WO2022158756A1 - Biomarker for predicting risk of developing food allergy and use thereof - Google Patents

Abstract

The present invention relates to a biomarker for predicting the risk of developing a food allergy and a use thereof. The composition according to one aspect of the present invention can simply and accurately predict the risk of developing a food allergy on the basis of serum lipopolysaccharide-binding protein (LBP) levels. Particularly, since the risk of developing a food allergy can be predicted in advance for a subject who does not have any symptoms of the food allergy, the present invention is expected to be useful not only for the effective prevention and management of the food allergy, but also for the establishment of a personalized course of treatment.

Description

Biomarkers for predicting food allergy risk and uses thereof

This application claims priority to Korean Patent Application No. 10-2021-0007673 filed on January 19, 2021, and the entire specification is a reference to this application.

It relates to a biomarker for predicting the risk of developing a food allergy and a use thereof.

Food Allergy (FA) is an adverse reaction caused by an immune response after exposure to a specific food. . Among them, IgE-mediated food allergy causes an immediate reaction and can cause anaphylaxis, a severe systemic reaction, so the importance is being emphasized. In recent 20 years, the incidence of food allergy has been rapidly increasing worldwide, and research related to the pathogenesis of food allergy has been actively conducted in foreign countries, but studies in Korea are still insignificant.

On the other hand, lipopolysaccharide-binding protein (LBP) is a 65 kDa water-soluble acute phase protein mainly produced in hepatocytes, intestinal epithelial cells and visceral adipocytes. , metabolic syndrome, type 2 diabetes, and atherosclerosis have been reported to have a positive correlation.

The present inventors confirmed that the level of lipopolysaccharide-binding protein is increased when a normal person with no symptoms or signs of food allergy is sensitized to a food antigen, and thus the expression level of the lipopolysaccharide-binding protein can be predicted and food allergy risk The present invention was completed by confirming that it can be used for the prevention of allergen sensitization.

One aspect is to provide a composition for predicting the risk of developing a food allergy, comprising an agent for measuring the expression level of lipopolysaccharide-binding protein (LBP).

Another aspect is to provide a kit for predicting the risk of developing a food allergy comprising the composition.

Another aspect is to provide an information providing method for predicting the risk of developing a food allergy.

Another aspect is to provide a method of screening for a food allergy antigen sensitization prophylaxis or a food allergy treatment agent.

One aspect provides a composition for predicting the risk of developing a food allergy, comprising an agent for measuring the lipopolysaccharide-binding protein (LBP) expression level.

As used herein, the term “lipopolysaccharide-binding protein (LBP)” is also referred to as LPS-binding protein or lipopolysaccharide-binding protein. The lipopolysaccharide-binding protein is a water-soluble acute-phase protein that is encoded by the LBP gene and induces an immune response by binding to bacterial lipopolysaccharide and presenting the lipopolysaccharide to important cell surface pattern recognition receptors such as CD14 and TLR4. The lipopolysaccharide-binding protein or a gene encoding the same can be obtained from a known database such as NCBI GenBank.

For example, but not limited thereto, the lipopolysaccharide-binding protein can be a protein encoded from a nucleic acid comprising the nucleotide sequence of NCBI Gene ID: 3929 in humans or a nucleic acid comprising the nucleotide sequence of EnsEMBL ID: ENSG00000129988. . In humans, the LBP gene encoding the lipopolysaccharide-binding protein is located on chromosome 20 (20q11.23) and is known to consist of about 15 exons.

As used herein, the term "biomarker" refers to a substance that can distinguish and diagnose a normal group individual from a food allergen sensitized individual or an individual at risk of developing a food allergy, and a food allergy antigen sensitized individual or food allergy It may include all organic biomolecules such as polypeptides, proteins, nucleic acids, genes, lipids, glycolipids, glycoproteins or sugars that show an increase or decrease in a subject at risk. In one embodiment, it may be a protein whose level is changed in an individual sensitized to a food allergen or a gene encoding the same, but is not limited thereto.

