JP7703161B2 - Method for obtaining data for predicting therapeutic effect of sublingual immunotherapy and use thereof - Google Patents

Description

The present invention relates to a method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy, and its use.

Allergen immunotherapy is known as a cure for allergic rhinitis (e.g., seasonal allergic rhinitis or perennial allergic rhinitis). Among these, sublingual immunotherapy, in which a therapeutic drug containing an allergen is administered sublingually, is widely used because of its simplicity.

Sublingual immunotherapy is known to be effective for, for example, cedar pollen allergy and house dust allergy rhinitis (Non-Patent Documents 1 and 2).

Okubo K, Gotoh M, Fujieda S, Okano M, Yoshida H, Morikawa H, et al. A randomized double-blind comparative study of sublingual immunotherapy for cedar pollinosis. Allergol Int 2008; 57:265-75. Feng B, Xiang H, Jin H, Gao J, Huang S, Shi Y, et al. Efficacy of Sublingual Immunotherapy for House Dust Mite-Induced Allergic Rhinitis: A Meta-Analysis of Randomized Controlled Trials. Allergy Asthma Immunol Res 2017; 9:220-8.

Although the above-mentioned sublingual immunotherapy requires long-term treatment (e.g., 2 to 5 years), the therapeutic effect cannot be predicted at the start of treatment. Furthermore, approximately 30% of patients who undergo treatment do not achieve therapeutic effect. For this reason, a technology that can predict the therapeutic effect of sublingual immunotherapy in advance has been desired.

The present invention has been made in consideration of the above problems, and aims to provide a method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy, and its use.

The inventors discovered that it is possible to predict the therapeutic effect of sublingual immunotherapy based on the genotype of at least one gene that constitutes the HLA gene complex, and thus completed the present invention.

That is, the present invention includes the following configurations.
<1> A method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy, comprising a detection step of detecting the genotype of at least one gene constituting an HLA gene complex in a sample collected from a subject.
<2> The method for obtaining data according to <1>, wherein the gene is a gene contained in the HLA-DP region, the HLA-DQ region, or the HLA-DR region.
<3> The method for acquiring data described in <1> or <2>, wherein in the detection step, the genotype is detected by one or more methods selected from the group consisting of genotyping by a PCR-rSSO/PCR-SSP (Luminex) method, genotyping by a tagSNP method, genotyping by Sanger sequencing, genotyping including an imputation method by genome-wide association study (GWAS), and genotyping by a next-generation sequencer.
<4> A predictive kit for predicting the therapeutic effect of sublingual immunotherapy, comprising an item for detecting the genotype of at least one gene that constitutes the HLA gene complex.
<5> The kit according to <4>, wherein the gene is a gene contained in the HLA-DP region, the HLA-DQ region, or the HLA-DR region.
<6> The kit described in <4> or <5>, wherein the above-mentioned item includes one or more items selected from the group consisting of a primer that specifically binds to the above-mentioned gene, a GWAS panel preparation reagent, and a next-generation sequencer library preparation reagent.

According to one aspect of the present invention, a method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy and its use can be provided.

FIG. 1 is a flowchart of an embodiment of the present invention. 1 is a graph showing the measurement results of tag SNP in an example of the present invention.

One embodiment of the present invention is described below, but the present invention is not limited to this. The present invention is not limited to each configuration described below, and various modifications are possible within the scope of the claims, and embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention. In addition, all academic literature and patent documents described in this specification are incorporated herein by reference. In addition, unless otherwise specified in this specification, "A to B" representing a numerical range is intended to mean "greater than or equal to A and less than or equal to B."

〔1. How to obtain data〕
A method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy according to one embodiment of the present invention includes a detection step of detecting the genotype of at least one gene constituting an HLA gene complex in a sample collected from a subject.

As shown in the examples described below, when comparing patients who benefited from sublingual immunotherapy with patients who did not, the genotype of at least one gene constituting the HLA gene complex was different between the two. Therefore, with the above configuration, it is possible to classify subjects (e.g., patients) according to their genotype, and to predict the therapeutic effect of sublingual immunotherapy before the start of treatment according to this classification.

For subjects for whom sublingual immunotherapy is thought to be effective, for example, treatment with sublingual immunotherapy can be actively continued. On the other hand, for subjects for whom sublingual immunotherapy is thought to be ineffective, for example, a treatment other than sublingual immunotherapy can be selected.

