Method for assisting in determining treatment plan for dog having skin condition

Abstract

Provided is a method for assisting in determining a treatment plan for a dog that has a skin condition like atopic dermatitis.　Provided is a method that includes the following steps: a) a step for measuring the TARC level included in a blood sample taken from a subject, and b) a step for determining that treatment for atopic dermatitis is useful when the TARC level is equal to or greater than a threshold value, and determining that treatment for another disease is useful when the TARC level is less than the threshold value.

Description

Method for assisting in determining treatment strategies for dogs with skin conditions

[0001] The present invention relates to a method for assisting in determining a treatment plan for dogs with canine atopic dermatitis-like skin symptoms, and a reagent to be used therefor.

[0002] Atopic dermatitis is a disease that often involves a predisposition to atopy and results in chronic, recurring eczematous skin lesions over a long period of time. Hematological findings often include abnormalities such as increased peripheral blood eosinophil counts and elevated serum IgE levels. Immunological and non-immunological abnormalities have been reported as the main causes of atopic dermatitis. In recent years, atopic dermatitis has become more prevalent not only in humans but also in pet animals such as dogs, creating a need for methods to detect and treat it.

[0003] It has been reported that human atopic dermatitis patients exhibit high blood TARC levels and that this correlates with the pathological condition (see Non-Patent Documents 1 and 2). On the other hand, there are also reports that TARC expression is observed in healthy areas of humans, and it cannot be said that the usefulness of TARC, for example, as a diagnostic marker or therapeutic target, has yet to be fully investigated.

[0004] The relationship between atopic dermatitis and TARC has been extensively studied using model mice and humans, but the mechanism of atopic dermatitis onset does not necessarily match between animals, and the establishment of a disease marker and diagnostic method specific to dogs has been desired. Patent Document 1 reports an antibody that specifically recognizes canine TARC, and Patent Document 2 reports a method for diagnosing canine atopic dermatitis using canine serum TARC as a biomarker.

[0005] In addition to atopic dermatitis, other known skin diseases in dogs include scabies, demodex, infectious diseases, and food allergies. Some of these diseases show skin findings similar to atopic dermatitis, making them difficult to differentiate. However, because effective treatments for each disease vary, it is desirable to accurately differentiate these diseases at an early stage in order to achieve early recovery.

[0006] International Publication No. 2017 / 065203 Japanese Patent Application Laid-Open No. 2019-191007

[0007] Takao Fujisawa et al., Journal of the Japanese Society of Pediatric Allergy, Vol. 19, No. 5, pp. 744-757 (2005); Kunihiko Tamaki et al., Journal of the Japanese Dermatological Association, Vol. 116, No. 1, pp. 27-39 (2006)

[0008] The present invention aims to provide a method for assisting in determining a treatment plan for dogs with atopic dermatitis-like skin symptoms using objective indicators obtained from blood tests, as well as reagents and kits for use therein.

[0009] The present invention is as follows: [1] A method for assisting in determining a treatment strategy for a canine subject with atopic dermatitis-like skin symptoms, comprising: a) measuring the amount of TARC contained in a blood sample taken from the subject; and b) determining that treatment for atopic dermatitis is effective if the TARC amount is equal to or greater than a threshold, and determining that treatment for another disease is effective if the TARC amount is less than the threshold. [2] The method of [1], wherein in step b), the other disease is one or more diseases selected from the group consisting of scabies, demodex, flea infestation, tick infestation, lice infestation, bacterial infection, fungal infection (including yeast infection), immune-mediated disease (including psoriasis and food allergy), stress-induced scratching, mast cell tumor, and cutaneous lymphoma. [3] The method of [2], wherein the other disease is scabies. [4] The method of any of [1] to [3], wherein in step a), the amount of TARC is measured using an antibody that specifically binds to canine TARC. [5] A reagent for assisting in determining a treatment plan for a canine subject with atopic dermatitis-like skin symptoms, the reagent having a function of measuring the amount of TARC contained in a blood sample taken from the subject, and the determination of the treatment plan includes determining that treatment for atopic dermatitis is effective when the TARC amount is equal to or greater than a threshold, and determining that treatment for another disease is effective when the TARC amount is less than a threshold. [6] The reagent according to [5], wherein the other disease is one or more diseases selected from the group consisting of scabies, demodex folliculorum, flea infestation, tick infestation, lice infestation, bacterial infection, fungal infection (including yeast infection), immune-mediated disease (including psoriasis and food allergy), stress-induced scratching, mast cell tumor, and cutaneous lymphoma. [7] The reagent according to [6], wherein the other disease is scabies. [8] The reagent according to any of [5] to [7], comprising an antibody that specifically binds to canine TARC.[9] A method for assisting in the differential diagnosis of atopic dermatitis in a canine subject, comprising: a) measuring the amount of TARC contained in a blood sample taken from the subject; and b) determining that the subject is suffering from atopic dermatitis if the amount of TARC is equal to or greater than a threshold, and determining that the subject is suffering from another disease if the amount of TARC is less than the threshold.

