JPH08508734A - Treatment of Kawasaki syndrome by immunomodulation - Google Patents

Abstract

  (57) [Summary] The present invention relates to various methods for the diagnosis of Kawasaki Syndrome. Various bacteria including TSST-1 producing Staphylococcus aureus and SREB and SREC producing streptococci have been found to be indicative of pathological conditions. It also describes an isolate of Staphylococcus aureus associated with Kawasaki syndrome and methods for preventing and treating this condition.

Description

Detailed Description of the Invention                   Treatment of Kawasaki syndrome by immunomodulation   The invention described herein is in part supported by a NIH grant (ML37260). It has been developed. Accordingly, the US Government has certain rights in this invention.   Area of invention   The present invention relates to the field of immunology. More specifically, skin mucosa lymph node syndrome From the observation of the causative factor of Kawasaki syndrome, also known as Kawasaki disease Regarding derived items obtained.   Background and prior art   Kawasaki syndrome (KS) is an acute multisystem vasculitis of unknown etiology. This disease is mainly It affects infants and children, ie young people under the age of 16. Kawasaki, Jpn.J.Allergol. 16: 178-222 (1967); Rauch et al., Pediatr. Infect. Dis. 4: 702-702 (1985). K S occurs worldwide, but is most common in children of Japanese and Japanese descent ． Early clinical signs included long-term fever, bilateral nonexudative conjunctivitis, limb induration and Erythema, lip and oropharyngeal inflammation, pleomorphic eruption, and cervical lymphadenopathy It is done. These signs are used in the clinical diagnosis of KS.   In Japan and the United States, KS is the most common cause of acquired heart disease in children It is one. Recent studies have shown that γ-globulin is present during the acute phase of the disease. Intravenous administration (IVGG) of patients It was clarified that the lesions of coronary arteries that occurred in 1% were significantly reduced. Newburger Et al., N. Engl. J. Med. 315: 341-6 (1986); Nagashima et al., J. Pediatr. 110: 710-2 (1987); Firisho et al., Lancet ii: 1055-57 (1984); Rowley et al., J. Pediatr. 113: 290-94 (1988); See Newburger et al., N. Eng. J. Med. 324: 1633-39 (1991). Therefore, this disease is effective. As with all other vasculitis diseases, early differential diagnosis is necessary for effective treatment. It is essential.   KS is characterized by acute as well as convalescent stages. In the acute phase In addition, marked activation of immunity is characteristic. Based on previous research, HLA-DR Increased numbers of circulating and infiltrating T cells with activating antigen, and serum soluble IL-2 Elevated receptor levels have been demonstrated. These phenomena are related to the activation of T cells It is an index. Leung et al., J. Clin. Invest. 79: 468-472 (1987); Terai et al., Hum. Pathol. 21: 231-234 (1990); Lang et al., J. Pediatr. 116: 592-596 (1990). More acute K S is IL-1β, TNF-α, IL-6, IL-2, and interferon With increased production of -γ (IFN-γ). For example, Matsubara et al., Clin.Immuno l. Immunopathol. 56: 29-36 (1990); Maury et al., J. Lab. Clin. Med. 113: 651-54 (1989). Lang et al., J. Pediatr. 115: 939-43 (1989); Leung et al., Lancet ii: 1928-1302 (1989). Rowley et al., Ped. Inf. Dis. J. 7: 663-67 (1988); Ueno et al., Clin. Exp. Immunol. 76:33. 7-342 (1989); Jordan et al., In Kawasaki (ed.), The Third International Kawasa See ki Disease Symposium 1989: 144-46. These cells with T cells and monocytes Tocaine production is due to their proinflammatory and prothrombotic effects on endothelial cells Thought to play an important role in the pathogenesis of vascular cell injury in the acute KS phase ． See Mantovani et al., Immunol. Today 10: 370-74 (1989). Blood in KS lesions Vascular endothelium is a cytokine-induced white that is known to be involved in the localization of inflammatory cells. It has been proven to express blood cell adhesion molecules. See Leung, supra. Acute KS Have anti-cytotoxic effects on IL-1β, TNF-α and INF-γ stimulated endothelial cells. It has been found to have a body but no antibodies to unstimulated cells. Leung Et al., J. Clin. Invest. 77: 1428-35 (1986); Leung et al., J. Exp. Med. 164: 1958-72 (1986). )reference.   Epidemiologic studies aimed at identifying potential environmental toxins, as well as known Despite clinical laboratory culture of body fluids for biological agents, KS The causative agent of Escherichia coli has not yet been found. Rauch et al., Ped.Infect.Dis.J.6: 1016-21 (1987). Acute self-limiting nature of the disease, geographical cluster of disease, measles, bug Clinical manifestations of fever and rash similar to symptoms and diseases like lax and scarlet fever Humoral immunity to this organism prematurely due to the pathology and the unique sensitivity of children. It has been suggested that it appears during the period. KS is rarely seen after the age of eight, This is because there is an asymptomatic infection caused by a universal factor, followed by the general population. This suggests that protective immunity is expressed.   From a systemic observation of KS, this disease is caused by a toxin called superantigen (SAg) Suggested to have some similarities with diseases characterized by response to quality Be done. Superantigens can have significant effects on the T cell repertoire A new group of antigens [Herman et al. (1991) Ann. Rev. Immunol. 9: 745]. Antigen presentation Unlike ordinary antigens, which have to be processed by cells, SAg It is irritating in its native conformation. In fact, ASg's mitoji En action is destroyed by normal antigen processing [Fraser (1989) Nature 339: 2. twenty one]. Major Histocompatibility Complex (MHC) molecules like normally processed antigens Rather than interacting with the T cell antigen receptor (TCR) by binding in the groove of ASg binds to the flanks of both TCR and MHC molecules, Form crosslinks between cell surface molecules. Selective activation of T cells by SAg is specific Of the TCR of responding cells, independent of the specificity of their receptor for MHC-treated antigens of It is uniquely determined by the Vβ element. Whether in vitro or in vivo Nevertheless, acute exposure to these antigens results in large-scale activation of T cells with the appropriate Vβ element. Often leads to sexualization. Chronic exposure to superantigens could be repeated injections or chronic infections. Or retroviral superantigens due to inheritance of active provirus, In addition, T cell activation is followed by elimination of T cells with the appropriate Vβ element.   All the superantigens identified to date are bacteria, mycoplasma or Is a protein produced by the virus. For example, for S. aureus Enterotoxin, staphylococcal or streptococcal exotoxin, mycoplasma joint Flame Mitogen (MAM) [Marrack & Kappler (1990) Science 248: 705; Herman Et al. (1991) Ann. Rev. Immunol. 9: 745; Cole & Atkin (1991) Immunol. Today 12; 271. Review], and the 3'-LRT of mouse mammary tumor virus (MTV) Products of open reading frames [Woodland et al. (1991) Nature 349: 529; Choi (1991) Nature 350: 203; Acha-Orbea et al. (1991) Nature 350: 207].   