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WO2021246199A1 - Method for predicting therapeutic effect on psoriasis, method for evaluating candidate substance for treating psoriasis, and method and kit for evaluating disease state of psoriasis - Google Patents

Method for predicting therapeutic effect on psoriasis, method for evaluating candidate substance for treating psoriasis, and method and kit for evaluating disease state of psoriasis Download PDF

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Publication number
WO2021246199A1
WO2021246199A1 PCT/JP2021/019347 JP2021019347W WO2021246199A1 WO 2021246199 A1 WO2021246199 A1 WO 2021246199A1 JP 2021019347 W JP2021019347 W JP 2021019347W WO 2021246199 A1 WO2021246199 A1 WO 2021246199A1
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Prior art keywords
psoriasis
amount
complex
keratinocytes
therapeutic effect
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PCT/JP2021/019347
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French (fr)
Japanese (ja)
Inventor
由紀子 松永
由起 橋本
晃 石河
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学校法人東邦大学
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Priority to JP2022528743A priority Critical patent/JPWO2021246199A1/ja
Publication of WO2021246199A1 publication Critical patent/WO2021246199A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor

Definitions

  • the present invention relates to a method for predicting a therapeutic effect on psoriasis, a method for evaluating a candidate substance for treatment of psoriasis, a method for evaluating the pathology of psoriasis, and a kit used for these methods.
  • Psoriasis is a disease in which erythema with well-defined white scales frequently occurs, and the epidermis is abnormally proliferated on the patient's skin. In Japan, it is said that there are about 400,000 to 500,000 patients, and the male-female ratio is about 2: 1 and there are many males. Abnormal activation of T cells is said to be involved in the pathology, but the cause is unknown. It is also said to be related to metabolic syndrome. Psoriasis is a lifelong disease once it develops and has the problem of significantly reducing the patient's quality of life (QOL).
  • QOL quality of life
  • Psoriasis vulgaris accounts for 90% of all psoriasis, and up to 9% of the remaining 10% is psoriatic arthritis.
  • vitamin D 3 ointment Treatment of psoriasis, (1) vitamin D 3 ointment, as begins topical therapies such as topical steroid (ointment), then the severity is high, (2) UVB therapy, ultraviolet such as PUVA therapy (light)
  • the treatment method is selected in the order of therapy, (3) PDE4 inhibitor, vitamin A derivative, oral therapy (drinking drug) such as immunosuppressant, and (4) injection therapy of biologics. Since the treatment method is selected based on the disease state and patient background, objective, accurate, and prompt diagnosis is very important.
  • PASI Psoriasis Area and Safety Index
  • the whole body is divided into four regions of head, trunk, upper limbs, and lower limbs, and the degree of erythema, infiltration, desquamation, and the degree of lesion range in each region are scored and judged.
  • this method is subjectively judged and scored, there is a problem that the score varies depending on the evaluator and a unified score cannot be obtained among the evaluators.
  • the PASI score is not sufficient as an index when selecting a treatment method.
  • the S100A8 / A9 complex (sometimes referred to as Calprojectin) is a heterodimer of the S100A8 protein and the S100A9 protein, and is a protein normally present in the cytoplasm.
  • the S100A8 / A9 complex has been reported to have antibacterial activity and to induce proliferation, inflammation, and angiogenesis of other cells via Toll-like receptors, but the detailed mechanism is unknown.
  • the present invention relates to a non-invasive, simple and objective method for predicting a therapeutic effect on psoriasis, a method for evaluating a candidate substance for treatment of psoriasis, a method for evaluating the pathology of psoriasis, and a method for evaluating psoriasis. It is intended to provide a kit used in the method.
  • the present inventors have found that the amount of S100A8 / A9 complex in keratinocytes is closely related to the pathology of psoriasis.
  • the present invention is based on the above-mentioned findings by the present inventors, and the means for solving the above-mentioned problems are as follows. That is, ⁇ 1> Prediction of therapeutic effect on psoriasis, which comprises predicting the therapeutic effect on psoriasis by the above-mentioned treatment using the amount of S100A8 / A9 complex in the keratinocytes of the treated subject as an index. The method. ⁇ 2> For the treatment of psoriasis, which comprises evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index.
  • ⁇ 3> A method for evaluating the pathology of psoriasis, which comprises evaluating the pathology of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
  • ⁇ 4> A kit used in the method according to any one of ⁇ 1> to ⁇ 3>. It is a kit characterized by containing a corneocyte collecting means and an antibody for detecting an S100A8 / A9 complex.
  • a method for predicting a therapeutic effect on psoriasis which can solve the above-mentioned problems in the past, achieve the above-mentioned object, and can be evaluated non-invasively, easily, and objectively, of psoriasis. It is possible to provide a method for evaluating a candidate substance for treatment, a method for evaluating the pathology of psoriasis, and a kit used for these methods.
  • FIG. 1A is a diagram showing the results of verifying the correlation between the PASI score of a psoriasis vulgaris patient and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 1B is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site.
  • FIG. 1A is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site.
  • FIG. 1B “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 1C is a diagram showing the results of verifying the correlation between the EASI score of a patient with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 2A is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriasis vulgaris and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 2B is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area.
  • FIG. 2A is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area.
  • FIG. 2B “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 2C is a diagram showing the results of verifying the correlation between the EASI score of a patient with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 3 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption part in Case 1 and the result of the PASI score.
  • amount of S100A8 / A9 complex represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 4 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the rash portion in Case 2 and the result of the PASI score.
  • FIG. 3 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption part in Case 1 and the result of the PASI score.
  • amount of S100A8 / A9 complex represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 4 is a diagram showing the amount of
  • FIG. 5 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 3.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 6A is a diagram showing changes in the PASI score before and after treatment of 12 cases.
  • FIG. 6B is a diagram showing changes in the amount of S100A8 / A9 complex contained in keratinocytes in the eruption region before and after treatment of 12 cases.
  • amount of S100A8 / A9 complex represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 7 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 4.
  • FIG. 6B is a diagram showing changes in the amount of S100A8 / A9 complex contained in keratinocytes in the eruption region before and after treatment of 12 cases.
  • amount of S100A8 / A9 complex represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 7 is a diagram showing the amount of S100A8 / A9
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 8 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 5.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 9 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the non-rash portion and the result of the PASI score in Case 6.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • FIG. 10 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the non-rash portion and the result of the PASI score in Case 7.
  • “amount of S100A8 / A9 complex” represents the amount (unit: ⁇ g) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
  • Method for predicting a therapeutic effect on psoriasis includes at least an evaluation step, and if necessary, further includes other steps such as a measurement step. ..
  • the evaluation step in the method for predicting the therapeutic effect is a step of predicting the therapeutic effect on psoriasis by the treatment using the amount of the S100A8 / A9 complex in the keratinocytes of the treated subject as an index.
  • psoriasis The type of psoriasis is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include psoriasis vulgaris, guttate psoriasis, pustular psoriasis, psoriatic erythema, and psoriatic arthritis. Among these, it can be suitably used for predicting the therapeutic effect on at least one of psoriasis vulgaris and psoriatic arthritis.
  • the subject is not particularly limited as long as it is a subject whose therapeutic effect on psoriasis is required to be predicted, and can be appropriately selected depending on the purpose.
  • the species of the subject is not particularly limited and may be appropriately selected depending on the intended purpose, but humans are preferable.
  • the sex and age of the subject are not particularly limited and can be appropriately selected according to the purpose.
  • the type of the treatment is not particularly limited and may be appropriately selected depending on the purpose.
  • external therapy ovaltment
  • ultraviolet (light) therapy ultraviolet
  • internal therapy drinking drug
  • biologics biologics
  • the injection therapy that was used can be mentioned.
  • the present invention can be suitably used for predicting the therapeutic effect of various therapies.
  • the external therapy is not particularly limited and may be appropriately selected depending on the purpose, for example, vitamin D 3 ointment, and the like topical steroid.
  • the ultraviolet light therapy is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include UVB therapy and PUVA therapy.
  • the oral therapy is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include PDE4 inhibitors, vitamin A derivatives and immunosuppressants.
  • the biologics for the injection therapy are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include brodalumab, secukinumab, ixekizumab and adalimumab.
  • the keratinocytes may be derived from the rash portion of the subject or may be derived from the non-rash portion, but those derived from the rash portion are preferable.
  • the method for preparing (collecting) the keratinocytes is not particularly limited, and a known method can be appropriately selected, but a method for collecting by the tape stripping method in that it can be collected non-invasively. Is preferable.
  • the corneocytes may be subjected to a treatment such as ultrasonic treatment, if necessary, in order to measure the amount of the S100A8 / A9 complex.
  • the S100A8 / A9 complex is a heterodimer of S100A8 protein and S100A9 protein, and is a protein present in cells.
  • the amount of the S100A8 / A9 complex in the keratinocytes of the treated subject used in the evaluation step may be measured during the evaluation step or may be measured separately. good.
  • the method for measuring the amount of the S100A8 / A9 complex in the keratinocytes is not particularly limited, and a known method can be appropriately selected.
  • an antibody that specifically recognizes the S100A8 / A9 complex can be selected. Examples thereof include a method of measuring by an immunological method.
  • the immunological method is not particularly limited, and a known method can be appropriately selected. For example, ELISA method, EIA method, RIA method, Western blotting method, immunohistochemical staining method, flow cytometry analysis and the like can be selected. Can be mentioned.
  • the antibody that specifically recognizes the S100A8 / A9 complex is not particularly limited, and an antibody prepared by a known method or a commercially available product can be used.
  • the antibody may contain a label of a fluorescent substance or the like, if necessary.
  • the type of the antibody is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a monoclonal antibody, a polyclonal antibody, a human antibody, a chimeric antibody, and a humanized antibody.
  • the evaluation (prediction) method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
  • the reference amount is not particularly limited and may be appropriately selected depending on the intended purpose.
  • S100A8 / in the keratinocytes of a subject or an individual suffering from psoriasis before treatment Amount based on the amount of A9 complex (hereinafter sometimes referred to as "disease value I")
  • a value based on may be referred to as “value I”) ”.
  • the amount based on the disease value I may be, for example, the same amount as the disease value I or an amount close to the disease value I.
  • the amount based on the normal value I may be, for example, the same amount as the normal value I or an amount close to the normal value I.
  • the reference amount is (I) an amount based on the disease value I
  • (i) the amount of S100A8 / A9 complex in the keratinocytes of the subject treated with the treatment is predicted to have a therapeutic effect on psoriasis by the treatment when the amount is less than the amount based on the disease value I.
  • the amount of is equal to or more than the amount based on the disease value I, it is predicted that the therapeutic effect of the treatment on psoriasis is ineffective or low.
  • the reference amount is based on (II) normal value I
  • (i) the amount of S100A8 / A9 complex in the keratin cells of the subject to be treated is the normal value I.
  • the amount is equal to or less than the amount based on, it is predicted that the therapeutic effect on psoriasis by the treatment is effective or high, and (ii) the amount of S100A8 / A9 complex in the keratin cells of the subject treated with the treatment is , If the amount is higher than the amount based on the normal value I, it is predicted that the therapeutic effect of the treatment on psoriasis is ineffective or low.
  • the reference amount may be measured and set when the method for predicting the therapeutic effect is carried out, or may be measured and set in advance.
  • the other steps in the method for predicting the therapeutic effect are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose. Examples thereof include a measurement step.
  • the measuring step in the method for predicting the therapeutic effect is a step of measuring the amount of the S100A8 / A9 complex in the keratinocytes of the treated subject.
  • the method for measuring the amount of the S100A8 / A9 complex in the keratinocytes is as described in the above-mentioned item-S100A8 / A9 complex-.
  • the method for predicting the therapeutic effect may be performed in combination with other methods.
  • the therapeutic effect can be predicted at an early stage in a non-invasive, simple and objective manner.
  • the method for evaluating a candidate substance for treatment of psoriasis of the present invention includes at least an evaluation step, and further includes other steps as necessary.
  • the evaluation step in the evaluation method of the candidate substance for treatment is a step of evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index. be.
  • psoriasis The type of psoriasis is the same as that described in the above-mentioned item of the method for predicting the therapeutic effect of the present invention-psoriasis.
  • the subject is not particularly limited as long as it can evaluate the therapeutic effect of the candidate substance on psoriasis, and can be appropriately selected depending on the intended purpose.
  • the species, sex, and age of the subject are the same as those described in the item-Subject-of the above-mentioned method for predicting the therapeutic effect of the present invention.
  • the candidate substance is not particularly limited and may be appropriately selected depending on the intended purpose.
  • the administration method, administration period, and dose of the candidate substance are not particularly limited and may be appropriately selected depending on the intended purpose.