[10] A reagent for assisting in the differential diagnosis of atopic dermatitis in a canine subject, the reagent having a function of measuring the amount of TARC contained in a blood sample taken from the subject, and the differential diagnosis comprising determining that the subject is suffering from atopic dermatitis if the amount of TARC is equal to or greater than a threshold, and determining that the subject is suffering from another disease if the amount of TARC is less than the threshold. This specification incorporates the disclosure of Japanese Patent Application No. 2022-148724, from which the present application claims priority.

[0010] According to the present invention, a method for assisting in determining a treatment plan for dogs with atopic dermatitis-like skin symptoms using objective indicators obtained from blood tests, as well as reagents and kits for use therein, can be provided.

[0011] 1. First Embodiment - Method for Assisting in Decision-Making of a Treatment Plan The first embodiment of the present invention is a method for assisting in decision-making of a treatment plan for a canine subject having atopic dermatitis-like skin symptoms, the method comprising the following steps a) and b): a) measuring the amount of TARC contained in a blood sample taken from the subject, and b) determining that treatment for atopic dermatitis is effective when the amount of TARC is equal to or greater than a threshold, and determining that treatment for another disease is effective when the amount of TARC is less than the threshold.

[0012] 1-1 Definition and Overview In the method of this embodiment, the subject for determining a treatment plan is a dog. In this specification, the term "dog" refers to all dogs belonging to the genus Canis lupus familiaris, and includes all breeds.

[0013] TARC (Thymus Activation-Regulated Chemokine, also referred to as "CCL17") is a chemokine that binds to CC chemokine receptor 4 (CCR4) and induces migration and infiltration of leukocytes in the body. The TARC measured in this embodiment is TARC contained in a canine blood sample. As used herein, TARC refers to canine-derived TARC. Specifically, it refers to a protein that comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence having 90% or more, preferably 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, and that has binding activity to CCR4.

[0014] Clinical symptoms of atopic dermatitis include skin conditions such as erythema, lichenification, inflammation, pigmentation, scratch marks, and alopecia, as well as itching. Many other diseases are known to exhibit similar clinical symptoms. These other diseases include, but are not limited to, parasitic diseases such as scabies, demodex, flea infestation, tick infestation, and lice infestation; bacterial infections (e.g., staphylococcal infections); fungal infections (including yeast (Malassezia) infections); immune-mediated diseases (including psoriasis and food allergies); stress-induced scratching; and tumors such as mast cell tumors and cutaneous lymphoma. Among these diseases, scabies in particular is difficult to distinguish from atopic dermatitis based on appearance, etc.

[0015] In dogs with atopic dermatitis-like skin symptoms, atopic dermatitis has been diagnosed using the following method. First, antibiotics or other drugs are administered to rule out parasitic or infectious diseases. Next, food allergies are ruled out using IgE tests and food challenge tests. Furthermore, the presence or absence of elevated IgE levels in response to environmental allergens is confirmed to rule out other immune-mediated diseases. As such, diagnosing dogs with atopic dermatitis involves a process of treating with drugs corresponding to other diseases and confirming their effectiveness, which has led to problems such as time-consuming disease identification, high costs due to ineffective treatment, and the risk of side effects.

