WO2004002520A1 - Method and kit for the tolerance induction to allergy-causing substances, and the tolerance achievment against the substances thereby - Google Patents

Method and kit for the tolerance induction to allergy-causing substances, and the tolerance achievment against the substances thereby Download PDF

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Publication number
WO2004002520A1
WO2004002520A1 PCT/KR2003/001270 KR0301270W WO2004002520A1 WO 2004002520 A1 WO2004002520 A1 WO 2004002520A1 KR 0301270 W KR0301270 W KR 0301270W WO 2004002520 A1 WO2004002520 A1 WO 2004002520A1
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Prior art keywords
tolerance
allergens
substances
allergy
unit
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Application number
PCT/KR2003/001270
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French (fr)
Inventor
Geun-Woong Noh
Original Assignee
Geun-Woong Noh
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Application filed by Geun-Woong Noh filed Critical Geun-Woong Noh
Priority to AU2003244262A priority Critical patent/AU2003244262A1/en
Publication of WO2004002520A1 publication Critical patent/WO2004002520A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/21—Interferons [IFN]
    • A61K38/217—IFN-gamma
    • A—HUMAN NECESSITIES
    • A63—SPORTS; GAMES; AMUSEMENTS
    • A63B—APPARATUS FOR PHYSICAL TRAINING, GYMNASTICS, SWIMMING, CLIMBING, OR FENCING; BALL GAMES; TRAINING EQUIPMENT
    • A63B57/00—Golfing accessories
    • A63B57/10—Golf tees
    • A—HUMAN NECESSITIES
    • A63—SPORTS; GAMES; AMUSEMENTS
    • A63B—APPARATUS FOR PHYSICAL TRAINING, GYMNASTICS, SWIMMING, CLIMBING, OR FENCING; BALL GAMES; TRAINING EQUIPMENT
    • A63B57/00—Golfing accessories
    • A63B57/20—Holders, e.g. of tees or of balls
    • A63B57/203—Tee holders
    • A—HUMAN NECESSITIES
    • A63—SPORTS; GAMES; AMUSEMENTS
    • A63B—APPARATUS FOR PHYSICAL TRAINING, GYMNASTICS, SWIMMING, CLIMBING, OR FENCING; BALL GAMES; TRAINING EQUIPMENT
    • A63B69/00—Training appliances or apparatus for special sports
    • A63B69/36—Training appliances or apparatus for special sports for golf

Definitions

  • the present invention relates to a method and kit for inducing tolerance to certain allergens, and a method for acquiring the tolerance using them.
  • the human immune system produces antibodies to remove antigens, such as foreign substances. Allergy attributes to an antigen-antibody response to certain substances that are oversensitive in any persons but harmless to most persons. IgE is known to be a main antibody mediating the allergic reaction.
  • Diseases caused by allergy include respiratory asthma, allergic nasitis, allergic conjunctivitis, atopic dermatitis and contact dermatitis.
  • the exact mechanism of allergic diseases is not yet established, and there is little or no fundamental treatment methods or solutions against the allergic diseases. Particularly in the case of foods, food allergy becomes a serious problem in that it greatly interferes with daily life.
  • Treatment methods that are widely used for allergic diseases include allergen avoidance, drug therapy and tolerance acquirement.
  • the allergen avoidance is a method where certain substances serving as an allergy-causing source are removed from the living of patients such that the patients are not exposed to the allergy-causing substances.
  • it is a negative solution in that it is not a fundamental method capable of treating allergy.
  • food avoidance there is a problem in that a growing child who exhibits allergy against relevant foods cannot safely ingest the foods although they have good nutrients.
  • the foods are essential foods that are always required in usual dietary life, persons should be continued to suffer from allergic diseases.
  • the drug therapy is a method where chemicals capable of inhibiting allergic response, such as anti-histamines, are administered.
  • This method is only a temporary treatment method since allergy frequently recurs. Also, this method often causes serious side effects due to the use of chemicals.
  • the tolerance acquirement is based on a theory where immune response is regulated by the balance between a helper T cell 1 (hereinafter, referred to as 'Thl cell') and a helper T cell 2 (hereinafter, referred to as 'Th2 cell').
  • 'Thl cell' helper T cell 1
  • 'Th2 cell' helper T cell 2
  • the . Thl cell is mainly involved in inflammatory reaction, and the Th2 cell in allergic reaction.
  • Interferon-gamma (hereinafter, referred to as IFN- ⁇ secreted by the Thl cell inhibits the growth of cells to inhibit the synthesis of interleukin-4 as a cell regulator involving in allergic reaction.
  • IFN- ⁇ secreted by the Thl cell inhibits the growth of cells to inhibit the synthesis of interleukin-4 as a cell regulator involving in allergic reaction.
  • IFN- ⁇ secreted by the Thl cell inhibits the growth of cells to inhibit the synthesis of interleukin-4 as a cell regulator involving in allergic reaction.
  • IFN- ⁇ is used to form an in vivo Thl cell predominant state and to induce tolerance to allergy-causing substances (hereinafter, also referred to as allergens), thereby acquiring tolerance to the allergens.
  • allergens allergy-causing substances
  • a tolerance induction method and kit that allow tolerance to certain allergens to be acquired lastingly using LFN- ⁇ , as well as a method for acquiring the tolerance using them.
  • an object of the present invention is to provide a method and kit for inducing tolerance to certain allergens using LFN- ⁇ , and a method for acquiring the tolerance using them.
  • the present invention relates to a method and kit for inducing tolerance to certain substances causing allergy, and a method for acquiring the tolerance using them.
  • the method for acquiring tolerance to allergy-causing substances comprises the steps of: confirmation of allergic diseases; maximum avoidance of allergens; allergen challenge; minimum avoidance of allergens; induction of tolerance using the inventive tolerance induction method and kit; and tolerance maintenance.
  • the allergy-causing substances can be all substances causing allergy, such as house dust mite, pollen, animal hair, mold, and certain chemicals, in addition to foods, such as milk, eggs, soybeans, wheat flour, pork, fowl, rice, and garlic.
  • FIG. 1 schematically shows a process for acquiring tolerance to milk according to the tolerance acquirement method of the present invention.
  • each step will be described on the basis of FIG. 1.
  • Food allergy will be described as an example.
  • allergy testing there can be used all the conventional allergy tests, including skin prick test, allergy patch test, blood eosinophil count, blood IgE concentration, and the identification of IgE specific to certain foods, as well as a detailed examination on the history of allergy.
  • the step of allergen challenge is then performed.
  • DBPCFC double-blind placebo-controlled food challenge
  • OFFC open food challenge
  • the double-blind placebo-controlled food challenge is performed with foods to be tested on the causing or non-causing of allergy, as well as foods causing no problems (i.e., placebo).