Staphylococcus aureus is a common human pathogen, SEA (staphylococcal enterotox A) to SEE (staphylococcal enterotoxin E) Produces several enterotoxins that can cause venom and sometimes shock in humans [Marrack & Kappler (1990) supra; Bohack et al. (1990) Crit. Rev. Mocrobio. 117: 251]. Some S. aureus isolates also have human toxic shock syndrome Shock syndrome toxin-1 that is believed to be involved in most cases of , And exfoliative crest associated with scalded skin syndrome Produces syn (ExTF). Streptococcus pyogenes or streptococcus group A is another common Toxins with similar superantigen-like properties in human skin and pharyngeal human pathogens [Abe et al. (1991) J. Immun. 146: 3747]. These are streptococcal erythrocytes It is named as SP (A to C) (SPEA to SPEC).   Staphylococcal enterotoxin and streptococcal red toxin (SPE) are monocytes Is a strong inducer of IL-1 and TNF-α from [Fast et al. (1989) Infect. and Immunity 57: 291]. One method that enables measurement of activated T cells Is due to their ability to produce the cytokine IL-2. Staphylococcus entero Stimulation of monocytes induced by toxins and SPE results in a cluster on the surface of monocytes. Results of positive signal binding and transmission by the SII major histocompatibility complex molecule. Is. In the presence of antigen-presenting cells, these bacterial toxins will not react with T cells in culture. It is a powerful stimulator of cell proliferation, which is a specific Vβ gene segment It is carried out by selectively stimulating T cells expressing a mutant [Marrack & Kapp ler (1990), supra]. For example, in vitro, staphylococcal TSST-1 Most of the T cells that were challenged specifically express Vβ2. More importantly, Circulating Vβ2 is found in peripheral blood T cells of patients with sick shock syndrome⁺T According to Choi et al. (1990) J.Exp.Med.244: 811 that significant expansion of cells is demonstrated. Is indicated. These studies are based on staphylococcal enterotoxins In vivo activation of T cells is associated with specific superantigens secreted in vitro. It suggests that it is related. Vasculitic diseases, especially KS and superantigen induction The similarities between diseases suggest that the same phenomenon may be involved in both. ．   Abe et al. (1992) Proc. Natl. Acad. Aci. USA 89: 4066 (this disclosure is incorporated by reference in its entirety). Experiments examining the T cell repertoire of patients with KS Is described. Variable region Vβ2⁺And, to a lesser extent, Vβ8⁺But these Was found to be enlarged compared to controls and other potential areas. This is mainly because streptococcal exotoxin or allogeneic exotoxin is involved in the pathogenesis of acute KS. It gives Zhang more support.   Significant research efforts have been made to diagnose autoimmune diseases, which are also superantigen-induced diseases. The focus has been on the development of treatment and treatment methods. For example, European Patent Publication No. 340,109. No. of Invention: Anti-T-cell receptor determinants as autoimmune disease treatment (anti-T cell receptor determinant as a treatment for autoimmune diseases) and US Xu 4,550,086, issued October 29, 1985, Reinherz et al., Title of invention: Monoclonal antibodies that recognize human T cells (Null antibody) is a specific array of T cell receptor variable region genes associated with a specific disease. A method for detecting a sequence and a method for treating the disease with an antibody against the sequence is described. ing. US Pat. No. 4,886,743, issued December 12, 1989, Hood et al., Title of invention: D iagnostic reagents based on unique sequences within the variable region of the T cell receptor and uses thereof Diagnostic reagents based on unique sequences and their use) are associated with specific diseases A method for diagnosing a disease based on the presence of T cells having a unique sequence in the Vβ region is described. Has been done. PCT patent application publication WO 90/06758 describes rheumatoid arthritis (RA). Detection of the specific Vβ regions with particularity Vβ3, Vβ9 and Vβ10 and Vβ3 A method of treating RA with a monoclonal antibody that recognizes Vβ9 and Vβ10 Has been described.   US Patent Application Serial No. 07 / 732,114 filed July 18, 1991, title of invention Name: Method for Identifying T Cells Disease Involved in Autoimmune Disease (Method for identifying T cell disease involved in autoimmune disease) (Particularly in the present specification for reference) In addition, it was confirmed that the specific Vβ element can be used for the diagnosis of autoimmune diseases. It is standing. In this reference, the high rate of Vβ14 in synovial fluid⁺Presence of RA diagnoses RA Was used for.   US Patent Application Serial No. 07 / 827,540, filed January 28, 1992, title of invention Name: Method for modifying T cell response This article is specifically incorporated herein by reference) for the treatment of superantigen-induced diseases. The law is described. Mutant superantigen molecule, eg Staphylococcus entero Pre-exposure of animals to toxin B (TSST-1) resulted in subsequent wild-type toxin A protective effect was imparted against exposure to adverse effects. Mutant superantigen molecule Modifies T cell response to wild-type toxin without modifying B cell response The PCT / US93 / 00839, filed January 28, 1993, title of invention: Protective effects  of mutated superantigens (protective effect of mutant superantigens) (for reference In particular, it is introduced in this specification). And their ability to selectively eliminate or inactivate specific T cell populations has been described.   Brief summary of the invention   The present invention produces the toxic shock syndrome toxin-1 associated with Kawasaki syndrome. It is premised on the identification of a unique strain of Staphylococcus aureus. Also in the present invention Includes biologically pure cultures of this unique strain of S. aureus.   We herein use the staphylococcal or streptococcal bacteria and their It is possible to diagnose Kawasaki Syndrome by the assay of these related antigens, especially TSST-1. Indicates that there is.   The present invention also includes therapeutic methodologies for the treatment and prevention of Kawasaki Syndrome, This is based on the discovery that Staphylococcus aureus is one of the causative factors. Toku In addition, the present invention relates to the specific Vβ and Vβ of the T cell receptor (TCR) associated with Kawasaki syndrome. Preventing toxic effects of TSST-1 by treatment with a molecule that interacts with two elements Include methods. In particular, these molecules induce mutations or modifications of TSST-1. It is composed of conductors. The pathological condition known as Kawasaki Syndrome is a mutation of the present invention. It was revealed that administration of superantigen molecule can prevent or treat.   The molecule of the present invention eliminates the Vβ region associated with Kawasaki syndrome, especially Vβ2 or It can function by guiding inactivation / desensitization.   Brief description of the drawings   Figure 1 compares Southern blots of isolates from patients with Kawasaki Syndrome with controls. Show.   Figure 2 shows a Western blot of isolates using anti-TSST antibody.   FIG. 3 shows SDS-of recombinant TSST-1 (24 kD) purified from E. coli. Figure 8 shows a PAGE analysis.   