  • keratinocytes and the method for preparing the keratinocytes are the same as those described in the above-mentioned item-Keratinocytes-in the method for predicting the therapeutic effect of the present invention.
  • the evaluation method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
  • the reference amount is not particularly limited and may be appropriately selected depending on the intended purpose.
  • An amount based on the amount of / A9 complex (hereinafter sometimes referred to as "disease value II"), (IV) the amount of S100A8 / A9 complex in the keratinocytes of an individual not suffering from psoriasis (hereinafter, "" A value based on "normal value II” may be mentioned. These may be used alone or in combination of two or more.
  • the amount based on the disease value II may be, for example, the same amount as the disease value II or an amount close to the disease value II.
  • the amount based on the normal value II may be, for example, the same amount as the normal value II or an amount close to the normal value II.
  • the reference amount is (III) an amount based on the disease value II
  • the amount is less than the amount based on the disease value II
  • the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered is equal to or more than the amount based on the disease value II, the candidate substance has no or low therapeutic effect on psoriasis. That is, it is evaluated that the candidate substance is not a candidate substance for treatment of psoriasis.
  • the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered is the normal value.
  • the amount is equal to or less than the amount based on II, it is evaluated that the candidate substance has a therapeutic effect on psoriasis or is high, that is, the candidate substance is a candidate substance for treatment of psoriasis, and (ii) the candidate substance is When the amount of S100A8 / A9 complex in the keratinocytes of the administered subject is higher than the amount based on the normal value II, the therapeutic effect of the candidate substance on psoriasis is ineffective or low, that is, the candidate substance is It is evaluated that it is not a candidate substance for the treatment of psoriasis.
  • the one measured and set when the evaluation method of the candidate substance for treatment is carried out may be used, or the one measured and set in advance may be used.
  • the evaluation method of the therapeutic candidate substance may be performed in combination with other methods.
  • a therapeutic candidate substance can be selected non-invasively, easily, and objectively.
  • the psoriasis disease evaluation method of the present invention includes at least an evaluation step, and further includes other steps as necessary.
  • the evaluation step in the disease evaluation method is a step of evaluating the disease state of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
  • psoriasis The type of psoriasis is the same as that described in the above-mentioned item of the method for predicting the therapeutic effect of the present invention-psoriasis.
  • the subject is not particularly limited as long as it is a subject for which evaluation of the pathological condition of psoriasis is required, and can be appropriately selected depending on the purpose.
  • the species, sex, and age of the subject are the same as those described in the item-Subject-of the above-mentioned method for predicting the therapeutic effect of the present invention.
  • keratinocytes and the method for preparing the keratinocytes are the same as those described in the above-mentioned item-Keratinocytes-in the method for predicting the therapeutic effect of the present invention.
  • the evaluation method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
  • the reference amount is not particularly limited and may be appropriately selected depending on the intended purpose.
  • the amount of the S100A8 / A9 complex in the keratinocytes of an individual suffering from (V) psoriasis (hereinafter, "disease”).
  • Amount based on (sometimes referred to as "value III"), (VI) amount of S100A8 / A9 complex in keratinocytes of individuals not suffering from psoriasis hereinafter sometimes referred to as "normal value III”
  • the value based on is mentioned. These may be used alone or in combination of two or more.
  • the amount based on the disease value III may be, for example, the same amount as the disease value III or an amount close to the disease value III.
  • the amount based on the normal value III may be, for example, the same amount as the normal value III or an amount close to the normal value III.
  • the reference amount is (V) an amount based on the disease value III
  • the amount of the S100A8 / A9 complex in the corneocyte of the subject is the disease value III.
  • the amount is less than the amount based on the disease value III, it is evaluated that the disease has declined, and (ii) when the amount of the S100A8 / A9 complex in the corneocytes of the subject is equal to or more than the amount based on the disease value III. It is evaluated that there is no change in the disease or the disease has progressed.
  • the reference amount is an amount based on the (VI) normal value III
  • the amount of the S100A8 / A9 complex in the corneocyte of the subject is the same as the amount based on the normal value III.
  • the amount is less than or equal to the amount, it is evaluated that the disease has declined, and (ii) there is no change in the disease when the amount of the S100A8 / A9 complex in the keratinocytes of the subject is larger than the amount based on the normal value III. Or it is evaluated that the disease has progressed.
  • the one measured and set at the time of carrying out the disease evaluation method may be used, or the one measured and set in advance may be used.
  • the disease evaluation method may be performed in combination with other methods.
  • the pathological condition of psoriasis can be evaluated non-invasively, easily and objectively.
  • the invention also relates to a method of collecting data for predicting a therapeutic effect on psoriasis, for evaluating a therapeutic candidate for psoriasis, or for assessing the pathology of psoriasis.
  • the method of collecting the data includes at least a measurement step and, if necessary, other steps.
  • the measurement step in the method for collecting the data includes the amount of S100A8 / A9 complex in the keratinocytes of the treated subject, the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance was administered.
  • it is a step of measuring the amount of the S100A8 / A9 complex in the corneocytes of the subject, and is performed in the same manner as the measurement step described in the item of ⁇ Other steps> in the above-mentioned method for predicting the therapeutic effect of the present invention. Can be done.
  • the other steps in the method for collecting the data are not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include a corneocyte preparation step.
  • useful data can be collected for predicting the therapeutic effect on psoriasis, for evaluating a candidate substance for treatment of psoriasis, or for evaluating the pathological condition of psoriasis.
  • kit The kit of the present invention is a kit used for at least one of the above-mentioned method for predicting the therapeutic effect of the present invention, the method for evaluating a candidate substance for treatment, and the method for evaluating the pathological condition of psoriasis, and is used as a means for collecting keratinocytes. , S100A8 / A9 complex detection antibody, and if necessary, further include other configurations.
  • the means for collecting corneocytes is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include adhesive tapes such as cellophane tape.
  • the antibody for detecting the S100A8 / A9 complex is not particularly limited and may be appropriately selected depending on the intended purpose.
  • the item of -S100A8 / A9 complex-in the above-mentioned method for predicting the therapeutic effect of the present invention The same as those described in.
  • kit of the present invention it is possible to predict the therapeutic effect on psoriasis, evaluate the candidate substance for the treatment of psoriasis, or evaluate the pathological condition of psoriasis in a non-invasive, simple and objective manner.
  • test examples of the present invention will be described below, but the present invention is not limited to these test examples.
  • Test Example 1 ⁇ Method> -Sampling from the subject- A cellophane tape (Nichiban Co., Ltd.) cut into 10 cm was attached to the skin surface of the covering part (mainly the trunk and lower leg) of the following target, and the surface of the stratum corneum was stripped several times to collect corneocytes.
  • psoriasis vulgaris patients, psoriatic arthritis patients, and atopic dermatitis patients collected samples from two sites, a lesion (rash) and a healthy area (rash).
  • the PASI score of the lesion was calculated by the attending physician according to the evaluation criteria. In this study, the same doctor determined the PASI score. Samples were taken before and after the intervention.
  • Psoriasis vulgaris patients 25 males (average age 49.4 years), 5 females (average age 55.2 years)
  • Psoriatic arthritis patients 20 males (average age 49.3 years), 3 females (average age 68.0 years)
  • Patients with atopic dermatitis (disease control) 6 males (average age 41.0 years), 2 females (average age 35.5 years) ⁇ Healthy person (control) 2 males (average age 51.5 years), 3 females (average age 48.0 years)
  • the amount of the S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in atopic dermatitis patients is 5.1 ⁇ g on average, which is about 1/10 of psoriasis patients. there were.
  • the amount of the S100A8 / A9 complex was significantly different between each psoriasis patient and atopic dermatitis patient, and between each psoriasis patient and a healthy person (P ⁇ 0.001 (T-test (T-test (T-test)). both sides))). No significant difference was observed between patients with psoriasis vulgaris and patients with psoriatic arthritis.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in patients with psoriasis vulgaris was compared between the sample derived from the lesion and the sample derived from the healthy part. It was 48.1 ⁇ g in the sample, and 2.9 ⁇ g on average in the sample derived from the healthy part.
  • the amount of S100A8 / A9 complex in the psoriasis vulgaris patient was significantly different between the lesioned part and the healthy part (P ⁇ 0.001 (T-test (two-sided))). It was also confirmed that the expression level of the S100A8 / A9 complex was higher than that of the healthy person even when measured in the healthy part.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in patients with psoriatic arthritis was compared between the sample derived from the lesion and the sample derived from the healthy part. It was 41.8 ⁇ g in the sample, and 2.5 ⁇ g on average in the sample derived from the healthy part.
  • the amount of the S100A8 / A9 complex was significantly different between the lesioned part and the healthy part as in the case of the psoriatic vulgaris patient (P ⁇ 0.001 (T). Test (both sides))). It was also confirmed that the expression level of the S100A8 / A9 complex was higher than that of the healthy person even when measured in the healthy part.
  • FIG. 1A A patient with psoriasis vulgaris is shown in FIG. 1A
  • FIG. 1B a patient with psoriatic arthritis is shown in FIG. 1B
  • FIG. 1C shows the results of verifying the correlation between the EASI score of patients with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site. As shown in FIGS.
  • FIG. 2A A patient with psoriasis vulgaris is shown in FIG. 2A, and a patient with psoriatic arthritis is shown in FIG. 2B.
  • FIG. 2C shows the results of verifying the correlation between the EASI score of patients with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash-free area. As shown in FIGS.
  • Case 1 is a case of a psoriasis vulgaris patient (56 years old, male). Case 1 is a prominent example of brodalumab (anti-IL-17 receptor A antibody: sometimes referred to as "BRO"), which is a biopharmacy.
  • FIG. 3 shows the PASI score (bar graph) measured 6 weeks before the introduction of brodalumab (before treatment) and 52 weeks after the introduction of brodalumab (after treatment), and S100A8 contained in 1 mg of total protein contained in the keratinocytes of the eruption site. The amount (solid line) of the / A9 complex is shown. Case 1 is an example in which the skin has healed to almost the same condition as that of a healthy person.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was 200 ⁇ g or more before the treatment, but became almost zero after the treatment.
  • the amount of the S100A8 / A9 complex reflected clinical symptoms very sensitively.
  • Case 2 is a case of a psoriasis vulgaris patient (61 years old, male).
  • the biopharmacy secukinumab (sometimes referred to as "SEC") became ineffective and was successfully transferred to another biopharmacy, ixekizumab (sometimes referred to as "IXE”).
  • SEC biopharmacy secukinumab
  • IXE biopharmacy secukinumab
  • FIG. 4 shows the PASI score (bar graph) measured 28 weeks after the introduction of secukinumab, 41 weeks, 82 weeks (at the time of introduction of ixekizumab), and 28 weeks after the introduction of ixekizumab, and 1 mg of total protein contained in the keratinocytes of the rash.
  • Case 3 is a case of a psoriatic arthritis patient (62 years old, male). Case 3 is a case in which the biopharmacy adalimumab (sometimes referred to as "ADA") was reintroduced and the effect was remarkable.
  • FIG. 5 shows the PASI score (bar graph) measured at the time of introduction of adalimumab, 8 weeks after reintroduction of adalimumab (with self-interruption of outpatient), and 37 weeks (with self-interruption of outpatient), and the total amount contained in the keratinocytes of the rash. The amount (solid line) of the S100A8 / A9 complex contained in 1 mg of protein is shown. Case 3 is an example in which the PASI score gradually increased, and then the effect improved significantly. As shown in FIG. 5, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes changed in parallel with the symptom.
  • FIG. 6A The change in PASI score before and after treatment of 12 cases is shown in FIG. 6A, and the change in the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site is shown in FIG. 6B.
  • FIG. 6A it can be seen that the PASI score was very high before the treatment, but it has improved significantly after the treatment.
  • FIG. 6B the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site also changed, and it was almost zero after the treatment.
  • the PASI score is a subjective evaluation, for example, it is not possible to simply compare which is more severe between a person with a score of 40 and a person with a score of 27.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes correlates with the PASI score even between cases, there is an advantage that the same result can be obtained objectively by anyone. There is. Therefore, for example, it is very useful data as a method used for evaluation of a treatment candidate substance.
  • Case 4 is a case of a psoriasis vulgaris and psoriasis liquid psoriasis patient (37 years old, male). Case 4 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
  • FIG. 7 shows the PASI score (bar graph) measured before, 6 weeks, 9 weeks, and 18 weeks after the introduction of Otezura, and the S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site. Shows body mass (solid line). Case 4 is a case in which the PASI score gradually improves and worsens.