  • the two kinds of foods must be indistinguishable from each other, and during the test, both a doctor and a patient cannot be aware of the kind of foods tested.
  • the placebo is confirmed as not causing allergy while allergy reaction caused by specific foods is shown, the foods are evaluated as allergy- causing foods.
  • the open food challenge is performed in a state where a patient is aware that any foods are administered.
  • the open food challenge is conducted for a patient who shows allergic reaction even to placebo in the double-blind placebo-controlled challenge.
  • the step of tolerance induction is performed using the tolerance induction method and kit of the present invention.
  • the tolerance induction step consists of: a first sub-step for tolerance induction where an IFN- ⁇ -containing composition and milk are administered together; and a second sub-step for confirmation of tolerance induction where only milk is administered.
  • each of the first and second induction sub- steps is preferably carried out for 7 days.
  • milk is administered in one unit twice a day for at least 7 days to confirm if tolerance to milk was acquired.
  • a specific allergic reaction is not shown even if milk is continued to administer to a patient or when the result of the food challenge with milk shows the disappearance of allergy, it is evaluated that tolerance to milk was acquired.
  • the tolerance induction method of the present invention consists of: a first step for tolerance induction where an LFN- ⁇ -containing composition and milk are administered together; and a second step for the confirmation of tolerance induction where only milk is administered.
  • each of the first step for tolerance induction and the second step for the confirmation of tolerance induction is preferably performed for 7 days.
  • the first step for tolerance induction comprises administering an LNF- ⁇ -containing composition to a patient with allergy against specific substances in a specific unit per body surface area and at the same time, administering the substances while gradually increasing the amount of the substances.
  • one dose of an LNF- ⁇ -containing composition is administered, and then immediately, allergy-causing substances is administered in 1/4 unit a day, at days 1 and 2.
  • one dose of the LNF- ⁇ -containing composition is administered, and then immediately, the allergy- causing substances are administered in 1/2 unit a day.
  • one dose of the LNF- ⁇ -containing composition is administered, and then immediately, the allergy- causing substances are administered in one unit a day.
  • one dose of the INF- ⁇ -containing composition is administered, and then immediately, the allergy- causing substances are administered in 1.5 units.
  • the second step for the confirmation of tolerance induction comprises administering only allergy- causing substances while gradually increasing the amount of the substances as in the tolerance induction step without administering the LFN- ⁇ -containing composition.
  • allergy- causing substances are administered in 1/4 unit a day at days 1 and 2.
  • the allergens are administered in 1/2 unit a day, and at days 5 and 6, the allergens are administered in one unit a day.
  • the allergens are administered in 1.5 units.
  • the present invention provides a tolerance induction kit allowing tolerance to allergy-causing substances to be acquired.
  • the tolerance induction kit of the present invention consists of a first step for tolerance induction and a second step for the confirmation of tolerance induction, in the same manner as the tolerance induction method of the present invention.
  • the tolerance induction kit of the present invention comprises an INF- ⁇ - containing composition of a quantity to be administered for the first tolerance induction step of a given period, allergy-causing substances of a quantity to be administered together with the composition, and allergy-causing substances of a quantity to be administered alone in the second step for the confirmation of tolerance induction.
  • the tolerance induction kit of the present invention consists of a first container unit containing the LNF- ⁇ -containing composition and a second container unit containing the allergens.
  • it may consist of a first container unit containing the composition and allergens to be used in the first step for toleration induction, and a second container unit containing the allergens to be used in the second step for the confirmation of tolerance induction.
  • each of the LFN- ⁇ -containing composition and the allergens is contained in one container or several different small containers.
  • the respective container units may have several small containers according to their arrangement method.
  • the tolerance induction kit of the present invention comprises the LFN- ⁇ -containing composition of a quantity to be administered in the first step for tolerance induction for 7 days, and the allergy-causing substances of a quantity to be administered therewith, and also the allergens of a quantity to be administered in the second step for the confirmation of tolerance induction for 7 days.
  • the construction of the kit will be described.
  • the first containers may contain seven doses of the LFN- ⁇ -containing composition
  • the second container may contain 10 units of the allergy-causing substances.
  • the first container unit may consist of one small container containing seven doses of the composition, or seven small containers each containing one dose of the composition.
  • the second container unit may consist of one small container containing 10 units of the allergy-causing substances, or it may consist of 4 containers each containing 1/4 unit of the allergens, 4 containers each containing 1/2 unit of the allergens, 4 containers each containing one unit of the allergens and 2 containers each containing 1.5 units of the allergens.
  • the first container unit may contain seven doses of the IFN- ⁇ - containing composition and 5 units of the allergy-causing substances
  • the second container unit may contain 5 units of the allergy-causing substances.
  • the first container unit may consist of one small container containing seven doses of the composition and one small container containing 5 units of the allergens, or it may consist of seven small containers each containing one dose of the composition, and two containers each containing 1/4 unit of the allergens, two containers each containing 1/2 units of the allergens, two containers each containing one unit of the allergens, and one containing 1.5 unit of the allergens.
  • the second container unit may consist of one small container containing 5 units of the allergens, or it may consist of two containers each containing 1/4 unit of the allergens, two containers each containing 1/2 unit of the allergens, two containers each containing one unit of the allergens, and one container containing 1.5 units of the allergens.
  • the tolerance induction kit of the present invention may comprise a syringe required for administration, and directions showing a method for the administration of each component, etc.
  • the allergy-causing substances are foods, they can be provided in a form that can be easily administered, such as powder, cake and the like.
  • the tolerance induction kit of the present invention may comprise one or more allergens.
  • the kit may comprise the LFN- ⁇ -containing composition of a quantity to be used in two tolerance induction processes, and milk and soybeans of quantities to be used in each of the first step for tolerance induction and the second step for the confirmation of tolerance induction.
  • the tolerance induction kit of the present invention may be used alone or in combination with hormone therapy, chemical therapy and other methods of inducing tolerance to allergy using biological response modifiers.
  • the LFN- ⁇ - containing composition may contain IFN- ⁇ , fragments of LFN- ⁇ with Thl cell- stimulating activity, or derivatives with functions similar with these substances, as an active ingredient, and if necessary, other active ingredients.
  • the IFN- ⁇ - containing composition may comprise pharmaceutically suitable, physiologically acceptable adjuvants, in addition to the active ingredient as described above. Examples of such adjuvants include excipients, disintegrants, sweetening agents, binders, coating agents, blowing agents, lubricants, flavoring agents, and solubilizing agents.
  • the IFN- ⁇ - containing composition may be preferably formulated as a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers for administration, in addition to the active ingredient as described above.
  • examples of the pharmaceutically acceptable carriers which can be used in formulating IFN- ⁇ into liquid solution, include saline solution, sterilized water,
  • Ringer's solution buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture thereof.