The agent capable of measuring the expression level of the lipopolysaccharide-binding protein includes, but is not limited to, an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the lipopolysaccharide-binding protein. (aptamer) may include one or more selected from the group consisting of.

As used herein, the term "measurement of protein level" is a process of confirming the expression level of LBP as a marker protein that can predict the risk of developing food allergy or an individual sensitized to a food allergen in an isolated biological sample, for example, the marker The amount of the protein can be confirmed by using an antibody that specifically binds to the protein.

The protein level measurement or comparative analysis method includes protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry analysis, radioimmunoassay, radioimmunodiffusion method, octeroni immunodiffusion method, rocket immunoelectrophoresis, immunohistochemistry analysis, immunocytochemistry analysis, complement fixation assay, 2D electrophoresis analysis, liquid chromatography-Mass Spectrometry (LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/Mass Spectrometry), Western blotting or ELISA (Enzyme-linked immunosorbent assay), etc. However, the present invention is not limited thereto.

As used herein, the term “antibody” refers to a specific protein molecule directed against an antigenic site. In one embodiment, the antibody refers to an antibody that specifically binds to a lipopolysaccharide-binding protein, and may include polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. It is apparent to those skilled in the art that the antibody can be easily prepared using techniques well known in the art.

In addition, the antibody may comprise a functional fragment of an antibody molecule as well as a complete form having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having at least an antigen-binding function, and may be, for example, Fab, F(ab'), F(ab')2 or Fv.

The agent capable of measuring the expression level of the gene encoding the lipopolysaccharide-binding protein, but is not limited thereto, is at least one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the gene. may include

The expression level of the gene can be confirmed by measuring the amount of mRNA or protein.

As used herein, the term "measuring mRNA level" is a process of confirming the mRNA expression level of LBP as a marker gene that can predict the risk of developing food allergy or a food allergy antigen sensitized individual or food allergy in an isolated biological sample. Measuring the amount of mRNA can be performed. Analysis methods for this include reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR), and RNase protection assay. , Northern blotting, or a DNA chip, but is not limited thereto.

As used herein, the term "primer" includes all combinations of primer pairs consisting of forward and reverse primers recognizing the target gene sequence, and preferably means a primer pair that provides analysis results with specificity and sensitivity. can

As used herein, the term "probe" refers to a material capable of specifically binding to a target material to be detected in a biological sample, and refers to a material capable of specifically confirming the presence of the target material in the sample through the binding. .

The probe is a material commonly used in the art, but is not limited thereto, and preferably PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA or DNA. In addition, the probe may be derived from or similar to a biological material as a biomaterial, or manufactured in vitro, and may be, for example, an enzyme, a protein, an antibody, a microorganism, an animal or plant cell and organ, a nerve cell, DNA or RNA. , The DNA includes cDNA, genomic DNA, and oligonucleotides, and the RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins include antibodies, antigens, enzymes, peptides, and the like.

As used herein, the term "antisense oligonucleotide" refers to DNA or RNA or a derivative thereof comprising a nucleic acid sequence complementary to a specific mRNA sequence, which binds to a complementary sequence in mRNA and inhibits the translation of mRNA into protein. do The antisense oligonucleotide sequence refers to a DNA or RNA sequence that is complementary to the mRNA of the genes and is capable of binding to the mRNA. This may inhibit the translation of the gene mRNA, translocation into the cytoplasm, maturation or any other essential activity for overall biological functions. The length of the antisense oligonucleotide may be 6 to 100 bases, preferably 8 to 60 bases, and more preferably 10 to 40 bases. The antisense oligonucleotide may be synthesized in vitro by a conventional method and administered in vivo, or the antisense oligonucleotide may be synthesized in vivo.

Another aspect provides a kit for predicting the risk of developing a food allergy comprising the composition.