  • Case 5 is a case of a psoriatic arthritis patient (58 years old, male). Case 5 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
  • FIG. 8 shows the PASI score (bar graph) measured before, 18 weeks, and 26 weeks after the introduction of Otezura, and the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the rash. Solid line) is shown.
  • Case 5 is a case in which the PASI score has decreased and then increased again. In Case 5, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes increased after remaining unchanged.
  • Case 6 is a case of a psoriatic arthritis patient (57 years old, female). Case 6 is a case in which methotrexate (MTX), which is an oral therapy, was introduced and a sufficient effect was not obtained.
  • FIG. 9 shows the PASI score (bar graph) measured at the time of MTX introduction, 2 weeks, 6 weeks, and 18 weeks after MTX introduction, and S100A8 / A9 contained in 1 mg of total protein contained in keratinocytes in the rash area. The amount of complex (solid line) is shown. In case 6, two weeks after the introduction of MTX, the amount of S100A8 / A9 complex was increased, whereas it was the same as that at the time of introduction of the PASI score. This increase in the amount of S100A8 / A9 complex is considered to have predicted the deterioration of the PASI score 6 weeks after the introduction.
  • Case 7 is a case of a patient with arthritis vulgaris (24 years old, male). Case 7 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
  • FIG. 10 shows the PASI score (bar graph) measured at the time of introduction of Otezura, 2 weeks, 6 weeks, and 13 weeks after the introduction of Otezura, and S100A8 / A9 contained in 1 mg of total protein contained in the keratinocytes of the rash area. The amount of complex (solid line) is shown.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was high in psoriasis patients and changed in parallel with the symptoms.
  • the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes is an objective index of the disease state, and may be useful for determining the effect of treatment candidates in developing a therapeutic drug for psoriasis. Shown. It was also shown that the amount of S100A8 / A9 complex contained per 1 mg of total protein contained in corneocytes may be useful in predicting the course of treatment. In particular, it is considered to be useful as a material for determining the continuation of other treatments, which take a long time to determine the effect, rather than the biopharmacy in which the therapeutic effect appears rapidly.
  • Examples of aspects of the present invention include the following.
  • Prediction of therapeutic effect on psoriasis which comprises predicting the therapeutic effect on psoriasis by the above-mentioned treatment using the amount of S100A8 / A9 complex in the keratinocytes of the treated subject as an index.
  • ⁇ 3> For the treatment of psoriasis, which comprises evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index.
  • This is an evaluation method for candidate substances.
  • a method for evaluating the pathology of psoriasis which comprises evaluating the pathology of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
  • ⁇ 5> The method according to any one of ⁇ 1> to ⁇ 4>, wherein the psoriasis is at least one of psoriasis vulgaris and psoriatic arthritis.
  • ⁇ 6> The method according to any one of ⁇ 1> to ⁇ 5>, wherein the keratinocytes are derived from the rash portion of the subject.
  • ⁇ 7> A kit used in the method according to any one of ⁇ 1> to ⁇ 6>. It is a kit characterized by containing a corneocyte collecting means and an antibody for detecting an S100A8 / A9 complex.

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Abstract

A method for predicting therapeutic effect on psoriasis, said method comprising predicting the therapeutic effect of a therapy on psoriasis using, as an index, the amount of S100A8/A9 complex in corneocytes of a subject having received the therapy.

Description

乾癬に対する治療効果の予測方法、乾癬の治療用候補物質の評価方法、及び乾癬の病勢評価方法、並びにキットMethods for predicting the therapeutic effect on psoriasis, methods for evaluating candidate substances for treatment of psoriasis, methods for evaluating the pathology of psoriasis, and kits.
　本発明は、乾癬に対する治療効果の予測方法、乾癬の治療用候補物質の評価方法、及び乾癬の病勢評価方法、並びにこれらの方法に用いられるキットに関する。 The present invention relates to a method for predicting a therapeutic effect on psoriasis, a method for evaluating a candidate substance for treatment of psoriasis, a method for evaluating the pathology of psoriasis, and a kit used for these methods.
　乾癬は、境界明瞭な白色鱗屑を付着する紅斑が多発する疾患であり、患者の皮膚では、表皮が異常増殖している。日本では、約４０～５０万人の患者がいるといわれており、男女比は約２：１で男性が多い。Ｔ細胞の異常活性化が病態に関与すると言われているが、その原因は不明である。また、メタボリックシンドロームとの関連性も言われている。乾癬は、一度発症すると生涯続く疾患であり、患者のクオリティ・オブ・ライフ（ＱＯＬ）を著しく低下させるという問題がある。 Psoriasis is a disease in which erythema with well-defined white scales frequently occurs, and the epidermis is abnormally proliferated on the patient's skin. In Japan, it is said that there are about 400,000 to 500,000 patients, and the male-female ratio is about 2: 1 and there are many males. Abnormal activation of T cells is said to be involved in the pathology, but the cause is unknown. It is also said to be related to metabolic syndrome. Psoriasis is a lifelong disease once it develops and has the problem of significantly reducing the patient's quality of life (QOL).
　乾癬には、尋常性乾癬、滴状乾癬、膿疱性乾癬、乾癬性紅皮症、及び乾癬性関節炎の５つの亜型が存在する。尋常性乾癬は乾癬全体の９０％を占めており、残りの１０％のうちの９％までが乾癬性関節炎である。 There are five subtypes of psoriasis: psoriasis vulgaris, guttate psoriasis, pustular psoriasis, psoriatic erythematosus, and psoriatic arthritis. Psoriasis vulgaris accounts for 90% of all psoriasis, and up to 9% of the remaining 10% is psoriatic arthritis.
　乾癬の治療は、（１）ビタミンＤ３外用薬、ステロイド外用薬などの外用療法（塗り薬）から始まり、次いで、重症度が高くなるにつれ、（２）ＵＶＢ療法、ＰＵＶＡ療法などの紫外線（光線）療法、（３）ＰＤＥ４阻害薬、ビタミンＡ誘導体、免疫抑制剤などの内服療法（飲み薬）、（４）生物学的製剤の注射療法の順番で、治療法を選択する。治療法は、病勢や患者背景を基に選択するため、客観的、正確、迅速な診断が非常に重要となる。 Treatment of psoriasis, (1) vitamin D 3 ointment, as begins topical therapies such as topical steroid (ointment), then the severity is high, (2) UVB therapy, ultraviolet such as PUVA therapy (light) The treatment method is selected in the order of therapy, (3) PDE4 inhibitor, vitamin A derivative, oral therapy (drinking drug) such as immunosuppressant, and (4) injection therapy of biologics. Since the treatment method is selected based on the disease state and patient background, objective, accurate, and prompt diagnosis is very important.
　現在、乾癬の主な評価方法としては、ＰＡＳＩ（Ｐｓｏｒｉａｓｉｓ　Ａｒｅａ　ａｎｄ　Ｓｅｖｅｒｉｔｙ　Ｉｎｄｅｘ）スコアが用いられている。この方法は、全身を頭部、体幹、上肢、及び下肢の４領域に分け、各領域の紅斑、浸潤、落屑の程度及び病巣範囲の程度を点数化し、判定する方法である。しかしながら、この方法は主観的に判断して点数化するものであるため、評価者によってばらつきが生じ、評価者間で統一されたスコアが得られないという問題がある。 Currently, the PASI (Psoriasis Area and Safety Index) score is used as the main evaluation method for psoriasis. In this method, the whole body is divided into four regions of head, trunk, upper limbs, and lower limbs, and the degree of erythema, infiltration, desquamation, and the degree of lesion range in each region are scored and judged. However, since this method is subjectively judged and scored, there is a problem that the score varies depending on the evaluator and a unified score cannot be obtained among the evaluators.
　そのため、ＰＡＳＩスコアは、治療法の選択の際の指標としては、十分とは言えないという問題がある。また、乾癬の治療薬を開発する際においても、治療候補物質の効果判定のためには、乾癬の病勢を客観的に評価することが必要である。 Therefore, there is a problem that the PASI score is not sufficient as an index when selecting a treatment method. In addition, when developing a therapeutic drug for psoriasis, it is necessary to objectively evaluate the pathological condition of psoriasis in order to determine the effect of a therapeutic candidate substance.
　Ｓ１００Ａ８／Ａ９複合体（カルプロテクチン（Ｃａｌｐｒｏｔｅｃｔｉｎ）と称することもある。）は、Ｓ１００Ａ８タンパク質とＳ１００Ａ９タンパク質のヘテロダイマーで、通常は細胞質に存在するタンパク質である。Ｓ１００Ａ８／Ａ９複合体は、抗菌作用やＴｏｌｌ様受容体を介した他の細胞の増殖や炎症、血管新生を惹起する作用が報告されているが、その詳細なメカニズムは分かっていない。 The S100A8 / A9 complex (sometimes referred to as Calprojectin) is a heterodimer of the S100A8 protein and the S100A9 protein, and is a protein normally present in the cytoplasm. The S100A8 / A9 complex has been reported to have antibacterial activity and to induce proliferation, inflammation, and angiogenesis of other cells via Toll-like receptors, but the detailed mechanism is unknown.
　これまでに、乾癬患者の表皮では、Ｓ１００Ａ８タンパク質、Ｓ１００Ａ９タンパク質が健常人より高発現していること、また、乾癬発症に関わるＪｕｎ及びＪｕｎｂの２種類の遺伝子をノックアウトした乾癬モデルマウスやイミキモド塗布乾癬モデルマウスでは、Ｓ１００Ａ９遺伝子をノックアウトすると、表皮増殖変化がキャンセルされることが報告されている（例えば、非特許文献１参照）。 So far, in the epidermis of psoriasis patients, S100A8 protein and S100A9 protein are more highly expressed than in healthy subjects, and psoriasis model mice and imikimod-coated psoriasis in which two genes, Jun and Jumb, involved in the onset of psoriasis have been knocked out. In model mice, it has been reported that knocking out the S100A9 gene cancels epidermal growth changes (see, for example, Non-Patent Document 1).
　しかしながら、非侵襲的で簡便に、かつ客観的に乾癬の病勢を評価することができる方法や、乾癬に対する治療効果の予測方法及び乾癬の治療用候補物質の評価方法は未だ提供されておらず、その速やかな提供が強く求められているのが現状である。 However, a method capable of non-invasively, easily and objectively evaluating the pathological condition of psoriasis, a method for predicting a therapeutic effect on psoriasis, and a method for evaluating a candidate substance for treatment of psoriasis have not yet been provided. The current situation is that prompt provision is strongly required.
　本発明は、前記従来における諸問題を解決し、以下の目的を達成することを課題とする。即ち、本発明は、非侵襲的で簡便に、かつ客観的に評価することができる乾癬に対する治療効果の予測方法、乾癬の治療用候補物質の評価方法、及び乾癬の病勢評価方法、並びにこれらの方法に用いられるキットを提供することを目的とする。 It is an object of the present invention to solve the above-mentioned conventional problems and to achieve the following objects. That is, the present invention relates to a non-invasive, simple and objective method for predicting a therapeutic effect on psoriasis, a method for evaluating a candidate substance for treatment of psoriasis, a method for evaluating the pathology of psoriasis, and a method for evaluating psoriasis. It is intended to provide a kit used in the method.
　本発明者らは、前記目的を達成するべく鋭意検討を行った結果、角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、乾癬の病勢と密接に関係していることを知見した。 As a result of diligent studies to achieve the above object, the present inventors have found that the amount of S100A8 / A9 complex in keratinocytes is closely related to the pathology of psoriasis.
　本発明は、本発明者らによる前記知見に基づくものであり、前記課題を解決するための手段としては、以下の通りである。即ち、
　＜１＞　治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記治療による乾癬への治療効果を予測することを含むことを特徴とする乾癬に対する治療効果の予測方法である。
　＜２＞　候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記候補物質による乾癬への治療効果を評価することを含むことを特徴とする乾癬の治療用候補物質の評価方法である。
　＜３＞　被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記被検体における乾癬の病勢を評価することを含むことを特徴とする乾癬の病勢評価方法である。
　＜４＞　前記＜１＞から＜３＞のいずれかに記載の方法において用いられるキットであって、
　角質細胞採取手段と、Ｓ１００Ａ８／Ａ９複合体検出用抗体とを含むことを特徴とするキットである。 The present invention is based on the above-mentioned findings by the present inventors, and the means for solving the above-mentioned problems are as follows. That is,
<1> Prediction of therapeutic effect on psoriasis, which comprises predicting the therapeutic effect on psoriasis by the above-mentioned treatment using the amount of S100A8 / A9 complex in the keratinocytes of the treated subject as an index. The method.
<2> For the treatment of psoriasis, which comprises evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index. This is an evaluation method for candidate substances.