  • other conventional additives such as antioxidants, buffer solution and bacteriostatics, may also be added.
  • diluents, dispersants, surfactants, binders and lubricants may be further added to formulate LNF- ⁇ as injection formulations, such as aqueous solution, suspension and emulsion, pills, capsules, granules and tablets.
  • injection formulations such as aqueous solution, suspension and emulsion, pills, capsules, granules and tablets.
  • it can be preferably formulated according to diseases or components, using a method described in Remington 's Pharmaceutical Science, Mack Publishing Company, Easton PA, which is a suitable method in the relevant field of art.
  • the IFN- ⁇ - containing composition can be formulated as pharmaceutical formulations, such as granules, powders, coated tablets, tablets, capsules, suppositories, syrup, juice, suspension, emulsion, drops, injectable liquid, and sustained-release preparations.
  • the IFN- ⁇ -containing composition of the present invention is relatively nontoxic.
  • the IFN- ⁇ - containing composition can be administered in the conventional manner via the subcutaneous, intravenous, intraarterial, intraabdominal, intramusclar, intrasternal, percutaneous, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal route.
  • the IFN- ⁇ - containing composition is administered at the amount of 2 x 10 6 U to 4 x 10 6 U per m of body surface area, and preferably 3 x 10 U per m of body surface area.
  • the amount of the composition administered can vary depending on various factors, including the severity of allergic diseases, the kind and content of an active ingredient and other components contained in the composition, the kind of a formulation, and patient's age, weight, general health condition, sex and diet, and administration time, administration route, the secretion % of the composition, administration period, and the kind of drugs used in combination, for administration by subcutaneous injection.
  • the dosage of the allergy-causing substances is expressed in a given unit for each substance.
  • one unit is defined as the average amount of the foods that an adult ingests in eating.
  • one unit is preferably is defined as the amount of 200 g per 1.71 m 2 of body surface area.
  • the induction of tolerance is achieved by a change in immune system caused by the administration of the IFN- ⁇ -containing composition and then the allergy-causing substances. This induction of tolerance is schematically shown in FIG. 2.
  • the IFN- ⁇ -containing composition administered in the first step for tolerance induction serves to convert the immune status of a subject into a Thl cell predominant state.
  • the administered allergens act as an antigen, and the dose of the allergens is gradually increased from small amount, and thus, a normal immune response other than an instant allergic reaction as in the prior art is induced.
  • the second step for the confirmation of tolerance induction only the allergens are administered to accumulate the immune response so that the acquired tolerance to the allergens can be maintained more securely.
  • the tolerance induction method and kit of the present invention allow tolerance to certain allergens to be effectively induced, and since then, allow the tolerance to be maintained lastingly.
  • This effect of tolerance acquirement of the present invention is a characteristic effect shown by only the tolerance induction method of the present invention, other than a residual effect also obtainable by the simple administration of IFN- ⁇ used in the prior art, or an effect of natural tolerance acquirement caused by continuous exposure to the allergens.
  • the tolerance induction method and kit of the present invention allow tolerance to allergens to be positively acquired, and at the same time, allow the acquired tolerance to be maintained lastingly, in significant improvements over the existing method for the inhibition of allergy, which remains at a negative level, such as allergen avoidance, or could not overcome side effects caused by chemical therapy. Accordingly, the present invention will be useful in the field of the prevention and inhibition of allergy.
  • FIG. 1 is a schematic view showing a process for acquiring tolerance to milk using a method and kit for inducing tolerance to allergy according to the present invention
  • FIG. 2 is a schematic view showing a change in immune system according to the administration of an LNF- ⁇ -containing composition and allergens
  • FIG. 3 is a schematic view showing methods for inducing tolerance to milk allergy using methods of the present invention and the prior art;
  • FIG. 4a shows clinical changes occurring when conducting a tolerance induction method of the present invention.
  • FIG. 4b shows clinical severity and an abnormal reaction on the skin according to time, which occur when conducting a tolerance induction method of the present invention.
  • Example 1 Acquirement of tolerance to milk in patients with atopic dermatitis
  • the test was carried out on 250 patients with atopic dermatitis, who visited
  • histamine hydrochloride (Bencard GmbH) that is a commercially available typical allergen was used as a positive control group, and physiological saline and distilled water were used as a negative control group.
  • reaction corresponded to two times or more of the positive control group.
  • the minimum reaction size in the positive reaction was 3 mm.
  • FAST test fluorescent allergo- sorbent-test
  • the patients were subjected to maximum avoidance as follows. All foods, which had been suspected to cause allergy through the history examination, the skin prick test, and the identification of allergen-specific IgE, were eliminated from diets for at least one week and thus avoided, and also foods containing them were strictly avoided.
  • dietary diaries were prepared for all the patients.
  • the respective dietary diaries were analyzed by professional dietitians, nurses, and physicians.
  • a food challenge test was carried out using the double-blind placebo- controlled food challenge in the case of milk, wheat and beans, and using the open food challenge in the case of beef and fowl. After the maximum avoidance of allergens was conducted, when allergic symptoms were clinically surely improved and this condition was lasting for at least two weeks, and when it was confirmed that foods to be tested were avoided from diet, a food challenge test was conducted. Dietary diaries were analyzed by a professional dietitian to analyze the above particulars while the respective patients were maintained at their usual living habit for the largest possible removal of environmental factors.
  • Excipient I consisted of a mixture of 5g of unhulled powder, 5g of glutinous rice powder and 5g of barley powder, and excipient
  • 15g of the excipient powder was mixed with 6g of foods to be challenged, and then administered to the patients.
  • 21g of a powder mixture having the same composition as the excipient powder was administered to the patients.
  • the excipient was administrated in cake form.
  • the patients were subjected to food challenge with the powder mixtures to be used as placebo, i.e., excipient I and excipient II, for three days, thereby confirming if the patients have tolerance to placebo or not.
  • excipient I was used as placebo and excipients in the test.
  • expicient II was administered for three days in order to confirm if the patients have tolerance to excipient II or not, so that whether the excipients can be used or not was determined.
  • the randomization and preparation of the food challenge test were conducted by dietitians for each clinical study unit. All the results of the food challenge test were added up according to a clinical severity scoring system given in Table 2 below.
  • the open food challenge was carried out for the same period as the double- blind placebo-controlled food challenge, and also conducted in such a manner that the test foods are administered while gradually increasing their amount daily, as shown in Table 3.
  • test foods were administered one time one day every morning for 4 days, and the results was evaluated.
  • results was evaluated.
  • one of the following cases was evaluated as a positive reaction: Patients show more than 20% increase in a severity score as compared to that prior to the test, they show an evaluated severity score higher than 100, and inflammation or itching is shown on the skin.
