WO2017188101A1 - アトピー性皮膚炎の皮膚局所の重症度及び治療効果の指標 - Google Patents
アトピー性皮膚炎の皮膚局所の重症度及び治療効果の指標 Download PDFInfo
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- 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
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- 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
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- 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
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/81—Protease inhibitors
- G01N2333/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- G01N2333/811—Serine protease (E.C. 3.4.21) inhibitors
- G01N2333/8121—Serpins
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- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/56—Staging of a disease; Further complications associated with the disease
Definitions
- the present invention relates to a squamous cell carcinoma-associated antigen (Squamous Cell Carcinoma Antigen-1, hereinafter referred to as “SCCA-1”) in skin horny layer cells as an index, and non-invasively determines the local pathology of atopic dermatitis Provide a way to evaluate.
- SCCA-1 squamous Cell carcinoma-associated antigen
- Atopic dermatitis is one of the most common skin diseases, and it occurs mainly in early childhood, but in recent years the number of patients after puberty has increased.
- Atopic dermatitis is a disease mainly caused by pruritus with pruritus that repeats exacerbations and remissions, and its onset mechanism is still unknown, but various environmental factors mainly based on genetic predisposition It is thought that it develops by adding.
- Non-patent Document 1 Genetic predisposition is 1) a family history or history of bronchial asthma, allergic rhinitis, conjunctivitis, atopic dermatitis, or 2) a predisposition to easily produce IgE antibodies (Japan Dermatological Association Atopy) Guideline for dermatitis (Non-patent Document 1)).
- atopic dermatitis is that rashes with different symptom levels coexist in the same patient, and changes or repetitions of remission / deterioration are observed due to various factors. For example, there are changes in symptoms due to the season, changes due to changes in the living environment, or severeness or intractability due to some factors. Treatment status also has a significant effect on the course.
- Atopic dermatitis is a chronic disease and it is difficult to achieve complete cure in a true sense. Therefore, in the treatment, we aim to maintain a remission period with no symptoms as the appearance and patient's consciousness, and not exacerbate it. In recent years, it has been suggested that atopic dermatitis is associated with other allergies such as food allergies, and particularly in the early childhood, it is necessary to perform appropriate treatment at an early stage to approach the remission period. Because it is a disease that is often treated for a long time regardless of whether it is an infant or an adult, it is a disease for which sufficient explanation to patients, compliance with treatment, and adherence should be considered.
- Non-patent Document 1 Severity Scoring of Atopic Dermatitis (SCORAD) and Eczema Area and Severity Index (EASI) by European Task Force on Atopic Dermatisis are commonly used worldwide for classification of severity of atopic dermatitis (Non-patent Document 1) . In both cases, severity is classified by highly objective scores, with the highest score being 103 for SCORAD and 72 for EASI.
- the skin symptom score is the area of the rash (percentage of the body surface area) and erythema, infiltration / papules, exudate / crust, scratches, lichenification, and dry rash
- a step of calculating is required.
- the calculation of SCORAD values has the disadvantage that it requires specialized skills by skilled doctors and takes time and effort. When the test subject is an infant, it is not desirable that the evaluation takes time.
- Non-patent Document 2 Non-patent Document 2
- the degree of symptom is determined by the ratio of the eruption to the body surface area.
- atopic dermatitis is not sufficient because eruptions with different symptom levels exist in the same patient.
- moisturizers for the purpose of supplementing dryness and lowering of barrier function and preventing relapse of inflammation when the severity of skin rash is minor.
- a moisturizer alone will not provide a sufficient effect.
- Topical steroids and tacrolimus ointment are used to suppress inflammation of atopic dermatitis.
- the strength and dosage form of topical steroids classified in order of strong clinical effect should be selected in consideration of the severity of each rash, application site, age, and the like. Since the strength of the effects of steroid topical drugs and the likelihood of side effects are generally balanced, it is possible to appropriately select drugs with the appropriate rank for each rash without selecting steroid topical drugs that are stronger than necessary. is important. Therefore, it is desired to provide an objective index of the degree of local symptom of atopic dermatitis.
- the severity of the skin rash is minor, it can be dealt with by skin care by using a moisturizer or protective agent that does not contain steroids or tacrolimus, but external treatment will continue even for minor skin symptoms If this is not done, inflammation may easily relapse, and it may be necessary to rely on the use of stronger topical steroids.
- the judgment of the severity of atopic dermatitis relies on the findings by the naked eye, so in the case that unnecessary treatment is continued even though medication is not actually required, or necessary treatment is interrupted. As a result, symptoms often relapse.
- proactive therapy in which treatment is performed with a weak anti-inflammatory agent and skin care before severe symptoms relapse can be adopted. For this reason, an objective local index that makes it possible to grasp a disease state that is difficult to judge with the naked eye is strongly desired.
- Substances that serve as indicators of the severity of atopic dermatitis include peripheral blood eosinophil count, serum total IgE level, LDH (lactate dehydrogenase) level, and serum Thymus and Activation-Regulated Chemokine (TARC) ( Sugawara et al., Allergy (2002) 57: 180-181: Non-Patent Document 4).
- TARC Thymus and Activation-Regulated Chemokine
- the amount of TARC in serum correlates with the severity of atopic dermatitis and can be an indicator of systemic disease status.
- atopic dermatitis sometimes develops throughout the body, it often develops locally and often has different symptom levels at each onset site.
- TARC in the stratum corneum (hereinafter referred to as “corneal layer TARC”) is insufficient as an index for objectively judging the local symptom. This is because there is no significant difference between the measured value of the stratum corneum TARC in healthy subjects and the value of the stratum corneum TARC derived from the non-rash area of patients with atopic disease (Morita et al., Allergy ( 2010) 65: 1166-1172: Fig. 2 of Non-Patent Document 5. In addition, since stratum corneum TARC is not significantly different between patients with moderate severity and severe patients (Fig. 3 of the non-patent document 4), the correlation with the severity is also low.
- TARC is used as an index of the severity of atopic disease
- it is only possible to measure TARC in serum it is not easy in terms of collecting serum samples, and it cannot be used for local diagnosis. It is not an effective indicator.
- invasive sampling such as blood collection is difficult for both patients and doctors, and it is desirable that it can be avoided.
- Squamous cell carcinoma-associated antigen is an antigen extracted from squamous cell carcinoma cells and shows high blood levels in squamous cell carcinomas of the cervix, lungs, esophagus, and skin. It is often used (H. Kato et al. Cancer 40: 1621-1628 (1977) Non-Patent Document 6; N. Mino et al. Cancer 62: 730-734 (1988): Non-Patent Document 7).
- the SCCA blood level correlates well with the progression stage, malignancy, tumor size, etc. of squamous cell carcinoma, so that not only early detection of cancer but also evaluation of cancer treatment effects and diagnosis of recurrence It is a particularly effective cancer marker.
- SCCA is also known to have increased expression in the upper layer of the psoriatic epidermis (Takeda A. et al., J. Invest. Dermatol. (2002) 118 (1), 147-154; Non-Patent Document 8) .
- Psoriasis is a skin disease, and there is psoriasis, which is a chronic, recurrent inflammatory keratosis characterized by abnormal proliferation and differentiation of epidermal cells and inflammatory cell infiltration. Psoriasis is thought to develop by adding various environmental factors to a genetic predisposition (Hopso-Havu et al. British Journal of Dermatology (1983) 109, 77-85: Non-patent document 9).
- Non-patent Document 10 discloses that SCCA in serum correlates with the severity of atopic dermatitis.
- the protein in serum and the same protein in stratum corneum are not always in a correlation.
- Epidermal keratinocytes proliferate in the basal layer, migrate to the upper layer, differentiate, become the stratum corneum, and finally peel off from the skin.
- This stratum corneum is keratinized by normal keratinization of the skin. It is generated through a characteristic state such as the phenomenon of “denucleation” in which the nucleus disappears.
- atopic dermatitis is a disease in which a plurality of rashes having different symptom degrees are mixed, and it is clear from the fact that doctor's findings are composed of a plurality of items.
- Patent Document 1 describes the prediction of keratinization caused by atopic dermatitis caused by SCCA of skin horny layer cells (hereinafter sometimes referred to as horny layer SCCA). It is disclosed that it becomes a parameter
- This publication uses the stratum corneum SCCA as an index and shows the knowledge that subjects who have not yet suffered from atopic disease can predict the risk of suffering from the disease in the future. Therefore, it is clear that stratum corneum SCCA is involved in the development of atopic dermatitis.
- stratum corneum SCCA is a causative factor for the onset of atopic dermatitis, it is not always an indicator of the severity after the onset of the disease.
- ⁇ -fetoprotein which is a tumor marker, is a diagnostic marker, but its value does not match the size of the tumor, and prognosis cannot be determined by the degree of increase in the numerical value.
- atopic dermatitis is a disease that repeats exacerbations and remissions, is not easy to cure completely, and it is important to control its symptoms during the course of treatment.