The kit may be a reverse transcription polymerase chain reaction (RT-PCR) kit, a DNA chip kit, an enzyme-linked immunosorbent assay (ELISA) kit, a protein chip kit, or a rapid kit, but is not limited thereto.

In addition, the kit may further comprise one or more other components, solutions or devices suitable for the assay method.

For example, the RT-PCR kit may include each pair of primers specific for a marker gene, in addition to a test tube or other suitable container, a reaction buffer (with varying pH and magnesium concentrations), deoxynucleotide ( dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNAse, RNAse inhibitor DEPC-water, sterile water, and the like.

The DNA chip kit may include a substrate to which cDNA or oligonucleotide corresponding to a gene or fragment thereof is attached, and reagents, agents, enzymes, etc. for preparing a fluorescent probe, wherein the substrate is a control gene or its It may contain cDNA or oligonucleotides corresponding to fragments.

The protein chip kit may be a kit in which one or more antibodies against a marker are arranged at a predetermined position on a substrate and immobilized at a high density. The protein chip can be used as a method of isolating a protein from a biological sample, hybridizing the isolated protein with a protein chip to form an antigen-antibody complex, and reading it to check the presence or expression level of the protein.

As used herein, the term “antigen-antibody complex” refers to a combination of a corresponding protein antigen in a biological sample and an antibody that recognizes it. The detection of the antigen-antibody complex can be detected using methods known in the art, for example, spectroscopic, photochemical, biochemical, immunochemical, electrical, optical, chemical and other methods.

The rapid kit may include an antibody specific for a protein and/or an antibody specific for a control protein. In addition, the rapid kit is a reagent capable of detecting the bound antibody, for example, a nitrocellulose membrane to which a specific antibody and a secondary antibody are immobilized, a membrane bound to an antibody bound bead, an absorption pad and a sample pad, etc. material may be included.

In the kit for predicting the risk of developing a food allergy according to an aspect, the prediction may be a prediction of the risk of developing a food allergy in a subject without food allergy symptoms or signs.

Another aspect provides an information providing method for predicting the risk of developing a food allergy, comprising measuring the expression level of a lipopolysaccharide-binding protein or a gene encoding the same from an isolated biological sample.

As used herein, the term "biological sample" refers to a sample, such as tissue, cell, whole blood, serum, plasma, saliva, cerebrospinal fluid or urine, with a difference in gene expression level or protein expression level due to food allergy antigen sensitization or food allergy outbreak. However, it may be preferably selected from the group consisting of blood, serum and plasma, and more preferably serum.

In one embodiment, it was confirmed that the median serum LBP level of the sensitized group was significantly higher than that of the non-sensitized group that was not sensitized to the food allergen. Therefore, the expression level of LBP or the expression level of the gene encoding LBP is measured from the sample isolated from the individual, and when the expression level is higher than the expression level of the gene encoding LBP or LBP of the normal control sample, the food allergen It can be judged as being sensitized. In addition, it can be determined that the subject is at risk of developing a food allergy.

An overlapping description of the agent capable of measuring the expression level of the lipopolysaccharide-binding protein or a gene encoding the same will be omitted.

Another aspect is a screening method for food allergy antigen sensitization prevention or food allergy treatment,

processing the candidate material in the isolated biological sample; and

When the expression level of the lipopolysaccharide-binding protein or the gene encoding the lipopolysaccharide-binding protein in the biological sample is reduced compared to the control group not treated with the candidate substance, the candidate substance is selected as a food allergy antigen sensitization prevention or food allergy treatment agent It provides a screening method comprising the steps.

As used herein, the term "candidate material" may be an individual nucleic acid, protein, other extract or natural product, or compound, which is estimated to have the potential for food allergy prevention or treatment, or randomly selected according to a conventional selection method.