<3> A method for evaluating the pathology of psoriasis, which comprises evaluating the pathology of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
<4> A kit used in the method according to any one of <1> to <3>.
It is a kit characterized by containing a corneocyte collecting means and an antibody for detecting an S100A8 / A9 complex.
　本発明によれば、従来における前記諸問題を解決し、前記目的を達成することができ、非侵襲的で簡便に、かつ客観的に評価することができる乾癬に対する治療効果の予測方法、乾癬の治療用候補物質の評価方法、及び乾癬の病勢評価方法、並びにこれらの方法に用いられるキットを提供することができる。 According to the present invention, a method for predicting a therapeutic effect on psoriasis, which can solve the above-mentioned problems in the past, achieve the above-mentioned object, and can be evaluated non-invasively, easily, and objectively, of psoriasis. It is possible to provide a method for evaluating a candidate substance for treatment, a method for evaluating the pathology of psoriasis, and a kit used for these methods.
図１Ａは、尋常性乾癬患者のＰＡＳＩスコアと皮疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図１Ａ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 1A is a diagram showing the results of verifying the correlation between the PASI score of a psoriasis vulgaris patient and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area. In FIG. 1A, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図１Ｂは、乾癬性関節炎患者のＰＡＳＩスコアと皮疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図１Ｂ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 1B is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site. In FIG. 1B, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図１Ｃは、アトピー性皮膚炎患者のＥＡＳＩスコアと皮疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図１Ｃ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 1C is a diagram showing the results of verifying the correlation between the EASI score of a patient with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site. In FIG. 1C, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図２Ａは、尋常性乾癬患者のＰＡＳＩスコアと無疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図２Ａ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 2A is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriasis vulgaris and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area. In FIG. 2A, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図２Ｂは、乾癬性関節炎患者のＰＡＳＩスコアと無疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図２Ｂ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 2B is a diagram showing the results of verifying the correlation between the PASI score of patients with psoriatic arthritis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area. In FIG. 2B, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図２Ｃは、アトピー性皮膚炎患者のＥＡＳＩスコアと無疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を示す図である。図２Ｃ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 2C is a diagram showing the results of verifying the correlation between the EASI score of a patient with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area. In FIG. 2C, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図３は、症例１における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図３中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 3 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption part in Case 1 and the result of the PASI score. In FIG. 3, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図４は、症例２における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図４中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 4 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the rash portion in Case 2 and the result of the PASI score. In FIG. 4, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図５は、症例３における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図５中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 5 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 3. In FIG. 5, “amount of S100A8 / A9 complex” represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図６Ａは、１２症例の治療前後におけるＰＡＳＩスコアの変化を示す図である。FIG. 6A is a diagram showing changes in the PASI score before and after treatment of 12 cases. 図６Ｂは、１２症例の治療前後における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量の変化を示す図である。図６Ｂ中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 6B is a diagram showing changes in the amount of S100A8 / A9 complex contained in keratinocytes in the eruption region before and after treatment of 12 cases. In FIG. 6B, "amount of S100A8 / A9 complex" represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図７は、症例４における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図７中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 7 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 4. In FIG. 7, “amount of S100A8 / A9 complex” represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図８は、症例５における皮疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図８中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 8 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the eruption site and the result of the PASI score in Case 5. In FIG. 8, “amount of S100A8 / A9 complex” represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図９は、症例６における無疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図９中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 9 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the non-rash portion and the result of the PASI score in Case 6. In FIG. 9, “amount of S100A8 / A9 complex” represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes. 図１０は、症例７における無疹部の角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量とＰＡＳＩスコアの結果を示す図である。図１０中、「Ｓ１００Ａ８／Ａ９複合体の量」は、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（単位：μｇ）を表す。FIG. 10 is a diagram showing the amount of S100A8 / A9 complex contained in the keratinocytes of the non-rash portion and the result of the PASI score in Case 7. In FIG. 10, “amount of S100A8 / A9 complex” represents the amount (unit: μg) of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes.
（乾癬に対する治療効果の予測方法）
　本発明の乾癬に対する治療効果の予測方法（以下、「治療効果の予測方法」と称することがある。）は、評価工程を少なくとも含み、必要に応じて更に、測定工程等のその他の工程を含む。 (Method of predicting therapeutic effect on psoriasis)
The method for predicting a therapeutic effect on psoriasis of the present invention (hereinafter, may be referred to as "method for predicting a therapeutic effect") includes at least an evaluation step, and if necessary, further includes other steps such as a measurement step. ..
＜評価工程＞
　前記治療効果の予測方法における評価工程は、治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記治療による乾癬への治療効果を予測する工程である。 <Evaluation process>
The evaluation step in the method for predicting the therapeutic effect is a step of predicting the therapeutic effect on psoriasis by the treatment using the amount of the S100A8 / A9 complex in the keratinocytes of the treated subject as an index.
－乾癬－
　前記乾癬の種類としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、尋常性乾癬、滴状乾癬、膿疱性乾癬、乾癬性紅皮症、乾癬性関節炎が挙げられる。これらの中でも、尋常性乾癬及び乾癬性関節炎の少なくともいずれかに対する治療効果の予測に好適に用いることができる。 -psoriasis-
The type of psoriasis is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include psoriasis vulgaris, guttate psoriasis, pustular psoriasis, psoriatic erythema, and psoriatic arthritis. Among these, it can be suitably used for predicting the therapeutic effect on at least one of psoriasis vulgaris and psoriatic arthritis.
－被検体－
　前記被検体としては、乾癬に対する治療効果の予測が求められる被検体であれば、特に制限はなく、目的に応じて適宜選択することができる。
　前記被検体の種としても、特に制限はなく、目的に応じて適宜選択することができるが、ヒトが好適に挙げられる。
　前記被検体の性別、年齢としても、特に制限はなく、目的に応じて適宜選択することができる。 -Subject-
The subject is not particularly limited as long as it is a subject whose therapeutic effect on psoriasis is required to be predicted, and can be appropriately selected depending on the purpose.
The species of the subject is not particularly limited and may be appropriately selected depending on the intended purpose, but humans are preferable.
The sex and age of the subject are not particularly limited and can be appropriately selected according to the purpose.
－治療－
　前記治療の種類としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、外用療法（塗り薬）、紫外線（光線）療法、内服療法（飲み薬）、生物学的製剤を用いた注射療法などが挙げられる。本発明は、各種療法の治療効果の予測に好適に用いることができる。
　前記外用療法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ビタミンＤ３外用薬、ステロイド外用薬などが挙げられる。
　前記紫外線療法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ＵＶＢ療法、ＰＵＶＡ療法などが挙げられる。
　前記内服療法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ＰＤＥ４阻害薬、ビタミンＡ誘導体、免疫抑制剤などが挙げられる。
　前記注射療法の生物学的製剤としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ブロダルマブ、セクキヌマブ、イキセキズマブ、アダリムマブなどが挙げられる。 -Treatment-
The type of the treatment is not particularly limited and may be appropriately selected depending on the purpose. For example, external therapy (ointment), ultraviolet (light) therapy, internal therapy (drinking drug), and biologics are used. The injection therapy that was used can be mentioned. The present invention can be suitably used for predicting the therapeutic effect of various therapies.
As the external therapy is not particularly limited and may be appropriately selected depending on the purpose, for example, vitamin D 3 ointment, and the like topical steroid.
The ultraviolet light therapy is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include UVB therapy and PUVA therapy.
The oral therapy is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include PDE4 inhibitors, vitamin A derivatives and immunosuppressants.
The biologics for the injection therapy are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include brodalumab, secukinumab, ixekizumab and adalimumab.
－角質細胞－
　前記角質細胞は、被検体の皮疹部由来のものであってもよいし、無疹部由来のものであってもよいが、皮疹部由来のものが好ましい。 -Keratinocytes-
The keratinocytes may be derived from the rash portion of the subject or may be derived from the non-rash portion, but those derived from the rash portion are preferable.
　前記角質細胞を調製する（採取する）方法としては、特に制限はなく、公知の方法を適宜選択することができるが、非侵襲的に採取することができる点で、テープストリッピング法により採取する方法が好ましい。
　前記角質細胞は、Ｓ１００Ａ８／Ａ９複合体の量を測定するために、必要に応じて超音波処理などの処理が施されてもよい。 The method for preparing (collecting) the keratinocytes is not particularly limited, and a known method can be appropriately selected, but a method for collecting by the tape stripping method in that it can be collected non-invasively. Is preferable.
The corneocytes may be subjected to a treatment such as ultrasonic treatment, if necessary, in order to measure the amount of the S100A8 / A9 complex.
－Ｓ１００Ａ８／Ａ９複合体－
　前記Ｓ１００Ａ８／Ａ９複合体は、Ｓ１００Ａ８タンパク質とＳ１００Ａ９タンパク質のヘテロダイマーであり、細胞内に存在するタンパク質である。 -S100A8 / A9 complex-
The S100A8 / A9 complex is a heterodimer of S100A8 protein and S100A9 protein, and is a protein present in cells.
　前記評価工程で用いる治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量は、前記評価工程を行う際に測定したものであってもよいし、別途測定したものであってもよい。 The amount of the S100A8 / A9 complex in the keratinocytes of the treated subject used in the evaluation step may be measured during the evaluation step or may be measured separately. good.
　前記角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を測定する方法としては、特に制限はなく、公知の方法を適宜選択することができ、例えば、前記Ｓ１００Ａ８／Ａ９複合体を特異的に認識する抗体を用いて、免疫学的手法により測定する方法などが挙げられる。 The method for measuring the amount of the S100A8 / A9 complex in the keratinocytes is not particularly limited, and a known method can be appropriately selected. For example, an antibody that specifically recognizes the S100A8 / A9 complex can be selected. Examples thereof include a method of measuring by an immunological method.
　前記免疫学的手法としては、特に制限はなく、公知の方法を適宜選択することができ、例えば、ＥＬＩＳＡ法、ＥＩＡ法、ＲＩＡ法、ウェスタンブロット法、免疫組織化学染色法、フローサイトメトリー解析などが挙げられる。
　前記Ｓ１００Ａ８／Ａ９複合体を特異的に認識する抗体としては、特に制限はなく、公知の方法により調製したものや市販品を用いることができる。前記抗体は、必要に応じて蛍光物質の標識などを含んでいてもよい。また、前記抗体の種類としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、モノクローナル抗体、ポリクローナル抗体、ヒト抗体、キメラ抗体、ヒト化抗体などが挙げられる。 The immunological method is not particularly limited, and a known method can be appropriately selected. For example, ELISA method, EIA method, RIA method, Western blotting method, immunohistochemical staining method, flow cytometry analysis and the like can be selected. Can be mentioned.
The antibody that specifically recognizes the S100A8 / A9 complex is not particularly limited, and an antibody prepared by a known method or a commercially available product can be used. The antibody may contain a label of a fluorescent substance or the like, if necessary. The type of the antibody is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a monoclonal antibody, a polyclonal antibody, a human antibody, a chimeric antibody, and a humanized antibody.
－評価－
　前記評価（予測）の方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、参照量と比較する方法が挙げられる。 -evaluation-
The evaluation (prediction) method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
　前記参照量としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、（Ｉ）治療が施される前の被検体又は乾癬に罹患している個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「疾患値Ｉ」と称することがある。）に基づく量、（ＩＩ）乾癬に罹患していない個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「正常値Ｉ」と称することがある。）に基づく値などが挙げられる。これらは、１種単独で使用してもよいし、２種以上を併用してもよい。
　前記疾患値Ｉに基づく量としては、例えば、前記疾患値Ｉと同量としたり、前記疾患値Ｉに近い量としたりすることができる。
　前記正常値Ｉに基づく量としては、例えば、前記正常値Ｉと同量としたり、前記正常値Ｉに近い量としたりすることができる。 The reference amount is not particularly limited and may be appropriately selected depending on the intended purpose. For example, (I) S100A8 / in the keratinocytes of a subject or an individual suffering from psoriasis before treatment. Amount based on the amount of A9 complex (hereinafter sometimes referred to as "disease value I"), (II) amount of S100A8 / A9 complex in keratinocytes of individuals not suffering from psoriasis (hereinafter "normal") A value based on (may be referred to as “value I”) ”. These may be used alone or in combination of two or more.
The amount based on the disease value I may be, for example, the same amount as the disease value I or an amount close to the disease value I.
The amount based on the normal value I may be, for example, the same amount as the normal value I or an amount close to the normal value I.