[0016] For parasitic diseases and infectious diseases, differentiation may be possible if the parasitic / infectious organism can be collected and identified; however, collecting and identifying the parasitic / infectious organism is not easy, and there are occasional cases where it cannot be collected or identified in some individuals, so this method is not necessarily capable of reliable differential diagnosis.

[0017] The treatment methods for many of the other diseases are different from those for atopic dermatitis. Known treatments for canine atopic dermatitis include, for example, administration of prednisolone (Asahi Kasei Pharma, Nipro, Towa Pharmaceutical, Kyorin Rimedio, Kyorin Pharmaceutical, Koaisei, Nippon Generic, Takeda Teva Pharmaceutical, Takeda Pharmaceutical Co., Ltd., Mylan Pharmaceuticals, Pfizer, Yoshindo, Shionogi & Co., Ltd.), antihistamines (Nichi-Iko, Takata, Isei, Sawai, Towa, Tsuruhara, Taiyo), cyclosporine (Elanco, Virbac, Asuka, Veterinary Medical Development), desensitization therapy (Nippon Zenyaku Kogyo), interferon gamma (Toray), oclacitinib (Zoetis), anti-IL-31 antibody drugs (Zoetis), and moisturizing shampoo therapy.

[0018] On the other hand, for diseases related to parasitism such as scabies, demodicosis, flea infestation, tick infestation, and lice infestation, treatment with ivermectin (product names: Cardomec (registered trademark) (Boehringer Ingelheim Animal Health), Panamectin (registered trademark) (Meiji Seika Pharma), Azabasca (registered trademark) (Nissin Pharmaceutical), Heartmectin (registered trademark) (Asuka Animal Health), Stromectol (registered trademark) (Maruho, MSD), etc.), imidaclobrid (product name: Avocate (trademark) (Bayer)), selamectin (product name: Revolution (registered trademark) (Zoetis)), sarolaner (product name: Simparica (registered trademark) (Zoetis)), filonil (product names: Frontline (registered trademark) and Frontline (registered trademark) Plus (Boehringer Ingelheim Animal Health)), etc. is considered effective.

[0019] Effective treatments for bacterial and yeast (Malassezia) infections include fusidic acid and betamethasone (trade name: Isaderm™ (Deca)), gentamicin sulfate (trade name: Gentakarm™ (Decra Vetti Lineary Products)), cephalexin (trade name: Cephaclear™ (Rikenbets Animal Pharma), cephaceptin (Vetoquinol™ (Nippon Zenyaku Kogyo), Telios™ CJ (Seba), Telios™ (DS Pharma Animal Health), Rilexipet™ (Virvac)), gentamicin sulfate (trade name: Tiasil™ (Virvac), Airsotic™ (Virvac), Otomax™ (MSD), Gentakarm™ (Decra)), amoxicillin and clavulanic acid (trade name: Clavamox™ (Zoetis)), and ciprofloxacin (Teva). It is said that the use of miconazole nitrate (product names: Malaseb (registered trademark) Shampoo (Dermacare), Slolan Ear Drops (trademark) (Elanco), Airsotic (trademark) (Virbac), etc.), polymyxin B sulfate (product name: Slolan Ear Drops (trademark) (Elanco)), hydrocortisone aceponate (product name: Slolan Ear Drops (trademark) (Elanco)), gentamicin sulfate (product name: Slolan Ear Drops (trademark) (Elanco)), etc. is known as an effective treatment for yeast (Malassezia) infections. The use of itraconazole (Nippon Chemiphar, Sawai Pharmaceutical, Nichi-Iko, Meiji Seika Pharma, Kobayashi Kako, Kaken Pharmaceutical), itrizole (Janssen Pharma) and the like is known as an effective treatment for fungal infections.

