: 248 into PBS and then it was further diluted with an equal volume of anti-IgE serum to the site.So the final dilutions for mouse IgE were 1: 124, 1: 248, and 1: 496 while the mouse anti-IgE was diluted 1 : 2. Control dilutions of IgE were prepared with only PBS as diluent IgE dilutions, with and without anti-IgE serum, were incubated for 1 hour at 27 ° C and 50 μL of each was taken for evaluation by reaction Heterologous PSA.

Results The 50 μl samples of diluted mouse IgE were injected intradermally into the shaved back of rats in a pattern that was a set of two rows of four injections. The rows were a row of three IgE controls diluted 1: 124, 1: 248 and 1: 496 in PBS only, in parallel with a row of serially diluted IgE incubated with site-specific anti-IgE. The fourth injection of each row was PBS only, as a control for tissue trauma. The pattern doubled in two rats.

After 24 hours, PSA reactions were induced by intravenous injection of 1 mg of DNP-Oa conjugate in 1% Evans blue dye. One hour later the rats were euthanized and peeled. The DNP-Oa allergen had mouse anti-Oa IgE bound to cross-linked receptor on the rat mast cells. The degranulation triggered by crosslinking increased the permeability of the Evans blue dye and the appearance of blue areas on the surface below the skin of the rats proportional to the degree of degranulation. However, wherever I g E has been drastically reduced by the site-specific morin anti-IgE, it was less available to sensitize the rat mast cells and the PCA reactions were suppressed. The PCA reactions were evaluated by measuring the diameters of the blue zones on the skin surface of the rats in two directions at right angles and taking the average. The results are shown in Table 7 for the double inhibition of PCA determinations in two rats. In their sensitivities inherent to the IgE rat of PCA reactions inhibited by anti-IgE and control should be compared only on the same rat. PCA reactions mediated by mouse I g E were inhibited by both rats by the murine antiserum with specificity for the target antigenic site on mouse IgE. Thus, the antibody response resulting from immunization by a peptide composition specific for the target antigenic site of a non-human IgE resulted in the suppression of the inflammatory response mediated by the same non-human I g E.

Table 1 Sequence 224 230 240 250 253b 260 Human e (Ident. Sec. No.:1) VCSRDFTPPTVKILQSS-CDGGGHF-PPTIQLLCLVSGYTPGTINI Dog e ACALNFIPPTVKLFHSS-CN-PVGDTHTTIQLLCLISGYVPGDMEV (Ident. Sec. No.:2) Rat and ARPVNITKPTVD LHSS-CD-PNAF-HSTIQLYCFVYGHIQNDVSI (Ident. Sec. No / 3) Mouse e VRPVTHSLSPPWSYSIHRCD-PNAF-HSTIQLYCFIYGHILNDVSV (Ident. Sec. No / 4) 270 280 290 300 310 Human e (Ident. Sec. No / 1) TWLEDGQ-VMDVDLSTA-STTQEGELASTQSELTLSQKHWLSDRTY Dog e (Ident. Sec. No / 2) IWLVDGQKATNIFPYTAPGTK-EGNVTSTHSELNITQGEWVSQKTY Rat e (Ident. Sec. No / 3) HWLMDDRKIYDTHAQNV-LIKEEGKLASTYSRLNITQQQWMSESTF Mouse e (Ident. Sec. No / 4) S LMDDREITDTLAQTV-LIKEEGKLASTCSKLNITEQQWMSESTF 320 330 340 350 Human e (Ident. Sec. No: 1) TCQV-TYQGHTFEDSTKKCADSNPRGVSAYLSRPSPFDLFIRKSPT Dog e (Ident. Sec. No: 2) TCQGFTFKDEARK CSESDPRGVTSYLSPPSPLDLYVHKAPK Rat e (Ident. Sec. No: 3) TCKV-TSQGENYWAHTRRCSDDEPRGVITYLIPPSPLDLYENGTPK Mouse e (Ident. Sec. No: 4) TCRV-TSQGCDYLAHTRRCPDHEPRGAITYLIPPSPLDLYQNGAPK Table 1 (Continued) Table 2 Table 2 (continued) . NT. N / 41) Table 2 (continued) Table 2 (continued) * = cyclized peptide t = numbers of amino acid residues of Table 1, SEC. FROM IDENT. No.1? = Cross-reactivity results are for a mixture of peptides "b" and "c" (C) = cysteine introduced into the native sequence for cyclisation C? S = Serine substituted by cysteine residue, D->? C = cysteine substituted by aspartic acid residue.

Table 3 Table 4A * klh = haemocyanin from slotted, chemically bonded limpet (see Example 1) fPAL = Palindromic Table 4B Immunogenicity of Representative Peptide Constructs of the Invention The guinea pigs, pigs and baboons were immunized with human IgE peptide constructs at 0, 3 and 6 weeks, with sera collected at 8 wpi for ELISA test by human IgE and inhibition of L.H. b The guinea pigs were immunized with dog IgE peptide constructs at 0, 2 and 4 weeks with sera collected at 6 wpi for dog IgE ELISA. c% average of L H. d% of inhibition of L H = control -% of L H / control x 100 GP Cobayo NT: Not tested Table 5 Amino Acid Sequences of Th Epitopes Derived from Strange Pathogens Table 6 Amino Acid Sequences of Representative Artificial Phytic Epitopes and SSAL Table 7 * very pale blue ± fi¿¿ ^ ii £ eM * ¿bMfaií? LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANT: UNITED BIOMEDICAL INC., Et al. (?) TITLE OF THE INVENTION: PEPTIDE COMPOSITION AS IMMUNOGENOUS FOR THE TREATMENT OF ALLERGY (ii) NUMBER OF SEQUENCES: 91 (iv) DIRECT CORRESPONDENCE: (A) RECIPIENT: Morgan & Finnegan (B) STREET: 345 Park Avenue (C) CITY: New York (D) STATE: NY (E) COUNTRY: USA (F) POSTAL CODE: 10154-0053 (v) READING FORM ON THE COMPUTER: (A) TYPE OF MEDIUM: Flexible Disk (B) COMPUTER: compatible with an IBM PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: WORD 8.0 (vi) DATA OF THE CURRENT APPLICATION: (A) NUMBER OF APPLICATION: To be assigned (B) DATE OF SUBMISSION: June 21, 1999 (C) CLASSIFICATION: (vii) DATA FROM THE PREVIOUS APPLICATION: (A) APPLICATION NUMBER: US 09 / 100,287 (B) DATE OF SUBMISSION: 20 June 1998 * Oe9fc, t * Jl * asJk'íÁÍÍ.M, '+ • **.