Figure 4 shows the nucleotide and wild-type toxic shock syndrome toxin-1. And the amino acid sequence.   Figure 5 shows the primers used for mutagenesis of TSST-1. Each plastic Immers 1 to 13 top have 99% of the correct nucleotide at each position as 1 nucleotide The% was made to have a mixture of all different combinations.   Figure 6 shows a schematic diagram of the synthesis of mutant TSST-1 molecules.   Detailed description of the preferred embodiment   Both the foregoing general description and the following detailed general description are merely exemplary and explanatory. It is understood that the description is provided for the sake of clarity and does not limit the claimed invention. Should be.   The present invention relates to toxic shock syndrome toxin- Kawasaki syndrome caused by the isolation and identification of a unique strain of Staphylococcus aureus that produces 1 Includes methods for group diagnosis, prevention and treatment. Further, according to the present invention, the yellow grape Included are biologically pure cultures of specific strains of cocci.   Methodologically, in general, samples taken from patients suspected of having KS And (ii) the presence of toxic shock syndrome toxin, (ii) culture White, toxic shock syndrome toxin-producing Staphylococcus aureus in culture The presence of (iii) streptococcal exotoxin B or C, or (iv) the listed streptococci Identifying at least one of streptococci that produce any of the coccoid exotoxins Is. Any of these "markers" in a patient are signs of KS. K If diagnosed with S, the method of the present invention is further known in the art. Mutant super antibody molecules according to the methods also described herein, especially TSST-1 Production of molecules, identification of antigen mutants that cannot bind either MHC or TCR , And testing for protection against exposure to nonmutant superantigens.   The molecule of the present invention is useful for treating diseases mediated or induced by antigens such as Kawasaki syndrome. It appears to exert effects in various ways in prevention or treatment. The present invention Of the various ways in which the molecules of E. coli are involved in modifying the T cell response and pathogenesis Production of antigens that provide protection.   In the present invention, the unique strain of S. aureus that produces TSST-1 is KS Was isolated and identified as a virulence factor of E. coli (Examples 1-4). Then TSST-1 mutation Bodies were produced and purified as described in Examples 5-9 below. Next, TSST- 1 mutant contains Vβ15 using human DR-expressing cell line as presentation cell Functional toxin was stimulated by stimulation of mouse T cell hybridomas with α / β receptors. Were examined for the presence of (Example 7).   A number of terms are used in this description, but it is important to define them. Seems useful. These definitions are given below, but in the examples below, these It should be remembered in case the term is used.   The key event of the immune response is complex formation due to the interaction of MHC molecules and antigens, It is the presentation to T cells. In general, T cell responses are quite specific and Is a very limited subpopulation of T cells that responds to a specific complex of It is only a group. The response is generally the phase of most or all components of the T cell receptor. Interaction is required. However, in some circumstances, the presented antigen is a receptor Requires interaction with only the Vβ element of, and all other elements are essentially irrelevant There are cases. This causes the antigen to react with much more T cell populations than usual It means that it is possible and actually reacting.   The molecule of the present invention shows that the Vβ element of the T cell receptor and T cells respond to superantigens. Interact in a manner that leads to modification of the method. The term "modification of T cell responsiveness" suddenly If the mutant molecule is induced by administration of the molecule, or at the same time, in advance Or may alter the response of the patient's T cells to subsequently administered antigens. It means that you can. For example, in the early stages of T cell development, certain subpopulations are offered. It is thought to interact with the indicated antigen and be eliminated. The molecule of the present invention is Mode, ie, at least one or two of the T cells presenting a particular Vβ element It can function by elimination of more than one subpopulation or induction of inactivation / desensitization. Wear.   In a particular embodiment of the invention, the molecule is normally present in the patient under consideration. It modifies T cell responses without altering these B cell responses. This type Substances are useful, for example, to provide passive immunity to patients, and to And is effective. If superantigen derivatives are used, these derivatives They are not early superantigens because they do not elicit special T cell responses, but It will still act as an antigen in generating the vesicle response. Super anti of the present invention The original derivative is able to induce normal antibody production against the superantigen protein. Wear.   Molecules of the invention can also be viewed as competitors to other antigens. This If the molecule described here interacts with MHC elements, it becomes an antigen or superantigen. In contrast to other cases where it is required to generate a response on the full scale of The response can be prevented or its degree can be reduced.   The molecules of the invention impair the T cell responsiveness of an individual due to any of a number of causes. Or, when it is weakened, it can be regarded as an “enhancer”. Included in the present invention Administration of these molecules can significantly expand an individual's T cell population.   As used herein, the term “modification of T cell responsiveness” always refers to the second element (or For example, the Vβ element as part of their T cell receptor. It always means the responsiveness of T cells that are presented as. Essential with other components of the receptor Irrelevant to.   The molecule of the present invention has at least an amino acid sequence of sufficient size for binding to an MHC molecule. Contains a column. The rest of the molecule may be composed of amino acids, carbohydrates and May contain a lipid structure.   The term "decreased responsiveness" also includes the elimination of the portion of T cells that express a particular Vβ element. Is the intention.   As used herein, the term “superantigen derivative” refers to its structure, at least Is also presented by superantigens or parts of superantigens, MHC or T cells Contains an amino acid sequence that is substantially identical to the amino acid sequence required for binding to either Means a molecule that does.   A "modified" superantigen derivative (or fragment) is a "mutated" sue. Different from per-antigen derivative (or fragment). The term "modified superantigen" Contains an amino acid sequence identical to that of the superantigen, It is defined to mean a molecule containing modifications not found in the original molecule itself. Tato For example, if the superantigen contains amino acids 1-250, then the "modified" superantigen The antigen derivative has a sequence identical to amino acids 50-75 to the native superantigen molecule. Is contained between a series of amino acids not found. Other modifications include, for example, green Differences or deletions in cosilation patterns, or glycosylation not present in normal molecules Ration is included.   