[0020] Prednisolone (Asahi Kasei Pharma, Nipro, Towa Pharmaceutical, Kyorin Rimedio, Kyorin Pharmaceutical, Koaisei, Nippon Generic, Takeda Teva Pharmaceutical, Takeda Pharmaceutical Co., Ltd., Mylan Pharmaceuticals, Pfizer, Yoshindo, Shionogi & Co.) is known as a treatment for immune-mediated diseases (psoriasis, food allergies). Among immune-mediated diseases, food allergies require identifying the allergen and eliminating it from the diet.

[0021] Known treatments for stress-induced scratching include tryptophan (product name: Vet's Best (registered trademark) (Hero Pet Brand)), valerian (product name: Vet's Best (registered trademark) (Hero Pet Brand)), dexmedetomidine (product name: Sileo (trademark) (Zoetis)), clomipramine hydrochloride (product name: Clomicalm (registered trademark) (Elanco)), and clomipramine hydrochloride (product name: Clofuranil (trademark) (Sun Pharma Laboratories)).

[0022] Known treatments for mast cell carcinoma include toceranib phosphate (trade name: Palladia (registered trademark) (Zoetis)), imatinib (trade name: Gleevec (registered trademark) (Novartis Pharma)), and imatinib (Daiichi Sankyo Espha, Daiichi Sankyo, Elmed, Nichi-Iko, Nippon Generic, Kyoso Mirai Pharma, Kobayashi Pharmaceutical, Nippon Kayaku, Ohara Pharmaceutical, Nippon Chemiphar, Sawai Pharmaceutical, Takeda Pharmaceutical, Takeda Teva Pharma, Towa Pharmaceutical, Kyowa Criticare, Nipro, Mylan Pharmaceuticals, Pfizer, Yakult Honsha, Takada Pharmaceutical, Meiji Seika Pharma, and Tatsumi Chemical). Known treatments for cutaneous lymphoma include lomustine (MediClone).

[0023] According to the method of this embodiment, it is possible to determine early on whether a treatment for atopic dermatitis is effective for a canine subject with atopic dermatitis-like symptoms, without having to go through a process of confirming the effectiveness of a therapeutic drug for another disease, thereby reducing the time and cost required for treatment and the risk of side effects.

[0024] 1-2 Step a) Step a) of the method of this embodiment is a step of measuring the amount of TARC contained in a blood sample taken from a canine subject.

[0025] As a blood sample, serum, plasma, or whole blood can be used, with serum being particularly preferred. The method for measuring the amount of TARC in a blood sample is not particularly limited, and any known method, such as immunological methods, high-performance liquid chromatography, or mass spectrometry, can be used. However, immunological methods using antibodies that specifically bind to canine TARC are particularly preferred. Examples of immunological methods include solid-phase immunoassays (RIA, EIA, FIA, CLIA, etc.), dot blotting, latex agglutination-turbidimetric immunoassay (LA), and immunochromatography. Among these, enzyme-linked immunosorbent assay (ELISA), a type of enzyme immunoassay (EIA), is preferred from the standpoint of quantitative performance.

[0026] In the ELISA method, a sample is added to a microwell plate immobilized with an anti-TARC antibody to allow an antigen-antibody reaction to occur, and then an enzyme-labeled anti-TARC antibody is added to allow the antigen-antibody reaction to occur. After washing, the sample is reacted with an enzyme substrate to develop color, and the absorbance is measured to detect TARC in the sample, and the TARC concentration in the sample can be calculated from the measured value. Alternatively, a fluorescently labeled anti-TARC antibody may be used to allow the antigen-antibody reaction to occur, followed by fluorescence measurement. The antigen-antibody reaction can be carried out at 4°C to 45°C, more preferably 20°C to 40°C, and even more preferably 25°C to 38°C. The reaction time is 10 minutes to 18 hours, more preferably 10 minutes to 1 hour, and even more preferably 30 minutes to 1 hour.

[0027] Although the ELISA method is not particularly limited, it is preferable to use a quantitative method, which can involve measuring a standard solution containing canine TARC of a known concentration simultaneously with the sample, and calculating the canine TARC concentration in the sample based on a calibration curve prepared based on the actual measured values ​​of the standard solution.