As mentioned above, the therapeutic effect of sublingual immunotherapy varies from person to person, with some patients seeing therapeutic effects after a certain period of time and others not seeing therapeutic effects even after a certain period of time has passed. In this specification, patients who see therapeutic effects at the time point when allergy symptoms peak two years after starting treatment with sublingual immunotherapy (for example, for cedar pollen allergy, 12 weeks from mid-February) can be defined as "responsive to sublingual immunotherapy," while patients who do not see therapeutic effects can be defined as "refractory to sublingual immunotherapy." In this specification, "seeing therapeutic effects" means that the visual assessment scale, which quantifies the overall allergy symptoms of the patient in the examples described below, has a value of 5 or less.

Conventionally, there was no indicator of whether a patient was responsive or refractory to sublingual immunotherapy, and it was not possible to know until the patient actually underwent treatment with sublingual immunotherapy. It is surprising that the present invention uses genotype as a biomarker to predict the therapeutic effect of sublingual immunotherapy before it is administered.

In one embodiment of the invention, the subject is a human. In one embodiment of the invention, the subject is a non-human mammal. Examples of non-human mammals include ungulates (cattle, boar, pig, sheep, goat, etc.), perissodactyls (horses, etc.), rodents (mice, rats, hamsters, squirrels, etc.), lagomorphs (rabbits, etc.), and carnivores (dogs, cats, ferrets, etc.). The non-human mammals mentioned above include wild animals as well as livestock or companion animals (pets).

In this specification, the term "sample" refers to a general "sample" collected from a subject. The "sample" also includes (i) biological samples (e.g., samples from which DNA (e.g., human DNA) can be collected, such as blood, saliva, buccal mucosa, and skin), pathological specimens prepared by fixing biological samples with a fixative (e.g., formalin), and (ii) solubilized products obtained by solubilizing the above (i) in a desired solution.

The allergen used in the sublingual immunotherapy of the present invention is not particularly limited and may be any allergen. Examples of allergens include pollen (e.g., cedar pollen, cypress pollen), house dust, dust mites, foods (e.g., apples, peaches, melons, wheat, eggs), and antigen peptides that constitute them (e.g., Cry j 1, Der p 1, PR-10 protein), etc. From the viewpoint of being common allergens used in sublingual immunotherapy, the allergen is preferably pollen or house dust. The sublingual immunotherapy of the present invention may be a sublingual immunotherapy that treats allergies caused by the above-mentioned allergens.

The specific method of sublingual immunotherapy for predicting the therapeutic effect is not particularly limited, and any known method can be used. Examples of such methods include a method of injecting a drug subcutaneously, a method of placing a drug sublingually, a method of injecting a drug into a lymph node, etc.

In one embodiment of the present invention, the number of genes whose genotypes are detected from the HLA gene complex (in other words, the number of genes whose genotypes are detected in the detection process) is at least one. The number of genes whose genotypes are detected may be two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve or more. The more genes whose genotypes are detected, the more accurately the therapeutic effect of sublingual immunotherapy can be predicted. There is no upper limit to the number of genes to be detected. From the viewpoint of simplifying the detection process, the upper limit to the number of genes to be detected may be 20, 15, 10, or 5.

In one embodiment of the present invention, the detection step may more specifically include a step of determining whether the patient is a carrier (homozygote), a carrier (heterozygote), or a non-carrier for the genotype of the detected gene. With the above configuration, the therapeutic effect of sublingual immunotherapy can be predicted in more detail.

The above genes may be one or more types of genes contained in any one of the HLA-DP region, the HLA-DQ region, and the HLA-DR region of the HLA gene complex. With this configuration, the therapeutic effect of sublingual immunotherapy can be predicted more accurately before the start of treatment. When there are multiple genes for which genotypes are detected, (i) all of the genes may be contained in one of the HLA-DP region, the HLA-DQ region, or the HLA-DR region, (ii) all of the genes may be contained in two of the HLA-DP region, the HLA-DQ region, or the HLA-DR region, or (iii) each gene may be contained in any one of the HLA-DP region, the HLA-DQ region, or the HLA-DR region.

Examples of genes contained in the HLA-DP region include HLA-DPA1 and HLA-DPB1. For example, HLA-DPA1 and HLA-DPB1 can be suitably used to predict the therapeutic effect of sublingual immunotherapy for hay fever (e.g., cedar pollen allergy, cypress pollen allergy).