　前記参照量と比較する方法では、前記参照量が（Ｉ）疾患値Ｉに基づく量の場合には、（ｉ）前記治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値Ｉに基づく量よりも少ない場合に、前記治療による乾癬への治療効果がある又は高いと予測し、（ｉｉ）前記治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値Ｉに基づく量と同量以上の場合に、前記治療による乾癬への治療効果がない又は低いと予測する。 In the method of comparison with the reference amount, when the reference amount is (I) an amount based on the disease value I, (i) the amount of S100A8 / A9 complex in the keratinocytes of the subject treated with the treatment is , The S100A8 / A9 complex in the keratinocytes of the subject treated with the treatment is predicted to have a therapeutic effect on psoriasis by the treatment when the amount is less than the amount based on the disease value I. When the amount of is equal to or more than the amount based on the disease value I, it is predicted that the therapeutic effect of the treatment on psoriasis is ineffective or low.
　また、前記参照量が、前記（ＩＩ）正常値Ｉに基づく量の場合には、（ｉ）前記治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値Ｉに基づく量と同量以下の場合に、前記治療による乾癬への治療効果がある又は高いと予測し、（ｉｉ）前記治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値Ｉに基づく量よりも多い場合に、前記治療による乾癬への治療効果がない又は低いと予測する。 When the reference amount is based on (II) normal value I, (i) the amount of S100A8 / A9 complex in the keratin cells of the subject to be treated is the normal value I. When the amount is equal to or less than the amount based on, it is predicted that the therapeutic effect on psoriasis by the treatment is effective or high, and (ii) the amount of S100A8 / A9 complex in the keratin cells of the subject treated with the treatment is , If the amount is higher than the amount based on the normal value I, it is predicted that the therapeutic effect of the treatment on psoriasis is ineffective or low.
　前記参照量は、前記治療効果の予測方法を実施する際に測定して設定したものを用いてもよいし、予め測定して設定したものを用いてもよい。 The reference amount may be measured and set when the method for predicting the therapeutic effect is carried out, or may be measured and set in advance.
＜その他の工程＞
　前記治療効果の予測方法におけるその他の工程としては、本発明の効果を損なわない限り、特に制限はなく、目的に応じて適宜選択することができ、例えば、測定工程などが挙げられる。 <Other processes>
The other steps in the method for predicting the therapeutic effect are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose. Examples thereof include a measurement step.
　前記治療効果の予測方法における測定工程は、治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を測定する工程である。
　前記角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を測定する方法は、上記した－Ｓ１００Ａ８／Ａ９複合体－の項目に記載したとおりである。 The measuring step in the method for predicting the therapeutic effect is a step of measuring the amount of the S100A8 / A9 complex in the keratinocytes of the treated subject.
The method for measuring the amount of the S100A8 / A9 complex in the keratinocytes is as described in the above-mentioned item-S100A8 / A9 complex-.
　前記治療効果の予測方法は、その他の方法と組み合わせて行ってもよい。 The method for predicting the therapeutic effect may be performed in combination with other methods.
　本発明の治療効果の予測方法によれば、非侵襲的で簡便に、かつ客観的に治療効果を早期に予測することができる。 According to the method for predicting the therapeutic effect of the present invention, the therapeutic effect can be predicted at an early stage in a non-invasive, simple and objective manner.
（乾癬の治療用候補物質の評価方法）
　本発明の乾癬の治療用候補物質の評価方法（以下、「治療用候補物質の評価方法」と称することがある。）は、評価工程を少なくとも含み、必要に応じて更にその他の工程を含む。 (Evaluation method of candidate substances for treatment of psoriasis)
The method for evaluating a candidate substance for treatment of psoriasis of the present invention (hereinafter, may be referred to as "method for evaluating a candidate substance for treatment") includes at least an evaluation step, and further includes other steps as necessary.
＜評価工程＞
　前記治療用候補物質の評価方法における評価工程は、候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記候補物質による乾癬への治療効果を評価する工程である。 <Evaluation process>
The evaluation step in the evaluation method of the candidate substance for treatment is a step of evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index. be.
－乾癬－
　前記乾癬の種類としては、上記した本発明の治療効果の予測方法の－乾癬－の項目に記載したものと同様である。 -psoriasis-
The type of psoriasis is the same as that described in the above-mentioned item of the method for predicting the therapeutic effect of the present invention-psoriasis.
－被検体－
　前記被検体としては、前記候補物質による乾癬への治療効果を評価することができる被検体であれば、特に制限はなく、目的に応じて適宜選択することができる。
　前記被検体の種、性別、年齢としては、上記した本発明の治療効果の予測方法の－被検体－の項目に記載したものと同様である。 -Subject-
The subject is not particularly limited as long as it can evaluate the therapeutic effect of the candidate substance on psoriasis, and can be appropriately selected depending on the intended purpose.
The species, sex, and age of the subject are the same as those described in the item-Subject-of the above-mentioned method for predicting the therapeutic effect of the present invention.
－候補物質－
　前記候補物質としては、特に制限はなく、目的に応じて適宜選択することができる。
　前記候補物質の投与方法、投与期間、投与量としては、特に制限はなく、目的に応じて適宜選択することができる。 -Candidate substance-
The candidate substance is not particularly limited and may be appropriately selected depending on the intended purpose.
The administration method, administration period, and dose of the candidate substance are not particularly limited and may be appropriately selected depending on the intended purpose.
－角質細胞－
　前記角質細胞及びその調製方法としては、上記した本発明の治療効果の予測方法の－角質細胞－の項目に記載したものと同様である。 -Keratinocytes-
The keratinocytes and the method for preparing the keratinocytes are the same as those described in the above-mentioned item-Keratinocytes-in the method for predicting the therapeutic effect of the present invention.
－Ｓ１００Ａ８／Ａ９複合体－
　前記Ｓ１００Ａ８／Ａ９複合体及びその量の測定方法としては、上記した本発明の治療効果の予測方法の－Ｓ１００Ａ８／Ａ９複合体－の項目に記載したものと同様である。 -S100A8 / A9 complex-
The method for measuring the S100A8 / A9 complex and its amount is the same as that described in the above-mentioned item-S100A8 / A9 complex-in the method for predicting the therapeutic effect of the present invention.
－評価－
　前記評価の方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、参照量と比較する方法が挙げられる。 -evaluation-
The evaluation method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
　前記参照量としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、（ＩＩＩ）候補物質が投与される前の被検体又は乾癬に罹患している個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「疾患値ＩＩ」と称することがある。）に基づく量、（ＩＶ）乾癬に罹患していない個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「正常値ＩＩ」と称することがある。）に基づく値などが挙げられる。これらは、１種単独で使用してもよいし、２種以上を併用してもよい。
　前記疾患値ＩＩに基づく量としては、例えば、前記疾患値ＩＩと同量としたり、前記疾患値ＩＩに近い量としたりすることができる。
　前記正常値ＩＩに基づく量としては、例えば、前記正常値ＩＩと同量としたり、前記正常値ＩＩに近い量としたりすることができる。 The reference amount is not particularly limited and may be appropriately selected depending on the intended purpose. For example, S100A8 in the keratinocytes of a subject or an individual suffering from psoriasis before administration of the candidate substance (III). An amount based on the amount of / A9 complex (hereinafter sometimes referred to as "disease value II"), (IV) the amount of S100A8 / A9 complex in the keratinocytes of an individual not suffering from psoriasis (hereinafter, "" A value based on "normal value II" may be mentioned. These may be used alone or in combination of two or more.
The amount based on the disease value II may be, for example, the same amount as the disease value II or an amount close to the disease value II.
The amount based on the normal value II may be, for example, the same amount as the normal value II or an amount close to the normal value II.
　前記参照量と比較する方法では、前記参照量が（ＩＩＩ）疾患値ＩＩに基づく量の場合には、（ｉ）前記候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値ＩＩに基づく量よりも少ない場合に、前記候補物質による乾癬への治療効果がある又は高い、即ち前記候補物質が乾癬の治療用候補物質であると評価し、（ｉｉ）前記候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値ＩＩに基づく量と同量以上の場合に、前記候補物質による乾癬への治療効果がない又は低い、即ち前記候補物質が乾癬の治療用候補物質ではないと評価する。 In the method of comparison with the reference amount, when the reference amount is (III) an amount based on the disease value II, (i) the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered. However, when the amount is less than the amount based on the disease value II, it is evaluated that the candidate substance has a therapeutic effect on psoriasis or is high, that is, the candidate substance is a candidate substance for treatment of psoriasis. When the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered is equal to or more than the amount based on the disease value II, the candidate substance has no or low therapeutic effect on psoriasis. That is, it is evaluated that the candidate substance is not a candidate substance for treatment of psoriasis.
　また、前記参照量が、前記（ＩＶ）正常値ＩＩに基づく量の場合には、（ｉ）前記候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値ＩＩに基づく量と同量以下の場合に、前記候補物質による乾癬への治療効果がある又は高い、即ち前記候補物質が乾癬の治療用候補物質であると評価し、（ｉｉ）前記候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値ＩＩに基づく量よりも多い場合に、前記候補物質による乾癬への治療効果がない又は低い、即ち前記候補物質が乾癬の治療用候補物質ではないと評価する。 When the reference amount is based on (IV) normal value II, (i) the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered is the normal value. When the amount is equal to or less than the amount based on II, it is evaluated that the candidate substance has a therapeutic effect on psoriasis or is high, that is, the candidate substance is a candidate substance for treatment of psoriasis, and (ii) the candidate substance is When the amount of S100A8 / A9 complex in the keratinocytes of the administered subject is higher than the amount based on the normal value II, the therapeutic effect of the candidate substance on psoriasis is ineffective or low, that is, the candidate substance is It is evaluated that it is not a candidate substance for the treatment of psoriasis.
　前記参照量は、前記治療用候補物質の評価方法を実施する際に測定して設定したものを用いてもよいし、予め測定して設定したものを用いてもよい。 As the reference amount, the one measured and set when the evaluation method of the candidate substance for treatment is carried out may be used, or the one measured and set in advance may be used.
＜その他の工程＞
　前記治療用候補物質の評価方法におけるその他の工程としては、本発明の効果を損なわない限り、特に制限はなく、目的に応じて適宜選択することができ、例えば、上記した本発明の治療効果の予測方法の＜その他の工程＞の項目に記載したものと同様のものが挙げられる。 <Other processes>
The other steps in the method for evaluating a therapeutic candidate substance are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose. The same as those described in the item of <Other steps> of the prediction method can be mentioned.
　前記治療用候補物質の評価方法は、その他の方法と組み合わせて行ってもよい。 The evaluation method of the therapeutic candidate substance may be performed in combination with other methods.
　本発明の治療用候補物質の評価方法によれば、非侵襲的で簡便に、かつ客観的に治療用候補物質を選別することができる。 According to the method for evaluating a therapeutic candidate substance of the present invention, a therapeutic candidate substance can be selected non-invasively, easily, and objectively.
（乾癬の病勢評価方法）
　本発明の乾癬の病勢評価方法（以下、「病勢評価方法」と称することがある。）は、評価工程を少なくとも含み、必要に応じて更にその他の工程を含む。 (Psoriasis disease evaluation method)
The psoriasis disease evaluation method of the present invention (hereinafter, may be referred to as “pathology evaluation method”) includes at least an evaluation step, and further includes other steps as necessary.
＜評価工程＞
　前記病勢評価方法における評価工程は、被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記被検体における乾癬の病勢を評価する工程である。 <Evaluation process>
The evaluation step in the disease evaluation method is a step of evaluating the disease state of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
－乾癬－
　前記乾癬の種類としては、上記した本発明の治療効果の予測方法の－乾癬－の項目に記載したものと同様である。 -psoriasis-
The type of psoriasis is the same as that described in the above-mentioned item of the method for predicting the therapeutic effect of the present invention-psoriasis.
－被検体－
　前記被検体としては、前記乾癬の病勢の評価が求められる被検体であれば、特に制限はなく、目的に応じて適宜選択することができる。
　前記被検体の種、性別、年齢としては、上記した本発明の治療効果の予測方法の－被検体－の項目に記載したものと同様である。 -Subject-
The subject is not particularly limited as long as it is a subject for which evaluation of the pathological condition of psoriasis is required, and can be appropriately selected depending on the purpose.
The species, sex, and age of the subject are the same as those described in the item-Subject-of the above-mentioned method for predicting the therapeutic effect of the present invention.
－角質細胞－
　前記角質細胞及びその調製方法としては、上記した本発明の治療効果の予測方法の－角質細胞－の項目に記載したものと同様である。 -Keratinocytes-
The keratinocytes and the method for preparing the keratinocytes are the same as those described in the above-mentioned item-Keratinocytes-in the method for predicting the therapeutic effect of the present invention.