[0028] The anti-TARC antibody used in the immunological technique may be any antibody capable of binding to TARC. The anti-TARC antibody may be a monoclonal antibody or a polyclonal antibody, and may be a Fab, Fab', or F(ab') fragment of a monoclonal antibody. ２ Functional fragments having binding activity to TARC, such as those described above, can also be used. The antibody is preferably immunoglobulin G (IgG). The animal from which the antibody is derived is not particularly limited, but is preferably an animal other than a dog, and may be, for example, a mouse, rat, rabbit, goat, cow, pig, sheep, cat, monkey, camel, alpaca, bird, fish, or the like. Preferably, the antibody is derived from a mouse. In this specification, the term "antibody" does not necessarily refer to an intact antibody, and may refer to Fab, Fab', F(ab') ２ Alternatively, the antibody may be a single-domain antibody. Single-domain antibodies are also called single-chain antibodies or nanobodies (registered trademark), and are antibodies consisting of the variable region of an antibody (immunoglobulin) composed only of a heavy chain, and known examples include VHHs found in mammals of the Camelidae family and VNARs found in sharks.

[0029] A suitable example of an anti-TARC antibody that can be used is the anti-TARC monoclonal antibody described in Patent Document 1. Specifically, the anti-TARC monoclonal antibody described in Patent Document 1 is an anti-canine TARC monoclonal antibody that binds to canine TARC, or a functional fragment thereof that binds to canine TARC, which comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 3.

[0030] 1-3 Step b) Step b) of the method of the present embodiment is a step of determining that a treatment for atopic dermatitis is useful when the amount of TARC measured in step a) is equal to or greater than a threshold, and determining that a treatment for another disease is useful when the amount of TARC is less than the threshold.

[0031] In the method of this embodiment, the "threshold" may be set in advance or may be set each time a negative control or standard is measured simultaneously with sample measurement, based on the measured value of the negative control or standard. For example, a sample collected from a healthy dog ​​or a dog suffering from one of the above-mentioned other diseases may be measured simultaneously as a negative control. Here, "healthy dog" refers to a dog not suffering from dermatitis. It is generally known that the TARC concentration in the blood of dogs with atopic dermatitis is significantly higher than that of healthy dogs (see, for example, Patent Document 2). Therefore, for example, the threshold can be set to 2-fold, 5-fold, or 10-fold the concentration in a negative control, particularly a blood sample from a healthy dog. In this embodiment, the TARC value in a sample is also referred to as the TARC concentration or TARC level.

[0032] When the threshold value is set in advance, for example, the amount of TARC in specimens from healthy dogs or dogs suffering from the other diseases may be measured in advance, and the threshold value may be determined based on the measured value.

[0033] The threshold can be determined, for example, by ROC (receiver operating characteristic curve) analysis. Furthermore, the accuracy of determination (sensitivity and specificity) by the method of the present invention can be determined by ROC analysis. ROC analysis is an analytical method in which TARC values ​​are measured in samples collected from multiple dogs suffering from canine atopic dermatitis and multiple dogs suffering from the other diseases, the sensitivity and specificity at each threshold are calculated, and a graph (curve) is drawn by plotting the values ​​on a coordinate system with "1-specificity" on the horizontal axis and "sensitivity" on the vertical axis. The threshold at which the curve is closest to the upper left corner of the graph is adopted as the effective threshold.

[0034] For example, the threshold for serum TARC concentration can be set at 500 to 1000 pg / mL, preferably as measured by ELISA.

[0035] If the TARC concentration in the blood sample is equal to or greater than the threshold, the test dog can be determined to be suffering from atopic dermatitis. On the other hand, if the TARC concentration in the blood sample is less than the threshold despite the dog having atopic dermatitis-like skin symptoms, the dog can be determined to be suffering from one of the other diseases. Based on this result, it can be determined that the former test dog would benefit from treatment for atopic dermatitis, and the latter test dog would benefit from treatment for the other disease. Clinicians can use this result to assist in determining the treatment strategy for dogs with atopic dermatitis-like skin symptoms.