A “mutant” superantigen is a molecule in which the actual amino acid sequence of the mutant is the It means a structure that has changed with respect to the active form. For example, if the superantigen is Mutant superantigens, provided they contain 1-250 mino acids, are Contains amino acids 50-68 and 72-75 that are identical to the nucleotide sequence, but amino acids 69-71 This is the case. The difference is that "substitution", in which different amino acids are used, more "Additions," which include amino acids and whose sequence is longer than the native form, or amino acids It can be one of the missing "deletions".   “Vaccine” is typically typical of a vaccine when administered to a subject. It means a preparation that induces the same type of response, for example, an active immunoprotective effect. Waku The tin may contain an adjuvant or other material.   The class of molecules known as superantigens is the specific Vβ region of the T cell receptor It is known to interact and lead to large-scale expansion of specific T cell subpopulations. This presumed interaction between the MHC molecule and the superantigen It is almost completely independent of other regions of the T cell receptor.   Regarding the interaction between MHC and peptides, MHC molecules have various "phenotypes". Available and specific for peptides and antigens with diverse phenotypes It should be noted that there is something. For example, HLA-DR is It is known to be related. That is, another MHC phenotype is directed against another antigen Although valuable, it is important to determine the HLA phenotype and the specific antigen or antigen The association with Millie's presentation is within the skill of the engineer in this field.   Accordingly, the present invention is directed to MHC molecules and at least one Vβ receptor on T cells. Including modification of the T cell response through administration to a patient of a molecule that interacts with both The This interaction can affect T cell responses in a number of ways. Response Probably the most basic way to influence the answer is that the molecule interacts with the MHC molecule, It would be a way to prevent the binding of other molecules to MHC. The competing molecule is modified, Or, if it does not cause the proliferation of the original T cells, normal antigens or soot Complexes with MHC that are required for a molecule such as per-antigen to trigger the proliferative response of T cells The inability to form the body will result in a diminished or eliminated response.   Another mode of modifying the T cell response is "desensitization", "inactivation" or T cell depletion. It is through “anergy”. This mechanism includes MHC molecules, antigens and T cell receptor interaction followed by T cell down-regulation or inactivation is involved. It is done. This mechanism is more commonly observed in mature subjects, with exclusion events occurring in fetal subjects. Koru. The latter phenomenon is a decrease mediated by the interaction of the three units, which leads to various T cell The population is actually removed from the organism.   Modification of T cell responses can also include stimulation of T cell subpopulations. With knowledge of the mechanism described here, technicians will be able to identify MHC and specific subpopulations. Administration of a substance that interacts with T cells to a patient causes proliferation of the T cell subpopulation. It is possible to make it. This approach uses specific Vβ subpopulation (s) Particularly desirable for treatment of conditions associated with pathological conditions, such as autoimmune diseases It is.   Overall, the immune response includes both B and T cell responses. And should be understood. In one aspect of the invention, without modifying the B cell response The use of molecules that modify the response of T cells is contemplated. Such materials are described below. Thus, it is particularly useful as a vaccine.   The molecules of the invention are preferably, but not exclusively, superanti-antibodies. It is the original derivative. These derivatives, as described above, are also suddenly modified It may be mutated. These or any other molecule used in the present invention is It is administered in an amount sufficient to modify the T cell response in the manner described above. Material used The amount of material will vary depending on the actual material, the desired response, and the problem of the patient being treated. Move.   This molecule can also serve as a vaccine. These vaccines Is the normal form of the molecule that does not normally produce the associated complete T cell response, It confers protective immunity to the patient in generating a B cell response. In this case, too The material selected for the process should be within a range that can be controlled by the knowledge of the engineer. Meters such as the condition being treated, the Vβ molecule being adjusted, etc. Within the skill range. Vaccines include other materials normally found in vaccine compositions. It is also possible to add additives such as adjuvants and carriers.   The modes of administration of the materials described herein are likewise intravenous, intraperitoneal and intramuscular injection. As well as variable, including all other standard methods of administering therapeutics to patients Can be done.   The present invention also provides a method for making certain mutants useful in the above method, as well as isolation. Nucleic acid sequences encoding the mutated mutants, transformed by these Cell lines and vectors and plasmids used therefor, and isolation Disclosed mutant molecules such as mutant superantigens.   Other applications of the invention described herein will be apparent to those of skill in the art. It seems that there is no need to reiterate.   The terms and expressions used have been used as descriptive terms. Use of such terms and expressions in connection with It is not intended to exclude terms equivalent to parts, and various modifications are possible within the scope of the present invention. It should be recognized that there is.   Cultures from 16 naive KS patients were identified to identify the causative agent of Kawasaki syndrome. Collected (Example 1). Analysis of this culture revealed that 13 of these patients had Vβ2 Toxin associated with expressing T cells, ie toxic shock syndrome toxin (TSST) and streptococcal pyrogenic exotoxins B and C Found [Choi et al., J.EXP.Med. 172: 981-984 (1990); Abe et al., J. Immunol. 146: 3 747-3750 (1991); Tomai et al., Infect. Immun. 60: 701-705 (1992); Drake et al., J. Clin. Immunol. 12: 149-162 (1992)]. On the contrary, only one disease control is Producing any of the toxins, in this single instance, the causative organism is a new feature of the invention. It was not the only identified Staphylococcus aureus.   The bacteria found in these cultures were analyzed. Out of 13 positive cultures Eleven cases were found to contain a strain of S. aureus, while the remaining two cultures Were found to contain streptococcal pyrogenic exotoxin (SPE) B and C (Table 1).   