- a marker that can be an index of local symptom and therefore an objective index in explaining a treatment policy to a patient is desired by both doctors and patients, but does not exist.
- the present inventor is a representative item suffering from atopic dermatitis, and the expression level of SCCA-1 in the stratum corneum of the subject undergoing treatment for the disease is an index of atopic dermatitis
- the survey was conducted.
- the value of stratum corneum SCCA-1 and the severity of the disease due to skin findings were highly correlated (FIG. 1).
- the skin findings also increased from remission to mild, moderate, and severe.
- even in the subject who self-determined that the disease was apparently completely cured by the patient's own findings there are many cases in which the doctor's findings have symptoms.
- SCCA-1 expression is higher than normal. For example, when it is determined that the patient is in remission, skin care with moisturizers, protective agents, etc. is performed, and the disease recurrence is caused by leaving it unattended.
- SCCA-1 in skin horny layer cells as an index, subjects undergoing treatment for atopic dermatitis, subjects who have received treatment in the past, and subjects who are currently in remission, It becomes possible to grasp the local pathological condition in a subject who looks healthy but has a possibility of developing atopy.
- the present application provides the following inventions.
- a diagnostic assistance method for determining the treatment policy in a subject undergoing treatment for atopic dermatitis wherein the expression value of SCCA-1 in the stratum corneum cells of the subject is measured.
- the expression level of the SCCA-1 is not statistically significantly higher than that of those without atopic dermatitis, it is determined that the treatment is discontinued and the disease does not need to be continued.
- a diagnostic assistance method for determining the treatment policy in a subject undergoing treatment for atopic dermatitis wherein the expression value of SCCA-1 in the skin horny layer cells of the subject is measured, ii) Classifying the severity of the subject's atopic dermatitis using the expression level of the SCCA-1 as an index, and deciding to apply the atopic dermatitis treatment method suitable for each class to the subject , A diagnostic assistance method to determine the treatment policy for atopic dermatitis.
- the diagnostic assistance method according to any one of (1) to (3) wherein the determination of the treatment policy is repeated over a long period of time.
- the symptoms of atopic dermatitis include erythema, oozing, papule, excoriation, lichenification, dryness / xerosis and so on.
- the method of assisting diagnosis according to any one of (1) to (4), wherein the method is one or more selected from the group consisting of itch.
- the diagnostic support method according to any one of (1) to (6), wherein the expression of SCCA-1 is performed by an immunological detection method using an antibody specific for SCCA-1.
- the immunological detection method includes enzyme immunoassay (including ELISA and EIA), fluorescence immunoassay, radioimmunoassay (RIA), luminescence immunoassay, surface plasmon resonance (SPR method) , Quartz crystal microbalance (QCM method), immunoturbidimetric method, latex agglutination immunoassay, latex turbidimetry, particle agglutination method, gold colloid method, capillary electrophoresis, graphene biosensor, amperometric biosensor (7)
- the diagnosis assisting method selected from the group consisting of laser spectroscopy and surface enriched immunoassay.
- the value of SCCA-1 in the stratum corneum makes it possible to non-invasively grasp the local pathology of atopic dermatitis. Furthermore, it is an objective index for determining an effective treatment policy for a subject suffering from atopic dermatitis and receiving treatment for the disease. Thus, the conviction necessary for deciding the appropriate treatment method according to the severity of the disease or for deciding the treatment policy for continuing remission as much as possible, and explaining the future treatment policy to the patient. Some objective indicators were provided. As a result, it has become possible to determine the treatment policy, such as whether or not to continue the steroid topical drug, and whether or not to continue the topical drug with anti-inflammatory effects other than steroids such as tacrolimus.
- the patient can self-assess himself without seeking a doctor's diagnosis. Therefore, it is possible to objectively determine whether or not the patient is in need of treatment even at home without going to the hospital. Or it becomes possible to grasp
- the difference in treatment due to the difference in doctor skills is reduced. Furthermore, since the method of the present invention uses SCCA-1 in the stratum corneum as an index, the method of the present invention does not require blood collection and is noninvasive. Considering that infants account for about 89% of the incidence of atopy, it is extremely effective in that there is no pain in blood collection for patients and there are no technical difficulties in medical practice.
- SCORAD is frequently used to classify the severity of atopic dermatitis by taking stratum corneum samples at multiple locations, for example, 3 or more locations per subject, and using the average SCCA-1 value as an index. It is possible to provide a simple and reliable method for classifying systemic symptoms of atopic dermatitis in place of EISA and EISA.
- stratum corneum SCCA-1 is highly correlated with the severity of atopic dermatitis (skin topical).
- a bar graph (skin local) showing that the value of stratum corneum SCCA-1 is highly correlated with the severity of atopic dermatitis.
- the bar graph which shows that the value of stratum corneum SCCA-1 has high correlation with each symptom degree according to the skin finding item of atopic dermatitis.
- the figure which shows that the value of stratum corneum TARC has a low correlation with the severity of atopic dermatitis (skin local).
- the figure which shows the discrepancy between the local skin finding and the self-finding of the severity of atopic dermatitis The figure which shows the tracking of the treatment course of the patient who suffers from atopic dermatitis which used SCCA-1 and the skin findings by a doctor as an index. The figure which shows the tracking of the treatment course of the patient who suffers from atopic dermatitis which used SCCA-1 and the skin findings by a doctor as an index. The figure which shows the tracking of the treatment course of the patient who suffers from atopic dermatitis which used SCCA-1 and the skin findings by a doctor as an index.
- SCCA-1 SCCA is encoded by two genes, SCCA-1 and SCCA-2, arranged in tandem on chromosome 18q21.3.
- the proteins encoded by them, SCCA-1 and SCCA-2, are both proteins with a molecular weight of about 45,000, exhibiting high homology, and the homology is 95% at the nucleic acid level.
- SCCA belong to the ovalbumin-serine protease inhibitor (ov-serpin) family.
- ov-serpin has unique features in the Serpin Super family. In general, serpin is secreted and works outside the cell, but ov-serpin is a protease inhibitor that works mainly inside the cell.
- SCCA-1 is a papain-like cysteine protease inhibitor, but SCCA-2 is a chymotrypsin-like serine protease inhibitor, and although it has high homology, the amino acid sequence of the reaction site is different, so it may have different characteristics. (Schick et al. J. Biol. Chem. (1997) 27213, 1849-55). The present inventor has found that SCCA-1 is an index for evaluating the severity of atopic dermatitis, particularly by noninvasive means.
- Atopic dermatitis The definitions and explanations of the following terms are based on the guidelines for clinical treatment of atopic dermatitis by the Japanese Dermatological Association in principle.
- ⁇ Atopic dermatitis> It is a disease mainly caused by itchy eczema that repeats exacerbations and remissions, and many patients have atopic predisposition (with different degrees of rash within the same person).
- ⁇ Diagnostic criteria for atopic dermatitis> Generally, atopic dermatitis is judged to satisfy the basic three items of pruritus, characteristic rash and distribution, and chronic / repetitive course (more than 2 months for infants and more than 6 months for infants).
- ⁇ Symptom of atopic dermatitis rash and selection of external preparation According to the Japanese Dermatological Association Guidelines for Atopic Dermatitis, the severity of atopic dermatitis rash can be classified into remission, minor, mild, moderate, and severe. The following shows the condition of each symptom and the treatment method to be selected. 1) Severity and severity of rash: Erythema with severe swelling, edema, infiltration or lichenification, frequent papules, high scale, crust attachment, small blisters, erosions, numerous scratches, rash nodules, etc. Mainly. ⁇ Selection of topical preparation: Belly strong or strong class steroid topical preparation with the necessary and sufficient effect is the first choice.
- Remission Regardless of whether it is permanent or temporary, it refers to a clinically controlled condition in which the symptoms of the disease have improved or almost disappeared. In other words, even if it is not completely cured in a general sense, the condition improves clinically to a “no problem” level. After that, it also includes the possibility of worsening. (It is different from healing and complete cure.) -Selection of external preparations: Select external preparations and humectants that do not contain steroids.
- -Eruption Lesions appearing on the skin are collectively referred to as skin rash or rash.
- Dermatitis Inflammation in the skin (occupies 1 / 3-1 / 5 of dermatology outpatients). The inflammatory response that occurs in the skin is expressed as eczema. Clinically, itching, redness, desquamation and papules occur.
- Eczema Synonymous with dermatitis.
- Erythema Red spots caused by vasodilation and hyperemia in the dermal papilla and lower papillae.
- ⁇ Papule A localized bulge change (skin bulge) with a diameter of 1 cm or less.
- -Nodule A nodule is a localized skin change similar to papules, with a diameter of 1 cm or more and about 2 to 3 cm. It extends deeper than the papules to the dermis or subcutaneous tissue.
- ⁇ Lichen Permanent papule.
- Infiltration (oozing) A state in which white blood cells or lymphocytes have entered the skin at the lesion site where dermatitis has occurred. It may also refer to a situation where the lesion gradually invades the surrounding normal tissue.