Examples of genes contained in the HLA-DQ region include HLA-DQA1 and HLA-DQB1. For example, HLA-DQA1 and HLA-DQB1 can be suitably used to predict the therapeutic effect of sublingual immunotherapy for hay fever (e.g., cedar pollen allergy, cypress pollen allergy). HLA-DQB1 can be suitably used to predict the therapeutic effect of sublingual immunotherapy for house dust allergy (e.g., house dust allergic rhinitis).

An example of a gene contained in the HLA-DR region is HLA-DRB1. HLA-DRB1 can be particularly useful for predicting the therapeutic effect of sublingual immunotherapy for house dust allergies (e.g., house dust allergic rhinitis).

For example, when the genotype of the HLA-DPB1 gene to be detected is HLA-DPB1 ^* 05:01, it can be said that there is a high probability that the subject is refractory to sublingual immunotherapy.

The genotype of the gene to be detected may be detected directly, or may be detected by identifying the genotype (haplotype) of another region that is linked to the genotype of the gene to be detected.

When HLA-DPB1 ^* 05:01 is detected, examples of the genotype of the other region include HLA-DPB1 ^* 02:02 and SNPs linked to HLA-DPB1 ^* 05:01, which will be described later.

In the detection step, it is preferable to detect the genotype by one or more methods selected from the group consisting of genotyping by the PCR-rSSO/PCR-SSP (Luminex) method, genotyping by the tagSNP method, genotyping by Sanger sequencing, genotyping by genome-wide association analysis (GWAS), and genotyping by a next-generation sequencer. It is more preferable to detect the genotype by genotyping by the tagSNP method, since it is inexpensive and simple. Note that the specific techniques of the above-mentioned methods are publicly known, and the detection step can be performed according to the publicly known techniques.

The tagSNP method uses single nucleotide polymorphisms (hereinafter simply referred to as SNPs) as markers for specific genotypes. It is believed that there are tens of thousands to millions of SNPs in the genome, and some correspond to specific genotypes. In other words, patients with a specific genotype necessarily also have the corresponding SNP. Therefore, by detecting the corresponding SNP, it is possible to confirm the corresponding genotype.

Examples of SNPs corresponding to each genotype in the HLA gene complex are shown in Tables 1 to 3. In the tables, the rs numbers of the SNPs indicate the registration numbers in the dbSNP database of the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/projects/SNP/). The relationship between genotypes and SNPs is also described in the literature "Genes and Immunity (2012) 13, 543-548." The entire literature is incorporated herein by reference. In the present invention, for example, by detecting the SNPs listed in the following Tables 1 to 3 (corresponding to Tables 4 to 6 in the above literature), the corresponding genotype can be confirmed.

<Example 1 of prediction of effect of sublingual immunotherapy>
Based on genes constituting the HLA gene complex (eg, HLA-DPB1 ^* 05:01), it is possible to predict the effect of sublingual immunotherapy (eg, cedar sublingual immunotherapy) before the start of treatment.

First, before treatment with sublingual immunotherapy, it is determined whether the patient is a carrier or non-carrier of "HLA-DPB1 ^* 05:01."

When patients with "HLA-DPB1 ^* 05:01" are treated with sublingual immunotherapy, (i) it can be predicted that approximately 80% of the patients will experience improvement or remission of symptoms, and (ii) it can be predicted that approximately 20% of the patients will be refractory.

On the other hand, when sublingual immunotherapy is administered to patients who do not have "HLA-DPB1 ^* 05:01", (i) it is predicted that approximately 96% of patients will experience improvement or remission of symptoms, and (ii) it is predicted that approximately 4% of patients will be refractory.

Comparing the predicted value of (i) above when a patient having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy and the predicted value of (i) above when a patient not having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, it can be predicted that when a patient not having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, the probability of symptom improvement or remission is relatively higher than when a patient having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy.

<Example 2 of prediction of effect of sublingual immunotherapy>
Based on genes constituting the HLA gene complex (eg, HLA-DPB1 ^* 05:01), it is possible to predict the effect of sublingual immunotherapy (eg, cedar sublingual immunotherapy) before the start of treatment.

First, before treatment with sublingual immunotherapy, it is examined whether the patient is a carrier (homozygous), a carrier (heterozygous), or a non-carrier of "HLA-DPB1 ^* 05:01."