(C) CLASSIFICATION: 514 (viii) EMPLOYEE / AGENT INFORMATION: (A) NAME: MARIA CHLIN (B) REGISTRATION NUMBER: 29,323 (C) REFERENCE / FILE NUMBER: 1151- 4153PC1 (ix) TELECOMMUNICATION INFORMATION : (A) TELEPHONE: 212-758-4800 (B) TELEFAX: 212-751-6849 (2) INFORMATION FOR SEC. FROM IDENT. NO / 1: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 325 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: protein (ix) CHARACTERISTIC: (A) NAME / KEY: ? IgE human chain (x) REFERENCE: Dorrington and Benních, Immunol Rev, 1978, 41: 3-25. (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT.

NO: 1: Val Cys Ser Arg Asp Phe Thr Pro Pro Thr Val Lys 1 5 10 lle Leu Gln Ser Ser Cys Asp Gly Gly Gly His Phe 15 20 a ^^ fe ^^ g ^^^^ Pro Pro Thr lle Gln Leu Leu Cys Leu Val Ser Gly 25 30 35 Tyr Thr Pro Gly Thr lle Asn lle Thr Trp Leu Glu 40 45 Asp Gly Gln Val Met Asp Val Asp Leu Ser Thr Ala 50 55 60 Ser Thr Thr Gln Glu Gly Glu Leu Wing Ser Thr Gln 65 70 Ser Glu Leu Thr Leu Ser Gln Lys His Trp Leu Ser 75 80 Asp Arg Thr Tyr Thr Cys Gln Val Thr Tyr Gln Gly 85 90 95 His Thr Phe Glu Asp Ser Thr Lys Lys Cys Wing Asp 100 105 Ser Asn Pro Arg Gly Val Ser Wing Tyr Leu Ser Arg 110 115 120 Pro Ser Pro Phe Asp Leu Phe lle Arg Lys Ser Pro 125 130 Thr lle Thr Cys Leu Val Val Asp Leu Ala Pro Ser 135 140 Lys Gly Thr Val Asn Leu Thr Trp Ser Arg Ala Ser 145 150 155 Gly Lys Pro Val Asn His Ser Thr Arg Lys Glu Glu 160 165 Lys Gln Arg Asn Gly Thr Leu Thr Val Thr Ser Thr 170 175 180 Leu Pro Val Gly Thr Arg Asp Trp lle Glu Gly Glu 185 190 Thr Tyr Gln Cys Arg Val Thr His Pro His Leu Pro 195 200 Arg Ala Leu Met Arg Ser Thr Thr Lys Thr Ser Gly 205 210 215 Pro Arg Wing Wing Pro Glu Val Tyr Wing Phe Wing Thr 220 225 Pro Glu Trp Pro Gly Ser Arg Asp Lys Arg Thr Leu 230 235 240 Wing Cys Leu lle Gln Asn Phe Met Pro Glu Asp lle 245 250 15 Ser Val Gln Trp Leu His Asn Glu Val Gln Leu Pro 255 260 Asp Ala Arg His Ser Thr Thr Gln Pro Arg Lys Thr 265 270 275 Lys Gly Ser Gly Phe Phe Val Phe Ser Arg Leu Glu 280 285 Val Thr Arg Ala Glu Trp Gln Glu Lys Asp Glu Phe 290 295 300 lle Cys Arg Ala Val His Glu Ala Ala Ser Pro Ser 305 310"^ - ** - * - * *» »« - Gln Thr Val Gln Arg Ala Val Ser Val Asn Pro Gly 315 320 Lys 325 (2) INFORMATION FOR SECTION ID NO.:2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 312 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (¡i) TYPE OF MOLECULE: protein (¡x) FEATURE: (A) NAME / KEY:? Dog chain IgE (x) REFERENCE: Patel et al., Immunogenetics, 1995; 41: 282-286. (xi) DESCRIPTION OF THE SEQUENCE: SEC.

NO: 2: Ala Cys Ala Leu Asn Phe lle Pro Pro Thr Val Lys 1 5 10 Leu Phe His Ser Ser Cys Asn Pro Val Gly Asp Thr 15 20 His Thr Thr lle Gln Leu Leu Cys Leu lle Ser Gly 25 30 35 Tyr Val Pro Gly Asp Met Glu Val Val Trp Leu Val 40 45 Asp Gly Gln Lys Wing Thr Asn Lle Phe Pro Tyr Thr 50 55 60 Wing Pro Gly Thr Lys Glu Gly Asn Val Thr Ser Thr 65 70 His Ser Glu Leu Asn lle Thr Gln Gly Glu Trp Val 75 80 Ser Gln Lys Thr Tyr Thr Cys Gln Gly Phe Thr Phe 85 90 95 Lys Asp Glu Wing Arg Lys Cys Ser Glu Ser Asp Pro 100 105 Arg Gly Val Thr Ser Tyr Leu Ser Pro Pro Ser Pro 110 115 120 Leu Asp Leu Tyr Val His Lys Wing Pro Lys lle Thr 125 130 Cys Leu Val Val Asp Leu Ala Thr Met Glu Gly Met 135 140 Asn Leu Thr Trp Tyr Arg Glu Ser Lys Glu Pro Val 145 150 155 Asn Pro Gly Pro Leu Asn Lys Lys Asp His Phe Asn 160 165 Gly Thr lle Thr Val Thr Ser Thr Leu Pro Val Asn 170 175 180 Thr Asn Asp Trp lle Glu Gly Glu Thr Tyr Tyr Cys 185 190 Arg Val Thr His Pro His Leu Pro Lys Asp lle Val 195 200 Arg Ser lle Ala Lys Ala Pro Gly Lys Arg Ala Pro 205 210 215 Pro Asp Val Tyr Leu Phe Leu Pro Pro Glu Glu Glu 220 225 Gln Gly Thr Lys Asp Arg Val Thr Leu Thr Cys Leu 230 235 240 lle Gln Asn Phe Phe Pro Wing Asp lle Ser Val Gln 245 250 Trp Leu Arg Asn Asp Ser Pro lle Gln Thr Asp Gln 255 260 Tyr Thr Thr Thr Gly Pro His Lys Val Ser Gly Ser 265 270 275 Arg Pro Ala Phe Phe lle Phe Ser Arg Leu Glu Val 280 285 Ser Arg Val Asp Trp Glu Gln Lys Asn Lys Phe Thr 290 295 300 Cys Gln Val Val His Glu Ala Leu Ser Gly Ser Arg 305 310 (2) INFORMATION FOR SEC. FROM IDENT. NO.:3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 313 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear * -s - & to. J. i,? . -t¿ ^ ^ ia ^ * ^ (ii) TYPE OF MOLECULE: protein (¡x) CHARACTERISTIC: (A) NAME / KEY: í rat chain IgE (x) REFERENCE: Steen et al., J Mol Biol, 1984; 177: 19-32. (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT.