Bacteria isolated from the culture were further tested (Example 2). Each is yellow Produces 3.2 ug / ml TSST-1 in a quantity characteristic of most TSSTs related to D. aureus did. All isolates were found to be tryptophan auxotrophs.   Of great interest is Staphylococcus aureus and other Staphylococcus aureus in cultures of KS There were differences between the strains. In particular, all KS isolates are Staphylococcus aureus white On the other hand, the color of most strains of coagulase-positive Staphylococcus aureus is It is gold as the previous shows. This difference was also found in KS isolates, singles from febrile control patients. Staphylococcus aureus isolates isolated from skin infections, and toxic It was also found among vaginal isolates in patients with Cuck syndrome. Officially recognized In other assays using the technique, KS-related isolates were coagulase-positive and The production of proteases, hemolysins and proteases is associated with skin infections and toxic shock. It is shown to be substantially lower than in cultures from patients with Cuck syndrome. Table 2 Summarizes the data for bacterial isolates.   Then, a series of fruits for examining the DNA of TSST-1 secreting S. aureus in the culture An experiment was conducted (Example 3). The probe is the entire TSST-0 gene, ie Lee et al., J.I. nfect.Dis. 165: 1056-1063 (1992). This remains The gene differs from the TSST-1 producing gene tst only by 14 nucleotides. Do.   From the Southern blot data from these experiments, the pathogenic strain was identified by Chu et al., Infec. mmunol. 56: 2702-2708 (1988) inserted into the tryptophan operon. It was revealed to have the tst gene (Fig. 1). This is TSST-1 production It is a typical example of Daucobacterium.   Western blot / immunoblot assay was also performed. This result (Fig. 2) Compared to positive and negative controls in Western blot studies, Western / Im Confirm that the noblotting method can be used to identify the microorganism of interest.   The data presented in Examples 1-4 show the S. aureus isolates found in KS samples. Is derived from a clone, and the virulent factor expressed by this organism is the host It discloses that it determines the right place for colonization. TSST yellow Most staphylococcal isolates are clonally derived tryptophan auxotrophs. Is Musser et al., Proc.Natl.Acad.Sci. 87: 225-229 (1990) Ming It had been made clear. This conclusion was based on electrophoretic analysis of bacterial enzymes. All of the KS isolates were tryptophan auxotrophs in the analysis of the above example. And Southern hybridization that predicts tryptophan auxotrophs. Pattern is proved. This isolate also shares other diverse phenotypic characteristics. ing.   Shlievert et al., Ann. Inter.Med. 96: 937-940 (1982), Staphylococcus aureus The TSST-positive isolate of not only TSST-1 but also the preferred attachment site to the host Other yellows in that it also produces other potential virulent factors in patterns that predict It has been suggested that it is different from the Staphylococcus aureus isolate. For example, from the mucosal surface TSST isolates are isolated from penetrating skin infections (eg Carbunkel). Do not make a large amount of lipase. In skin infections, lipase production is required for skin invasion. It seems that life is needed. Such isolates tend to produce highly inflammatory lesions. On the other hand, mucosal TSST isolates are usually, if not at all, flares. Almost no disease occurs.   The KS isolate described here below, referred to as "Kawasaki Syndrome I," was found in this strain. Functional regulatory gene regulator, a global regulator of virulent factor production Most closely resembles a Staphylococcus aureus mutant lacking the agr [Recsel et al., Mol. Gen. Genet. 202: 58-61 (1986)]. KS-similar to detachment, arg- The mutant produces only small amounts of lipase, hemolysin and protease and is white [Peng et al., J. Bacteriol. 170: 4365-4372 (1989)]. However, in contrast In addition, the arg-mutant was compared to the amount produced by KS-related isolates in TSST-1. Produces almost no (less than 0.1 ug / ml).   Previous studies have shown that environmental conditions that control the production of TSST-1 by Staphylococcus aureus Is being analyzed. Schlievert et al., J. Infect. Dis. 147: 236-242 (1983); Todd et al. , Infect.Immunol. 45: 339-344 (1984), and Kass et al., J. Infect. Dis. 158: 44-5 1 (1988) showed high levels of animal protein, neutral pH, oxygen and low glucose environment. It was shown to be required for toxin production.   The above example provides a new method for the diagnosis of Kawasaki Syndrome. This method is KS Including assays of samples taken from patients suspected of having (I) presence of toxic shock syndrome toxin, (ii) white crested ibis in culture Existence of Staphylococcus aureus producing sick shock syndrome toxin, (iii) ren S. aureus exotoxin B or C or (iv) any of the listed streptococcal exotoxins Determine at least one of the streptococci produced. These "markers" Both indicate KS in the target.   Staphylococcus aureus, toxic shock syndrome toxin or streptococcus It is also known to indicate the state. In this regard, some You must emphasize the points. Patient populations commonly associated with KS, namely children, Mainly Oriental children, especially Japanese, are vulnerable to toxic shock syndrome. It does not match the size of the group. Moreover, as pointed out, KS has several other There is a diagnostic marker. Finally, TSST-1-producing Staphylococcus aureus associated with disease If is white, all other pathological conditions involving Staphylococcus are of standard gold color. It should be considered that it is due to bacteria. That is, white Staphylococcus aureus It is a specific marker of disease.   The method of measuring the KS index can be changed according to the wishes of researchers. Toki When assaying for syn, an immunoassay, such as the immunodiffusion assay described above, is used. preferable. Any with anti-toxin polyclonal or monoclonal antibody Standard immunoassays such as immunoblot, ELISA, RIA, Deutsch assay etc. can be used. The target molecule is TSST-1, SPEB or It can be SPEC.   If it is desired to culture the sample for bacteria, use any standard used for bacterial culture. Semi-medium, such as the blood agar medium described above, can be used. Then, of Staphylococcus aureus Visual inspection of cultures for white microbes with phenotypic and biological characteristics It can be implemented. Some of these features are mentioned above, others are skilled It is well known to the technicians who did this, and it seems that it is not necessary to list it here.   Cultured from samples taken from KS patients that meet the criteria set forth herein A specific strain of TSST-1-producing Staphylococcus aureus is American Type Culture C. Deposited to the ollection on March 24, 1993 and accepted as accession number A.T.C.C.55049 It was. This culture is used, for example, as an immunogen for the preparation of strain-specific antibodies. Substances for nucleic acids used in probe assays and as potential therapeutic agents Can be used for screening and / or development of. Normal level Of this virulent factor at low levels The object is useful for further studying the onset of KS.   