[0036] 2. Second Embodiment - Reagent for Assisting in Treatment Policy Decision-Making The reagent of the second embodiment of the present invention is a reagent for assisting in determining a treatment policy for a canine subject having atopic dermatitis-like skin condition, characterized in that the reagent has a function of measuring the amount of TARC contained in a blood sample taken from the subject, and the treatment policy decision includes determining that treatment for atopic dermatitis is effective when the amount of TARC is equal to or greater than a threshold, and determining that treatment for another disease is effective when the amount of TARC is less than the threshold. More specifically, the reagent etc. of this embodiment is a reagent etc. used for the method of the first embodiment. Unless otherwise specified, the definitions of terms in this embodiment are the same as those described in the first embodiment.

[0037] As used herein, a "reagent" may be a single composition, a combination of two or more compositions (e.g., a reagent set), or a combination of one or more compositions and an instrument (e.g., a kit). That is, the term "reagent" as used herein encompasses reagent sets and kits. For example, in the case of a reagent for measuring the amount of TARC by ELISA, the reagent may include a microwell plate on which an anti-TARC antibody is immobilized, a solution containing a labeled anti-TARC antibody, a standard solution, a diluent, a washing solution, etc. If necessary, an instruction manual, etc. may also be provided.

[0038] The reagent of this embodiment has a function of measuring the amount of TARC contained in a blood sample. The function of measuring the amount of TARC in a blood sample is not particularly limited, and can be a function that enables known measurement methods such as immunological methods, high-performance liquid chromatography, mass spectrometry, etc. In particular, as a function that enables immunological methods, it is preferable to have an antibody that specifically binds to canine TARC. The immunological method is not particularly limited, but it is preferable to use an ELISA method from the viewpoint of quantitativeness.

[0039] 3. Third and Fourth Embodiments - Method and Reagent for Aiding Differential Diagnosis A third embodiment of the present invention is a method for aiding in the differential diagnosis of atopic dermatitis in a canine subject, comprising the steps of: a) measuring the amount of TARC contained in a blood sample taken from the subject, and b) determining that the subject is suffering from atopic dermatitis if the amount of TARC is equal to or greater than a threshold, and determining that the subject is suffering from another disease if the amount of TARC is less than the threshold.

[0040] The method of this embodiment enables differential diagnosis of whether a dog with skin symptoms has atopic dermatitis or another disease through a blood test. The configuration of the method of this embodiment, the definitions of each term, and the like are the same as those of the first embodiment unless there is a particular contradiction.

[0041] A fourth embodiment of the present invention is a reagent for assisting in the differential diagnosis of atopic dermatitis in a canine subject, characterized in that the reagent has a function of measuring the amount of TARC contained in a blood sample taken from the subject, and the differential diagnosis includes determining that the subject is suffering from atopic dermatitis if the amount of TARC is equal to or greater than a threshold, and determining that the subject is suffering from another disease if the amount of TARC is less than the threshold.

[0042] The reagent of this embodiment is a reagent that can be used to perform a differential diagnosis of whether a dog with skin symptoms has atopic dermatitis or another disease through a blood test. More specifically, the reagent of this embodiment is a reagent for use in the method of the third embodiment. The configuration of the reagent of this embodiment, the definitions of each term, and the like are the same as those of the first and second embodiments unless otherwise specified.

[0043] The present invention will be specifically explained by the following examples, but the present invention is not limited to these examples.