NO: 3: Ala Arg Pro Val Asn Lle Thr Lys Pro Thr Val Asp 1 5 10 Leu Leu His Ser Ser Cys Asp Pro Asn Wing Phe His 15 20 Being Thr lle Gln Leu Tyr Cys Phe Val Tyr Gly His 25 30 35 lle Gln Asn Asp Val Ser Lle His Trp Leu Met Asp 40 45 Asp Arg Lys lle Tyr Asp Thr His Wing Gln Asn Val 50 55 60 Leu lle Lys Glu Glu Gly Lys Leu Ala Ser Thr Tyr 65 70 Ser Arg Leu Asn lle Thr Gln Gln Gln Trp Met Ser 75 80 Glu Ser Thr Phe Thr Cys Lys Val Thr Ser Gln Gly 85 90 95 Glu Asn Tyr Trp Wing His Thr Arg Arg Cys Ser Asp 100 105 ^^ s ^ Asp Glu Pro Arg Gly Val lle Thr Tyr Leu lle Pro 110 115 120 Pro Ser Pro Leu Asp Leu Tyr Glu Asn Gly Thr Pro 125 130 Lys Leu Thr Cys Leu Val Leu Asp Leu Glu Ser Glu 135 140 Glu Asn lle Thr Val Thr Trp Val Arg Glu Arg Lys 145 150 155 Lys Ser lle Gly Ser Ala Ser Gln Arg Ser Thr Lys 160 165 His His Asn Ala Thr Thr Ser lle Thr Ser lle Leu 170 175 180 Pro Val Asp Ala Lys Asp Trp lle Glu Gly Glu Gly 185 190 Tyr Gln Cys Arg Val Asp His Pro His Phe Pro Lys 195 200 Pro lle Val Arg Ser lle Thr Lys Ala Leu Gly Leu 205 210 215 Arg Ser Ala Pro Glu Val Tyr Val Phe Leu Pro Pro 220 225 Glu Glu Glu Glu Lys Asn Lys Arg Thr Leu Thr Cys 230 235 240 Leu lle Gln Asn Phe Phe Pro Glu Asp lle Ser Val 245 250 Gln Trp Leu Gln Asp Ser Lys Leu lle Pro Lys Ser 255 260 Gln His Ser Thr Thr Thr Pro Leu Lys Thr Asn Gly 265 270 275 Ser Asn Gln Arg Phe Phe lle Phe Ser Arg Leu Glu 280 285 Val Thr Lys Ala Leu Trp Thr Gln Thr Lys Gln Phe 290 295 300 Thr Cys Arg Val lle His Glu Ala Leu Arg Glu Pro 305 310 Arg (2) INFORMATION FOR SEC. FROM IDENT. NO.:4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 313 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (ix) CHARACTERISTIC: (A) NAME / KEY: ? mouse chain IgE (x) REFERENCE: Ishida et al., EMBO, 1982; 1: 1117-1123 (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 4: Val Arg Pro Val Thr His Ser Leu Ser Pro Pro Trp 1 5 10 . * & * gj ^ ^ aíj Ser Tyr Ser lle His Arg Cys Asp Pro Asn Ala Phe 20 His Ser Thr lle Gln Leu Tyr Cys Phe lle Tyr Gly 25 30 35 His lle Leu Asn Asp Val Ser Val Ser Trp Leu Met 40 45 Asp Asp Arg Glu lle Thr Asp Thr Leu Ala Gln Thr 50 55 60 Val Leu lle Lys Glu Glu Gly Lys Leu Ala Ser Thr 65 70 Cys Ser Lys Leu Asn lle Thr Gllu Gln Gln Trp Met 75 80 Ser Glu Ser Thr Phe Thr Cys Arg Val Thr Ser Gln 85 90 95 Gly Cys Asp Tyr Leu Wing His Thr Arg Arg Cys Pro 100 105 Asp His Glu Pro Arg Gly Ala lle Thr Tyr Leu lle 110 115 120 Pro Pro Pro Pro Leu Asp Leu Tyr Gln Asn Gly Wing 125 130 Pro Lys Leu Thr Cys Leu Val Val Asp Leu Glu Ser 135 140 Glu Lys Asn Val Asn Val Thr Trp Asn Gln Glu Lys 145 150 155 Lys Thr Ser Val Ser Wing Ser Gln Trp Tyr Thr Lys 160 165 Hís His Asn Asn Ala Thr Thr Ser Lle Thr Ser lle 170 175 180 Leu Pro Val Val Ala Lys Asp Trp lle Glu Gly Tyr 185 190 Gly Tyr Gln Cys lle Val Asp Arg Pro Asp Phe Pro 195 200 Lys Pro lle Val Arg Ser lle Thr Lys Thr Pro Gly 205 210 215 Gln Arg Ser Ala Pro Glu Val Tyr Val Phe Pro Pro 220 225 Pro Glu Glu Glu Ser Glu Asp Lys Arg Thr Leu Thr 230 235 240 Cys Leu lle Gln Asn Phe Phe Pro Glu Asp lle Ser 245 250 Val Gln Trp Leu Gly Asp Gly Lys Leu lle Ser Asn 255 260 Ser Gln His Ser Thr Thr Thr Pro Leu Lys Ser Asn 265 270 275 Gly Asn Gln Gly Phe Phe Phe Ser Arg Leu Glu 280 285 Val Wing Lys Thr Leu Trp Thr Gln Arg Lys Gln Phe 290 295 300 Thr Cys Gln Val lle His Glu Ala Leu Gln Lys Pro 305 310 Arg (2) INFORMATION FOR SEC. FROM IDENT. NO.:5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT.

NO: 5: Cys Gly Glu Thr Tyr Gln Ser Arg Val Thr His Pro 1 5 10 His Leu Pro Arg Ala Leu Met Arg Ser Thr Thr Lys 15 20 Cys 25 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO 6: - ^ ¡- ^^^^ ii ^^ & aita ^^^ Cys Gly Glu Thr Tyr Tyr Ser Arg Val Thr His Pro 1 5 10 His Leu Pro Lys Asp lle Val Arg Ser lle Ala Lys 15 20 Cys 25 ( 2) INFORMATION FOR SEC. FROM IDENT. NO.:7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 7: Cys Gly Glu Gly Tyr Gln Ser Arg Val Asp His Pro 1 5 10 His Phe Pro Lys Pro val Val Arg Ser lle Thr Lys 15 20 Cys 25 (2) INFORMATION FOR SEC. FROM IDENT. NO.:8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear ? ^ »G s ^^^ > Toxicity (i) TYPE OF MOLECULE: peptide (xi) SEQUENCE DESCRIPTION: SEC. FROM IDENT.