From the DNA analysis of Staphylococcus aureus described above, KS is a nucleic acid-based assay. For example, it is clear that diagnosis can also be performed by performing Southern blotting It was done. Other assays in this area include labeled probes such as radiolabels, bios. Assay with tin or other labeled oligonucleotide, for tst gene Polymerase chain reaction using corresponding oligonucleotides.   The invention also contemplates systems, such as kits, for performing the assay. In the case of a DNA assay, for example, the kit may be used to immobilize sample nucleic acids. Support means, such as nitrocellulose, and a small amount that hybridizes to the target. Includes at least one probe. Kit includes other optional buffers, hive A redissolution solution such as SSC or a washing solution can also be included. In the case of immunoassays, the kit may also be specific for the receptor, eg target molecule. A support, such as a membrane (eg, nitro Cellulose), beads, spheres, test tubes, rods, etc. can be included. Ki Also contains a second receptor, such as labeled antibody or labeled conjugated antibody fragment. Can also be included. Kits are toxins or toxin-presenting bacteria It can also be used in a sandwich assay to detect. For competitive assays A kit is also envisioned. Such a kit could, for example, be a sample of the toxin to be detected. Solid phase to which pull binds, and toxin-specific antibody or antibody fragment composition Includes parts. The binding receptor portion of the kit may be provided in another part of the kit, It may be bound in advance to a solid phase to which the toxin is bound. Such a system Can be used, for example, in displacement assays. Required for such a kit The element of the cell is the part where the factor that is the index of KS can be detected, and the factor directly. Or it is a solid phase that binds indirectly.   Polymerase chain reaction (PCR) and standard molecular biology methods described in Example 5 The method was used for the construction and expression of recombinant TSST-1. TSST-1 suddenly Mutants were generated as described in Example 6. In this method, mature TSST-1 Random mutations were introduced in about 17 residues of the protein. TSST-1 suddenly The first confirmation of the possibility of the mutant was the lysate from the transformant, Of mouse T cell hybridomas with various Vβ elements in a human DR-expressing cell line The presence of functional toxin was tested by stimulation. Implicit in the stimulation of T cell hybridomas The soluble lysate indicates the presence of TSST-1 as a monoclonal antibody against TSST-1. Tested by use of body (mAb) (Example 7). Produces non-functional TSST-1 The transformants were sequenced and the mutations were confirmed. mutation Transformants producing TSST-1 are propagated and mutated TSST- 1 was purified as described in Example 8. Then, the position of the mutation and the effect Was analyzed. That is, the mutant superantigen of the present invention is a wild-type superantigen. Used to selectively stimulate only a portion of the T cell population stimulated by Hara be able to.   Example 1.  Identification of bacterial cultures from patients with Kawasaki disease   The cultures consisted of 16 untreated acute KS patients and 15 fever and / or rashes. They were obtained from the pharynx, axilla, groin, and rectum of disease-controlling patients with. Cultures are all It was collected within 10 days after the onset of the disease in the patient. The criteria for diagnosing patients with acute KS are: American Heart Association Committee on Reumatic Fever Rheumatic fever committee), JAMA 44: 1218-1219 (1990). The culture is a cotton swab Was collected from all of the above sites. Swab swabs from all parts Incubate on top plate, while swabs from the groin and rectum are phenylethyl It was also cultured on alcoholic sheep blood agar. Both culture methods are well-known techniques Is. Bacterial cultures were identified and all isolated Staphylococcus and β-hemolytic For Streptococcus, see Schlivert et al., J. Infect. Dis. 147: 236-242 (1983) This disclosure is incorporated by reference in its entirety). The secretion of xin was screened.   Example 2.  Inspection of bacterial culture   Bacteria isolated from the culture of Example 1 were further examined. Most of each yellow The typical amount of staphylococcal TSST-1 produced was 3.2 ug / ml TSST-1. I was born. The amount of TSST-1 was measured as described in Example 1, while 10⁸Per bacterium Both lipase and hemolysin expressed in units are described by Schlivert et al., Ann. Intern. Med. 96: 937-940 (1992) (this disclosure is incorporated by reference in its entirety). It was. The amount of protease was also determined by Hynes et al., J. Microbiol. Meth. According to 4: 25-31 (1985) 10⁸Expressed in units of bacteria.   Example 3.  Southern blot analysis   Samples from all 11 positive cultures identified in Example 1, Chu et al., Infec. mmunol. 56: 2702-2708 (1988), tryptophan (H) and tyrosine. (4282) Collected with auxotrophic strains. All Bacteria in Todd Hewitt Broth Cultured. According to Chu et al., Supra, chromosomal DNA after treatment with lysostaphin Was isolated. The DNA sample is then digested with the restriction endonuclease Clal. And then the fragments are electrophoretically separated through agarose and then the DNA , Southern, J. Mol. Biol. According to the classical paper of 98: 503-517 (1975), Southern Transferred to nitrocellulose by rotting. Hybridization and detection All production was done using the Genius Kit from Boehringer Mannheim Corporation I followed the instructions.   Example 4.  Western / immunoblotting analysis   The isolated organism was grown on sheep blood agar as described in Example 1. It was. A disk of nitrocellulose filter paper is placed on top of the growing organism and then blood It was cultured on a liquid agar plate for 24 hours. Then remove the disc, Was operated. In particular, nitrocellulose is 3% gelatin (TBS buffer Liquid: 0.02 M Tris in 4 L of water, 0.5 M NaCl, pH 7.5, 3 g in 100 ml) 200 ml coated It was. This is then placed in 200 ml of 0.05% TBS / Tween (1 ml Tween / 2L TBS) at 37 ° C for 30-4. Incubated for 5 minutes. Following this, add nitrocellulose filter paper to 50 ml of TBS / Twe. Treated with rabbit polyclonal anti-TSST (25 ul) at room temperature in en. Incidentally , Wash it twice in TBS / Tween for 5 minutes each time, then anti-rabbit immunoglob Phosphorus-alkaline phosphatase conjugate (25ul per filter paper) and incubate for 1.5 hours I got it. Then, twice in TBS / Tween, then twice in Tween, all 5 minutes Washed for a while. Then, the developing solution [2 mg of 5-bromo-4-chloroindolyl Phosphate, 100 ul N, N-dimethylformamide, 18 ml barbital buffer (0.15M in acetic acid, pH 9.2), 2ml of 0.1% nitroblue tetrazolium, and 40u l of 2M MgCl₂-6H₂O] was added.   Example 5.  Construction and expression of recombinant TSST-1   Polymerase chain reaction (PCR)   PCR [Saiki et al. (1988) Science 239: 487] is based on AmpliTaq recombinant Taq polymerase. And a DNA thermal cycler from Perkin Elmer Cetus (New York, CT). gave. Denaturate at 95 ° C for 1 minute, anneal at 55 ° C and extend for 2 minutes for 35 cycles. I did Curu.   