As used herein, the term "control group" is an isolated cell, isolated tissue, or animal other than a human that has not been treated with the candidate substance, and is an isolated cell, isolated tissue, or human belonging to the group treated with the candidate substance in a parallel relationship. animals except for

The composition according to an aspect may simply and accurately predict the risk of developing a food allergy based on the level of lipopolysaccharide-binding protein (LBP). In particular, since it is possible to predict in advance the risk of developing food allergy in subjects who have no symptoms or symptoms of food allergy, it is expected to be useful not only for effective prevention and management of food allergy but also for establishing individual treatment directions.

1 is a diagram showing the serum LBP levels of non-sensitized (n = 107), single-sensitized (n = 160) and multi-sensitized (n = 89) children.

2 shows the correlation between serum LBP levels and sensitization to individual allergens. The correlations were expressed as adjusted Odds Ratios for gender, age, body mass index z-score, parental asthma, allergic rhinitis, atopic dermatitis and household income (95% confidence interval).

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these examples.

Experimental method

Target

From June to July 2015, 356 children aged 5-15 years (average: 9.6 years old, 95% CI: 9.4-9.8 years old) from three kindergartens and six elementary schools in Seongnam, Gyeonggi-do, Korea were included. Demographic characteristics and allergic status were investigated by distributing structured questionnaires to the parents of the subjects, and the subjects reported obvious signs of infection and/or symptoms such as fever, cough, runny nose, sputum, vomiting and/or diarrhea before or at the time of enrollment. Children shown were excluded.

Specifically, the presence or absence of food allergy signs and/or symptoms in subjects and the diagnosis of allergic diseases were determined using the modified Korean version of the ISAAC. Allergic disease was diagnosed by the presence or absence of related symptoms in the past 12 months. In addition, the questionnaire was used to investigate clinical factors that may affect serum lipopolysaccharide-binding protein (LBP) levels. BMI (Body Mass Index)-z scores were based on age and gender standardized measures of children according to World Health Organization (WHO) growth standards.

Lipopolysaccharide-binding protein (LBP)

Serum concentration of LBP was performed using a commercially available Enzyme-linked Immunosorbent Assay (ELISA) kit (Duoset, R & D Systems, Minneapolis, MN, USA) in duplicate according to the manufacturer's instructions. Before analysis, serum samples were diluted 1000-fold and used. No significant cross-reactivity or interference with LBP was observed. LBP levels at A450 nm were assessed using an Infinite 2000 Pro Multimode Plate Reader (Tecan, Vienna, VA, USA).

skin prick test

To evaluate allergic sensitization, extract and control solutions of Lofarma (Milan, Italy) were used to use 6 common atmospheric allergens (2 dust mites [ Dermatophagoides (D.) farinae and D. pteronyssinus ]; 3 tree pollen [Own work]) wood, oak, elm] and Japanese hop; 10 types of plant foods (apples, peaches, kiwis, oranges, tomatoes, strawberries, celery, peanuts, walnuts, wheat); and 6 types of type I food allergens (eggs) , milk, cod, pork, mussels, shrimp) were subjected to Skin Prick Test (SPT) In case of at least one positive reaction to an allergen (wheal diameter > 3 mm) Subjects were considered "sensitized".

All subjects were either nonsensitized, monosensitized (positive SPT to single antigen or multiple cross-reactive antigens without other positive tests) or polysensitized (positive SPT to antigens of different classes). ) were classified.