  • the number of foods causing abnormal reaction after food challenge was 2.3
  • tolerance induction methods varying with the groups were carried out.
  • a method of administering only IFN- ⁇ or milk was used, and as a control group, a method of administering placebo was used.
  • the patients of group I were administered with 1/4 unit of milk just following the administration of IFN- ⁇ , at days 1 and 2. At days 3 and 4, 1/2 unit of milk was administered just after administering IFN- ⁇ . At days 5 and 6, one unit of milk was administered just after administering IFN- ⁇ . At day 7, two units of milk were administered after administering IFN- ⁇ .
  • the patients of group II were administered with only IFN- ⁇ throughout the tolerance induction period, and the patients of group III were administered with only milk while gradually increasing the amount of milk according to the same program as group I.
  • the patients of group IV were administered with only placebo.
  • the patients of groups I and III were administered with milk while gradually increasing the amount of milk according to a program, thereby conducting the second step for the confirmation of tolerance induction.
  • IFN- ⁇ for tolerance induction recombinant IFN- ⁇ (specific activity: 2 x 10 6 U/mg), which is commercially available from Intermax gamma Co., was used and injected subcutaneously at the amount of 10 U/m .
  • IFN- ⁇ for tolerance induction
  • recombinant IFN- ⁇ (specific activity: 2 x 10 6 U/mg), which is commercially available from Intermax gamma Co., was used and injected subcutaneously at the amount of 10 U/m .
  • 10 mg/kg of oral acetaminophen was administered at one hour before administration and at 4 hours after administration. In this case, the maximum dosage of acetaminophen was 600 mg.
  • ketotifen/ ⁇ -linoleic acid 1% hydrocortisone and the oral administration of ketotifen/ ⁇ -linoleic acid were accepted from one month before the test start to one month after the test start.
  • the condition of the patients was observed for at least 4 weeks.
  • results are expressed in mean ⁇ standard deviation, and a statistical analysis was carried out according to a Wilcoxon matched-pair signed-ranks test.
  • the tolerance induction method and kit of the present invention and the method for acquiring tolerance using them, advantageously allow tolerance to certain allergens to be effectively induced, and since then, allow the tolerance to be maintained lastingly even upon the ordinary administration of the allergens.
  • the tolerance induction method and kit of the present invention and the method for acquiring tolerance using them, will be useful in the field of the prevention and inhibition of allergy.

Abstract

The present invention relates to a method and kit for inducing tolerance to certain substances causing allergy, and a method for acquiring the tolerance using them. The tolerance induction method and kit comprises a first step for the induction of tolerance and a second step for the confirmation of tolerance induction. In the tolerance induction method, the first step for tolerance induction comprises administering the IFN-y-containing composition to the patient with allergy to certain substances in a given unit per body surface area, and at the same time, administering the substances while gradually increasing the amount of the substances. The second step for the confirmation of tolerance induction comprises administering only the allergens during a given period while increasing the amount of the allergens as in the first step for tolerance induction, without administering the IFN-y-containing composition. The tolerance induction kit comprises an INF-y-containing composition of a quantity to be administered in the first step for the induction of tolerance, allergy-causing substances of a quantity to be administered together with the INF-y-containing composition in the first step for tolerance induction, and allergy-causing substances of a quantity to be administered in a second step for the confirmation of tolerance induction. The tolerance induction method and kit of the present invention, and the method for acquiring tolerance using them, advantageously allow tolerance to certain allergens to be effectively induced, and since then, allow the tolerance to be maintained lastingly even upon the ordinary administration of the allergens.

Description

METHOD AND KIT FOR THE TOLERANCE INDUCTION TO
ALLERGY-CAUSING SUBSTANCES, AND THE TOLERANCE
ACHIEVMENT AGAINST THE SUBSTANCES THEREBY
Technical Field
The present invention relates to a method and kit for inducing tolerance to certain allergens, and a method for acquiring the tolerance using them.
Background Art
The human immune system produces antibodies to remove antigens, such as foreign substances. Allergy attributes to an antigen-antibody response to certain substances that are oversensitive in any persons but harmless to most persons. IgE is known to be a main antibody mediating the allergic reaction. Diseases caused by allergy include respiratory asthma, allergic nasitis, allergic conjunctivitis, atopic dermatitis and contact dermatitis. The exact mechanism of allergic diseases is not yet established, and there is little or no fundamental treatment methods or solutions against the allergic diseases. Particularly in the case of foods, food allergy becomes a serious problem in that it greatly interferes with daily life.
Treatment methods that are widely used for allergic diseases include allergen avoidance, drug therapy and tolerance acquirement.
The allergen avoidance is a method where certain substances serving as an allergy-causing source are removed from the living of patients such that the patients are not exposed to the allergy-causing substances. However, it is a negative solution in that it is not a fundamental method capable of treating allergy. Particularly, in food avoidance, there is a problem in that a growing child who exhibits allergy against relevant foods cannot safely ingest the foods although they have good nutrients. Furthermore, if the foods are essential foods that are always required in usual dietary life, persons should be continued to suffer from allergic diseases.
Meanwhile, the drug therapy is a method where chemicals capable of inhibiting allergic response, such as anti-histamines, are administered. This method is only a temporary treatment method since allergy frequently recurs. Also, this method often causes serious side effects due to the use of chemicals.
The tolerance acquirement is based on a theory where immune response is regulated by the balance between a helper T cell 1 (hereinafter, referred to as 'Thl cell') and a helper T cell 2 (hereinafter, referred to as 'Th2 cell').
In the immune system, the. Thl cell is mainly involved in inflammatory reaction, and the Th2 cell in allergic reaction. Interferon-gamma (hereinafter, referred to as IFN-γ secreted by the Thl cell inhibits the growth of cells to inhibit the synthesis of interleukin-4 as a cell regulator involving in allergic reaction. Thus, when an LFN-γ and Thl cell predominant state is achieved, allergic symptoms are reduced and normal immune response occurs. However, if the regulation between the Thl cell and the Th2 cell is broken such that IFN-γ is relatively and absolutely insufficient, allergic symptoms will be caused. Considering this point, there is made an attempt in which IFN-γ is used to form an in vivo Thl cell predominant state and to induce tolerance to allergy-causing substances (hereinafter, also referred to as allergens), thereby acquiring tolerance to the allergens. Although it was recently reported that desensitization therapy with LFN-γ was conducted to induce tolerance to house dust mite, many studies still need to be carried out in order to examine if the tolerance lasts. Moreover, attempts to induce tolerance to certain foods causing allergy by means of IFN-γ were often conducted, but there have been no results showing that the tolerance was effectively acquired. Accordingly, there are demands for inventions of a tolerance induction method and kit that allow tolerance to certain allergens to be acquired lastingly using LFN-γ, as well as a method for acquiring the tolerance using them.