- Crust, crusting A solidified keratin, exudate, blood, pus or necrotic tissue. This is the so-called scab.
- ⁇ Pruritus Itching.
- the absolute value of the expression level of the stratum corneum SCCA-1 will vary depending on the measurement system of the stratum corneum SCCA-1. Therefore, in order for the test subject to be assigned to an appropriate symptom according to the horny layer SCCA-1 expression value, the horny layer SCCA-1 expression value and the symptom It is necessary to create a calibration curve that shows the relationship.
- skin findings are obtained in advance by a doctor's examination. There are various findings such as dry symptoms, erythema, scales, papules, scratch marks, swelling, edema, clogging, small blisters, erosion, and nodules.
- the evaluation criteria for each finding are, for example, 0: none, 1: mild, 2: moderate, and 3: severe.
- the number of subjects for creating a calibration curve is preferably as large as possible, and may be, for example, 50 or more, but is not limited. Then, the score of each of these items is summed up, for example, from 0 points (that is, when all items are evaluation criteria 0) to the highest point (that is, when all items are evaluation criteria 3) with the horizontal axis, the stratum corneum SCCA A calibration latitude line can be created with the measured value of -1 as the vertical axis.
- the horizontal axis is appropriately classified, for example, from the one with a small score, it can be assigned to the severity of normal (normal), remission, mild, moderate, severe, etc.
- SCCA-1 value When the subject's SCCA-1 value is obtained, it is applied to the calibration curve, and it is understood that the severity of the affected area of the subject corresponds to any severity level.
- the treatment policy suitable for each symptom can be determined according to, for example, the guideline for atopic dermatitis.
- the first choice should be steroid strong, belly strong or strong class steroids be able to.
- external preparations and humectants that do not contain steroids can be selected according to the same guidelines.
- Atopic dermatitis has a so-called “remission” state that is judged to be disease-free by appearance.
- “remission” even if eczema is once improved by, for example, an anti-inflammatory topical drug, in the histological image, inflammatory cell infiltration by lymphocytes around the blood vessel at a low level, swelling of endothelial cells, mild lesion with basement membrane thickening (Fiset PO, Leung DY. JACI; 118: 287-290, 2006) (Non-patent Document 2). Judgment of pathophysiology in remission is extremely difficult by appearance, so traditionally, the treatment method was switched to skin care using moisturizers etc.
- the state of remission can be evaluated in more detail, which is extremely advantageous.
- the value of the stratum corneum SCCA-1 of a plurality of healthy persons not suffering from atopic dermatitis is measured.
- the number of healthy persons to be measured is not particularly limited, but the larger the number, the more preferably, for example, 10 or more, more preferably 20 or more, and most preferably 50 or more, but is not limited thereto.
- the value of the horny layer SCCA-1 of a healthy person is known, and when the value of the horny layer SCCA-1 of the subject is statistically significantly higher than this value, the atopic dermatitis significantly relapses. It turns out that it is in an easy state.
- the present invention periodically performs repetitive application of weak anti-inflammatory agents and treatments with skin care such as moisturizers and protective agents in remission, and switches to only skin care with moisturizers and protective agents when treatment is unnecessary. It can be used for proactive therapy (prophylactic intermittent application method).
- the expression value of stratum corneum SCCA-1 is measured using a kit for measuring stratum corneum SCCA-1 in which the above calibration curve has been prepared in advance, such as a commercially available kit.
- a kit for measuring stratum corneum SCCA-1 in which the above calibration curve has been prepared in advance such as a commercially available kit.
- the creation of a calibration curve can be omitted, and by measuring the expression value of the subject's stratum corneum SCCA-1 with the measurement system, it is determined which symptom is appropriate, and A suitable treatment strategy can be established.
- the measurement of the expression of SCCA-1 according to the present invention can be carried out quantitatively or qualitatively according to any method capable of measuring SCCA-1.
- an immunoassay method using an antibody specific for SCCA-1 for example, an ELISA method using an enzyme label, an RIA method using a radioactive label, an immunoturbidimetric method, a Western blot method, a latex agglutination method, Various methods such as hemagglutination method, immunochromatography method, resonance plasmon resonance method, SAW (Surface Acoustic Wave) device, electronic device method and the like can be mentioned.
- immunoassay methods include competitive methods and sandwich methods.
- the expression level of SCCA-1 can also be determined by measuring the amount of the gene encoding it expressed in the cell.
- the expression of SCCA-1 is determined by measuring the amount of mRNA encoding SCCA-1 in the cell. Extraction of mRNA and quantitative or qualitative measurement of the amount thereof are also well known in the art, and can be performed by various well-known methods such as PCR, 3SR, NASBA, and TMA.
- SCCA-1 expression is qualitatively determined through in situ hybridization and measurement of its biological activity.
- the sample of the skin stratum corneum serving as the subject can be collected by any method, but the tape stripping method is preferable from the viewpoint of simplicity.
- Tape stripping is a method of collecting a stratum corneum sample by applying a piece of adhesive tape to the skin surface layer, peeling it off, and attaching the skin stratum corneum to the peeled adhesive tape. If the tape stripping method is used, it is possible to measure the expression of SCCA-1 just by collecting a single layer of the stratum corneum, and it becomes possible to grasp the pathological condition of atopic dermatitis using SCCA-1 as an index.
- the preferred method of tape stripping is to first remove the sebum, dirt, etc.
- the adhesive tape may be a commercially available cellophane tape or the like, for example, Scotch Superstrength Mailing Tape (3M), cellophane tape (cello tape (registered trademark); Nichiban Co., Ltd.) or the like.
- a suitable extract such as Tris-buffer (pH 8.0) (0.1M Tris-HCl, 0.14M NaCl, 0.1% Tween-20). It can be isolated and extracted from the tape by dipping and extracting the stratum corneum.
- Tris-buffer pH 8.0
- SCCA-1 is measured by an immunoassay method such as ELISA.
- the antibody specific for SCCA-1 used in the ELISA may be a monoclonal antibody or a polyclonal antibody.
- Methods for producing monoclonal antibodies and polyclonal antibodies are well known to those skilled in the art.For example, Lunstrum et el., J Biol. Chem. 1986, 261: 9042-9048; Hurle et al. J Cell Science 1994, 107: 2623-2634 Are listed.
- the sandwich immunoassay method is particularly preferable.
- the sandwich immunoassay method can be performed, for example, as follows.
- One of the two types of SCCA-1 specific antibodies is immobilized on a carrier as a primary antibody.
- a carrier a solid carrier is preferable.
- any solid carrier commonly used in immunoassays may be used.
- a polymer carrier such as styrene or polystyrene formed into an arbitrary size and shape
- Examples include reaction vessels formed of these appropriate materials, such as the inner walls of wells of ELISA plates.
- the primary antibody can be immobilized on a carrier according to a conventional method.
- the primary antibody is dissolved in a buffer solution such as phosphate buffered saline (PBS) or borate buffer, and adsorbed on the carrier.
- PBS phosphate buffered saline
- an antibody that binds to the primary antibody or another protein, such as protein C may be immobilized on a carrier in advance and then contacted with the primary antibody.
- an appropriate blocking agent such as PBS-BSA or a commercially available blocking agent such as Block Ace (Dainippon Pharmaceutical) is added to the carrier on which the primary antibody is immobilized in this manner.
- Blocking is preferably performed by incubation at 4 to 40 ° C., preferably 20 to 37 ° C., for 5 minutes to several days, preferably 10 minutes to 24 hours, more preferably 10 minutes to 3 hours.
- the other antibody specific to the above two types of SCCA-1 is used as a secondary antibody and labeled.
- the label include an enzyme label, a radioisotope label, and a fluorescent label.
- the enzyme can be directly bound to the secondary antibody for labeling, or indirectly labeled with an enzyme via an interactive protein such as avidin-biotin.
- the enzyme can be bound to the antibody or the like by, for example, introducing a thiol group into each of the enzyme and the antibody to be labeled using a commercially available thiol-introducing group reagent and then bonding them to each other through an SS bond. .
- the enzyme examples include horseradish peroxidase, alkaline phosphatase, ⁇ -D-galactosidase and the like.
- the enzyme can be detected using a substrate specific for the enzyme.
- TMB 3,3 ′, 5,5′-tetramethylbenzidine
- ABTS 2,2′-azine-di [3-ethylbenzthiazolinesulfonate]
- Such immunoassay is performed by mixing the carrier immobilized with the primary antibody, the labeled secondary antibody, and the test sample, and incubating the SCCA-1 in the test sample to the primary antibody immobilized on the carrier. Then, a labeled secondary antibody is bound to the SCCA-1 molecule.
- the labeled antibody is immobilized on the carrier through the primary antibody immobilized on the carrier and the SCCA-1 derived from the sample in an amount reflecting the amount of SCCA-1 in the sample.