When patients with "HLA-DPB1 ^* 05:01" (homozygous) are treated with sublingual immunotherapy, (i) it is predicted that approximately 74% of patients will experience improvement or remission of symptoms, and (ii) it is predicted that approximately 24% of patients will be refractory.

On the other hand, when patients with "HLA-DPB1 ^* 05:01" (heterozygotes) are treated with sublingual immunotherapy, (i) it is predicted that approximately 83% of the patients will experience improvement or remission of symptoms, and (ii) it is predicted that approximately 17% of the patients will be refractory.

On the other hand, when sublingual immunotherapy is administered to patients who do not have "HLA-DPB1 ^* 05:01", (i) it is predicted that approximately 96% of patients will experience improvement or remission of symptoms, and (ii) it is predicted that approximately 4% of patients will be refractory.

^From a comparison of the predicted value of (i) above when a patient (homo) having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, the predicted value of (i) above when a patient (hetero) having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, and the predicted value of (i) above when a patient not having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, it can be predicted that the probability of symptom improvement or remission is relatively the highest when a patient not having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy, and the probability of symptom improvement or remission is relatively the second highest when a patient (hetero) having "HLA-DPB1 ^* 05:01" is treated with sublingual immunotherapy. It can be predicted that when sublingual immunotherapy is administered to patients (homozygous patients) with "05:01", the probability of symptom improvement or remission is relatively lowest.

2. Kits
A prediction kit for predicting the therapeutic effect of sublingual immunotherapy according to one embodiment of the present invention includes an item for detecting the genotype of at least one gene constituting an HLA gene complex.

Items already mentioned in [1. Data Acquisition Method] will not be explained below.

As used herein, a "kit" may be any combination of reagents, etc., for any purpose. The purpose may be a medical purpose or a testing purpose. Specifically, the purpose may be a diagnostic purpose.

The gene to be detected may be one or more genes contained in the HLA-DP region, the HLA-DQ region, or the HLA-DR region.

The kit may, for example, include items for directly detecting a gene of interest, or may include items for detecting a SNP corresponding to the gene.

The above-mentioned article may include one or more types selected from the group consisting of a primer that specifically binds to the above-mentioned gene, a GWAS panel preparation reagent, and a next-generation sequencer library preparation reagent.

In addition, if it is possible to detect a gene of interest, in addition to the above-mentioned items, the device may further include an item for detecting a protein derived from the gene, or an item for detecting mRNA and a protein derived from the gene.

Examples of products for detecting mRNA derived from the above genes include probes that hybridize to the mRNA of the above genes (e.g., polynucleotides that hybridize to the mRNA), primers for detecting the expression of the above genes by PCR, reagents for preparing GWAS panels, and reagents for preparing next-generation sequencer libraries.

Examples of reagents for preparing the GWAS panel include the Infinium Asian Screening Array (manufactured by Illumina) and the Infinium Global Screening Array (manufactured by Illumina).

An example of the next-generation sequencer library preparation reagent is TruSight (registered trademark) HLA v2 sequencing panel (manufactured by Illumina).

Examples of products for detecting proteins derived from the above genes include antibodies that specifically bind to the above proteins, fluorescent dyes, and secondary antibodies labeled with enzymes.

The above-mentioned article may be, for example, (i) one that emits a detectable signal (e.g., fluorescence) by itself, (ii) one that binds to another structure that emits a detectable signal (e.g., fluorescence), or (iii) one that binds to an mRNA or protein that is bound to another structure that emits a detectable signal (e.g., fluorescence).

In one embodiment, the kit may include reagents and/or ancillary materials for use in diagnosis. In one embodiment, the kit may include one or more containers (boxes, bottles, dishes, etc.) for storing the reagents and/or ancillary materials.

[3. Other]
An embodiment of the present invention can also be configured as follows.
<1> A method for predicting the therapeutic effect of sublingual immunotherapy, comprising a detection step of detecting the genotype of at least one gene constituting an HLA gene complex in a subject (e.g., in a sample collected from the subject).
<2> The method for predicting the therapeutic effect of sublingual immunotherapy described in <1>, wherein the gene is a gene contained in the HLA-DP region, the HLA-DQ region, or the HLA-DR region.
<3> The method for predicting the therapeutic effect of sublingual immunotherapy described in <1> or <2>, in which in the detection step, the genotype is detected by one or more methods selected from the group consisting of genotyping by the PCR-rSSO/PCR-SSP (Luminex) method, genotyping by the tag SNP method, genotyping by Sanger sequencing, genotyping including an imputation method by genome-wide association analysis (GWAS), and genotyping by a next-generation sequencer.