Disclosure of Invention
Therefore, the present invention has been made to satisfy the above demands, and an object of the present invention is to provide a method and kit for inducing tolerance to certain allergens using LFN-γ, and a method for acquiring the tolerance using them. The present invention relates to a method and kit for inducing tolerance to certain substances causing allergy, and a method for acquiring the tolerance using them.
The method for acquiring tolerance to allergy-causing substances (hereinafter, also referred to as allergens) according to the present invention comprises the steps of: confirmation of allergic diseases; maximum avoidance of allergens; allergen challenge; minimum avoidance of allergens; induction of tolerance using the inventive tolerance induction method and kit; and tolerance maintenance.
In the tolerance acquirement method according to the present invention, the allergy-causing substances can be all substances causing allergy, such as house dust mite, pollen, animal hair, mold, and certain chemicals, in addition to foods, such as milk, eggs, soybeans, wheat flour, pork, fowl, rice, and garlic.
FIG. 1 schematically shows a process for acquiring tolerance to milk according to the tolerance acquirement method of the present invention. Hereinafter, each step will be described on the basis of FIG. 1. Food allergy will be described as an example.
In the step of confirmation of allergy disease, general allergy testing on a patient with suspected allergy is performed. In the allergy testing, there can be used all the conventional allergy tests, including skin prick test, allergy patch test, blood eosinophil count, blood IgE concentration, and the identification of IgE specific to certain foods, as well as a detailed examination on the history of allergy.
In the step of maximum avoidance of allergens, all factors suspected to cause allergy through the history examination, the skin prick test and the blood test, etc. (PI, P2 and P3 for foods), are eliminated from diets. In this case, in order to prevent unbalanced nutrition caused by the avoidance of these foods, substitute foods (NI, N2 and N3) that have similar nutrition with these foods while not causing allergy are administered.
After the maximum avoidance step, if atopic dermatitis symptoms are improved and a state where this condition no loner becomes worse is continued for at least one week, the step of allergen challenge is then performed.
Currently, the confirmation of allergy is possible only by an allergen challenge test, and in the case of foods, a food challenge test is performed for the purpose of confirming allergy-causing foods such that other foods confirmed as not causing allergy can be freely administered. The food challenge test is divided into a double-blind placebo-controlled food challenge (DBPCFC) and an open food challenge (OFC), according to the kind of foods.
The double-blind placebo-controlled food challenge is performed with foods to be tested on the causing or non-causing of allergy, as well as foods causing no problems (i.e., placebo). The two kinds of foods must be indistinguishable from each other, and during the test, both a doctor and a patient cannot be aware of the kind of foods tested. When the placebo is confirmed as not causing allergy while allergy reaction caused by specific foods is shown, the foods are evaluated as allergy- causing foods.
The open food challenge is performed in a state where a patient is aware that any foods are administered. The open food challenge is conducted for a patient who shows allergic reaction even to placebo in the double-blind placebo-controlled challenge.
When foods (P2) which were avoided are confirmed as not causing allergic reaction as a result of the food challenge, they are administered again and only foods (milk, PI and P3) confirmed as causing allergic reaction are continued to eliminate from diets, thereby performing the step of minimum avoidance of allergens.
After the minimum avoidance step, when a state where patient's symptoms no longer become worse is continued for at least two weeks, the step of tolerance induction is performed using the tolerance induction method and kit of the present invention.
The tolerance induction step consists of: a first sub-step for tolerance induction where an IFN-γ-containing composition and milk are administered together; and a second sub-step for confirmation of tolerance induction where only milk is administered. In the tolerance induction step, each of the first and second induction sub- steps is preferably carried out for 7 days.
After the tolerance induction step, milk is administered in one unit twice a day for at least 7 days to confirm if tolerance to milk was acquired. When a specific allergic reaction is not shown even if milk is continued to administer to a patient or when the result of the food challenge with milk shows the disappearance of allergy, it is evaluated that tolerance to milk was acquired.
Hereinafter, the tolerance induction method and kit of the present invention will be described in further detail.
The tolerance induction method of the present invention consists of: a first step for tolerance induction where an LFN-γ-containing composition and milk are administered together; and a second step for the confirmation of tolerance induction where only milk is administered.
In the tolerance induction method of the present invention, each of the first step for tolerance induction and the second step for the confirmation of tolerance induction is preferably performed for 7 days.
In the tolerance induction method of the present invention, the first step for tolerance induction comprises administering an LNF-γ-containing composition to a patient with allergy against specific substances in a specific unit per body surface area and at the same time, administering the substances while gradually increasing the amount of the substances.
When the first step for tolerance induction in the tolerance induction method of the present invention is carried out for 7 days, one dose of an LNF-γ-containing composition is administered, and then immediately, allergy-causing substances is administered in 1/4 unit a day, at days 1 and 2. At days 3 and 4, one dose of the LNF-γ-containing composition is administered, and then immediately, the allergy- causing substances are administered in 1/2 unit a day. At days 5 and 6, one dose of the LNF-γ-containing composition is administered, and then immediately, the allergy- causing substances are administered in one unit a day. At day 7, one dose of the INF-γ-containing composition is administered, and then immediately, the allergy- causing substances are administered in 1.5 units.
In the tolerance induction method of the present invention, the second step for the confirmation of tolerance induction comprises administering only allergy- causing substances while gradually increasing the amount of the substances as in the tolerance induction step without administering the LFN-γ-containing composition. When the second step for the confirmation of tolerance induction in the tolerance induction method of the present invention is performed for 7 days, allergy- causing substances are administered in 1/4 unit a day at days 1 and 2. At days 3 and 4, the allergens are administered in 1/2 unit a day, and at days 5 and 6, the allergens are administered in one unit a day. At day 7, the allergens are administered in 1.5 units.
Moreover, the present invention provides a tolerance induction kit allowing tolerance to allergy-causing substances to be acquired.
The tolerance induction kit of the present invention consists of a first step for tolerance induction and a second step for the confirmation of tolerance induction, in the same manner as the tolerance induction method of the present invention.
The tolerance induction kit of the present invention comprises an INF-γ- containing composition of a quantity to be administered for the first tolerance induction step of a given period, allergy-causing substances of a quantity to be administered together with the composition, and allergy-causing substances of a quantity to be administered alone in the second step for the confirmation of tolerance induction.
The tolerance induction kit of the present invention consists of a first container unit containing the LNF-γ-containing composition and a second container unit containing the allergens. Alternatively, it may consist of a first container unit containing the composition and allergens to be used in the first step for toleration induction, and a second container unit containing the allergens to be used in the second step for the confirmation of tolerance induction.
In order to increase the convenience and portability of the tolerance induction kit according to the present invention, each of the LFN-γ-containing composition and the allergens is contained in one container or several different small containers. Thus, the respective container units may have several small containers according to their arrangement method.