- a suitable buffer eg, PBS, at about 4-40 ° C., preferably 20-37 ° C., for 5 minutes to several days, preferably 10 minutes to 24 hours, more preferably 10 minutes to 3 hours.
- an operation of separating unbound labeled antibody from the carrier is performed.
- the carrier is a solid carrier
- this separation operation can be easily performed by solid-liquid separation.
- the label bound to the carrier and / or unbound label can be measured.
- an arbitrary labeled antibody is used, the label bound to the carrier is detected and measured.
- detection is preferably performed after the carrier is washed with a washing solution, for example, a buffer containing an appropriate surfactant, for example, PBS-Tween 20, to remove unbound labeled antibody. .
- Detection can be performed according to a conventional method depending on the type of label.
- the present invention will be described more specifically with specific examples. In addition, this invention is not limited by this.
- Evaluation and observation items (1) Physician's local findings at each site (A) erythema, (B) oozing, (C) papule, (D) excoriation, (E) Lichenification, (F) Dry / xerosis, (G) Pruritus (itch) were observed, and the severity was entered on the case card.
- the evaluation criteria were 0: none, 1: mild, 2: moderate, 3: severe. A healthy site, a rash site, and a rash site from which the stratum corneum was collected are shown in the whole body diagram.
- stratum corneum tape Collection and measurement of stratum corneum tape
- the stratum corneum at the test site was collected using cello tape (registered trademark) (2 cm x 6 cm).
- the obtained stratum corneum sample was immersed in a protein extract (0.1 M Tris-HCl (pH 8.0), 0.14 M NaCl, 0.1% Tween 20) and extracted with a bioraptor. Centrifugation was performed and the supernatant was used for ELISA.
- Monoclonal anti-SCCA1 antibody (Santa Cruz) was used as a secondary antibody and reacted at 37 ° C for 1 h. Then, horseradish peroxidase-conjugated anti-mouse F (ab ′) 2 (GE Healthcare) was added, and color was developed with TMB peroxidase EIA substrate kit (Bio-Rad). The reaction was stopped with 1 M H2SO4 and measured at 450 nm.
- Exclusion criteria for atopic dermatitis are 1) those who have received oral treatment including steroids or immunosuppressants for 1 month or more before the start of the study, 2) those for external use of steroids at the test site for 1 week or more before the start of the study, Those who have been treated with tacrolimus hydrate ointment, non-steroidal anti-inflammatory agents, and other external preparations (including over-the-counter drugs). Or those who are unable to use the drug on the test site for at least one week before the start of the study. However, moisturizers can be applied. 3) Those who have skin diseases other than atopic dermatitis at the test site, 4) Those who are pregnant or breastfeeding, 5) Investigator / student-sharing physician / study cooperation Doctors were deemed to be inappropriate for participation in the study.
- SCCA-1 values were measured for healthy individuals and patients with atopic dermatitis.
- the skin findings at the test site were (A) erythema, (B) oozing, (C) papule, (D) excoriation, ( E) Lichenification, (F) Dryness / xerosis, (G) Pruritus (itch) were evaluated according to the evaluation criteria described in the column ⁇ Skin local findings ''.
- the severity of the test site was determined from the total value of each item of skin findings.
- a graph in which the total value of each subject is plotted on the horizontal axis and the SCCA-1 value on the vertical axis is shown in FIG.
- Figure 1 shows the relationship between skin severity and stratum corneum SCCA-1 value in atopic dermatitis, and its Spearman correlation coefficient was as high as 0.753 (p ⁇ 0.001). SCCA1 showed a significantly strong correlation.
- the total score of skin findings for patients with atopic dermatitis is classified into 0, 1-7, 8-14, 15-21, and the average value of SCCA-1 in healthy individuals is calculated.
- the average of SCCA-1 values for each class was also calculated.
- a patient whose total value is 0 is a patient who suffered from atopic dermatitis, but none of the findings items were observed at the corresponding site.
- the average value of SCCA-1 was 19.1 ng / mg for healthy subjects, 86.3 ng / mg for skin findings total 0 (remission), 415.7 ng / mg for skin findings 1 to 7 (mild), skin
- the total findings (moderate) 8-14 were 1380.5 ng / mg, and the total skin findings 15-21 (severe) were 1930.9 ng / mg.
- Fig. 2 shows the average value of the stratum corneum SCCA-1 values of each class as a bar graph. It can be seen that the higher the severity according to the doctor's skin findings, the higher the value of stratum corneum SCCA-1. Therefore, by using SCCA-1 as an index, the degree of symptom of the patient can be objectively determined, which can be useful for determining a treatment policy.
- the mean value of SCCA-1 in the group of patients with a total doctor's skin findings of 0 was statistically significantly higher than the mean value of healthy individuals. That is, there are cases where the pathological condition can be evaluated by using SCCA-1 as an index even in a place where there is no apparent symptom. Therefore, by using SCCA-1 as an index, even a doctor may be able to grasp a pathological condition that is difficult to judge. As a result, it may be possible to prevent the patient from stopping treatment and control symptoms.
- Figures 3-1 and 2 show the average value of SCCA-1 values of the patient group according to the score of each item of the above skin findings (A) to (G).
- the horizontal axis represents the score of each item, and the vertical axis represents the SCCA. It is the average of -1 and is shown in a bar graph.
- the value of SCCA-1 was highly correlated with the severity of each finding. Especially for papules and dry / desquamation, there was a significant difference (p ⁇ 0.001) in the SCCA-1 level between the group of patients with a finding of 0 and healthy individuals. Therefore, by using SCCA-1 as an index, it is possible to objectively determine the degree of local skin symptom of the patient in all the general findings of symptoms of atopic dermatitis, and the treatment suitable for each finding It can be used to make policy decisions.
- TARC measurement calibration curve As with SCCA-1, the value of corneum TARC and its correlation with skin findings were examined.
- the stratum corneum TARC was measured by subjecting the obtained stratum corneum extract to ELISA. Quantikine ELISA Human CCL17 / TARC (R & D Systems) was used for ELISA. The result is shown in FIG.
- the Spearman correlation coefficient between skin severity and stratum corneum TARC1 in atopic dermatitis is as low as 0.2750, indicating that stratum corneum TARC cannot be an indicator of the severity of atopic dermatitis.
- the evaluation criteria were 0: none, 1: mild, 2: moderate, 3: severe.
- a healthy site, a rash site, and a rash site from which the stratum corneum was collected are shown in the whole body diagram. The result is shown in FIG. The self-finding of the two subjects was clearly lower than that of the doctor.
- atopic skin diseases patients tend to underestimate their own findings, especially when the severity is mild.
- atopic dermatitis is a disease in which symptom relapse is likely to occur by discontinuing or discontinuing treatment even though treatment is necessary.
- the stratum corneum SCCA-1 can be collected by a very simple and noninvasive operation such as tape stripping.
- SCCA-1 measurement can be done at home instead of hospitals and laboratories by developing a simple measurement kit, so it is an excellent index according to the evaluation of the severity of atopic skin disease and the current treatment effect It becomes.
- SCCA-1 measurement can be done at home instead of hospitals and laboratories by developing a simple measurement kit, so it is an excellent index according to the evaluation of the severity of atopic skin disease and the current treatment effect It becomes.
- there are many infants with atopic skin disease and it is extremely advantageous to be able to determine the symptom of their own disease at home simply for subjects who cannot complain of symptoms themselves and who cannot easily go to hospital.
- Patient TA04-1 Test site 1 of patient TA04 had a total of 10 skin findings and was in a moderate category. During the treatment observation period, observations and horny layer collection were performed at each visit. The internal drug used for treatment, its administration period, the name of the external preparation for administration and the administration test site were confirmed.
- the SCCA-1 value at the start of treatment was 6510 ng / mg, a fairly high value.
- the patient was given Allelock (registered trademark) OD, which is an antihistamine, and 100 mg of Flomox (registered trademark) tablets.
- the SCCA-1 value measured 28 days after the start of treatment was 415.9 ng / mg, which was significantly lower than that at the start of treatment, and the result was linked to the skin findings (total value 1). The result is shown in FIG.
- Patient TB03 had a skin appearance of 4 and was in a mild category. During the treatment observation period, observations and horny layer collection were performed at each visit. The internal drug used for treatment, its administration period, the name of the external preparation for administration and the administration test site were confirmed. The value of SCCA-1 at the start of treatment is 392.5 ng / mg, which is higher than that of healthy people. In this patient, the topical steroid Antepate (registered trademark) ointment was used continuously. 42 days after the start of treatment, the SCCA-1 value was 112.6 ng // mg, which was significantly lower than that at the start of treatment, and the result was linked to the skin findings (total value 0). The result is shown in FIG.