NO: 8: Cys Gly Tyr Gly Tyr Gln Ser Val val Asp Arg Pro 1 5 10 Asp Phe Pro Lys Pro val Val Arg Ser lle Thr Leu 15 20 Cys 25 (2) INFORMATION FOR SEC. FROM IDENT. NO.:9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 9: Lys Lys Lys lle lle Thr lle Thr Arg lle lle Thr 1 5 10 lle lle Thr Thr lle Asp 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid J * J ^ - - ^^ - ^^^^^^^^ S & ^^ L ^^^^^^ Lj ^ V ^^^^^ (D) TOPOLOGY: linear (¡i) TYPE OF MOLECULE : peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 6 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 10 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 11 -and- "-" (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = " Lle, Met or Leu "(ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 14 (D) OTHER INFORMATION: / note =" Gly or Thr "(ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 15 (D) OTHER INFORMATION: / note = "lle, Met or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 10: Xaa Xaa Glu lle Xaa Xaa Val lle Val Xaa Xaa Xaa 1 5 10 Glu Xaa Xaa 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide to ,. J (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Site modified (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 7 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 8 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY : Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 13 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: ^^^^^ f ^ ^ ^ ^ ^ ^ ^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ KEY: Modified site (B) LOCATION: 14 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 16 (D) ANOTHER INFORMATION: / note = "Gly or Thr" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 17 (D) OTHER INFORMATION: / note = "lle, Met or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 11: lle Ser Xaa Xaa Glu lle Xaa Xaa Val lle Val Xaa 1 5 10 Xaa Xaa Glu Xaa Xaa Leu Phe 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (li) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 12 Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO.:13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 16 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 13: Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala 1 5 10 Thr Tyr Gln Phe 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:14: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids (B) TYPE 'amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 14: Lys Lys Lys lle lle Thr lle Thr Arg lle lle Thr 1 5 10 lle lle Thr Thr lle Asp Gly Gly Cys Gly Glu Thr 15 20 Tyr Gln Ser Arg Val Thr His Pro His Leu Pro Arg 25 30 35 Ala Leu Met Arg Ser Thr Thr Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO.:15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 63 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 15: Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Wing 1 5 10 Thr Tyr Gln Phe Gly Gly Lys Lys Lys lle lle Thr 15 20 lle Thr Arg llelle Thr lle lle Thr Thr lle Asp 25 30 35 Gly Gly Cys Gly Glu Thr Tyr Gln Ser Arg Val Thr 40 45 His Pro His Leu Pro Arg Ala Leu Met Arg Ser Thr 50 55 60 Thr Lys Cys "- ^ .J t (2) INFORMATION FOR ID SECTION NO.:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear ( I) TYPE OF MOLECULE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: Pro Pro Xaa Pro Xaa Pro 1 5 (2) INFORMATION FOR SECTION ID: 17: ( i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 59 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO. 17. Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 Gly Gly Lys Lys Lys lle lle Thr lle Thr Arg lle 15 20 lle Thr lle lle Thr Thr lle Asp Gly Gly Cys Gly 25 30 35 ^^^^^ s ^ k¿ & ^^^ > ^^^^^^ Glu Thr Tyr Gln Ser Arg Val Thr His Pro His Leu 40 45 Pro Arg Ala Leu Met Arg Ser Thr Thr Lys Cys 50 55 (2) INFORMATION FOR SEC. FROM IDENT. NO.:18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 46 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 7 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 8 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 12 »& dk. * Fc« 8? TteL iKEan .'- gv., (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 13 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Gly or Thr "(xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT.

NO: 18: lle Ser lle Xaa Glu lle Xaa Xaa Val lle Val Xaa 1 5 10 Xaa lle Glu Xaa lle Leu Phe Gly Gly Cys Gly Glu 15 20 Thr Tyr Gln Ser Arg Val Thr His Pro His Leu Pro 25 30 35 Arg Ala Leu Met Arg Ser Thr Thr Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 63 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 21 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 24 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 25 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 29 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 30 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 33 (D) OTHER INFORMATION: / note = "Gly or Thr" ^ ggj ^ a ^ (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 19: Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Wing 1 5 10 Thr Gln Phe Gly Gly lle Ser lle Xaa Glu lle Xaa 15 20 Xaa Val lle Val Xaa Xaa lle Glu Xaa lle Leu Phe 25 30 35 Gly Gly Cys Gly Glu Thr Tyr Gln Ser Arg Val Thr 40 45 His Pro His Leu Pro Arg Ala Leu Met Arg Ser Thr 50 55 60 Thr Lys Cys (2) INFORMATION FOR SEC. FROM IDENT. NO.:20: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 18 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 21 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 22 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 26 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 27 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 30 (D) OTHER INFORMATION: / note = "Gly or Thr" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 20: Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 Gly Gly lle Ser lle Xaa Glu lle Xaa Xaa Val lle 15 20 Val Xaa Xaa lle Glu Xaa lle Leu Phe Gly Gly Cys 25 30 35 Gly Glu Thr Tyr Gln Ser Arg Val Thr His Pro His 40 45 Leu Pro Arg Ala Leu Met Arg Ser Thr Thr Lys Cys 50 55 60 (2) INFORMATION FOR SEC. FROM IDENT. NO.:21: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 42 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 6 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 10 ( D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 11 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 14 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 15 (D) OTHER INFORMATION: / note = "lle, Met or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 21: Xaa Xaa Glu lle Xaa Xaa Val lle Val Xaa Xaa Xaa 1 5 10 Glu Xaa Xaa Gly Gly Cys Gly Glu Thr Tyr Gln Ser 15 20 Arg Val Thr His Pro His Leu Pro Arg Ala Leu Met 30 35 Arg Ser Thr Thr Lys Cys 40 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 22: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 19 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (¡x) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 20 (D) ) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 23 s á ^ w-sfei.

(D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 24 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 28 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) ) LOCATION: 29 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 30 (D) OTHER INFORMATION: / nota = "lle, Met or Leu "(ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 32 (D) OTHER INFORMATION: / note =" Gly or Thr "(ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 33 (D) OTHER INFORMATION: / note = "lle, Met or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 22: Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala 1 5 10 Thr Tyr Gln Phe Gly Gly Xaa Xaa Glu lle Xaa Xaa 15 20 Val lle Val Xaa Xaa Xaa Glu Xaa Xaa Gly Gly Cys 25 30 35 Gly Glu Thr Tyr Gln Ser Arg Val Thr His Pro His 40 45 Leu Pro Arg Ala Leu Met Arg Ser Thr Thr Lys Cys 50 55 60 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 56 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 15 (D) OTHER INFORMATION: / note = "lle, Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 19 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 20 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) ) LOCATION: 24 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 25 (D) OTHER INFORMATION: / note = "Lys or Arg "(¡x) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 26 (D) OTHER INFORMATION: / note =" lle, Met or Leu "(ix) FEATURE: (A) NAME / KEY : Modified site (B) LOCATION: 28 (D) OTHER INFORMATION: / note = "Gly or Thr" (¡x) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 29 (D) OTHER INFORMATION: / note = "lle, Met, or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT.