Preferably, the tolerance induction kit of the present invention comprises the LFN-γ-containing composition of a quantity to be administered in the first step for tolerance induction for 7 days, and the allergy-causing substances of a quantity to be administered therewith, and also the allergens of a quantity to be administered in the second step for the confirmation of tolerance induction for 7 days. Hereinafter, the construction of the kit will be described.
The first containers may contain seven doses of the LFN-γ-containing composition, and the second container may contain 10 units of the allergy-causing substances. In this case, the first container unit may consist of one small container containing seven doses of the composition, or seven small containers each containing one dose of the composition. Similarly, the second container unit may consist of one small container containing 10 units of the allergy-causing substances, or it may consist of 4 containers each containing 1/4 unit of the allergens, 4 containers each containing 1/2 unit of the allergens, 4 containers each containing one unit of the allergens and 2 containers each containing 1.5 units of the allergens.
Alternatively, the first container unit may contain seven doses of the IFN-γ- containing composition and 5 units of the allergy-causing substances, and the second container unit may contain 5 units of the allergy-causing substances. In this case, the first container unit may consist of one small container containing seven doses of the composition and one small container containing 5 units of the allergens, or it may consist of seven small containers each containing one dose of the composition, and two containers each containing 1/4 unit of the allergens, two containers each containing 1/2 units of the allergens, two containers each containing one unit of the allergens, and one containing 1.5 unit of the allergens. Similarly, the second container unit may consist of one small container containing 5 units of the allergens, or it may consist of two containers each containing 1/4 unit of the allergens, two containers each containing 1/2 unit of the allergens, two containers each containing one unit of the allergens, and one container containing 1.5 units of the allergens.
If necessary, the tolerance induction kit of the present invention may comprise a syringe required for administration, and directions showing a method for the administration of each component, etc.
In the tolerance induction kit of the present invention, if the allergy-causing substances are foods, they can be provided in a form that can be easily administered, such as powder, cake and the like.
The tolerance induction kit of the present invention may comprise one or more allergens. For example, in the case of a patient who shows a positive allergic reaction to both milk and soybeans, the kit may comprise the LFN-γ-containing composition of a quantity to be used in two tolerance induction processes, and milk and soybeans of quantities to be used in each of the first step for tolerance induction and the second step for the confirmation of tolerance induction.
The tolerance induction kit of the present invention may be used alone or in combination with hormone therapy, chemical therapy and other methods of inducing tolerance to allergy using biological response modifiers.
In the tolerance induction method and kit of the present invention, the LFN-γ- containing composition may contain IFN-γ, fragments of LFN-γ with Thl cell- stimulating activity, or derivatives with functions similar with these substances, as an active ingredient, and if necessary, other active ingredients. In the tolerance induction method and kit of the present invention, the IFN-γ- containing composition may comprise pharmaceutically suitable, physiologically acceptable adjuvants, in addition to the active ingredient as described above. Examples of such adjuvants include excipients, disintegrants, sweetening agents, binders, coating agents, blowing agents, lubricants, flavoring agents, and solubilizing agents.
In the tolerance induction method and kit of the present invention, the IFN-γ- containing composition may be preferably formulated as a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers for administration, in addition to the active ingredient as described above. In the tolerance induction composition and kit of the present invention, examples of the pharmaceutically acceptable carriers, which can be used in formulating IFN-γ into liquid solution, include saline solution, sterilized water,
Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture thereof. If necessary, other conventional additives, such as antioxidants, buffer solution and bacteriostatics, may also be added. Moreover, diluents, dispersants, surfactants, binders and lubricants may be further added to formulate LNF-γ as injection formulations, such as aqueous solution, suspension and emulsion, pills, capsules, granules and tablets. Furthermore, it can be preferably formulated according to diseases or components, using a method described in Remington 's Pharmaceutical Science, Mack Publishing Company, Easton PA, which is a suitable method in the relevant field of art.
In the tolerance induction method and kit of the present invention, the IFN-γ- containing composition can be formulated as pharmaceutical formulations, such as granules, powders, coated tablets, tablets, capsules, suppositories, syrup, juice, suspension, emulsion, drops, injectable liquid, and sustained-release preparations. The IFN-γ-containing composition of the present invention is relatively nontoxic.
In the tolerance induction method and kit of the present invention, the IFN-γ- containing composition can be administered in the conventional manner via the subcutaneous, intravenous, intraarterial, intraabdominal, intramusclar, intrasternal, percutaneous, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal route.
In the tolerance induction method and kit of the present invention, the IFN-γ- containing composition is administered at the amount of 2 x 106U to 4 x 106 U per m of body surface area, and preferably 3 x 10 U per m of body surface area. The amount of the composition administered can vary depending on various factors, including the severity of allergic diseases, the kind and content of an active ingredient and other components contained in the composition, the kind of a formulation, and patient's age, weight, general health condition, sex and diet, and administration time, administration route, the secretion % of the composition, administration period, and the kind of drugs used in combination, for administration by subcutaneous injection.
In the tolerance induction method and kit of the present invention, the dosage of the allergy-causing substances is expressed in a given unit for each substance. In the case of foods, one unit is defined as the average amount of the foods that an adult ingests in eating. For milk, one unit is preferably is defined as the amount of 200 g per 1.71 m2 of body surface area.
In the tolerance induction method and kit of the present invention, the induction of tolerance is achieved by a change in immune system caused by the administration of the IFN-γ-containing composition and then the allergy-causing substances. This induction of tolerance is schematically shown in FIG. 2.
As shown in FIG. 2, the IFN-γ-containing composition administered in the first step for tolerance induction serves to convert the immune status of a subject into a Thl cell predominant state. Under this condition, the administered allergens act as an antigen, and the dose of the allergens is gradually increased from small amount, and thus, a normal immune response other than an instant allergic reaction as in the prior art is induced. Then, in the second step for the confirmation of tolerance induction, only the allergens are administered to accumulate the immune response so that the acquired tolerance to the allergens can be maintained more securely.
Thus, the tolerance induction method and kit of the present invention, and the method for acquiring tolerance using them, allow tolerance to certain allergens to be effectively induced, and since then, allow the tolerance to be maintained lastingly.
This effect of tolerance acquirement of the present invention is a characteristic effect shown by only the tolerance induction method of the present invention, other than a residual effect also obtainable by the simple administration of IFN-γ used in the prior art, or an effect of natural tolerance acquirement caused by continuous exposure to the allergens.