- Patient TB05 The 21-year-old male patient TB05 had a total of 5 skin findings and was in a mild category. During the treatment observation period, observations and horny layer collection were performed at each visit. The internal drug used for treatment, its administration period, the name of the external preparation for administration and the administration test site were confirmed. The SCCA-1 value at the start of treatment was 560.9 ng / mg. The patient was given 120 mg of Allegra (registered trademark), which is an antihistamine, and continued to use Mizer (registered trademark) cream. The SCCA-1 value measured 50 days after the start of treatment was 64.7 ng / mg, which was lower than that at the start of the treatment, and the results were linked to extremely good skin findings (total value 0).
- Allegra registered trademark
- Mizer registered trademark
- the SCCA-1 value measured 72 days and 113 days after the start of treatment, although the cause was unknown, increased to 125.0 ng / mg and 176.6 ng / mg, respectively, and the skin findings were also compared to the total value of 0 after 50 days of treatment. The total increased to 1, and the SCCA-1 level and skin findings were linked. The result is shown in FIG.
- SCCA-1 is linked to the severity of atopic skin disease by the doctor.
- the stratum corneum SCCA-1 can be collected by a very simple and noninvasive operation such as tape stripping.
- SCCA-1 measurement can be done at home instead of hospitals and laboratories by developing a simple measurement kit, so it is an excellent index according to the evaluation of the severity of atopic skin disease and the current treatment effect It becomes.
- SCCA-1 as an indicator, there are many infants, especially those with atopic skin disease, who are unable to complain of symptoms themselves, and who are not easy to go to the hospital without attendance. It is extremely advantageous to be able to objectively determine the symptom of the disease and to objectively evaluate the therapeutic effect.
- Patient AW1-1 Test site No. 1 of patient AW1 is the site diagnosed by the doctor as being in remission. At the start of observation, there was no skin symptom, and the skin findings (total value) by the doctor were zero. During the observation period, observations and stratum corneum collection were performed at each visit. The patient has not been treated for atopic skin disease during the observation period. The SCCA-1 value at the start of treatment was about 302.4 ng / mg, which was significantly higher than that of healthy subjects (19.1 ng / mg), which was not negligible. The skin findings by the doctors were 0 until about 15 days after the start of observation, but the skin findings increased to 1 as the SCCA- value increased and began to worsen. The value of self-finding by the patient did not change during the observation period and remained at 0. The observation result is shown in FIG.
- Patient AW1-2 Test site No2 of patient AW1 is a site diagnosed as mild by a doctor at the start of observation. At the start of observation, there was no skin symptom, and the doctor's skin findings (total value) were 0, but the doctor's findings increased with time. Although the self-assessment slightly increased (increased from 0 to 1), the value of the self-examination was lower than that of the doctor. The patient has not been treated for atopic skin disease during the observation period. The SCCA-1 value at the start of treatment was approximately 227.0 ng / mg. The skin findings by the doctors were also 0 until about 40 days after the start of observation. However, as the SCCA- value increased, the skin findings also increased to 1 and began to deteriorate. The value of the accident findings by the patient remained unchanged during the observation period. The observation result is shown in FIG.
- Patient AW1-5 Test site No. 5 of patient AW1 is also a site diagnosed by the doctor as being in remission.
- the skin findings total value
- the SCCA-1 value at the start of treatment was approximately 313.4 ng / mg.
- the doctor's skin findings were also 0 until about 30 days after the start of observation, but with the increase in SCCA- values, the skin findings also increased to 1 and began to deteriorate.
- the value of the accident findings by the patient remained unchanged during the observation period.
- the observation result is shown in FIG.
- Patient AW3-7 Test site No. 7 of patient AW3 showed no skin symptoms at the start of observation.
- the SCCA-1 value at the start of the treatment was 50.7 ng / mg, which was slightly lower but significantly higher than that of healthy individuals (19.1 ng / mg), and was not negligible.
- the SCCA-1 value was 250.6 ng / mg, which was significantly increased compared to the start of the observation, and the result was linked to the doctor's skin findings (total value 7).
- the increase in self-examination was slight throughout the observation period (total value 1). The result is shown in FIG.