Determination of total eosinophil count and 25-OH vitamin D

Blood samples were used to determine total eosinophil count (TEC) and serum vitamin D (25-hydroxyvitamin D or 25-OH vitamin D: 25[OH]D) concentrations. TEC was measured using an automated analyzer (Phadia AB, Uppsala, Sweden) and serum 25[OH]D levels were measured using a LIASON Chemiluminescence Immunoassay Analyzer (DiaSorin, Stillwater, Minnesota). Subjects were divided into two groups according to TEC levels (<4% or ≥4%) and three groups according to serum 25[OH]D levels (<20.0 ng/mL, deficient, 20.0-29.9 ng/mL, deficient; or ≧30.0 ng/mL, sufficient).

statistical analysis

Normality of the LBP level distribution was tested using the Kolmogorov-Smirnov test, and the results were expressed as the median (interquartile range [IQR]). The significance of differences between continuous variables was assessed using Mann-Whitney U test, Student's t -test, or ANOVA as appropriate. Categorical variables are expressed as absolute numbers and ratios. The significance of the ratio difference was assessed using a chi-square test. Correlation analysis by Pearson or Spearman was used to determine associations between serum LBP levels and various clinical factors. For multiple comparisons, Holm's method was used.

Partial correlations between factors obtained from structured questionnaires and LBP among sub-directory data constituting questionnaire responses were analyzed. Variables were included in the model if the P value was less than 0.1 or constituted a priori confounding factors (gender, age, and BMI z-score) in the multivariate analysis. According to partial correlation analysis, it was confirmed that household income, breastfeeding status, number of floors, and type of residence were significantly correlated with LBP ( P < 0.1). Generalized linear regression analysis using logit function was performed to identify factors independently and significantly correlated with allergic disease and allergic susceptibility. The analysis results were determined after adjusting for gender, age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis, household income, breastfeeding, number of floors and residence type. in each model. All statistical analyzes were performed using IBM SPSS Statistics ver 25.0 (IBM Co., Armonk, NY, USA). All statistical analyzes were performed with a two-tailed test and P < 0.05 was considered as a significant value.

Experiment result

Example 1. Characteristics of Subject Subjects

The demographic and clinical characteristics of 356 people who participated in the experiment are shown in Table 1 below.

characteristic	number of people	%
gender
other	177	49.7
female	179	50.3
BMI
Obesity (≥95 percent)	9	2.5
overweight (85-95 percent)	34	9.6
Normal weight group (<85 percent)	313	87.9
allergic disease
Asthma	19	5.3
Allergic rhinitis	167	47.2
Atopic dermatitis	80	22.5
25-OH vitamin D (25-hydroxyvitamin D)
normal	11	3.1
Insufficiency	172	48.3
Deficiency	173	48.6
Total eosinophil count
<4%	219	61.5
≥4%	137	38.5
Atopic sensitization
Nonsensitization	107	30.1
Monosensitization	160	44.9
Polysensitization	89	25.0

As shown in Table 1, a total of 160 (44.9%) subjects were single-sensitized, 89 (25.0%) multi-sensitized, and 107 (30.1%) non-sensitized subjects. A total of 233 children (65.4%) were sensitized to inhaled allergens such as house dust mites, 45 children (12.6%) were sensitized to plant food allergens, and type I food allergens (shrimp, pork, mussels, eggs, A total of 22 children (6.2%) were sensitized to cod, milk).

Example 2. Allergic sensitization and LBP

serum LBP levels; And the results of analyzing the correlation with continuous variables or categorical variables are shown in FIG. 1 and Table 2 below.