Before and after acquiring tolerance using the tolerance induction method and kit of the present invention, skin response to milk, the ratio of eosinophils in leukocytes, blood IgE concentration, and the concentration of IgE specific to allergens, show a statistically insignificant change pattern. This result disagrees with the existing concept that eosinophils and IgE antibodies are increased in allergic diseases. From this point, it is presumed that the effect of tolerance acquirement according to the tolerance induction method of the present invention is achieved by mechanisms, such as cell immunity, other than the mediation of IgE antibodies. The tolerance induction method and kit of the present invention, and the method for acquiring tolerance using them, allow tolerance to allergens to be positively acquired, and at the same time, allow the acquired tolerance to be maintained lastingly, in significant improvements over the existing method for the inhibition of allergy, which remains at a negative level, such as allergen avoidance, or could not overcome side effects caused by chemical therapy. Accordingly, the present invention will be useful in the field of the prevention and inhibition of allergy.
Brief Description of Drawings
FIG. 1 is a schematic view showing a process for acquiring tolerance to milk using a method and kit for inducing tolerance to allergy according to the present invention;
FIG. 2 is a schematic view showing a change in immune system according to the administration of an LNF-γ-containing composition and allergens; FIG. 3 is a schematic view showing methods for inducing tolerance to milk allergy using methods of the present invention and the prior art;
FIG. 4a shows clinical changes occurring when conducting a tolerance induction method of the present invention; and
FIG. 4b shows clinical severity and an abnormal reaction on the skin according to time, which occur when conducting a tolerance induction method of the present invention.
Best Mode for Carrying Out the Invention
The present invention will hereinafter be described in further detail by examples. It should however be borne in mind that the present invention is not limited to or by the examples.
Example 1 : Acquirement of tolerance to milk in patients with atopic dermatitis In order to confirm if tolerance to milk allergy is acquired by the tolerance induction method of the present invention or not, the following test was carried out. The test was carried out on 250 patients with atopic dermatitis, who visited
Seoul Allergy Clinic, Korea. In the test subjects, the ratio between the male and the female was 137:113, and mean age was 8.3 ± 6.0 years. 1) Conformation of Atopic Dermatitis
According to the Hanifin and Rajka criteria, the patients were subjected to a detailed examination of history on food allergy, a skin prick test and the identification of IgE specific to certain foods, and as a result, confirmed as having atopic dermatitis. 1. Skin Prick Test The skin prick test was carried out by applying the back of all the patients with foods used in the FAST test.
In this case, histamine hydrochloride (Bencard GmbH) that is a commercially available typical allergen was used as a positive control group, and physiological saline and distilled water were used as a negative control group.
At 15 minutes after application, the size of formed urticaria or flare was measured to evaluate the results of the reaction. The evaluated results were determined by comparison with the control groups and graded as follows:
0: no reaction; 1+: reaction occurred as compared to the negative control group but corresponded to less than 1/2 of the positive control group; 2+: reaction corresponded to 1/2 of the positive control; 3+: reaction equal to the positive control group; and 4+: reaction corresponded to two times or more of the positive control group. The minimum reaction size in the positive reaction was 3 mm.
2. Identification of IgE specific to certain foods
In order to identify food-specific IgE, a FAST test (fluorescent allergo- sorbent-test) was carried out on all the patients.
In this test, the following foods were used: soybeans, peaches, almonds, malt, hazels, parsley, pork, tomato, barley, shellfishes, salmons, shrimps, tunas, fowl, beef, mushrooms, sesame seeds, mackerels, celery, lettuce, baker's yeast, cheese, chocolate, eggs, milk, rye, wheat, codfishes, lobsters, walnuts, rice, peas, potatoes, mutton, apples, bananas, oranges, strawberries, grapes, lemons, onions, cabbage, spinach, carrots, herrings, sardines, Hippoglossoides dubius Schmidt, crabs, sea mussels, tea, oysters and coffee. These foods were subjected to the FAST test according to a method described in Allergenetics form 146, 1982. The test results were graded as follows: 0: a case where IgE concentration in the foods was identified as less than 0.02 IU/ml; 1: a case where IgE concentration in the foods was identified as 0.02-0.1 IU/ml; 2: a case where IgE concentration in the foods was identified as 0.1-0.5 IU/ml; 3: a case where IgE concentration in the foods was identified as 0.5-2.5 IU/ml; 4: a case where IgE concentration in the foods was identified as 2.5-12.5 IU/ml; 5: a case where IgE concentration in the foods was identified as 12.5-62.5 IU/ml; and 6: a case where IgE concentration in the foods was identified as more than 62.5 IU/ml.
2) Maximum avoidance
The patients were subjected to maximum avoidance as follows. All foods, which had been suspected to cause allergy through the history examination, the skin prick test, and the identification of allergen-specific IgE, were eliminated from diets for at least one week and thus avoided, and also foods containing them were strictly avoided.
For nutrition balance, foods substituting for the avoided foods were supplied as given in Table 1 below.
Table 1:
Figure imgf000018_0001
Meanwhile, in order to confirm if the foods identified as allergens are exactly avoided from a diet for the patients and at the same time, in order to suitably admimster foods, dietary diaries were prepared for all the patients. The respective dietary diaries were analyzed by professional dietitians, nurses, and physicians.
After the maximum avoidance of allergens was conducted, when the symptoms of atopic dermatitis are improved and a state where this condition no loner become worse was lasting for at least one week, a food challenge test was conducted.
3) Food Challenge Test
A food challenge test was carried out using the double-blind placebo- controlled food challenge in the case of milk, wheat and beans, and using the open food challenge in the case of beef and fowl. After the maximum avoidance of allergens was conducted, when allergic symptoms were clinically surely improved and this condition was lasting for at least two weeks, and when it was confirmed that foods to be tested were avoided from diet, a food challenge test was conducted. Dietary diaries were analyzed by a professional dietitian to analyze the above particulars while the respective patients were maintained at their usual living habit for the largest possible removal of environmental factors.
In the double-blind placebo-controlled food challenge, two kinds of powder mixtures were used as excipients. Excipient I consisted of a mixture of 5g of unhulled powder, 5g of glutinous rice powder and 5g of barley powder, and excipient
II consisted of a mixture of 3g of banana powder, lg of Perilla japonica, 8 g of corn and 3g of chestnuts.
15g of the excipient powder was mixed with 6g of foods to be challenged, and then administered to the patients. In a placebo test, 21g of a powder mixture having the same composition as the excipient powder was administered to the patients. For patients having a dislike to powders, the excipient was administrated in cake form.
To eliminate the case of reaction to placebo, the patients were subjected to food challenge with the powder mixtures to be used as placebo, i.e., excipient I and excipient II, for three days, thereby confirming if the patients have tolerance to placebo or not. For example, when patients were confirmed as having tolerance to excipient I, excipient I was used as placebo and excipients in the test. When the patients showed a positive reaction to excipient I, expicient II was administered for three days in order to confirm if the patients have tolerance to excipient II or not, so that whether the excipients can be used or not was determined. The randomization and preparation of the food challenge test were conducted by dietitians for each clinical study unit. All the results of the food challenge test were added up according to a clinical severity scoring system given in Table 2 below.