〔material and method〕
<Test subjects>
Patients were enrolled in the study who were diagnosed with cedar pollen allergy according to the Japanese guidelines for allergic rhinitis, i.e., (i) had nasal-ocular symptoms during the cedar pollen season (mid-February to April) and (ii) had cedar pollen-specific IgE antibodies and a blood IgE antibody concentration of >0.7 _{U A} /mL as determined by the ImmunoCAP test (Phadia, Tokyo, Japan). Informed consent was obtained from all patients.

Figure 1 shows a flow chart showing the test period. For example, in the Chubu Hokuriku region, cedar pollen dispersion usually lasts for 12 weeks from mid-February. Please note that the cedar pollen dispersion period differs depending on the region. Therefore, patients who started sublingual immunotherapy between October and December of the years shown in Figure 1 (2014-2017) were used as test subjects, and allergy symptoms were measured. The measurement period was the second cedar pollen season after starting sublingual immunotherapy (for example, if sublingual immunotherapy was started in 2014, the measurement period was until April 2016). Among the above patients, those who had developed asthma attacks, pregnant women, and those who were being treated for a different allergy (for example, house dust allergy) were excluded from the test subjects in accordance with the above-mentioned guidelines. All patients were standardized with cedar pollen extract according to the immunotherapy protocol. The amount of cedar pollen dispersion was measured using the Duraham method.

<Visual assessment scale>
The therapeutic effect of sublingual immunotherapy was evaluated using a visual assessment scale (VAS) that quantifies comprehensive allergic symptoms (general allergic symptoms such as sinusitis, stuffy nose, sneezing, and itchy eyes). The score range was from 0 (no symptoms) to 10 (severe symptoms). The score value of each patient was determined by having the patient report their symptoms. In each test result, at the time when allergic symptoms peak two years after treatment, patients with a VAS value of 5 or less were defined as responsive patients, and patients with a VAS value of more than 5 were defined as refractory patients.

<DNA extraction and HLA-DPB1 genotyping>
Peripheral blood samples were collected from each patient during treatment. DNA was extracted from leukocytes in peripheral blood using an automated DNA extraction system (QuickGene-610 L, Fujifilm) with a QuickGene DNA whole blood kit (Wako Pure Chemical Industries, Ltd.) according to the protocol. The genotype of the HLA-DPB1 gene in leukocytes was genotyped by the polymerase chain reaction sequence-specific oligonucleotide probe (PCR-SSOP) method using a WAKFlow HLA typing kit (Wakunaga Pharmaceutical Co., Ltd.) and Multiple Analyte Profiling (xMAP) technology (Luminwx System; Luminex Corporation).

<HLA-DPB1 ^* 05:01 tag SNP>
The tag-single nucleotide polymorphism (tagSNP) was used to verify the genotyping results of HLA-DPB1 genotype. Based on previous studies, rs9378177, a tagSNP in the HLA-DPB1 intron, with HLA-DPB1 ^* 05:01 genotype was selected. Each genotyping was performed by TaqMan genotyping system (Thermo Fisher Scientific) on ABI PRISM 7900HT sequence detection system (Thermo Fisher Scientific). PCR was performed in 384-well format, and automated genotype calling was performed using ABI PRISM 7900HT data collection and analysis software version 2.2.2 (Thermo Fisher Scientific).

<Statistical analysis>
The difference in age between responsive and non-responsive patients was analyzed using a t-test. The difference in total cedar pollen and IgE serum up to the second season between responsive and non-responsive patients was analyzed using a Wilcoxon rank-sum test. The difference in gender and the start time of sublingual immunotherapy between responsive and non-responsive patients was analyzed using Fisher's exact test. The frequency analysis of HLA genotypes of patients in this study was performed using Fisher's exact test, using the publicly available data on Japanese subjects at http://hla.or.jp/med/frequency_search/en/allele/ as a control. The data of HLA-DPB1 gene was analyzed including 4-digit genotypes in a logistic regression model, where each genotype is coded as a digenic marker (0, 1, or 2 copies of the genotype). Based on the hypothesis that patients with certain genotypes are more likely to be refractory to sublingual immunotherapy, odds ratios were calculated, adjusting for the total amount of Japanese cedar pollen during the study period. q values based on false discovery rates (FDRs) were calculated using the Benjamin-Hochberg (BH) method for multiple comparisons. Significance was defined in this study as a P value of less than 0.05 and a q value of less than 0.05. All analyses were performed using R software version 3.5.1 (R foundation for Statistical Computing).