Construction of TSST-1   The gene for superantigen TSST-1 was overexpressed in E. coli as follows. It was. Bacteria, Staphylococcus aureus TSST-1⁺And TSST isolates from Denver C Obtained from Dr. James Todd of hildren's Hospital. Bacteria are cultured and Genome D NA was isolated. PCR was performed with primers Atop and Bbot (Figure 5) It was. The primer sequences were published by Richard Novick & Patrick Schlivert. [Blomster-Hautamaa et al. (1986) J. Biol. Chem. 261: 15783] ． The 5'primer was as follows (SEQ ID NO: 2).   This primer contains a KpnI site and contains plasmid pTZ18R (Pharmacia TSST-1 gene when cloned into Fine Chemicals, Piscataway, NJ) Is placed in frame with the LacZ gene at the above site. This oligonucleo The tide primer is the starting point of the LacZ gene fragment and the TSST-1 gene. It also contains an S / D site to add GAG between, and TSST-1 also has its own GAG. It can be easily transferred to another plasmid. 3'primer is TSST -1 gene contains an XbaI site after the stop codon (SEQ ID NO: 3).   The PCR fragment was digested with KpnI and XbaI to give KpnI / XbaI. Ligated into pTZ18R digested with. E. coli XL1-Blue (Stratagen, LaJ olla, CA) was transformed with the above plasmid. Recombinant TSST-1 ( SDS-PAGE of the 24 kD) product is shown in FIG. This product is human Vβ^{twenty one}T cells It was specifically stimulated in a superantigen-like manner. This is a pure T It was functionally and immunologically identical to SST-1. Then the PCR amplification gene The sequence was determined (Fig. 4). The box shows the difference from the published sequence. public The sequence represented was incorrect and has now been corrected [Lee et al. (1992) J. Infect. Di. s. 165: 1056]. That is, this gene has the same sequence as the wild-type gene.   TSST- of at least one Staphylococcus aureus culture grown from KS patients One gene was also sequenced. This gene had the same sequence (results shown Absent).   Example 6.  Generation of TSST-1 mutants   Figure 6 shows a schematic diagram of the synthesis of mutant TSST-1 molecules. O shown in FIG. Rigonucleotide primers 1top to 13top have three incorrect bases at each position. It was synthesized so that each contained 1%. These mutant oligonucleotides It was effective as a primer in the PCR reaction (Example 5).   Using the mutagenic PCR primer “9top” as an example (FIG. 6), the first Use 9top (mutagenesis primer) and Bbot for PCR reaction, and The region of the TSST-1 gene existing between them was amplified (the primer itself was Introduced between the products). The carrot symbol in 9top matches the wild type sequence Represents a non-nucleotide. If 9top is incorporated into the PCR product, The mismatch of is introduced with it. During the reaction, all the nuclei within 9top There is a mismatch at the otide position but not on another molecule. Mutagenic primer Is 51 nucleotides long and encodes 17 amino acids in the protein.   The product from PCR1 was gel purified and used as a megaprimer in the second PCR reaction. And then paired with primer Atop (Fig. 5) [Sarker & Dommer ( 1990) BioTech. 8: 404]. As above, both primers are combined in the amplified DNA product. It becomes a full-length TSST-1 gene this time. Formed by PCR2 Many different DNA molecules contain nucleotides altered throughout the 9top region. It has (amino acid residues 120 to 136). These molecules are then cloned and tested , Sequenced (Examples 7 and 8). (1) possible mutants in each library Presence of soluble proteins, (2) Tan with native configuration Presence of protein, (3) mouse Vβ15⁺Ability to stimulate cells or human peripheral blood cells Were screened for lack of.   DNA sequencing   Plasmid insert is Sequenase (U.S. Biochemical Corp., Cleveland, OH) And improved double-stranded supercoiled plasmid template [Wickert & Chambliss (1989) i n Editorial Comments, U.S. Biochemical Corp., Cleveland, OH, pages 5-6] Sanger et al. (1977) Proc. Natl. Acad. Sci. USA 74: 5463 dideoxy nucleoti The sequence was determined directly by the Do method.   Example 7.  Transformant screener for TSST-1 mutants Long   Anti-TSST-1 monoclonal antibody (mAb)   Ten mAbs specific for at least 5 epitopes of TSST-1 were labeled with TS It was prepared by standard methods from B10.Q (βBR) immunized multiple times with ST-1. Two of these antibodies, 327H1 and 477L1, were labeled as TSST-1 and TSST-1. Natural mutants were used for both quantification and immunoaffinity purification. 327H1 is the first Has a high affinity for TSST-1 in the examination of the characteristics of Binds equally well to a monofunctional mutant, and the MHC class II complex is TSST-1 Selected from interacting with the TSST-1 toxin in the region that binds to It was. 477L1 has a high affinity for TSST-1 in the first study, It binds to all TSST-1 functional mutants equally well, and the toxin receives T cell receptor. Because it interacts with the TSST-1 toxin in the region that interacts with the body Selected (data not shown).   TSST-1 ELISA   The amount of TSST-1 in the preparation was quantified by ELISA. My Crotter wells were made into natural TSST-1 (Sigma Chemical Co., St. Louis, MO). Coated with 6 μg / ml solution overnight. Wells are then incubated with 25% FCS And washed thoroughly. Inhibits known and unknown TSST-1 preparations As well as adding a fixed amount of anti-TSST-1 antibody to the wells at various concentrations. [Polyclonal rabbit anti-TSST-1 (Toxin Technology, Madison, WI)] Was added in all experiments. After 1 hour, the wells were thoroughly washed and bound antibody was removed. Goat anti-rabbit IgG (Sigma Chemical Co.) or p-nitrofe as substrate Use alkaline phosphatase coupled to one of the nil phosphates It was detected by a standard technique. The OD of the reaction at 405 nm is the data Dose of inhibitor and TSST in unknown samples as assessed by data analysis It was proportional to 1 concentration.   Initial screening for possible mutations of TSST-1   Can contain mutant TSST-1 gene for primary screening Total lysates were prepared as described from each potential transformant. For each aliquot of lysate, using the human DR-expressing cell line as the presentation cell, Stimulation of a mouse T cell hybridoma carrying an α / β receptor containing Vβ15 The presence of functional toxin was tested. Stimulate any of these hybridomas The deficient lysate can cause sudden changes that affect the level or full length of TSST-1 produced. Assays were performed in the presence of TSST-1 protein to exclude differences.   Example 8.  Preparation of recombinant TSST-1   For the initial screening, the transformant removed from the agar plate 96-w containing 100 ul of 2XYT and carbenicillin. Transfer to the wells of a microtiter plate with ell. Add 1 mM IPTG to the medium A second plate was prepared in the same manner except that it was included. Each plate of the first plate 50 μl of glycerol was added to the mixture, mixed and then stored at -70 ° C. For preparation of TSST-1 containing lysates, add 3 mg / ml resuspension to each well of the second plate. HNM buffer containing zozyme and 300 ug / ml DNase I (10 mM Hepes, p H7.0, 30 mM NaCl, 5 mg MgCl₂) 50ul was added. Incubate the plate at 37 ° C for 15 minutes. After incubation, freeze-thawing was repeated 3 times and centrifugation was performed to pellet cell debris. Transfer the supernatant to a new plate and perform both ELISA and T cell hybridoma stimulation. The presence of TSST-1 was tested by the individual. 