NO: 23: Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 Gly Gly Xaa Xaa Glu lle Xaa Xaa Val lle Val Xaa 15 20 Xaa Xaa Glu Xaa Xaa Gly Gly Cys Gly Glu Thr Tyr 25 30 35 Gln Ser Arg Val Thr His Pro His Leu Pro Arg Wing 40 45 Leu Met Arg Ser Thr Thr Lys Cys 50 55 (2) INFORMATION FOR SEC. FROM IDENT. NO.:24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 46 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Ser or Thr" (¡x) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 7 (D) OTHER INFORMATION : / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 8 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) ) NAME / KEY: Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 13 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Gly or Thr" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 24: lle Ser lle Xaa Glu lle Xaa Xaa Val lle Val Xaa 1 5 10 i- Xaa lle Glu Xaa lle Leu Phe Gly Gly Cys Gly Tyr 15 20 Gly Tyr Gln Ser Val val Asp His Pro Asp Phe Pro 25 30 35 Lys Pro lle Val Arg Ser lle Thr Lys Cys 40 45 (2) INFORMATION FOR SEC . FROM IDENT. NO .: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 25: Lys Lys Lys lle lle Thr lle Thr Arg lle lle Thr 1 5 10 lle lle Thr Thr lle Asp Gly Gly Cys Gly Tyr Gly 15 20 Tyr Gln Ser lle Val Asp Hís Pro Asp Phe Pro Lys 25 30 35 Pro lle Val Arg Ser lle Thr Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO.:26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids - (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 26: Lys Lys Lys lle lle Thr lle Thr Arg lle lle Thr 1 5 10 lle lle Thr Thr lle Asp Gly Gly Cys Gly Glu Thr 15 20 Tyr Tyr Ser Arg Val Thr His Pro His Leu Pro Lys 25 30 35 Asp lle Val Arg Ser lle Ala Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 46 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note = "Met or Leu" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION : / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 7 (D) OTHER INFORMATION: / note = "Arg" (ix) FEATURE: (A) NAME / KEY : Modified site (B) LOCATION: 8 (D) OTHER INFORMATION: / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 13 (D) OTHER INFORMATION: / note = "Arg" (ix) FEATURE: (A) NAME / KEY: Modified site ( B) LOCATION: 14 (D) OTHER INFORMATION: / note = "Met or Leu" (ix) CHARACTERISTICS: ( A) NAME / KEY: Modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 17 (D) OTHER INFORMATION: / note = "Met or Val" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 27: Lle Ser Glu Lle Lys Gly Val Val Val 1 5 10 Lys lle Glu Gly Lle Leu Phe Gly Gly Cys Gly Glu 15 20 Thr Tyr Tyr Ser Arg Val Thr His Pro His Leu Pro 25 30 35 Lys Asp lle Val Arg Ser lle Ala Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO.:28: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 49 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 28: Cys Wing Asp Ser Asn Pro Arg Gly Val Ser Wing Tyr 1 5 10 Leu Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg 15 20 Lys Ser Pro Thr lle Thr Ser Leu Val Val Asp Leu 25 30 35 Ala Pro Ser Lys Gly Thr Val Asn Leu Thr Trp Ser 40 45 Arg (2) INFORMATION FOR SEC. OF LDENT. NO.:29: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 29: Gln Gly His Thr Phe Glu Asp Ser Thr Lys Lys Cys 1 5 10 Wing Asp Ser Asn Pro Arg Gly Val Ser Wing Tyr Leu 15 20 Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg Lys 25 30 35 Ser Pro Thr Lle Thr Ser Leu Val Val Asp Leu Wing 40 45 Pro Ser Lys Gly Thr Val Asn Leu Thr Trp Ser Arg 50 55 60 .áj afc j (2) INFORMATION FOR SEC. FROM IDENT. NO.:30: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 64 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 30: Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 1 5 10 Thr Lys Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 15 20 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 25 30 35 Phe lle Arg Lys Ser Pro Thr Lle Thr Ser Leu Val 40 45 Val Asp Leu Pro Wing Pro Lys Gly Thr Val Asn Leu 50 55 60 Thr Trp Ser Arg (2) INFORMATION FOR SEC. FROM IDENT. NO.:31: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 76 amino acids (B) TYPE-amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. OF FENT. NO: 31: Gln Lys His Trp Leu Ser Asp Arg Thr Tyr Thr Ser 1 5 10 Gln Val Thr Tyr Gln Gly Hís Thr Phe Glu Asp Ser 15 20 Thr Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 25 30 35 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 40 45 Phe lle Arg Lys Ser Pro Thr lle Thr Ser Leu Val 50 55 60 Val Asp Leu Pro Wing Pro Lys Gly Thr Val Asn Leu 65 70 Thr Trp Ser Arg 75 (2) INFORMATION FOR THE SEC. FROM IDENT. NO.:32: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 35 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 32: Cys Ala Asp Ser Asn Pro Arg Gly Val Ser Ala Tyr 1 5 10 Leu Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg 15 20 Lys Ser Pro Thr lle Thr Ser Leu Val Val Asp 25 30 35 (2) INFORMATION FOR SEC . FROM IDENT. NO .: 33: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 46 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 33: Gln Gly His Thr Phe Glu Asp Ser Thr Lys Lys Cys 1 5 10 Wing Asp Ser Asn Pro Arg Gly Val Ser Wing Tyr Leu 15 20 Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg Lys 25 30 35 Ser Pro Thr Lle Thr Ser Leu Val Val Asp 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO.:34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 50 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 34: Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 1 5 10 Thr Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 15 20 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 25 30 35 Phe lle Arg Lys Ser Pro Thr Lle Thr Ser Leu Val 40 45 Val Asp 50 (2) INFORMATION FOR SEC. FROM IDENT. NO.:35: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 62 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 35: Gln Lys His Trp Leu Ser Asp Arg Thr Tyr Thr Ser 1 5 10 Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 15 20 Thr Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 25 30 35 Ser Wing Tyr Leu Ser Arg Pro Be Pro Phe Asp Leu 40 45 Phe lle Arg Lys Ser Pro Thr lle Thr Ser Leu Val 50 55 60 Val Asp (2) INFORMATION FOR SEC. FROM IDENT. NO.:36: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 36: Cys Wing Asp Ser Asn Pro Arg Gly Val Ser Wing Tyr 1 5 10 Leu Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg 15 20 Lys Ser Pro Thr lle 25 (2) INFORMATION FOR SEC. FROM IDENT. NO.:37: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 40 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT.