〔result〕
A total of 219 Japanese cedar pollen allergy patients were included in the study, and the test results for 203 of them were available for the second season, when symptoms peak. Patient information is shown in Table 4. The average score for patients before treatment was approximately 7.5, regardless of genotype.

From Table 4, in the second season, the patients were divided into 176 responsive patients and 27 refractory patients. The amount of Japanese cedar pollen dispersion was 2508/ ^cm3 in 2015, 3505/ ^cm3 in 2016, 2570/ ^cm3 in 2017, 5041/ ^cm3 in 2018, and 10933/ ^cm3 in 2019 (from Figure 1). The values of Japanese cedar pollen dispersion were adjusted by statistical multivariate logistic regression analysis.

Table 5 shows the frequency of HLA-DPB1 genotypes in the patient population studied in this study and in the general Japanese population (1,483 people). Data from http://hla.or.jp/med/frequency_search/en/allele/ was used as a Japanese control.

As shown in Table 5, the most frequent genotype among the HLA-DPB1 genes was HLA-DPB1 ^* 05:01 (39.9%), followed by HLA-DPB1 ^* 02:01 (24.4%) and then HLA-DPB1 ^* 09:01 (9.9%).

Furthermore, among the HLA-DPB1 genotypes listed in Table 5, patients with genotypes with a frequency of more than 5% were classified into patients who were responsive or refractory to sublingual immunotherapy based on the VAS. Furthermore, statistical multivariate logistic regression analysis adjusted for the amount of cedar pollen dispersed was performed on the obtained values. The results are shown in Table 6.

Table 6 shows that patients with the HLA-DPB1 genotype HLA-DPB1 ^* 05:01 were significantly more likely to be refractory to sublingual immunotherapy than those with other genotypes (P = 0.006, q = 0.032, odds ratio = 2.28, 95% confidence interval = 1.27-4.20).

Finally, the accuracy of genotyping of HLA-DPB1 ^* 05:01 was confirmed using the single nucleotide polymorphism (SNP) rs9378177 present in the intron of the HLA-DPB1 gene. As a method of identification, first, the samples were examined by the PCR-SSO method, and the patients were classified into carriers (homozygotes), carriers (heterozygotes), and non-carriers for the HLA-DPB1 ^* 05:01 genotype. Furthermore, the samples were genotyped using the TaqMan real time PCR method to examine the genotype. The results are shown in FIG. 2.

The horizontal axis of Figure 2 indicates single-base mutations in HLA-DPB1 ^* 05:01 (Allele 1), and the vertical axis indicates single-base mutations in other HLA-DPB1 genotypes (Allele 2). As shown in Figure 2, the results of genotype identification by SNP measurement were completely consistent with the results of identification by the PCR-SSO method in all patients.

The present invention can be used to predict the therapeutic effect of sublingual immunotherapy.

Claims (4)

A detection step of detecting the genotype of at least one gene constituting an HLA gene complex in a sample collected from a subject,
The method for obtaining data for predicting the therapeutic effect of sublingual immunotherapy, wherein the gene is HLA-DP B1 . The data acquisition method according to claim 1, wherein in the detection step, the genotype is detected by one or more methods selected from the group consisting of genotyping by PCR-rSSO/PCR-SSP (Luminex) method, genotyping by tagSNP method, genotyping by Sanger sequencing, genotyping including imputation method by genome-wide association study (GWAS), and genotyping by next-generation sequencer. an article for detecting the genotype of at least one gene that constitutes an HLA gene complex;
A prediction kit for predicting the therapeutic effect of sublingual immunotherapy, wherein the gene is HLA-DP B1 . The kit according to claim 3, wherein the article includes one or more selected from the group consisting of a primer that specifically binds to the gene, a GWAS panel preparation reagent, and a next-generation sequencer library preparation reagent.

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