0.3 and 10 ub / ml with this method A preparation containing TSST-1 was prepared.   To prepare the purified mutant TSST-1, transformants were prepared at -70 ° C. It was recovered from the stored 96-well plate. Overnight culture solution containing IPTG (1 Bacteria) by centrifugation, 1-2 mg / ml lysozyme and 10 ug / ml Resuspended in 1:10 volume of HNM buffer containing DNase I and freeze-thawed three times. Iterated. Centrifuge suspension at 15,000g for 20 minutes to remove bacterial cell debris The supernatant was harvested and filtered (0.2u). The filtrate is a mAb (B 344) A solution containing 1:50 volumes of Sepharose 4B beads coupled with 2-3 mg / ml. I passed the ram. Wash the beads thoroughly with PBS and remove the toxin with 0.1 M glycine. Elute with hydrochloric acid salt (pH 2.7) and 1M Na₂CO₃Neutralized with. Concentrate TSST-1 to 1mg / ml The buffer solution from Centricon 10 microconcentrator (Amicon Corp., Denv ers, MA). In this method, 3 to 10 per 1 L of bacterial culture mg toxin was obtained. TSST-1 manufactured by this method and its sudden change The variants showed> 95% purity as assessed by SDS-PAGE. Those features Some of the altered TSST-1 mutants are listed in Table 3. Placed throughout the molecule There are nearly 500 non-functional mutants with mutations, which are still functional It is characterized by All mutations listed are for A to G or T to C Including nucleotide substitutions in. This is due to the methodology used to generate them Naturally, it is expected.   The mutants shown in Table 2 are all mouse Vβ15⁺T cells (TSST-1 response Responsive mouse subset) is not stimulated. Some mutants stimulated human T cells Absent.   Example 9.  In vitro T cell stimulation assay   TSST-1 is a member of the "superantigen" protein family. This These proteins are secreted by quiescent, inactive T cells to initiate their extensive proliferation. It is characterized by powerful ability. They also expand and begin to carry out some metabolic events. I will. When this occurs, the T cell is "activating". Measure activated T cells One determinable method is by their ability to produce the cytokine IL-1. is there. Therefore, the superantigen assay used here is the IL-2 assay. B. Mouse T cell hybridoma cells having a Vβ15 T cell receptor Varying doses of mutant TSST-1 protein candidates and LG-1 antigen presentation Mix with cells. Perform this assay in a 96-well plastic dish. 4 pieces E. coli extract containing wild-type TSST-1 was added to the E. coli extract without TSST-1 was added to 4 wells of Do. This leaves 88 wells for testing candidate E. coli clones. Pre Incubate the cells overnight. If the superantigen protein is active, T cell hybrid Dorma cells will make IL-2. Next day, inject the culture supernatant from the plate Carefully collect and transfer to a new plate. These supernatants are then diluted 1: 2 several times Do. Then the IL-2 dependent cell line is added. These cells produce IL-2 If you don't, you'll die. After overnight culture, measure the number of IL-2 dependent cells in each culture The Such clones are then sequenced to determine the altered nucleosides in the gene. Determine the position of the tide and identify the altered amino acid residue in the protein.   The numbers in parentheses indicate the numbers of the sample tested. TSST-1 is described above And lipase and hemolysin were both measured by Schlievert et al., Ann. Intern. Measured according to .Med., Unit / 10⁸Shown with bacteria. [Sequence list] SEQ ID NO: 1 Sequence length: 194 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Sequence features   Other Information: Toxic Shock Syndrome Toxin-1 Nucleotides and Ami Acid sequence Array SEQ ID NO: 2 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 3 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 4 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 5 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 6 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 7 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 8 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 9 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 10 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 11 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 12 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 13 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 14 Sequence length: 51 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 15 Sequence length: 42 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 16 Sequence length: 42 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 17 Sequence length: 42 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array SEQ ID NO: 18 Sequence length: 42 Sequence type: Nucleic acid Number of chains: Single chain Topology: linear Array

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁶ Identification number Internal reference number FI C12P 21/02 9162-4B C12N 15/00 ZNAA (C12N 15/09 ZNA C12R 1:01) (C12P 21/02) (C12R 1:19) (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AT, AU, BB, BG, BR, BY, CA, CH, CN, CZ, DE, DK , ES, FI, GB, HU, JP, KP, KR, KZ, LK, LU, MG, MN, MW, NL, NO, NZ, PL, PT, RO, RU, SD , SE, SK, UA, US, VN (72) Inventor Coupler, John United States 80207 Denver, Colorado USA Montview Avenue 4350 (72) Inventor Leung, Donald United States 80111 Englewood, Colorado Es. Geneva Street 5267

Claims (1)

[Claims]   1. A sufficient amount of superantigen or superantigen to modify the T cell response in the patient A method of preventing the onset of Kawasaki syndrome, which comprises administering an antigen derivative.   2. Superantigen or superantigen derivative is a modified superantigen or The method of claim 1 which is a modified superantigen derivative.   3. Superantigens or superantigen derivatives are mutant superantigens or The method according to claim 1, which is a naturally occurring superantigen derivative.   4. Superantigens or superantigen derivatives are S. aureus superantigens The method of claim 1 which is a conductor.   5. S. aureus superantigen is toxic shock syndrome toxin-1 A certain "claim 4" method.   6. A molecule consisting of a mutated TSST-1 superantigen.   7. A molecule consisting of a modified TSST-1 superantigen.   8. Superantigens that induce an immune response against TSST-producing S. aureus Or T-cell responses in patients in need of superantigen derivatives. A method for treating Kawasaki syndrome, which comprises administering a sufficient amount to decorate.   9. Superantigen or superantigen derivative is a modified superantigen or The method of claim 8 which is a modified superantigen fragment.   10. The superantigen or superantigen derivative is a mutant superantigen or The method of claim 8 which is a mutant superantigen fragment.   11. Superantigen or superantigen derivative is Staphylococcus aureus superantigen The method of claim 8 which is a derivative.   12. Staphylococcus aureus superantigen is toxic shock syndrome toxin-1 The method of claim 8 which is   13. Superantigens that induce an immune response against TSST-producing Staphylococcus aureus Or useful for controlling Kawasaki syndrome consisting of superantigen derivative and adjuvant Vaccine.   14. Superantigen or superantigen derivative is modified or mutated TSST The method according to claim 13 which is -1.