[0044] Blood samples were collected from 30 dogs (10 dogs in the atopic dermatitis (cAD) group, 10 dogs in the scabies group, and 10 dogs in the healthy control group), and serum TARC levels were measured. Anti-TARC monoclonal antibody CT-1 (see Patent Document 1) was immobilized on a 96-well microplate using 0.05 mol / L sodium carbonate buffer (pH 9.6). After washing, the plate was blocked at 37°C for 2 hours using Block Ace (registered trademark) (manufactured by Megmilk Snow Brand Co., Ltd.). After washing, each serum sample diluted with a diluent containing TARC-negative canine serum was dispensed into the wells and allowed to react at 37°C for 1 hour. After washing the plate, rabbit anti-canine TARC polyclonal antibody was added as the primary antibody and allowed to react at 37°C for 1 hour. The plate was then washed again, and horseradish peroxidase (HRP)-labeled anti-rabbit IgG antibody was added as the secondary antibody and allowed to react at 37°C for 1 hour. After adding 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate, color development was allowed to occur at 37°C for 20 minutes, and 0.5 mol / L sulfuric acid solution was added to stop the color development. The absorbance at 450 nm of each well was measured using a plate reader. In addition to the serum samples, similar measurements were performed on multiple recombinant canine TARC solutions with known concentrations, and a calibration curve was created based on the known concentrations and the resulting absorbance. The amount of TARC in each blood sample was calculated based on this calibration curve.

[0045] The amount of allergen-specific IgE was measured for the same serum samples. The recombinant mite antigen Derf2 was diluted with 0.05 mol / L sodium carbonate buffer (pH 9.6) and immobilized on a 96-well microplate. After washing the plate, a buffer containing bovine serum albumin was added for blocking. After washing the plate, each serum sample diluted with a buffer containing bovine serum albumin was dispensed into wells and incubated at 37°C for 1 hour. After washing the plate, horseradish peroxidase (HRP)-labeled anti-dog IgE antibody was added and incubated at 37°C for 1 hour. 3,3',5,5'-tetramethylbenzidine (TMB) was added as a substrate, followed by color development at 37°C for 30 minutes, and the color development was stopped by adding 0.5 mol / L sulfuric acid solution. The absorbance at 450 nm of each well was measured using a microplate reader.

[0046] Table 1 shows the distribution of TARC amounts in serum samples from each group. A Mann-Whittney test was performed to test for significance between the groups, resulting in p=0.06 between the cAD group and the healthy group, and p=0.03 between the cAD group and the scabies group. This indicates that a significantly higher TARC amount was obtained in the cAD group compared to the scabies group. Table 2 shows the distribution of IgE amounts in serum samples from each group. As shown in Table 2, it was shown that there was no significant difference in IgE amounts between the groups.

[0047]

[0048]

[0049] The amino acid sequences represented by SEQ ID NOs: 1 to 3 in this disclosure are shown in Table 3.

[0050] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. 1. A method for aiding in treatment decisions for a canine subject having an atopic dermatitis-like skin condition, comprising: a) measuring the amount of TARC contained in a blood sample taken from the subject; b) determining that a treatment for atopic dermatitis is useful when the amount of TARC is equal to or greater than a threshold, and determining that a treatment for another disease is useful when the amount of TARC is less than a threshold; A method comprising:

2. The method according to claim 1, wherein in step b), the other disease is one or more diseases selected from the group consisting of scabies, demodicosis, flea infestation, tick infestation, lice infestation, bacterial infection, fungal infection (including yeast infection), immune-mediated disease (including psoriasis and food allergy), stress-induced scratching, mast cell tumor and cutaneous lymphoma.

3. The method of claim 2, wherein the other disease is scabies.

4. The method of claim 1 , wherein in step a), the amount of TARC is measured using an antibody that specifically binds to canine TARC.

5. A reagent for aiding in determining a treatment plan for a canine subject having an atopic dermatitis-like skin condition, comprising: the reagent has a function of measuring the amount of TARC contained in a blood sample taken from the subject, The treatment plan determination includes determining that a treatment for atopic dermatitis is effective when the amount of TARC is equal to or greater than a threshold, and determining that a treatment for another disease is effective when the amount of TARC is less than a threshold. reagent.

6. The reagent according to claim 5, wherein the other disease is one or more diseases selected from the group consisting of scabies, demodicosis, flea infestation, tick infestation, lice infestation, bacterial infection, fungal infection (including yeast infection), immune-mediated disease (including psoriasis and food allergies), stress-induced scratching, mast cell tumor, and cutaneous lymphoma.

7. The method of claim 6, wherein the other disease is scabies.

8. The reagent of claim 5 , comprising an antibody that specifically binds to canine TARC.