NO: 37: Gln Gly His Thr Phe Glu Asp Ser Thr Lys Lys Cys 1 5 10 Wing Asp Ser Asn Pro Arg Gly Val Ser Wing Tyr Leu 15 20 Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg Lys 25 30 35 Ser Pro Thr lle 40 (2) INFORMATION FOR SEC. FROM IDENT. NO.:38: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 38: Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 1 5 10 Thr Lys Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 15 20 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 25 30 35 Phe lle Arg Lys Ser Pro Thr lle 40 (2) INFORMATION FOR SEC. FROM IDENT. NO.:39: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 56 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO-39. Gln Lys His Trp Leu Ser Asp Arg Thr Tyr Thr Ser 1 5 10 Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 15 20 Thr Lys Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 25 30 35 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 40 45 Phe lle Arg Lys Ser Pro Thr lle 50 55 (2) INFORMATION FOR SEC. FROM IDENT. NO.:40: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 76 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 40: Gln Lys His Trp Leu Ser Asp Arg Thr Tyr Thr Cys 1 5 10 Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 15 20 Thr Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 25 30 35 Ser Wing Tyr Leu Ser Arg Pro Ser Pro Phe Asp Leu 40 45 Phe lle Arg Lys Ser Pro Thr lle Thr Cys Leu Val 50 55 60 Val Asp Leu Ala Pro Ser Lys Gly Thr Val Asn Leu 65 70 Thr Trp Ser Arg 75 (2) INFORMATION FOR THE SEC. FROM IDENT. NO.:41: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 41: Cys Lys Gln Arg Asn Gly Thr Leu Thr Cys 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO.:42: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC . FROM IDENT. NO: 42: Gln Lys His Trp Leu Ser Asp Arg Thr Tyr Thr Cys 1 5 10 Gln Val Thr Tyr Gln Gly His Thr Phe Glu Asp Ser 15 20 Thr Lys Cys Wing Asp Ser Asn Pro Arg Gly Val 25 30 35 Ser Wing Tyr Leu Ser Arg Pro Ser Pro 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 43: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 34 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT.

NO: 43: Cys Pro Ser Lys Gly Thr Val Asn Leu Thr Trp Ser 1 5 10 Arg Wing Ser Gly Lys Pro Val Asn His Ser Thr Arg 15 20 Lys Glu Glu Lys Gln Arg Asn Gly Thr Cys 25 30 (2) INFORMATION THE SEC. FROM IDENT. NO.:44: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 33 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 44: Cys Pro Val Gly Thr Arg Asp Trp lle Glu Gly Glu 1 5 10 Thr Tyr Gln Cys Arg Val Thr His Pro His Leu Pro 15 20 Arg Ala Leu Met Arg Ser Thr Thr Cys 25 30 (2) INFORMATION FOR SEC. FROM IDENT. NO.:45: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 14 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (¡i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 45: Ser Thr Thr Lys Thr Ser Gly Pro Arg Ala Wing Pro Glu Val 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 46: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 14 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 46 Cys Trp Ser Arg Ala Ser Gly Lys Pro Val Cys Asn His Ser 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO.:47: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE of amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 47: Cys Ser Arg Pro Ser Pro Phe Asp Leu Phe lle Arg 1 5 10 Lys Ser Pro Thr lle Thr Cys 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 48: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 13 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 48: Cys Val Gly Thr Arg Asp Trp lle Glu Gly Glu Pro Cys 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO.:49: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 49: Cys Pro Pro Val Gly Thr Arg Asp Trp lle Glu Gly 1 5 10 Glu Pro Cys 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:50: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 16 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 50: Cys Lys Glu Glu Lys Gln Arg Asn Gly Thr Leu Thr 1 5 10 Val Thr Ser Cys 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 51: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 51: Lys Glu Glu Lys Gln Arg Asn Gly 1 5 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 52: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 11 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 52: Cys Trp Ser Arg Wing Ser Gly Lys Pro Val Cys 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 53: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 53: Pro Thr lle Thr Cys Leu Val Leu Asp Leu Ala Pro 1 5 10 Ser Lys Gly Thr Val Asn Leu Thr Cys 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 54: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 16 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 54: Pro Thr lle Thr Cys Leu Val Leu Asp Leu Ala Pro 1 5 10 Ser Lys Gly Thr 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:55: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 55: Thr Ser Thr Leu Pro Val Gly Thr Arg Asp Trp lle '1 5 10 Glu Gly Glu Thr Tyr Gln Cys Arg Val Thr His Pro 15 20 Hís 25 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 56: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 16 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 56: Pro Thr lle Thr Ser Leu Val Leu Cys Leu Ala Pro 1 5 10 Ser Lys Gly Cys 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 57: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 23 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ií) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 57: Cys Val Asn Leu Thr Trp Ser Arg Wing Ser Gly Lys 1 5 10 Pro Val Asn His Ser Thr Arg Lys Glu Glu Cys 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 58: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 53 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 58: Cys Thr Trp Ser Arg Wing Ser Gly Lys Pro Val Asn 1 5 10 His Ser Thr Arg Lys Glu Glu Lys Gln Arg Asn Gly 15 20 Thr Leu Thr Val Thr Ser Thr Leu Pro Val Gly Thr 25 30 35 Arg Asp Trp lle Glu Gly Glu Thr Tyr Gln Cys Arg 40 45 Val Thr His Pro His 50 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 59: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 59: i. »..3 Lys Thr Lys Gly Ser Gly Phe Phe Val Phe 1 5 10 (2) INFORMATION FOR SEC. FROM IDENT. NO.:60: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Ser or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 7 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 8 (D) OTHER INFORMATION: / note = "Gly or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 12 (D) OTHER INFORMATION: / note = "His or Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) ) LOCATION: 13 (D) OTHER INFORMATION: / note = "Lys or Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Gly or Thr "(xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 60: lle Ser lle Xaa Glu lle Xaa Xaa Val lle Val Xaa 1 5 10 Xaa lle Glu Xaa lle Leu Phe 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 61: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT.