－Ｓ１００Ａ８／Ａ９複合体－
　前記Ｓ１００Ａ８／Ａ９複合体及びその量の測定方法としては、上記した本発明の治療効果の予測方法の－Ｓ１００Ａ８／Ａ９複合体－の項目に記載したものと同様である。 -S100A8 / A9 complex-
The method for measuring the S100A8 / A9 complex and its amount is the same as that described in the above-mentioned item-S100A8 / A9 complex-in the method for predicting the therapeutic effect of the present invention.
－評価－
　前記評価の方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、参照量と比較する方法が挙げられる。 -evaluation-
The evaluation method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of comparing with a reference amount.
　前記参照量としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、（Ｖ）乾癬に罹患している個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「疾患値ＩＩＩ」と称することがある。）に基づく量、（ＶＩ）乾癬に罹患していない個体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量（以下、「正常値ＩＩＩ」と称することがある。）に基づく値などが挙げられる。これらは、１種単独で使用してもよいし、２種以上を併用してもよい。
　前記疾患値ＩＩＩに基づく量としては、例えば、前記疾患値ＩＩＩと同量としたり、前記疾患値ＩＩＩに近い量としたりすることができる。
　前記正常値ＩＩＩに基づく量としては、例えば、前記正常値ＩＩＩと同量としたり、前記正常値ＩＩＩに近い量としたりすることができる。 The reference amount is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the amount of the S100A8 / A9 complex in the keratinocytes of an individual suffering from (V) psoriasis (hereinafter, "disease"). Amount based on (sometimes referred to as "value III"), (VI) amount of S100A8 / A9 complex in keratinocytes of individuals not suffering from psoriasis (hereinafter sometimes referred to as "normal value III"). The value based on is mentioned. These may be used alone or in combination of two or more.
The amount based on the disease value III may be, for example, the same amount as the disease value III or an amount close to the disease value III.
The amount based on the normal value III may be, for example, the same amount as the normal value III or an amount close to the normal value III.
　前記参照量と比較する方法では、前記参照量が（Ｖ）疾患値ＩＩＩに基づく量の場合には、（ｉ）前記被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値ＩＩＩに基づく量よりも少ない場合に、病勢が衰えたと評価し、（ｉｉ）前記被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記疾患値ＩＩＩに基づく量と同量以上の場合に、病勢に変化が無い又は病勢が進行したと評価する。 In the method of comparison with the reference amount, when the reference amount is (V) an amount based on the disease value III, (i) the amount of the S100A8 / A9 complex in the corneocyte of the subject is the disease value III. When the amount is less than the amount based on the disease value III, it is evaluated that the disease has declined, and (ii) when the amount of the S100A8 / A9 complex in the corneocytes of the subject is equal to or more than the amount based on the disease value III. It is evaluated that there is no change in the disease or the disease has progressed.
　また、前記参照量が、前記（ＶＩ）正常値ＩＩＩに基づく量の場合には、（ｉ）前記被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値ＩＩＩに基づく量と同量以下の場合に、病勢が衰えたと評価し、（ｉｉ）前記被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量が、前記正常値ＩＩＩに基づく量よりも多い場合に、病勢に変化が無い又は病勢が進行したと評価する。 When the reference amount is an amount based on the (VI) normal value III, (i) the amount of the S100A8 / A9 complex in the corneocyte of the subject is the same as the amount based on the normal value III. When the amount is less than or equal to the amount, it is evaluated that the disease has declined, and (ii) there is no change in the disease when the amount of the S100A8 / A9 complex in the keratinocytes of the subject is larger than the amount based on the normal value III. Or it is evaluated that the disease has progressed.
　前記参照量は、前記病勢評価方法を実施する際に測定して設定したものを用いてもよいし、予め測定して設定したものを用いてもよい。 As the reference amount, the one measured and set at the time of carrying out the disease evaluation method may be used, or the one measured and set in advance may be used.
＜その他の工程＞
　前記病勢評価方法におけるその他の工程としては、本発明の効果を損なわない限り、特に制限はなく、目的に応じて適宜選択することができ、例えば、上記した本発明の治療効果の予測方法の＜その他の工程＞の項目に記載したものと同様のものが挙げられる。 <Other processes>
The other steps in the disease evaluation method are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose. Other steps> The same as those described in the item can be mentioned.
　前記病勢評価方法は、その他の方法と組み合わせて行ってもよい。 The disease evaluation method may be performed in combination with other methods.
　本発明の病勢評価方法によれば、非侵襲的で簡便に、かつ客観的に乾癬の病勢を評価することができる。 According to the pathological condition evaluation method of the present invention, the pathological condition of psoriasis can be evaluated non-invasively, easily and objectively.
　また、本発明は、乾癬に対する治療効果を予測するため、乾癬の治療用候補物質を評価するため、又は乾癬の病勢を評価するためのデータを収集する方法にも関する。前記データを収集する方法は、測定工程を少なくとも含み、必要に応じて更にその他の工程を含む。
　前記データを収集する方法における測定工程は、治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量、候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量、又は被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を測定する工程であり、上記した本発明の治療効果の予測方法における＜その他の工程＞の項目に記載した測定工程と同様にして行うことができる。
　前記データを収集する方法におけるその他の工程としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、角質細胞調製工程などが挙げられる。 The invention also relates to a method of collecting data for predicting a therapeutic effect on psoriasis, for evaluating a therapeutic candidate for psoriasis, or for assessing the pathology of psoriasis. The method of collecting the data includes at least a measurement step and, if necessary, other steps.
The measurement step in the method for collecting the data includes the amount of S100A8 / A9 complex in the keratinocytes of the treated subject, the amount of S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance was administered. Alternatively, it is a step of measuring the amount of the S100A8 / A9 complex in the corneocytes of the subject, and is performed in the same manner as the measurement step described in the item of <Other steps> in the above-mentioned method for predicting the therapeutic effect of the present invention. Can be done.
The other steps in the method for collecting the data are not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include a corneocyte preparation step.
　前記データを収集する方法によれば、乾癬に対する治療効果を予測するため、乾癬の治療用候補物質を評価するため、又は乾癬の病勢を評価するために有用なデータを収集することができる。 According to the method of collecting the data, useful data can be collected for predicting the therapeutic effect on psoriasis, for evaluating a candidate substance for treatment of psoriasis, or for evaluating the pathological condition of psoriasis.
（キット）
　本発明のキットは、上記した本発明の治療効果の予測方法、治療用候補物質の評価方法、及び乾癬の病勢評価方法の少なくともいずれかの方法に用いられるキットであって、角質細胞採取手段と、Ｓ１００Ａ８／Ａ９複合体検出用抗体とを少なくとも含み、必要に応じて更にその他の構成を含む。 (kit)
The kit of the present invention is a kit used for at least one of the above-mentioned method for predicting the therapeutic effect of the present invention, the method for evaluating a candidate substance for treatment, and the method for evaluating the pathological condition of psoriasis, and is used as a means for collecting keratinocytes. , S100A8 / A9 complex detection antibody, and if necessary, further include other configurations.
＜角質細胞採取手段＞
　前記角質細胞採取手段としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、セロハンテープ等の粘着テープなどが挙げられる。 <Means for collecting keratinocytes>
The means for collecting corneocytes is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include adhesive tapes such as cellophane tape.
＜Ｓ１００Ａ８／Ａ９複合体検出用抗体＞
　前記Ｓ１００Ａ８／Ａ９複合体検出用抗体としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、上記した本発明の治療効果の予測方法における－Ｓ１００Ａ８／Ａ９複合体－の項目に記載したものと同様のものが挙げられる。 <Antibody for detecting S100A8 / A9 complex>
The antibody for detecting the S100A8 / A9 complex is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the item of -S100A8 / A9 complex-in the above-mentioned method for predicting the therapeutic effect of the present invention. The same as those described in.
＜その他の構成＞
　前記その他の構成としては、本発明の効果を損なわない限り、特に制限はなく、目的に応じて適宜選択することができ、例えば、免疫学的測定法に用いる試薬、キットの取扱説明書などが挙げられる。 <Other configurations>
The other configurations are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose. Can be mentioned.
　本発明のキットによれば、非侵襲的で簡便に、かつ客観的に乾癬に対する治療効果の予測、乾癬の治療用候補物質の評価、又は乾癬の病勢評価を行うことができる。 According to the kit of the present invention, it is possible to predict the therapeutic effect on psoriasis, evaluate the candidate substance for the treatment of psoriasis, or evaluate the pathological condition of psoriasis in a non-invasive, simple and objective manner.
　以下に本発明の試験例を説明するが、本発明は、これらの試験例に何ら限定されるものではない。 The test examples of the present invention will be described below, but the present invention is not limited to these test examples.
（試験例１）
＜方法＞
－対象からのサンプルの採取－
　下記対象の被覆部（主に体幹や下腿）の皮膚表面に１０ｃｍに切ったセロハンテープ（ニチバン株式会社）を貼付し、角層表面を数回ストリッピングし、角質細胞を採取した。
　下記対象のうち、尋常性乾癬患者、乾癬性関節炎患者、及びアトピー性皮膚炎患者は、病変部（皮疹部）及び健常部（無疹部）の２か所からサンプルを採取した。また、同時に、病変部のＰＡＳＩスコアを主治医が評価基準に沿って算出した。なお、本試験においては同じ医師がＰＡＳＩスコアを判定するものとした。
　サンプルの採取は、治療介入前と後に行った。 (Test Example 1)
<Method>
-Sampling from the subject-
A cellophane tape (Nichiban Co., Ltd.) cut into 10 cm was attached to the skin surface of the covering part (mainly the trunk and lower leg) of the following target, and the surface of the stratum corneum was stripped several times to collect corneocytes.
Among the following subjects, psoriasis vulgaris patients, psoriatic arthritis patients, and atopic dermatitis patients collected samples from two sites, a lesion (rash) and a healthy area (rash). At the same time, the PASI score of the lesion was calculated by the attending physician according to the evaluation criteria. In this study, the same doctor determined the PASI score.
Samples were taken before and after the intervention.
　対象の内訳は、以下の通りである。
　・　尋常性乾癬患者
　　　男性　２５人（平均年齢４９．４歳）、女性　５人（平均年齢５５．２歳）
　・　乾癬性関節炎患者
　　　男性　２０人（平均年齢４９．３歳）、女性　３人（平均年齢６８．０歳）
　・　アトピー性皮膚炎患者（疾患コントロール）
　　　男性　　６人（平均年齢４１．０歳）、女性　２人（平均年齢３５．５歳）
　・　健常人（コントロール）
　　　男性　　２人（平均年齢５１．５歳）、女性　３人（平均年齢４８．０歳） The breakdown of the subjects is as follows.
・ Psoriasis vulgaris patients 25 males (average age 49.4 years), 5 females (average age 55.2 years)
・ Psoriatic arthritis patients 20 males (average age 49.3 years), 3 females (average age 68.0 years)
・ Patients with atopic dermatitis (disease control)
6 males (average age 41.0 years), 2 females (average age 35.5 years)
・ Healthy person (control)
2 males (average age 51.5 years), 3 females (average age 48.0 years)
－タンパク質の抽出と定量－
　ストリッピングした２枚目のテープを細切りしたのち超音波破砕機（ＢＩＯＲＵＰＴＯＲ、ソニックバイオ株式会社）にかけ、角質細胞からタンパク質を抽出した。抽出用のバッファーには、０．１Ｍ　Ｔｒｉｓ－ＨＣｌ緩衝液（ｐＨ　８．０）を用いた。
　抽出液中の総タンパク質の量は、ＤＣ　Ｐｒｏｔｅｉｎ　Ａｓｓａｙ　Ｒｅａｇｅｎｔ（Ｂｉｏ　Ｒａｄ）を用いて定量した。また、抽出液中のＳ１００Ａ８／Ａ９複合体の量は、Ｈｕｍａｎ　Ｓ１００Ａ８／Ｓ１００Ａ９　Ｈｅｔｅｒｏｄｉｍｅｒ　Ｑｕａｎｔｉｋｉｎｅ　ＥＬＩＳＡ　Ｋｉｔ（Ｒ＆Ｄ　ｓｙｓｔｅｍｓ）を用いて定量した。これらの定量の結果から、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量を求めた。 -Protein extraction and quantification-
The stripped second tape was cut into small pieces and then subjected to an ultrasonic crusher (BIORUPTOR, Sonic Bio Co., Ltd.) to extract proteins from the keratinocytes. A 0.1 M Tris-HCl buffer (pH 8.0) was used as the extraction buffer.
The amount of total protein in the extract was quantified using DC Protein Assay Reagent (Bio Rad). The amount of S100A8 / A9 complex in the extract was quantified using Human S100A8 / S100A9 Heterodimer Quantine ELISA Kit (R & D systems). From the results of these quantifications, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was determined.