		asthma			allergic rhinitis			atopic dermatitis			Allergen sensitization
continuous variable		OR (95% CI)	P value		OR (95% CI)	P value		OR (95% CI)	P value		OR (95% CI)	P value
LBP	crude	1.013 (0.963-1.065)	0.615		1.027 (1.003-1.052)	0.028		0.971 (0.943-1.000)	0.049		1.052 (1.020-1.085)	0.001
	Adjusted †	0.985 (0.922-1.052)	0.644		1.012 (0.982- 1.043)	0.438		0.984 (0.952-1.017)	0.350		1.037 (1.003-10.72)	0.033
	Adjusted ‡	0.974 (0.913-1.040)	0.435		1.012 (0.983-1.043)	0.416		0.993 (0.962-1.026)	0.691		1.040 (1.005-1.075)	0.023
	Adjusted §	0.996 (0.931-1.066)	0.916		1.015 (0.985-1.045)	0.334		0.991 (0.959-1.024)	0.599		1.041 (1.007-1.076)	0.016
categorical variable		OR (95% CI)	P value		OR (95% CI)	P value		OR (95% CI)	P value		OR (95% CI)	P value
LBP	1 Q †	Ref			Ref			Ref			Ref
	2 Q	1.307 (0.173-9.845)	0.795		1.021 (0.466-2.238)	0.959		0.480 (0.199-1.155)	0.101		1.391 (0.665-2.910)	0.381
	3 Q	1.749 (0.239-12.819)	0.582		1.715 (0.786-3.742)	0.176		0.556 (0.236-1.311)	0.180		2.744 (1.217-6.185)	0.015
	4 Q	1.432 (0.191-10.760)	0.727		1.974 (0.917-4.246)	0.082		0.855 (0.384-1.906)	0.702		3.599 (1.576-8.222)	0.002

- Results for 356 subjects (19 asthma, 167 allergic rhinitis, 80 atopic dermatitis, 249 allergen sensitization), OR was calculated using generalized linear regression model with logit function (CI, confidence) Interval; OR, Odds Ratio; Adjusted Odds Ratio; LBP, Lipopolysaccharide-binding protein). Allergen sensitization is considered sensitized when one or more positive reactions (rash diameter > 3 mm) to all allergens of the SPT.

- † Multivariate logistic regression analysis (Model I) adjusted for gender, age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis and household income.

- ‡ Multivariate logistic regression analysis adjusted for gender, age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis and residence type (Model II).

- § Multivariate logistic regression analysis adjusted for gender, age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis and number of residence floors (Model III).

Referring to FIG. 1 , it was confirmed that the median serum LBP level of the sensitized group (20.3 [IQR: 14.8-25.8]) was significantly higher than that of the non-sensitized group (25.5 ng/mL [IQR: 20.3-30.7]). can be ( P = 0.308). There were no significant differences between the single-sensitized and multi-sensitized groups.

Referring to Table 2, LBP levels were positively correlated with higher risk of allergic sensitization (OR = 1.052, 95% CI = 1.020-1.085, P = 0.001). It remained significant even after exclusion of confounding factors (adjusted OR [aOR]=1.041, 95% CI=1.007-1.076, P =0.016). In addition, it can be seen that children with LBP levels in the highest quartile have a significantly increased risk of allergen sensitization compared to children in the lowest quartile (aOR=3.599, 95% CI=1.576). -8.222, P = 0.002).

Furthermore, it can be seen that children with self-reported allergic rhinitis and atopic dermatitis had significantly higher median serum LBP than controls ( P = 0.028 and P = 0.049, respectively). However, the correlation no longer showed a significant difference after adjusting for gender, age, BMI z-score, type of residence or number of floors. The children with asthma and the control group had similar LBP levels.

Additionally, serum LBP levels; And the results of analyzing the correlation with specific blood components are shown in Table 3 below.

blood component	LBP (ng/mL)
	Crude β (SE)	P value		Adjusted β (SE)	P value
25-OH Vitamin D 3 (ng/mL)	-0.003 (-0.011 to 0.005)	0.429		-0.002 (-0.011 to 0.007)	0.617
Hemoglobin, g/L	0.000 (-0.052 ~ 0.051)	0.986		-0.014 (-0.070 to 0.042)	0.621
White blood cell count, Х10 9 /L	0.015 (-0.015 to 0.044)	0.338		0.010 (-0.020 ~ 0.040)	0.499
Neutrophil count, Х10 9 /L	-0.001 (-0.006 to 0.003)	0.561		-0.003 (-0.008 to 0.002)	0.295
Eosinophil count, %	-0.009 (-0.023 to 0.006)	0.239		-0.010 (-0.025 to 0.005)	0.213

Referring to Table 3, the serum LBP level did not show a significant correlation with 25[OH]D, hemoglobin level, or the number of leukocytes, neutrophils or total eosinophils.