NO: 61: Leu Ser Glu lle Lys Gly Val lle Val His Arg Leu 1 5 10 Glu Gly Val 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 62: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 62: Gly lle Leu Glu Ser Arg Gly lle Lys Ala Arg lle 1 5 10 Thr His Val Asp Thr Glu Ser Tyr 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 63: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 63: Lys Lys Gln Tyr lle Lys Wing Asn Ser Lys Phe lle 1 5 10 Gly lle Thr Glu Leu 15 (2) INFORMATION FOR SEC. FROM IDENT. DO NOT. : 64: t r xiakñ & £ j &tiiiÍ * t? . J. - and ^ .. and tr ^ ^ ^ ^ L ^ L ___ ¿^ jj ^ fa ^: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 64: Lys Lys Phe Asn Asn Phe Thr Val Ser Phe Trp Leu 1 5 10 Arg Val Pro Lys Val Ser Wing Ser His Leu 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 65: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 30 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 65: Lys Lys Leu Arg Arg Leu Leu Tyr Met lle Tyr Met 1 5 10 Ser Gly Leu Ala Val Arg Val His Val Ser Lys Glu 15 20 Glu Gln Tyr Tyr Asp Tyr 25 30 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 66: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 27 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 66: Tyr Asp Pro Asn Tyr Leu Arg Thr Asp Ser Asp Lys 1 5 10 Asp Arg Phe Leu Gln Thr Met Val Lys Leu Phe Asn 15 20 Argle Lys 25 (2) INFORMATION FOR SEC. FROM IDENT. NO.:67: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 67: Gly Wing Tyr Wing Arg Cys Pro Asn Gly Thr Arg Wing 1 5 10 Leu Thr Val Wing Glu Leu Arg Gly Asn Wing Glu Leu 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 68: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 68: Phe Phe Leu Leu Thr Arg lle Leu Thr lle Pro Gln 1 5 10 Ser Leu Asp 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:69: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 69: Val Ser Phe Gly Val Trp lle Arg Thr Pro Pro Ala 1 5 10 Tyr Arg Pro Pro Asn Ala Pro lle Leu 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:70: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 70 Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 1 5 10 Thr Ala Ser Ala Leu Tyr Arg Glu 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:71: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. OF DENT. NO: 71 Pro His His Thr Ala Leu Arg Gln Ala lle Leu Cys 1 5 10 ^ .. Adjusters Trp Gly Glu Leu Met Thr Leu Wing 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:72: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 72: Trp Val Arg Asp lle lle Asp Asp Phe Thr Asn Glu 1 5 10 Ser Ser Gln Lys Thr 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 73: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 73: Arg Ala Gly Arg Ala lle Leu His lle Pro Thr Arg 1 5 10 He Arg Gln Gly Leu Glu Arg 15 (2) INFORMATION FOR SEC. FROM IDENT. NO .: 74: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 74: Wing Val Wing Glu Gly Thr Asp Arg Val lle Glu Val 1 5 10 Leu Gln Arg Ala Gly Arg Ala lle Leu 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:75: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 75: Ala Leu Asn lle Trp Asp Arg Phe Asp Val Phe Ser 1 5 10 Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn 15 20 Ser 25 (2) INFORMATION FOR SEC. FROM IDENT. DO NOT. : 76: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 76: Asp Ser Glu Thr Wing Asp Asn Leu Glu Lys Thr Val 1 5 10 Ala Ala Leu Ser lle Leu Pro Gly His Gly 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:77: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 77: "* -" *. »**** & * / Glu Glu lle Val Ala Gln Ser lle Ala Leu Ser Ser 1 5 10 Leu Met Val Ala Gln Ala lle Pro Leu Val Gly Glu 15 20 Leu Val Asp lle Gly Phe Ala Ala Thr Asn Phe Val 25 30 35 Glu Ser Cys (2) INFORMATION FOR SEC. FROM IDENT. NO.:78: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 78: Asp lle Glu Lys Lys lle Wing Lys Met Glu Lys Wing 1 5 10 Ser Ser Val Phe Asn Val Val Asn Ser 15 20 (2) INFORMATION FOR SEC. FROM IDENT. DO NOT. : 79: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 79: Lys Trp Phe Lys Thr Asn Wing Pro Asn Gly Val Asp 1 5 10 Glu Lys lle Arg lle 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:80: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 14 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO-80: Gly Leu Gln Gly Lys lle Wing Asp Wing Val Lys Wing 1 5 10 Lys Gly (2) INFORMATION FOR SEC. FROM IDENT. NO.:81: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 81: Gly Leu Wing Wing Gly Leu Val Gly Met Wing Wing Asp 1 5 10 Wing Met Val Glu Asp Val Asn 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:82: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (i) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC . FROM IDENT. NO: 82: Ser Thr Glu Thr Gly Asn Gln His His Tyr Gln Thr 1 5 10 Arg Val Val Ser Asn Wing Asn Lys 15 20 (2) INFORMATION FOR SEC. FROM IDENT. NO.:83: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 83: Cys Pro Ser Pro Phe Asp Leu Phe lle Arg Lys Ser 1 5 10 Pro Thr Cys 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:84: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 84: Cys Gly Glu Thr Tyr Lys Ser Thr Val Ser His Pro 1 5 10 Asp Leu Pro Arg Glu Val Val Arg Ser lle Ala Lys 15 20 Cys 25 (2) INFORMATION FOR SEC. FROM IDENT. NO.:85: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 18 (D) OTHER INFORMATION: / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 21 (D) OTHER INFORMATION: / note = "Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 22 (D) OTHER INFORMATION: / note = "Thr" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 26 (D) OTHER INFORMATION: / note = "Thr" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 27 (D) OTHER INFORMATION: / note = "Arg" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 30 (D) OTHER INFORMATION: / note = "Thr" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. : Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 Gly Gly lle Ser lle Ser Glu lle Lys Gly Val lle 15 20 Val His Lys lle Glu Gly lle Leu Phe Gly Gly Cys 25 30 35 Gly Gly Thr Tyr Gln Ser Arg Val Thr His Pro His 40 45 Leu Pro Arg Ala Leu Met Arg Ser Thr Thr Lys Cys 50 55 60 (2) INFORMATION FOR SEC. FROM IDENT. NO.:86: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO.86: Lys Trp Phe Lys Thr Asn Ala Pro Asn Gly Val Asp 1 5 10 Glu Lys lle Arg lle 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:87: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 62 amino acids 1 ^ ^ 2! Gfc ^? Rf! Tj¡tí ^ ¡ijAi ^^ ¿^^^? T ^^^^^ _ ^^ i = i _ ^^ ¿^ _ ^^^^^^ ßi (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 87: Lys Trp Phe Lys Thr Asn Wing Pro Asn Gly Val Asp 1 5 10 Glu Lys lle Arg lle Lys Lys Lys Lys lle lle Thr 15 20 lle Thr Arg lle lle Thr lle lle Thr Yhr lle Asp 25 30 35 Lys Cys Gly Glu Thr Tyr Tyr Ser Arg Val Thr His 40 45 Pro His Leu Pro Lys Asp lle Val Arg Ser lle Ala 50 55 60 Lys Cys (2) INFORMATION FOR SEC. FROM IDENT. NO.:88: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 57 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 88: Thr lle Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 Lys Lys Lys Lys lle lle Thr lle Thr Arg lle lle 15 20 Thr lle lle Thr Tyr lle Asp Lys Cys Gly Glu Thr 25 30 35 Tyr Tyr Ser Arg Val Thr His Pro His Leu Pro Lys 40 45 Asp lle Val Arg Ser lle Ala Ala Cys 50 55 (2) INFORMATION FOR SEC. FROM IDENT. NO.:89: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 89: lle Ser Leu Thr Glu lle Arg Thr Val lle Val Thr 1 5 10 Arg Leu Glu Thr Val Leu Phe 15 (2) INFORMATION FOR SEC. FROM IDENT. NO.:90: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids ry- "j, (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT. NO: 90: lle Ser Leu Thr Glu lle Arg Thr Val lle Val Thr 1 5 10 Arg Leu Glu Thr Val Leu Phe Lys Cys Gly Glu Thr 15 20 Tyr Tyr Ser Arg Val Thr His Pro His Leu Pro Lys 25 30 35 Asp lle Val Arg Ser lle Ala Lys Cys 40 45 (2) INFORMATION FOR SEC. FROM IDENT. NO.:91: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 63 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEC. FROM IDENT.