＜結果＞
　角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、尋常性乾癬患者では平均で４８．１μｇ、乾癬性関節炎患者では平均で４１．８μｇであった（いずれも病変部由来のサンプル）。
　一方、健常人における角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は平均で０．０４μｇであり、ほとんど検出されなかった。アトピー性皮膚炎患者（病変部由来のサンプル）における角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれる前記Ｓ１００Ａ８／Ａ９複合体の量は平均で５．１μｇであり、乾癬患者の約１／１０であった。
　前記Ｓ１００Ａ８／Ａ９複合体の量は、各乾癬患者とアトピー性皮膚炎患者との間、各乾癬患者と健常人との間では、有意差が認められた（Ｐ＜０．００１（Ｔ検定（両側）））。なお、尋常性乾癬患者と乾癬性関節炎患者との間では、有意差は認められなかった。 <Result>
The average amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was 48.1 μg in patients with psoriasis vulgaris and 41.8 μg in patients with psoriatic arthritis (both lesions). Origin sample).
On the other hand, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in healthy subjects was 0.04 μg on average, and was hardly detected. The amount of the S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in atopic dermatitis patients (samples derived from lesions) is 5.1 μg on average, which is about 1/10 of psoriasis patients. there were.
The amount of the S100A8 / A9 complex was significantly different between each psoriasis patient and atopic dermatitis patient, and between each psoriasis patient and a healthy person (P <0.001 (T-test (T-test (T-test)). both sides))). No significant difference was observed between patients with psoriasis vulgaris and patients with psoriatic arthritis.
　尋常性乾癬患者における角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量について、病変部由来のサンプルと健常部由来のサンプルとで比較したところ、病変部由来のサンプルでは平均で４８．１μｇ、健常部由来のサンプルでは平均で２．９μｇであった。
　前記尋常性乾癬患者におけるＳ１００Ａ８／Ａ９複合体の量は、病変部と健常部との間で有意差が認められた（Ｐ＜０．００１（Ｔ検定（両側）））。なお、健常部で測定した場合でも、健常人よりもＳ１００Ａ８／Ａ９複合体の発現量が高いことも確認された。 The amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in patients with psoriasis vulgaris was compared between the sample derived from the lesion and the sample derived from the healthy part. It was 48.1 μg in the sample, and 2.9 μg on average in the sample derived from the healthy part.
The amount of S100A8 / A9 complex in the psoriasis vulgaris patient was significantly different between the lesioned part and the healthy part (P <0.001 (T-test (two-sided))). It was also confirmed that the expression level of the S100A8 / A9 complex was higher than that of the healthy person even when measured in the healthy part.
　乾癬性関節炎患者における角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量について、病変部由来のサンプルと健常部由来のサンプルとで比較したところ、病変部由来のサンプルでは平均で４１．８μｇ、健常部由来のサンプルでは平均で２．５μｇであった。
　前記乾癬性関節炎患者においても、尋常性乾癬患者の場合と同様に、Ｓ１００Ａ８／Ａ９複合体の量は、病変部と健常部との間で有意差が認められた（Ｐ＜０．００１（Ｔ検定（両側）））。また、健常部で測定した場合でも、健常人よりもＳ１００Ａ８／Ａ９複合体の発現量が高いことも確認された。 The amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes in patients with psoriatic arthritis was compared between the sample derived from the lesion and the sample derived from the healthy part. It was 41.8 μg in the sample, and 2.5 μg on average in the sample derived from the healthy part.
In the psoriatic arthritis patient as well, the amount of the S100A8 / A9 complex was significantly different between the lesioned part and the healthy part as in the case of the psoriatic vulgaris patient (P <0.001 (T). Test (both sides))). It was also confirmed that the expression level of the S100A8 / A9 complex was higher than that of the healthy person even when measured in the healthy part.
　次に、乾癬患者のＰＡＳＩスコアと皮疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した。尋常性乾癬患者の場合を図１Ａに、乾癬性関節炎患者の場合を図１Ｂに示す。また、アトピー性皮膚炎患者のＥＡＳＩスコアと皮疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を図１Ｃに示す。
　図１Ａ～１Ｃに示されたように、尋常性乾癬患者及び乾癬性関節炎患者の皮疹部におけるＳ１００Ａ８／Ａ９複合体の量と、ＰＡＳＩスコアとの間には、正の相関があることが確認された。一方、アトピー性皮膚炎患者の皮疹部におけるＳ１００Ａ８／Ａ９複合体の量と、ＥＡＳＩスコアとの間では、正の相関は認められなかった。 Next, the correlation between the PASI score of psoriasis patients and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area was examined. A patient with psoriasis vulgaris is shown in FIG. 1A, and a patient with psoriatic arthritis is shown in FIG. 1B. In addition, FIG. 1C shows the results of verifying the correlation between the EASI score of patients with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the eruption site.
As shown in FIGS. 1A-1C, it was confirmed that there is a positive correlation between the amount of S100A8 / A9 complex in the eruption of psoriasis vulgaris patients and psoriatic arthritis patients and the PASI score. rice field. On the other hand, no positive correlation was found between the amount of S100A8 / A9 complex in the eruption area of patients with atopic dermatitis and the EASI score.
　また、乾癬患者のＰＡＳＩスコアと無疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した。尋常性乾癬患者の場合を図２Ａに、乾癬性関節炎患者の場合を図２Ｂに示す。また、アトピー性皮膚炎患者のＥＡＳＩスコアと無疹部における角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関を検証した結果を図２Ｃに示す。
　図２Ａ～２Ｃに示されたように、尋常性乾癬患者及び乾癬性関節炎患者の無疹部におけるＳ１００Ａ８／Ａ９複合体の量と、ＰＡＳＩスコアとの間でも、正の相関があることが確認された。一方、アトピー性皮膚炎患者の無疹部におけるＳ１００Ａ８／Ａ９複合体の量と、ＥＡＳＩスコアとの間では、正の相関は認められなかった。 We also examined the correlation between the PASI score of psoriasis patients and the amount of S100A8 / A9 complex contained in keratinocytes in the rash area. A patient with psoriasis vulgaris is shown in FIG. 2A, and a patient with psoriatic arthritis is shown in FIG. 2B. In addition, FIG. 2C shows the results of verifying the correlation between the EASI score of patients with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes in the rash-free area.
As shown in FIGS. 2A-2C, it was confirmed that there is also a positive correlation between the amount of S100A8 / A9 complex in the rash-free area of psoriasis vulgaris patients and psoriatic arthritis patients and the PASI score. rice field. On the other hand, no positive correlation was found between the amount of S100A8 / A9 complex in the rash-free area of atopic dermatitis patients and the EASI score.
　次に、乾癬患者におけるＰＡＳＩスコアと角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関係数と、アトピー性皮膚炎患者におけるＥＡＳＩスコアと角質細胞に含まれるＳ１００Ａ８／Ａ９複合体の量との相関係数を算出した結果を下記に示す。
　・　尋常性乾癬患者
　　　　皮疹部　０．２０８、無疹部　０．５０３
　・　乾癬性関節炎患者
　　　　皮疹部　０．３５８、無疹部　０．１３７
　・　アトピー性皮膚炎
　　　　皮疹部　０．２４０、無疹部　０．２０３ Next, the correlation coefficient between the PASI score in patients with psoriasis and the amount of S100A8 / A9 complex contained in keratinocytes, and the EASI score in patients with atopic dermatitis and the amount of S100A8 / A9 complex contained in keratinocytes. The result of calculating the correlation coefficient of is shown below.
・ Patients with psoriasis vulgaris rash 0.208, rash 0.503
・ Patients with psoriatic arthritis Rash part 0.358, Rashless part 0.137
・ Atopic dermatitis Rash part 0.240, Rashless part 0.203
　以下、個々の症例について記載する。 The individual cases will be described below.
－症例１－
　症例１は、尋常性乾癬患者（５６歳、男性）についてのケースである。症例１は、生物学的製剤であるブロダルマブ（抗ＩＬ－１７受容体Ａ抗体：「ＢＲＯ」と称することがある。））の著効例である。
　図３に、ブロダルマブ導入６週間前（治療前）と、ブロダルマブ導入５２週間後（治療後）に測定したＰＡＳＩスコア（棒グラフ）と、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例１は、ほぼ健常人の皮膚と同じ状態まで治った例である。図３に示したように、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、治療前は２００μｇ以上あったが、治療後はほぼゼロになった。前記Ｓ１００Ａ８／Ａ９複合体の量は、非常に鋭敏に臨床症状を反映していた。なお、ＰＡＳＩスコアも治療前は２５を超えていたものが、ほぼゼロになっており、改善を反映していた。 -Case 1-
Case 1 is a case of a psoriasis vulgaris patient (56 years old, male). Case 1 is a prominent example of brodalumab (anti-IL-17 receptor A antibody: sometimes referred to as "BRO"), which is a biopharmacy.
FIG. 3 shows the PASI score (bar graph) measured 6 weeks before the introduction of brodalumab (before treatment) and 52 weeks after the introduction of brodalumab (after treatment), and S100A8 contained in 1 mg of total protein contained in the keratinocytes of the eruption site. The amount (solid line) of the / A9 complex is shown.
Case 1 is an example in which the skin has healed to almost the same condition as that of a healthy person. As shown in FIG. 3, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was 200 μg or more before the treatment, but became almost zero after the treatment. The amount of the S100A8 / A9 complex reflected clinical symptoms very sensitively. The PASI score, which exceeded 25 before the treatment, became almost zero, reflecting the improvement.
－症例２－
　症例２は、尋常性乾癬患者（６１歳、男性）についてのケースである。症例２は、生物学的製剤であるセクキヌマブ（「ＳＥＣ」と称することがある。）が無効となり、別の生物学的製剤であるイキセキズマブ（「ＩＸＥ」と称することがある。）に移行し奏功した症例である。
　図４に、セクキヌマブ導入２８週間後、４１週間後、８２週間後（イキセキズマブ導入時）、及びイキセキズマブ導入２８週間後に測定したＰＡＳＩスコア（棒グラフ）と、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例２は、ＰＡＳＩスコアが徐々に上がって、その後著効して良くなった例である。図４に示したように、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、ＰＡＳＩスコア以上に敏感に悪化を反映し、改善に伴ってほぼゼロになった。 -Case 2-
Case 2 is a case of a psoriasis vulgaris patient (61 years old, male). In case 2, the biopharmacy secukinumab (sometimes referred to as "SEC") became ineffective and was successfully transferred to another biopharmacy, ixekizumab (sometimes referred to as "IXE"). This is a case of Secukinumab.
FIG. 4 shows the PASI score (bar graph) measured 28 weeks after the introduction of secukinumab, 41 weeks, 82 weeks (at the time of introduction of ixekizumab), and 28 weeks after the introduction of ixekizumab, and 1 mg of total protein contained in the keratinocytes of the rash. The amount (solid line) of the S100A8 / A9 complex contained in the above is shown.
Case 2 is an example in which the PASI score gradually increased, and then the effect improved significantly. As shown in FIG. 4, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes more sensitively reflected the deterioration than the PASI score, and became almost zero with the improvement.
－症例３－
　症例３は、乾癬性関節炎患者（６２歳、男性）についてのケースである。症例３は、生物学的製剤であるアダリムマブ（「ＡＤＡ」と称することがある。）を再導入し、著効となった症例である。
　図５に、アダリムマブ導入時、アダリムマブ再導入８週間後（通院自己中断あり）、及び３７週間後（通院自己中断あり）に測定したＰＡＳＩスコア（棒グラフ）と、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例３は、ＰＡＳＩスコアが徐々に上がって、その後著効して良くなった例である。図５に示したように、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、症状と平行して推移した。 -Case 3-
Case 3 is a case of a psoriatic arthritis patient (62 years old, male). Case 3 is a case in which the biopharmacy adalimumab (sometimes referred to as "ADA") was reintroduced and the effect was remarkable.
FIG. 5 shows the PASI score (bar graph) measured at the time of introduction of adalimumab, 8 weeks after reintroduction of adalimumab (with self-interruption of outpatient), and 37 weeks (with self-interruption of outpatient), and the total amount contained in the keratinocytes of the rash. The amount (solid line) of the S100A8 / A9 complex contained in 1 mg of protein is shown.
Case 3 is an example in which the PASI score gradually increased, and then the effect improved significantly. As shown in FIG. 5, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes changed in parallel with the symptom.
　１２症例の治療前後におけるＰＡＳＩスコアの変化を図６Ａに、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量の変化を図６Ｂに示す。
　図６Ａに示したように、治療前はＰＡＳＩスコアが非常に高かったのが、治療後には非常に改善している様子がわかる。また、図６Ｂに示したように、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量も同様に変化しており、治療後はほぼゼロに向かっていた。 The change in PASI score before and after treatment of 12 cases is shown in FIG. 6A, and the change in the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site is shown in FIG. 6B.