- the calculation of the SCORAD value has the disadvantage that it requires specialized work by a skilled doctor and takes time. Therefore, by using the stratum corneum SCCA-1 value as an index, a simpler and objective method for classifying the severity can be provided in place of SCORAD.
- the present invention provides a diagnostic assistance method for simple and objective evaluation of the treatment policy in a subject undergoing treatment for atopic dermatitis by using SCCA-1 in the stratum corneum as an index. Enable.
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Abstract
Description
以上のように、皮膚角層細胞中のSCCA-1を指標とすることで、アトピー性皮膚炎の治療を受療中の対象者、過去に受療し、現在は寛解期にある対象者、更には健常と見えるがアトピーの発症の可能性を持っている対象者などにおいて局所病態を把握することが可能となる。
(1)アトピー性皮膚炎の治療を受療中の対象者において当該治療方針を決定するための診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
i)当該SCCA-1の発現値がアトピー性皮膚炎を有しない者の値に比べ、統計学的に有意に高くない場合、当該治療を中断し当該疾患は治療を継続する必要はないと決定する、診断補助方法。
(2)アトピー性皮膚炎の治療を受療中の対象者において当該治療方針を決定する診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
ii)当該SCCA-1の発現値を指標として当該対象者のアトピー性皮膚炎の症度を階級分けし、各階級に合ったアトピー性皮膚炎治療方法を当該対象者に適用することを決定する、アトピー性皮膚炎の治療方針を決定する診断補助方法。
(3)アトピー性皮膚炎の治療の要否判断が必要な対象者あるいは治療を受療中の対象者において、当該治療の要否判断あるいは当該治療を必要と判断した場合の治療方針を決定する診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
i)当該SCCA-1の発現値がアトピー性皮膚炎を有しない者の値に比べ、統計学的に有意に高くない場合、当該治療は不要と判断するあるいは受療中の場合は当該治療により当該疾患は治療を継続する必要がないと決定し、統計学的に有意に高い場合、治療を開始する判断あるいは当該治療を継続する必要があると決定した場合、さらに、
ii)当該SCCA-1の発現値を指標として当該対象者のアトピー性皮膚炎の症度を階級分けし、各階級に合ったアトピー性皮膚炎治療方法を当該対象者に適用することを決定する、アトピー性皮膚炎の治療の要否判断あるいは治療方針を決定する診断補助方法。
(4)前記治療方針の決定を長期にわたり繰り返し行う、(1)~(3)のいずれかの診断補助方法。
(5)アトピー性皮膚炎の症度の項目が、紅斑(erythema)、浸潤(oozing)、丘疹(papule)、掻破痕(excoriation)、苔癬化(lichenification)、乾燥・落屑(xerosis)及びそう痒感(itch)から成る群から選ばれる1または複数である、(1)~(4)のいずれかの診断補助方法。
(6)前記皮膚細胞がテープストリッピングにより非侵襲的に採取される、(1)~(5)のいずれかの診断補助方法。
(7)前記SCCA-1の発現をSCCA-1に特異的な抗体を使用する免疫学的検出方法により実施する、(1)~(6)のいずれかの診断補助方法。
(8)前記免疫学的検出方法が酵素免疫測定法(ELISA法、EIA法を含む)、蛍光免疫測定法、放射免疫測定法(RIA)、発光免疫測定法、表面プラズモン共鳴法(SPR法)、水晶振動子マイクロバランス(QCM法)、免疫比濁法、ラテックス凝集免疫測定法、ラテックス比濁法、粒子凝集反応法、金コロイド法、キャピラリー電気泳動法、グラフェンバイオセンサ、アンペロメトリックバイオセンサ、レーザー分光法及び表面濃縮免疫測定法から成る群から選ばれる、(7)の診断補助方法。
(9)前記皮膚細胞の採取及び前記SCCA-1の発現の測定が在宅で行われる、(7)又は(8)の診断補助方法。
(10)前記テープストリッピングによる採取を、対象者当たり3箇所以上の皮膚部位にて行う、(1)~(9)のいずれかの診断補助方法。
SCCAは染色体18q21.3上にタンデムに並んでいる二つの遺伝子SCCA-1及びSCCA-2遺伝子によりコードされる。それらによりコードされるタンパク質、SCCA-1及びSCCA-2は共に分子量約45,000のタンパクであり、高い相同性を示し、そのホモロジーは核酸レベルで95%である。これらのSCCAはovalbumin‐serine protease inhibitor(ov-serpin)ファミリーに属している。ov-serpinはセルピンス-パーファミリーの中でもユニークな特徴を有している。一般にセルピンは分泌されて細胞外で働くとされているが、ov-serpinは主に細胞内でも働くprotease inhibitorである。
SCCA-1はパパイン様システインプロテアーゼ阻害剤であるが、SCCA-2はキモトリプシン様セリンプロテアーゼ阻害であり、相同性が高いにも関わらず反応部位のアミノ酸配列が異なるため、異なった特性を有することもある(Schick et al.J.Biol.Chem.(1997)27213,1849-55)。本発明者は、特にSCCA-1が非侵襲的手段により、アトピー性皮膚炎の症度の評価の指標となることを見出した。
以下の用語の定義や説明は、原則として日本皮膚科学会のアトピー性皮膚炎診療ガイドラインに基づく。
<アトピー性皮膚炎>
増悪・寛解を繰り返す、そう痒のある湿疹を主病原とする疾患であり、患者の多くは、アトピー素因を持つ(同一人内に異なる症度の皮疹をもつ)。
<アトピー性皮膚炎の診断基準>
一般に、そう痒、特徴的皮疹と分布、慢性・反復的経過(乳児では2か月以上、その他6か月以上)、の基本3項目を満たすものをアトピー性皮膚炎と判断する。
日本皮膚科学会のアトピー性皮膚炎診療ガイドラインなどによれば、アトピー性皮膚炎の皮疹の症度は、寛解、軽微、軽症、中症、重症に階級分けすることができる。以下、各症度の状態及び選択すべき治療法を示す。
1)重症
・皮疹の重症度:高度の腫脹・浮腫・浸潤または苔癬化を伴う紅斑、丘疹の多発、高度の鱗屑、痂皮の付着、小水疱、びらん、多数の掻破痕、痒疹結節などを主体とする。
・外用剤の選択:必要かつ十分な効果のあるベリーストロングまたはストロングクラスのステロイド外用剤を第一選択とする。痒疹結節でベリーストロングでも十分な効果が得られない場合はその部位に限定してストロンゲストクラスの使用もある。
2)中症
・皮疹の重症度:中程度までの紅斑、鱗屑、少数の丘疹、掻破痕などを主体とする。
・外用剤の選択:ストロングまたはミディアムクラスのステロイド外用薬を第一選択とする。
3)軽症
・皮疹の重症度:乾燥および軽度の紅斑、鱗屑などを主体とする。
・外用剤の選択:ミディアム以下のステロイド外用剤を第一選択とする。
4)軽微:炎症症状に乏しい乾燥症状主体。
・外用剤の選択:ステロイドを含まない外用剤を選択する。
5)寛解:永続的であるか一時的であるかを問わず、病気による症状が好転または、ほぼ消失し、臨床的にコントロールされた状態を指す。すなわち、一般的な意味で完治せずとも、臨床的に「問題ない程度」にまで状態がよくなる。その後、増悪する可能性も含む状態。(治癒、完治とは異なる。)
・外用剤の選択:ステロイドを含まない外用剤、保湿剤などを選択する。
・皮疹(eruption):皮膚に現れる病変を総称して皮疹又は発疹という。
・皮膚炎(dermatitis):皮膚に生じている炎症のこと (皮膚科外来の1/3-1/5を占める)。皮膚で生じた炎症反応が湿疹として表現される。臨床的には、そう痒や発赤、落屑、丘疹を生じる。
・湿疹(eczema):皮膚炎と同義。
・紅斑(erythema):真皮乳頭および乳頭下層での血管拡張,充血により生じる紅色の斑のこと。
・丘疹(papule):直径1cm以下の限局性の隆起変化(皮膚の隆起)をいう。
・結節(nodule):結節は丘疹と同様の限局性の皮膚変化で直径1 cm以上のもので2~3cm程度のもの。丘疹より深く真皮あるいは皮下組織に及ぶ。
・苔癬 (lichen): 丘疹の永続したもの。
・浸潤(oozing):皮膚炎が起こった病変部位の皮膚内に、白血球細胞やリンパ球が侵入した状態。病巣が周囲に正常組織へと徐々に侵入していく状況を指す場合もある。
・痂皮(crust, crusting):角質、滲出液、血液、膿または壊死組織が凝固したもの。いわゆる、瘡蓋である。
・そう痒(pruritus):痒み。
かかる症度の客観的な指標を患者に提示することで、例えばステロイド薬に対する誤った知識をもつ患者に対しても、それを使用した治療方針が必要であることを納得させる材料となり、その患者の症度に合った有効な治療方針を適用することを可能にする。アトピー性皮膚炎は治療が長期にわたる疾患であり、医者と患者との信頼関係や患者の受療に対するモチベーションの維持が重要であることから、本指標は有効であると考える。
従前の方法では、アトピー性皮膚炎が、治療が必要かどうかを評価する客観的な指標は存在しなかったため、常に一部の熟練した医師による所見を仰いでいた。本発明によれば、寛解の状態をより詳細に評価することが可能となり、極めて有利である。そのためには、まずアトピー性皮膚炎に罹ってない複数の人数の健常人の角層SCCA-1の値を測定する。測定する健常人の数が特に限定されないが、多いほど好ましくは、例えば10人以上、より好ましくは20人以上、最も好ましくは50人以上とするが、それらに限定されるものではない。これにより、健常人の角層SCCA-1の値がわかり、この値に対し被検者の角層SCCA-1の値が統計学的に有意に高い場合、アトピー性皮膚炎が有意に再燃しやすい状態にあることがわかる。
本発明は寛解において定期的に、弱い抗炎症剤の間欠塗布と、保湿剤、保護剤などのスキンケアによる治療を行い、治療が不要な段階に至ったら保湿剤、保護剤によるスキンケアのみに切り替える、といったプロアクティブ療法(予防的な間欠塗布法)に採用することができる。
2種類のSCCA-1に特異的な抗体の一方を一次抗体として担体に固定化する。担体としては固体担体が好ましく、例えば固体担体として免疫測定法において常用される任意のものを使用してよく、例えば任意の大きさ、形状に成形されたスチレンやポリスチレンなどの高分子担体のほか、これらの適当な材料で成形した反応容器、例えばELISAプレートのウェルの内壁などが挙げられる。
以下、具体例を挙げて、本発明を更に具体的に説明する。なお、本発明はこれにより限定されるものではない。
(1)医師による皮膚局所の所見
各部位で、(A)紅斑(erythema)、(B)浸潤(oozing)、(C)丘疹(papule)、(D)掻破痕(excoriation)、(E)苔癬化(lichenification)、(F)乾燥・落屑(xerosis)、(G)そう痒感(itch)を観察し、重症度をケースカードに記入した。