NO: 91: Lys Trp Phe Lys Thr Asn Wing Pro Asn Gly Val Asp 1 5 10 Glu Lys lle Arg lle Lys lle Ser Leu Thr Glu lle 15 20 Arg Thr Val lle Val Thr Arg Leu Glu Thr Val Leu 30 35 Phe Lys Cys Gly Glu Thr Tyr Tyr Ser Arg Val Thr 40 45 His Pro His Leu Pro Lys Asp lle Val Arg Ser lle 50 55 60 Ala Lys Cys j ^ j ^ - ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^

Claims (28)

1. CLAIMS 1. An IgG-CH3 domain antigen peptide between about 25 and about 29 amino acids in length containing two cysteine residues separated by approximately 23 amino acid residues, selected from the group is characterized in that it consists of SEC. FROM IDENT. NO: 5, SEC. FROM IDENT. NO: 6, SEC. FROM IDENT. NO.7, SEC. FROM IDENT. NO: 8, and SEC. FROM IDENT. NO: 84, homologous sequences of the epsilon heavy chain of IgE-CH3, and cross-reactive and immunologically functional analogues thereof.
2. 2. The confoirmity IgE-CH3 domain antigen peptide with claim 1, characterized in that it is selected from the group consisting of SEC. FROM IDENT. NO: 5, SEC. FROM IDENT. NO: 6, SEC NO.7, SEC. FROM IDENT. NO: 8, and SEC. FROM IDENT. NO: 84.
3. 3. The synthetic peptide of about 50 to about 90 amino acids, characterized in that it comprises: (a) an attendant T cell epitope (Th), (b) an IgG-CH3 domain antigen peptide according to claim 1; and (c) an immunostimulatory invasin domain.
4. 4. The peptide conjugate characterized in that it comprises an attached T cell epitope sequence (Th) - < * »•• * covalently to an IgG-CH3 domain antigen peptide according to claim 1.
5. 5. The peptide conjugate represented by the formula (A) n- (IgE-CH3 antigen domain) - (B) 0- (Th) mX or (A) n- (Th) m- (B) 0- (IgE-CH3 antigen domain) -X characterized in that each A is independently an amino acid or a general immunostimulatory sequence; each B is selected from the group consisting of amino acids, -NHCH (X) CH2SCH2CO-, -NHCH (X) CH2SCH2CO (eN) Lys-, -NHCH (X) CH2S-succinimidyl (eN) Lys-, and -NHCH (X) CH2S- (succinimidyl) -; each Th is independently a sequence of amino acids constituting epistebuses of the attendant T cell, or an immunomodulatory analog or segment thereof; the IgE-CH3 domain antigen represents the sequence of a IgE-CH3 domain antigen peptide according to claim 1; X is an amino acid a-COOH or -CONH2; n is from 0 to about 10; m is from 1 to about 4; I is from 0 to approximately 10.
6. 6. The peptide conjugate represented by the formulas (IgE-CH3 antigen domain) - (B) 0- (Th) mX (Th) m- (B) 0- (IgE-CH3 domain antigen ) - (A) nX characterized in that each A is independently an amino acid or a general immunostimulatory sequence, each B is selected from the group consisting of amino acids -NHCH (X) CH2SCH2CO-, -NHCH (X) CH2SCH2CO (eN) Lys-, -NHCH (X) CH2S-succinimidyl (eN) Lys-, and -NHCH (X) CH2S- (succinimidyl) -; each Th is independently a sequence of amino acids that constitute an assistant cell epitope, or an immunomodulatory analog or segment thereof; the IgE-CH3 domain antigen represents the sequence of an IgE-CH3 domain antigen peptide according to claim 1; X is an amino acid a-COOH or a-CONH2; n is from 0 to about 10; m is from 1 to about 4; and o is from 0 to approximately 10. The peptide conjugate according to any of claims 4-6, characterized in that the Th is SSAL. 8. The peptide conjugate according to any of claims 4-6, characterized in that the IgG-CH3 domain antigen peptide has an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NO: 5, «* # - *? SEC. FROM IDENT. NO: 6, SEC NO.7, SEC. FROM IDENT. NO: 8, and SEC. FROM IDENT. NO: 84 9. The peptide conjugate according to claim 7, characterized in that the IgG-CH3 domain antigenic peptide has an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NO: 5, SEC. FROM IDENT. NO: 6, SEC NO.
7. 7, SEC. FROM IDENT. NO: 8, and SEC. FROM IDENT. NO: 84. 10. The peptide conjugate according to any of claims 4-6, characterized in that the Th has an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NOS: 9-12, and SEC. FROM IDENT. NOS: 61-82 and 84. 11. The peptide conjugate according to claim 7, characterized in that the Th has an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NO: 9-12, and SEC. FROM IDENT. NO: 61-82 and 84. 12. The peptide conjugate according to claim 8, characterized in that the Th has an amino acid sequence selected from the group consisting of SEp. FROM IDENT. NOS: 9-12, and SEC. FROM IDENT. NOS: 61-82 and 84. 13. The peptidic conjugate according to claim 9, characterized in that the Th has an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NOS: 9-12, and SEC. FROM IDENT. NOS: 61-82 and 84. 14. A peptide characterized in that it comprises an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NOS: 14, 15, 17-27, 85, 87, 88, 90, 91. 15. A peptide conjugate according to claim 5 or 6, characterized in that at least one A is an invasin domain. 16. A peptide conjugate according to claim 5 or 6, characterized in that n is 3, and (A) 3 is (invain domain) -Gly-Gly. 17. The peptide conjugate according to claim 15, characterized in that the invasin domain has the amino acid sequence of SEQ. FROM IDENT. NO: 13 18. The peptide conjugate according to claim 16, characterized in that the invasin domain has the amino acid sequence of SEQ. FROM IDENT. NO: 13 19. The peptide conjugate characterized in that it comprises a carrier protein covalently linked to one or more IgG-CH3 domain antigen peptides according to claim 1. 20. The peptide conjugate according to claim 19, characterized in that the carrier protein is hemokinin of slotted limpet. 21. A peptide characterized in that it comprises an amino acid sequence selected from the group consisting of SEC. FROM IDENT. NOS: 14, 15, 26, 90. 22. The branched polymer characterized in that it comprises a nucleus of lysine, trilisine, or heptalisin, covalently linked to two, four, or eight peptide conjugates, respectively, according to any of claims 4-6 or 14. 23. The polymer characterized in that it comprises one or more peptide conjugates according to any of claims 4-6 or 14, crosslinked by a bifunctional crosslinking agent 24. The pharmaceutical composition characterized in that it comprises an immunologically effective amount of a peptide or peptide conjugate according to any of the claims 4-6 or 14, and a pharmaceutically acceptable carrier 25. The pharmaceutical composition according to claim 23, characterized in that the immunologically effective amount of the peptide or peptide conjugate is between about 0.5 μg and about 1 mg per kilogram of body weight per dose. . 26. The method for inducing the production of anti-IgE antibody in a mammal characterized in that it comprises administering to the mammal a composition according to claim 23. 27. The method for inducing the production of anti-IgE antibody in a mammal characterized in that it comprises We will administer to the mammal a pharmaceutical composition according to claim 24. 28. The nucleic acid characterized in that it comprises a sequence which encodes a peptide according to any of claims 1-6. ** - > * - yyU ~~ yy,. *? r