Example 3. Types of Allergens and LBP

serum LBP levels; And the results of analyzing the correlation with each type of allergen sensitization are shown in Table 4 below.

dependent variable (allergens)	Lipopolysaccharide-binding protein
	OR (95% CI)	P value		aOR (95% CI) *	P value		aOR (95% CI) †	P value
non-sensitizing ‡	Ref			Ref			Ref
single-sensitization	1.019 (0.985 ~ 1.055)	0.274		1.015 (0.979 to 1.052)	0.419		1.020 (0.986 to 1.056)	1.000
multi-sensitization	1.100 (1.055 to 1.147)	<0.001		1.110 (1.059 to 1.164)	<0.001		1.105 (1.056 to 1.156)	0.002
allergen
any inhaled allergen	1.017 (0.990 to 1.045)	0.227		1.010 (0.982 to 1.040)	0.467		1.017 (0.989 ~ 1.046)	1.000
any food allergen	1.363 (1.156 to 1.607)	<0.001		1.080 (1.029 ~ 1.133)	<0.001		1.326 (1.206 to 1.457)	<0.001

- Allergen sensitization is considered sensitized if one or more positive reactions (rash diameter > 3 mm) to all allergens of the SPT are observed.

- * Multivariate logistic regression adjusted for gender (male), age, BMI z-score and household income.

- † Multivariate logistic regression adjusted for gender (male), age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis and household income. Holm's method is used for multiple comparisons.

- ‡ P values for correlation between serum LBP levels and allergen sensitization (non-sensitized, single-sensitized and multi-sensitized) were analyzed by polynomial regression.

- § Multivariate logistic regression analysis adjusted for gender, age, BMI z-score, parental asthma, allergic rhinitis, atopic dermatitis and number of residence floors (Model III).

Referring to Table 4, it can be seen that the multi-sensitized children had significantly higher median serum LBP than the non-sensitized children (OR = 1.10, 95% CI = 1.055-1.147, Po0.001 ). The correlation remained significant even after exclusion of a priori confounding factors (aOR = 1.110, 95% CI = 1.059-1.164, Po0.001 ).

LBP levels were sensitized to all food allergens (aOR = 1.080, 95% CI = 1.029-1.133, P = 0.002), as well as sensitization to plant food allergens (aOR = 1.256, P = 0.001) and to type I food allergens. It was confirmed that it had a significant correlation with all of the sensitization factors (aOR = 1.128, P = 0.043). On the other hand, the rise in LBP level did not show a significant correlation with sensitivity to inhalation allergens including house dust mites.

Furthermore, the results of analyzing the correlation between serum LBP levels and individual food allergen sensitization are shown in FIG. 2 .

Referring to Figure 2, for plant allergens, LBP levels were found in kiwi (aOR = 1.014, P = 0.018), orange (aOR = 1.005, P = 0.039), celery (aOR = 1.064, P <0.001), peanut (aOR = 1.054, 95% CI = 1.135-1.223, P = 0.001), walnut (aOR = 1.043, 95% CI = 1.126-1.216, P = 0.003) and wheat (aOR = 1.04, 95% CI = 1.151-1.274, P = 0.007), it can be seen that there is a significant correlation. In addition, LBP levels were found in shrimp (aOR = 1.034, 95% CI = 1.115-1.204, P = 0.005), mussels (aOR = 1.013, 95% CI = 1.099-1.194, P = 0.024) and eggs (aOR = 1.011, P ). = 0.027) and also showed a significant correlation.

Example 4. Skin prick test

The results of the skin prick test are shown in Table 5 below.