評価基準は、0:なし(absence)、1:軽症(mild)、2:中症(moderate)、3:重症(severe)とした。角層を採取した健常部位、皮疹部位、無疹部位を全身図に記載した。
被験部位の角層をセロテープ(登録商標)(2 cm x 6cm)を用いて採取した。得られた角層サンプルは、タンパク質抽出液( 0.1M Tris-HCl (pH8.0), 0.14M NaCl ,0.1% Tween 20 )に浸漬し、バイオラプターにて抽出した。遠心を行い、その上清をELISAに用いた。
リコンビナントヒトSCCA-1はSCCA-1 cDNAをpQE30 ベクターに挿入して得た。Ni-Agaroseにてリコンビナントを精製した後に、Mono Q chromatographyで更に精製を行い、ELISAアッセイのスタンダードとしたSCCA-1測定方法を示す。Rabbit antiSCCA-1 polyclonal antibody(資生堂作製)を、一次抗体として96-wellプレートに固相化して用いた。ImmunoBlock(DS Pharma Biomedical, Osaka, Japan)でブロッキング後 、100 μl の角層抽出液をwellに添加し、プレートを1 h 、37oCで反応させた。Monoclonal anti-SCCA1 antibody (Santa Cruz) を二次抗体として用いて、1 h 、37oCで反応させた。その後、horseradish peroxidase-conjugated anti-mouse F(ab’)2 (GE Healthcare)を添加し、TMB peroxidase EIA substrate kit (Bio-Rad)で発色させた。 反応は1 M H2SO4で停止し、450 nmで測定した。
健常人52人およびアトピー性皮膚炎患者189人を対象として試験を実施した。健常人は、医師によりアレルギー性疾患(アトピー性皮膚炎を含む)を有さないと診断された者とした。アトピー性皮膚炎患者の適格条件は、1)過去あるいは現在、医師よりアトピー性皮膚炎と診断された者、2)試験開始前から1週間以上、被験部位に試験開始前から1週間以上、被験部位に上記薬剤を使用しないことが可能な者、3)20歳以上の者とした。
アトピー性皮膚炎の除外基準は、1)試験開始前から1ヶ月以上、ステロイドもしくは免疫抑制剤を含む内服治療を受けた者、2)試験開始前から1週間以上、被験部位にステロイド外用剤、タクロリムス水和物軟膏、非ステロイド系抗炎症剤、その他外用剤(一般用医薬品を含む)等による治療を行った者。もしくは試験開始前から1週間以上、被験部位に上記薬剤を使用しないことが不可能な者。ただし、保湿剤の塗布は可とする、3)被験部位にアトピー性皮膚炎以外の皮膚疾病を有する者、4)妊娠中、授乳中の者、5)試験責任医師・試験分担医師・試験協力医師に、試験への参加が不適当と判断された者とした。
検量線の作成
SCCA-1と同様、角層TARCについてもその値と皮膚所見と相関を調べた。角層TARCの測定は、得られた角層抽出液をELISAに供することで行った。ELISAにはQuantikine ELISA Human CCL17/TARC(R&D Systems)を用いた。
その結果を図4に示す。アトピー性皮膚炎における皮膚の重症度と角層TARC1値のSpearman相関係数は0.2750と低く、角層TARCはアトピー性皮膚炎の症度の指標にはなれないことを示す。
アトピー性皮膚炎の症度の判別は熟練した医師でさえも困難な場合があり、ましてや医療経験のない一般人が症度を判別するのは極めて困難である。本実施例では、アトピー性皮膚炎の診察について熟練した医師と、患者自身とによる皮膚所見の乖離について調べた。
アトピー性皮膚炎の症度が比較的軽い2名の被検者について、各部位で、(A)紅斑(erythema)、(B)浸潤(oozing)、(C)丘疹(papule)、(D)掻破痕(excoriation)、(E)苔癬化(lichenification)、(F)乾燥・落屑(xerosis)、(G)そう痒感(itch)を、医師及び被検者自身が各々観察し、重症度をケースカードに記入した。評価基準は、0:なし(absence)、1:軽症(mild)、2:中症(moderate)、3:重症(severe)とした。角層を採取した健常部位、皮疹部位、無疹部位を全身図に記載した。
その結果を図5に示す。2名の被検者の自己所見は共に、医師所見に比べ、明らかに低い値を示した。したがって、アトピー性皮膚疾患については、特に症度が軽い場合、患者は自己所見を低く評価しがちであることがわかる。その結果アトピー性皮膚炎は、治療が必要な状態であるにもかかわらず、治療を中止・中断することで症状の再燃が起きやすい疾患であることがわかる。上述のとおり、角層SCCA-1は例えばテープストリッピングといった極めて簡単且つ非侵襲的な操作で採取可能である。SCCA-1の測定は簡易測定キットの開発により病院や検査施設でなく、在宅でも可能となる得るため、アトピー性皮膚疾患の症度や現在施されている治療効果の評価にとおり極めて優れた指標となる。特にアトピー性皮膚疾患の患者には乳幼児が多く、自ら症状を訴えることができず、また通院も容易でない対象者にとって簡単に在宅で自己の疾患の症度を判別できることは極めて有利である。
治療経過の追跡1-1
患者TA04-1
患者TA04の被験部位1は皮膚所見の合計値が10と、症度としては中症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1値は6510ng/mgと、かなり高い値を呈していた。同患者に抗ヒスタミン剤であるアレロック(登録商標)ODを連用すると共に、フロモックス(登録商標)錠100mgを服用させた。治療開始28日後に測定したSCCA-1値は415.9 ng/mgと、治療開始時に比べ顕著に低下し、その結果は皮膚所見(合計値1)と連動していた。その結果を図6(A)に示す。
患者TB03
患者TB03は皮膚所見が4と、症度としては軽症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1の値は392.5ng/mgと、健常人に比べ高めの値を呈し。同患者にはステロイド外用剤アンテペート(登録商標)軟膏を連用した。治療開始42日後に測定板SCCA-1値は112.6ng//mgと、治療開始時に比べ顕著に低下し、その結果は皮膚所見(合計値0)と連動していた。その結果を図6(B)に示す。
患者TA03
28才女性患者TA-03は皮膚所見の合計値が7と、症度としては軽症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1値は1745.6ng//mgと、かなり高い値を呈していた。同患者には56日後からアンテベート(登録商標)軟膏を連用させた。治療開始56日後に測定したSCCA-1値は39.6 ng//mgと、治療開始時に比べ顕著に低下し、その結果は良好な皮膚所見(合計値1)と連動していた。その結果を図6(C)に示す。
患者TA04-2
27才男性患者TA04の被験部位2は皮膚所見の合計値が21と、症度としては重症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1値は9119.9ng//mgと、かなり高い値を呈していた。同患者に抗ヒスタミン剤であるアレロック(登録商標)ODを連用すると共に、フロモックス(登録商標)錠100mgを服用させた。治療開始28日後に測定したSCCA-1値は36.6 ng/mgと、治療開始時に比べ顕著に低下し、その結果は良好な皮膚所見(合計値1)と連動していた。その結果を図6(D)に示す。
患者TB03-2
55才男性患者TB03の被験部位2は皮膚所見の合計値が4と、症度としては軽症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1値は392.5ng/mgとの値を呈していた。同患者にはアンテベート(登録商標)軟膏を連用させた。治療開始42日後に測定したSCCA-1値は112.6 ng/mgと、治療開始時に比べ低下し、その結果は極めて良好な皮膚所見(合計値0)と連動していた。その結果を図6(E)に示す。
患者TB05
21才男性患者TB05は皮膚所見の合計値が5と、症度としては軽症の部類に入っていた。治療観察期間の間、来院毎に観察及び角層の採取を行った。治療に使用した投与内服薬剤及びその投与期間、投与外用剤の名称及び投与被験部位を確認した。治療開始時のSCCA-1値は560.9ng/mgとの値を呈していた。同患者に抗ヒスタミン剤であるアレグラ(登録商標)120mgを服用させると共に、マイザー(登録商標)クリームを連用させた。治療開始50日後に測定したSCCA-1値は64.7 ng/mgと、治療開始時に比べ低下し、その結果は極めて良好な皮膚所見(合計値0)と連動していた。治療開始72日後及び113日後に測定したSCCA-1値は、原因は不明であるがそれぞれ125.0 ng/mg及び176.6 ng/mgへと上昇し、皮膚所見も治療開始50日後の合計値0に比べ合計値1へと上昇し、SCCA-1値と皮膚所見は連動していた。その結果を図6(F)に示す。
経過の追跡2-1-1
患者AW1-1
患者AW1の被験部位No.1は医師により寛解期と診断された部位である。観察開始時は皮膚症状を呈しておらず、医師による皮膚所見値(合計値)は0であった。観察期間の間、来院毎に観察及び角層の採取を行った。なお、同患者は観察の期間の間アトピー性皮膚疾患の治療は受けていない。治療開始時のSCCA-1値は約302.4 ng/mgと健常人に比べ優位に高く(19.1 ng/mg)無視できない程度の値を示していた。医師による皮膚所見は、観察を開始してから15日目ぐらいまでは値は0であったが、SCCA-の値の上昇と共に、皮膚所見値が1へと上昇し、悪化し始めた。患者による自己所見の値は観察期間中変動せず、0のままであった。この観察結果を図7(A)に示す。
患者AW1-2
患者AW1の被験部位No2は医師により観察開始時には軽症と診断された部位である。観察開始時は皮膚症状を呈しておらず、医師による皮膚所見値(合計値)は0であったが、医師による所見は時間経過とともに上昇した。自己所見はわずかながらも上昇したが(0から1に上昇)、医師の所見に比べると、自己所見の値は低かった。なお、同患者は観察の期間の間アトピー性皮膚疾患の治療は受けていない。治療開始時のSCCA-1値は約227.0 ng/mgであった。医師による皮膚所見も、観察を開始してから40日目ぐらいまでは値は0であったが、SCCA-の値の上昇と共に、皮膚所見値も1へと上昇し、悪化し始めた。なお、患者による事故所見の値は観察期間中変動せず、0のままであった。この観察結果を図7(B)に示す。
患者AW1-5
患者AW1の被験部位No.5も医師により寛解期と診断された部位である。観察開始時は皮膚症状を呈しておらず、医師による皮膚所見値(合計値)は0であった。観察期間の間、来院毎に観察及び角層の採取を行った。なお、同患者は観察の期間の間アトピー性皮膚疾患の治療は受けていない。治療開始時のSCCA-1値は約313.4 ng/mgであった。医師による皮膚所見も、観察を開始してから30日目ぐらいまでは値は0であったが、SCCA-の値の上昇と共に、皮膚所見値も1へと上昇し、悪化し始めた。なお、患者による事故所見の値は観察期間中変動せず、0のままであった。この観察結果を図7(C)に示す。