As shown in FIG. 6A, it can be seen that the PASI score was very high before the treatment, but it has improved significantly after the treatment. In addition, as shown in FIG. 6B, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site also changed, and it was almost zero after the treatment.
　ＰＡＳＩスコアは主観的な評価であるため、例えば、スコアが４０の人と２７の人とでどちらが重症であるのかを単純に比較することができない。一方、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、症例間においてもＰＡＳＩスコアと相関があるため、客観的に、誰が測定しても同じ結果が出るというメリットがある。そのため、例えば、治療候補物質の評価に用いる方法として、非常に有用なデータとなる。 Since the PASI score is a subjective evaluation, for example, it is not possible to simply compare which is more severe between a person with a score of 40 and a person with a score of 27. On the other hand, since the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes correlates with the PASI score even between cases, there is an advantage that the same result can be obtained objectively by anyone. There is. Therefore, for example, it is very useful data as a method used for evaluation of a treatment candidate substance.
－症例４－
　症例４は、尋常性乾癬及び液状乾癬患者（３７歳、男性）についてのケースである。症例４は、内服療法であるＰＤＥ４阻害薬（オテズラ）を導入し、十分な効果が得られなかった症例である。
　図７に、オテズラ導入前、オテズラ導入６週間後、９週間後、及び１８週間後に測定したＰＡＳＩスコア（棒グラフ）と、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例４は、ＰＡＳＩスコアが徐々に良くなって、また悪化したケースである。症状改善が十分ではなかった症例４では、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、最初徐々に下がるが、オテズラ導入９週間後に上昇に転じた。これは、オテズラ導入１８週間後の症状の悪化を予測した可能性がある。即ち、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、ＰＡＳＩスコアの悪化を事前に予測できる可能性がある。 -Case 4-
Case 4 is a case of a psoriasis vulgaris and psoriasis liquid psoriasis patient (37 years old, male). Case 4 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
FIG. 7 shows the PASI score (bar graph) measured before, 6 weeks, 9 weeks, and 18 weeks after the introduction of Otezura, and the S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the eruption site. Shows body mass (solid line).
Case 4 is a case in which the PASI score gradually improves and worsens. In Case 4, where the improvement in symptoms was not sufficient, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes gradually decreased at first, but turned to an increase 9 weeks after the introduction of Otezura. This may have predicted worsening of symptoms 18 weeks after the introduction of Otezura. That is, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes may be able to predict the deterioration of the PASI score in advance.
－症例５－
　症例５は、乾癬性関節炎患者（５８歳、男性）についてのケースである。症例５は、内服療法であるＰＤＥ４阻害薬（オテズラ）を導入し、十分な効果が得られなかった症例である。
　図８に、オテズラ導入前、オテズラ導入１８週間後、及び２６週間後に測定したＰＡＳＩスコア（棒グラフ）と、皮疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例５は、ＰＡＳＩスコアが下がった後、また上がったケースである。症例５では、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、下がらずに推移した後に上がった。これは、症状の悪化を予測した可能性がある。即ち、この症例からも、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量が、ＰＡＳＩスコアの悪化を事前に予測できる可能性が示された。 -Case 5-
Case 5 is a case of a psoriatic arthritis patient (58 years old, male). Case 5 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
FIG. 8 shows the PASI score (bar graph) measured before, 18 weeks, and 26 weeks after the introduction of Otezura, and the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in the keratinocytes of the rash. Solid line) is shown.
Case 5 is a case in which the PASI score has decreased and then increased again. In Case 5, the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes increased after remaining unchanged. This may have predicted worsening of symptoms. That is, from this case as well, it was shown that the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in corneocytes may be able to predict the deterioration of the PASI score in advance.
－症例６－
　症例６は、乾癬性関節炎患者（５７歳、女性）についてのケースである。症例６は、内服療法であるメトトレキサート（ＭＴＸ）を導入し、十分な効果が得られなかった症例である。
　図９に、ＭＴＸ導入時、ＭＴＸ導入２週間後、６週間後、及び１８週間後に測定したＰＡＳＩスコア（棒グラフ）と、無疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例６では、ＭＴＸ導入２週間後では、ＰＡＳＩスコア導入時と変わらなかったのに対し、Ｓ１００Ａ８／Ａ９複合体の量は増加していた。このＳ１００Ａ８／Ａ９複合体の量の増加は、導入６週間後でのＰＡＳＩスコアの悪化を予測したと考えられる。 -Case 6-
Case 6 is a case of a psoriatic arthritis patient (57 years old, female). Case 6 is a case in which methotrexate (MTX), which is an oral therapy, was introduced and a sufficient effect was not obtained.
FIG. 9 shows the PASI score (bar graph) measured at the time of MTX introduction, 2 weeks, 6 weeks, and 18 weeks after MTX introduction, and S100A8 / A9 contained in 1 mg of total protein contained in keratinocytes in the rash area. The amount of complex (solid line) is shown.
In case 6, two weeks after the introduction of MTX, the amount of S100A8 / A9 complex was increased, whereas it was the same as that at the time of introduction of the PASI score. This increase in the amount of S100A8 / A9 complex is considered to have predicted the deterioration of the PASI score 6 weeks after the introduction.
－症例７－
　症例７は、尋常性関節炎患者（２４歳、男性）についてのケースである。症例７は、内服療法であるＰＤＥ４阻害薬（オテズラ）を導入し、十分な効果が得られなかった症例である。
　図１０に、オテズラ導入時、オテズラ導入２週間後、６週間後、及び１３週間後に測定したＰＡＳＩスコア（棒グラフ）と、無疹部の角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量（実線）を示す。
　症例７では、オテズラ導入２週間後では、導入時よりもＰＡＳＩスコアは下がったのに対し、Ｓ１００Ａ８／Ａ９複合体の量はあまり下がらなかった。このＳ１００Ａ８／Ａ９複合体の量はあまり下がらなかったことは、導入６週間後でのＰＡＳＩスコアの悪化を予測したと考えられる。 -Case 7-
Case 7 is a case of a patient with arthritis vulgaris (24 years old, male). Case 7 is a case in which a PDE4 inhibitor (Otezura), which is an oral therapy, was introduced and a sufficient effect was not obtained.
FIG. 10 shows the PASI score (bar graph) measured at the time of introduction of Otezura, 2 weeks, 6 weeks, and 13 weeks after the introduction of Otezura, and S100A8 / A9 contained in 1 mg of total protein contained in the keratinocytes of the rash area. The amount of complex (solid line) is shown.
In case 7, two weeks after the introduction of Otezura, the PASI score was lower than that at the time of introduction, whereas the amount of S100A8 / A9 complex was not so low. It is considered that the fact that the amount of this S100A8 / A9 complex did not decrease so much predicted the deterioration of the PASI score 6 weeks after the introduction.
　以上のように、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は乾癬患者で高く、症状と平行して推移することが確認された。角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は病勢の客観的な指標となり、乾癬の治療薬を開発する際における治療候補の効果の判定などに有用であることが示された。また、角質細胞に含まれる総タンパク質１ｍｇ当たりに含まれるＳ１００Ａ８／Ａ９複合体の量は、治療経過の予測に役立つ可能性があることも示された。特に、治療効果が速やかに現れる生物学的製剤よりも、効果判定に時間のかかる他の治療の継続を判定する材料としての有用性が考えられる。 As described above, it was confirmed that the amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes was high in psoriasis patients and changed in parallel with the symptoms. The amount of S100A8 / A9 complex contained in 1 mg of total protein contained in keratinocytes is an objective index of the disease state, and may be useful for determining the effect of treatment candidates in developing a therapeutic drug for psoriasis. Shown. It was also shown that the amount of S100A8 / A9 complex contained per 1 mg of total protein contained in corneocytes may be useful in predicting the course of treatment. In particular, it is considered to be useful as a material for determining the continuation of other treatments, which take a long time to determine the effect, rather than the biopharmacy in which the therapeutic effect appears rapidly.
　本発明の態様としては、例えば、以下のものなどが挙げられる。
　＜１＞　治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記治療による乾癬への治療効果を予測することを含むことを特徴とする乾癬に対する治療効果の予測方法である。
　＜２＞　前記治療が、内服療法、紫外線療法、外用療法、及び生物学的製剤を用いた注射療法からなる群から選択される少なくとも１種である前記＜１＞に記載の方法である。
　＜３＞　候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記候補物質による乾癬への治療効果を評価することを含むことを特徴とする乾癬の治療用候補物質の評価方法である。
　＜４＞　被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記被検体における乾癬の病勢を評価することを含むことを特徴とする乾癬の病勢評価方法である。
　＜５＞　前記乾癬が、尋常性乾癬及び乾癬性関節炎の少なくともいずれかである前記＜１＞から＜４＞のいずれかに記載の方法である。
　＜６＞　前記角質細胞が、前記被検体の皮疹部由来のものである前記＜１＞から＜５＞のいずれかに記載の方法である。
　＜７＞　前記＜１＞から＜６＞のいずれかに記載の方法において用いられるキットであって、
　角質細胞採取手段と、Ｓ１００Ａ８／Ａ９複合体検出用抗体とを含むことを特徴とするキットである。 Examples of aspects of the present invention include the following.
<1> Prediction of therapeutic effect on psoriasis, which comprises predicting the therapeutic effect on psoriasis by the above-mentioned treatment using the amount of S100A8 / A9 complex in the keratinocytes of the treated subject as an index. The method.
<2> The method according to <1>, wherein the treatment is at least one selected from the group consisting of oral therapy, ultraviolet light therapy, external therapy, and injection therapy using a biologic.
<3> For the treatment of psoriasis, which comprises evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index. This is an evaluation method for candidate substances.
<4> A method for evaluating the pathology of psoriasis, which comprises evaluating the pathology of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
<5> The method according to any one of <1> to <4>, wherein the psoriasis is at least one of psoriasis vulgaris and psoriatic arthritis.
<6> The method according to any one of <1> to <5>, wherein the keratinocytes are derived from the rash portion of the subject.
<7> A kit used in the method according to any one of <1> to <6>.
It is a kit characterized by containing a corneocyte collecting means and an antibody for detecting an S100A8 / A9 complex.

Claims (7)

  1. 　治療が施された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記治療による乾癬への治療効果を予測することを含むことを特徴とする乾癬に対する治療効果の予測方法。 A method for predicting a therapeutic effect on psoriasis, which comprises predicting a therapeutic effect on psoriasis by the above-mentioned treatment using the amount of S100A8 / A9 complex in the keratinocytes of a treated subject as an index.
  2. 　前記治療が、内服療法、紫外線療法、外用療法、及び生物学的製剤を用いた注射療法からなる群から選択される少なくとも１種である請求項１に記載の方法。 The method according to claim 1, wherein the treatment is at least one selected from the group consisting of internal therapy, ultraviolet light therapy, external therapy, and injection therapy using a biologic.
  3. 　候補物質が投与された被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記候補物質による乾癬への治療効果を評価することを含むことを特徴とする乾癬の治療用候補物質の評価方法。 A candidate substance for the treatment of psoriasis, which comprises evaluating the therapeutic effect of the candidate substance on psoriasis using the amount of the S100A8 / A9 complex in the keratinocytes of the subject to which the candidate substance is administered as an index. Evaluation method.
  4. 　被検体の角質細胞におけるＳ１００Ａ８／Ａ９複合体の量を指標として、前記被検体における乾癬の病勢を評価することを含むことを特徴とする乾癬の病勢評価方法。 A method for evaluating the pathology of psoriasis, which comprises evaluating the pathology of psoriasis in the subject using the amount of the S100A8 / A9 complex in the keratinocytes of the subject as an index.
  5. 　前記乾癬が、尋常性乾癬及び乾癬性関節炎の少なくともいずれかである請求項１から４のいずれかに記載の方法。 The method according to any one of claims 1 to 4, wherein the psoriasis is at least one of psoriasis vulgaris and psoriatic arthritis.
  6. 　前記角質細胞が、前記被検体の皮疹部由来のものである請求項１から５のいずれかに記載の方法。 The method according to any one of claims 1 to 5, wherein the keratinocytes are derived from the rash portion of the subject.
  7. 　請求項１から６のいずれかに記載の方法において用いられるキットであって、
    　角質細胞採取手段と、Ｓ１００Ａ８／Ａ９複合体検出用抗体とを含むことを特徴とするキット。     A kit used in the method according to any one of claims 1 to 6.
    A kit comprising a corneocyte collecting means and an antibody for detecting an S100A8 / A9 complex.
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