	LBP
	Rho	P value
atmospheric allergens
Dermatophagoides farinae	0.103	0.054
Dermatophagoides pteronyssinus	0.087	0.102
birch	0.088	0.097
kind of oak	0.021	0.700
elm	0.099	0.062
Ginseng vine (Japanese hop)	0.143	0.007
plant food allergens
apologize	0.167	0.002
peach	0.186	<0.001
Kiwi	0.157	0.003
Orange	0.178	0.001
tomato	0.136	0.010
Strawberry	0.094	0.077
salary	0.287	<0.001
peanut	0.260	<0.001
Walnut	0.273	<0.001
wheat	0.215	<0.001
Type I food allergens
egg	0.141	0.008
milk	0.125	0.018
Daegu (Cod)	0.066	0.220
Pork	0.003	0.954
mussel	0.149	0.005
shrimp	0.224	<0.001
Histamine (positive control)	0.030	0.575
Saline (negative control)	0.051	0.342

Referring to Table 5, serum LBP levels and rash diameters for almost all food allergens (apples, peaches, kiwis, oranges, tomatoes, celery, peanuts, walnuts, wheat, eggs, milk, mussels, shrimp) were significantly higher. It can be seen that there is a correlation.

Through the above results, it was confirmed that by measuring the LBP level, it was possible to differentiate a subject sensitized to a food allergen from a subject without symptoms of a food allergy. Therefore, the LBP level can be usefully used for predicting the risk of developing a food allergy and preventing food allergen sensitization, and furthermore, by using it for the development of intestinal preparations, foods, or biologics containing anti-LBP antibodies to LBP, food allergens It is expected that it can be used for prevention or treatment of sensitization and further food allergy.

The above description is for illustration, and those of ordinary skill in the art to which the present invention pertains will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (12)

1. A composition for predicting the risk of developing a food allergy, comprising an agent capable of measuring the expression level of a lipopolysaccharide-binding protein (LBP) or a gene encoding the same.
2. The method according to claim 1,

   The agent capable of measuring the expression level of the lipopolysaccharide-binding protein is selected from the group consisting of an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the lipopolysaccharide-binding protein. The composition comprising one or more selected.
3. The method according to claim 1,

   The agent capable of measuring the expression level of the gene encoding the lipopolysaccharide-binding protein comprises at least one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the gene. .
4. A kit for predicting the risk of developing a food allergy comprising the composition according to claim 1.
5. 5. The method according to claim 4,

   The kit is a RT-PCR (Reverse transcription polymerase chain reaction) kit, a DNA chip kit, an ELISA (Enzyme-linked immunosorbent assay) kit, a protein chip kit or a rapid kit, the kit.
6. 5. The method according to claim 4,

   The kit, wherein the prediction is a prediction of the risk of developing a food allergy in a subject without food allergy symptoms or signs.
7. A method of providing information for predicting the risk of developing a food allergy, comprising measuring the expression level of a lipopolysaccharide-binding protein or a gene encoding the same from the isolated biological sample.
8. 8. The method of claim 7,

   Wherein the biological sample is selected from the group consisting of blood, serum and plasma.
9. 8. The method of claim 7,

   The agent capable of measuring the expression level of the lipopolysaccharide-binding protein is selected from the group consisting of an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the lipopolysaccharide-binding protein. The method of providing information, including one or more selected.
10. 8. The method of claim 7,

    The agent capable of measuring the expression level of the gene encoding the lipopolysaccharide-binding protein comprises at least one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the gene. How to provide.
11. As a screening method for food allergy antigen sensitization prevention or food allergy treatment agent,

    processing the candidate material in the isolated biological sample; and

    When the expression level of the lipopolysaccharide-binding protein or a gene encoding the same in the biological sample is decreased compared to the control group not treated with the candidate substance, the candidate substance is selected as a food allergy antigen sensitization prevention or food allergy treatment agent A screening method comprising the steps.
12. 12. The method of claim 11,

    The biological sample is selected from the group consisting of blood, serum and plasma.

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