患者AW3-4
患者AW3の被験部位No.4は、観察開始時も皮膚症状を呈さなかった。観察期間の間、来院毎に観察及び角層の採取を行った。治療開始時のSCCA-1値は43.9 ng/mgと、やや低めながらも健常人に比べ優位に高く(19.1 ng/mg)、無視できない程度の値を示していた。観察開始43日後にSCCA-1値は184.7 ng/mgと、観察開始時に比べ顕著に増加し、その結果は医師による皮膚所見(合計値3)と連動していた。一方、自己所見の上昇は観察期間を通してわずかであった(合計値1)。その結果を図8(A)に示す。
患者AW3の被験部位No.6は、観察開始時も皮膚症状を呈さなかった。観察期間の間、来院毎に観察及び角層の採取を行った。治療開始時のSCCA-1値は82.5 ng/mgと、健やや低めながらも健常人に比べ優位に高く(19.1 ng/mg)、無視できない程度の値を示していた。観察開始85日後にSCCA-1値は425.9ng/mgと、観察開始時に比べ顕著に増加し、その結果は医師による皮膚所見(合計値7)と連動していた。一方、自己所見の上昇は観察期間を通してわずかであった(合計値1)。その結果を図8(B)に示す。
患者AW3の被験部位No.7は、観察開始時も皮膚症状を呈さなかった。観察期間の間、来院毎に観察及び角層の採取を行った。治療開始時のSCCA-1値は50.7 ng/mgと、やや低めながらも健常人に比べ優位に高く(19.1 ng/mg)、無視できない程度の値を示していた。観察開始85日後にSCCA-1値は250.6ng/mgと、観察開始時に比べ顕著に増加し、その結果は医師による皮膚所見(合計値7)と連動していた。一方、自己所見の上昇は観察期間を通してわずかであった(合計値1)。その結果を図8(C)に示す。
アトピー性皮膚炎の重症度の分類に頻用されているのはSCORAD値である(非特許文献1)。対象者の皮膚症状を、皮疹の面積(A)(体表面積に占める割合)および紅斑、浸潤/皮疹、滲出液/痂皮、掻破痕、苔癬化、乾燥の各皮疹につき、その程度(0=なし、1=軽度、2=中程度、3=重度)の合計(B)を求め、被験者による自覚症状をVASによって評価(C)し、A/5+7XB/2+CによりSCORAD値として算出した。
SCORAD値を求めた各対象者について、テープストリッピングにより角層試料を採譜し、SCCA-1値も測定した。その際、皮膚の採取は対象者によりを1箇所のみ行った者(N=1)、2箇所行った者(N=2)、3箇所行なった者(N=3)とに分け、2箇所、3箇所採取した場合の対象者のSCCA-1値はそれぞれ平均値を求めた。次に、各対象者のSCORAD値と、N=1、N=2、N=3の各対象者のSCCA-1値との相関を求めた。
以下にその結果をまとめた。
Claims (10)
- アトピー性皮膚炎の治療を受療中の対象者において当該治療方針を決定するための診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
i)当該SCCA-1の発現値がアトピー性皮膚炎を有しない者の値に比べ、統計学的に有意に高くない場合、当該治療を中断し当該疾患は治療を継続する必要はないと決定する、診断補助方法。 - アトピー性皮膚炎の治療を受療中の対象者において、当該治療方針を決定する診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
ii)当該SCCA-1の発現値を指標として当該対象者のアトピー性皮膚炎の症度を階級分けし、各階級に合ったアトピー性皮膚炎治療方法を当該対象者に適用することを決定する、アトピー性皮膚炎の治療方針を決定する診断補助方法。 - アトピー性皮膚炎の治療の要否判断が必要な対象者あるいは治療を受療中の対象者において、当該治療の要否判断あるいは当該治療を必要と判断した場合の治療方針を決定する診断補助方法であって、当該対象者の皮膚角層細胞中のSCCA-1の発現値を測定し、
i)当該SCCA-1の発現値がアトピー性皮膚炎を有しない者の値に比べ、統計学的に有意に高くない場合、当該治療は不要と判断するあるいは受療中の場合は当該治療により当該疾患は治療を継続する必要がないと決定し、統計学的に有意に高い場合、治療を開始する判断あるいは当該治療を継続する必要があると決定した場合、さらに、
ii)当該SCCA-1の発現値を指標として当該対象者のアトピー性皮膚炎の症度を階級分けし、各階級に合ったアトピー性皮膚炎治療方法を当該対象者に適用することを決定する、アトピー性皮膚炎の治療の要否判断あるいは治療方針を決定する診断補助方法。 - 前記治療方針の決定を長期にわたり繰り返し行う、請求項1~3のいずれか1項に記載の診断補助方法。
- アトピー性皮膚炎の症度の項目が、紅斑(erythema)、浸潤(oozing)、丘疹(papule)、掻破痕(excoriation)、苔癬化(lichenification)、乾燥・落屑(xerosis)及びそう痒感(itch)から成る群から選ばれる1または複数である、請求項1~4のいずれか1項に記載の診断補助方法。
- 前記皮膚細胞がテープストリッピングにより非侵襲的に採取される、請求項1~5のいずれか1項に記載の診断補助方法。
- 前記SCCA-1の発現の測定をSCCA-1に特異的な抗体を使用する免疫学的検出方法により実施する、請求項1~6のいずれか1項に記載の診断補助方法。
- 前記免疫学的検出方法が酵素免疫測定法(ELISA法、EIA法を含む)、蛍光免疫測定法、放射免疫測定法(RIA)、発光免疫測定法、表面プラズモン共鳴法(SPR法)、水晶振動子マイクロバランス(QCM法)、免疫比濁法、ラテックス凝集免疫測定法、ラテックス比濁法、粒子凝集反応法、金コロイド法、キャピラリー電気泳動法、グラフェンバイオセンサ、アンペロメトリックバイオセンサ、レーザー分光法及び表面濃縮免疫測定法から成る群から選ばれる、請求項7に記載の診断補助方法。
- 前記皮膚細胞の採取及び前記SCCA-1の発現の測定が在宅あるいは検診で行われる、請求項6~8のいずれか1項に記載の診断補助方法。
- 前記テープストリッピングによる採取を、対象者当たり3箇所以上の皮膚部位にて行う、請求項1~9のいずれか1項に記載の診断補助方法。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006098523A1 (ja) * | 2005-03-18 | 2006-09-21 | Shiseido Company, Ltd. | 扁平上皮細胞癌関連抗原を指標とする皮膚性状の評価方法 |
WO2007046463A1 (ja) * | 2005-10-21 | 2007-04-26 | Fancl Corporation | アトピー性皮膚炎マーカーとその利用技術 |
JP2009115813A (ja) * | 2008-12-26 | 2009-05-28 | Tokiwa Yakuhin Kogyo Kk | アトピー性皮膚炎の局所病態の非侵襲的評価方法 |
JP2012177601A (ja) * | 2011-02-25 | 2012-09-13 | Fancl Corp | アトピー性皮膚炎の検査方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016179043A1 (en) * | 2015-05-01 | 2016-11-10 | Dermtech, Inc. | Non-invasive skin collection system |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006098523A1 (ja) * | 2005-03-18 | 2006-09-21 | Shiseido Company, Ltd. | 扁平上皮細胞癌関連抗原を指標とする皮膚性状の評価方法 |
WO2007046463A1 (ja) * | 2005-10-21 | 2007-04-26 | Fancl Corporation | アトピー性皮膚炎マーカーとその利用技術 |
JP2009115813A (ja) * | 2008-12-26 | 2009-05-28 | Tokiwa Yakuhin Kogyo Kk | アトピー性皮膚炎の局所病態の非侵襲的評価方法 |
JP2012177601A (ja) * | 2011-02-25 | 2012-09-13 | Fancl Corp | アトピー性皮膚炎の検査方法 |
Non-Patent Citations (6)
Title |
---|
KENJI IDEHARA: "Allergy Shikkan no Transcriptome Kaiseki II Shinki Allergy Shikkan Marker to shite no Henpei Johi Saibo Gan Kogen (SCCA) no Dotei", CLINICAL TESTING, vol. 49, no. 7, 15 July 2005 (2005-07-15), pages 769 - 772, XP009516413, ISSN: 0485-1420 * |
See also references of EP3450978A4 * |
SHOICHIRO OTA, KENJI IDEHARA: "III. Atopy-sei Hifuen no Atarashii Biomarker = [III. New Biomarkers for Atopic Dermatitis]", ALLERGOLOGY & IMMUNOLOGY, vol. 23, no. 2, 15 January 2016 (2016-01-15), pages 190 - 196, XP009512842, ISSN: 1344-6932 * |
TAKASHI ABE: "Skincare Keshohin no Kenkyu Kaihatsu Doko (2007 Nen", FRAGRANCE JOURNAL, vol. 36, no. 4, 15 April 2008 (2008-04-15), pages 78 - 83, XP009516412, ISSN: 0288-9803 * |
YUKO MATSUMOTO ET AL.: "Nyuyoji Atopi-sei Hifuen o Taisho to shita Tei Shigekisei Skincare no Koka to Kakusochu no SCCA1 Tanpakushitsu no Hendo", THE ANNUAL MEETING OF THE JAPANESE SOCIETY OF PEDIATRIC DERMATOLOGY PROGRAM . SHOROKUSHU, vol. 36, 2012, pages 112, XP009516414 * |
YUKO MATSUMOTO ET AL.: "Nyuyoji no Atopy-sei Hifuen to SCCA1 Oyobi S 100A8 /A9 no Kanrensei", JAPANESE JOURNAL OF PEDIATRIC ALLERGY AND CLINICAL IMMUNOLOGY, vol. 25, no. 3, 20 August 2011 (2011-08-20), pages 545, XP009513082, ISSN: 0914-2649, DOI: 10.3388/jspaci.25.531 * |
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