Table 2:
Figure imgf000020_0001
In the case of patients with food-specific IgE or patients with a secure history where severe allergy to foods was shown, the food challenge test was carefully carried out. Here, the standard of the secure history was one having experience where allergic reaction occurred immediately after administering the test foods alone over recent two years, so that emergency treatment by a doctor was required.
The open food challenge was carried out for the same period as the double- blind placebo-controlled food challenge, and also conducted in such a manner that the test foods are administered while gradually increasing their amount daily, as shown in Table 3.
Table 3:
Figure imgf000021_0001
Namely, the test foods were administered one time one day every morning for 4 days, and the results was evaluated. In this evaluation, one of the following cases was evaluated as a positive reaction: Patients show more than 20% increase in a severity score as compared to that prior to the test, they show an evaluated severity score higher than 100, and inflammation or itching is shown on the skin.
Foods, which had been confirmed as causing the positive reaction, were avoided, and if symptoms became severe, the next challenge test was deferred for at least 4 days until a condition prior to challenge was recovered. Similarly, when foods to be avoided were administered, the challenge test was deferred for 4 days.
4) Minimum Avoidance
According to the result of the food challenge test, minimum avoidance was carried out while avoiding only the foods, which had been confirmed as causing abnormal reaction in the respective patients.
After minimum avoidance, the identification of IgE specific to certain foods, and food challenge test, were carried out to examine foods causing allergic reaction. Here, the standard of a positive allergy reaction was the case where the itching grade or skin inflammation grade is higher than grade 3.
In 250 patients who had been tested in the above example, clinical severity was greatly improved from 540 ± 50 to 150 ± 80 through the maximum avoidance, food challenge and minimum avoidance steps, although several patients showed worse allergic symptoms in the food challenge test. The identification of IgE specific to certain foods showed that the number of allergy-causing foods per patient was 3.5 ± 2.1, and the skin prick test showed that the number of allergy-causing foods per patient 7.5 ± 4.3.
The number of foods causing abnormal reaction after food challenge was 2.3
± 3.5. During the minimum avoidance, 88 patients reaching 35.2% showed symptoms, such as skin inflammation or itching reaction to milk, as a result of the double-blind placebo-controlled food challenge, and were subjected to tolerance induction. Then, after a state where patient's symptoms no longer become worse lasted for at least two weeks, a tolerance induction test was conducted.
5) Tolerance Induction Test
88 patients who showed abnormal reaction to milk were divided into four groups as shown in Table 5 below, and then a test program was made.
Table 4:
Figure imgf000023_0001
As shown in Table 4, tolerance induction methods varying with the groups were carried out. In this case, as a group for comparison with the tolerance induction method of the present invention, a method of administering only IFN-γ or milk was used, and as a control group, a method of administering placebo was used.
Namely, in the first step for tolerance induction, the patients of group I were administered with 1/4 unit of milk just following the administration of IFN-γ, at days 1 and 2. At days 3 and 4, 1/2 unit of milk was administered just after administering IFN-γ. At days 5 and 6, one unit of milk was administered just after administering IFN-γ. At day 7, two units of milk were administered after administering IFN-γ.
On the other hand, the patients of group II were administered with only IFN- γ throughout the tolerance induction period, and the patients of group III were administered with only milk while gradually increasing the amount of milk according to the same program as group I. The patients of group IV were administered with only placebo.
After the first step for tolerance induction, the patients of groups I and III were administered with milk while gradually increasing the amount of milk according to a program, thereby conducting the second step for the confirmation of tolerance induction.
As IFN-γ for tolerance induction, recombinant IFN-γ (specific activity: 2 x 106 U/mg), which is commercially available from Intermax gamma Co., was used and injected subcutaneously at the amount of 10 U/m . In order to reduce side effects, such as symptoms similar with myalgia, pyrexia and influenza, 10 mg/kg of oral acetaminophen was administered at one hour before administration and at 4 hours after administration. In this case, the maximum dosage of acetaminophen was 600 mg.
To eliminate the effect of other drugs, the administration of systemic steroid and other drugs which has been used in the prior are were gradually reduced and then stopped at two weeks before the start of the test. However, the local application of
1% hydrocortisone and the oral administration of ketotifen/γ-linoleic acid were accepted from one month before the test start to one month after the test start.
6) Confirming if tolerance to milk was acquired after tolerance induction In order to confirm if tolerance to milk was acquired after tolerance induction, one unit of milk was administered to group I twice a day for at least 7 days.
The condition of the patients was observed for at least 4 weeks.
The results are expressed in mean ± standard deviation, and a statistical analysis was carried out according to a Wilcoxon matched-pair signed-ranks test.
1. Confirming if tolerance to milk was acquired
On the respective groups subjected to the tolerance induction process, whether tolerance to milk was acquired or not was confirmed. The results are given in Table 5 below.
Table 5:
Figure imgf000025_0001
As shown in Table 5, all the patients of Group I acquired tolerance to milk as a result of the induction of tolerance to milk. Even when one unit of milk was administered twice a day for 7 days after the tolerance induction period, the patients of Group I no longer showed abnormal reaction to milk. This tolerance to milk was maintained even after six months, and maintained for 16.1 ± 3.7 months on the average. Meanwhile, clinical change during the tolerance induction period was observed, and the results are shown in FIGS. 4a and 4b.
As shown in FIG. 4a, 32 patients among 66 patients of group I showed abnormal reactions, including itching and skin inflammation such as epidermic eruption, during the tolerance induction period. The degree of the abnormal reactions was most severe at 3 and 4 days after the start of tolerance induction, but such symptoms were rapidly improved at day 6 and then substantially not shown at day 7. Even after the tolerance induction period, such symptoms were no longer developed.
Meanwhile, as shown in FIG. 4b, itching and skin inflammation were developed in different patterns, and the higher the clinical severity, the patients showed increased itching. The progress pattern of allergic symptoms, which were temporarily shown during the tolerance induction period, approximately coincided with that of itching.
On the other hand, none of the patients of groups II, III and IV acquired tolerance to milk. Also, within one day after administration of milk, they showed itching, urticaria, acute redness, epidermic eruption, excoriation or more than 20% increase in clinical severity. In addition, these patients showed pyrexia and rigor (53%), mylgia (32%), headache (18%), and stomachache (9%).
2. Skin Prick Test
A skin prick test was carried out on the respective groups subjected to the tolerance induction procedure. The test results are given in Table 6 below.
Table 6:
Group I Group II Group III Group IV
Wound size 3.2 +1.1 to 3.1 + 1.2 3.0 ± 1.1 to 2.9 + 3.1 ± 1.0 to 3.2 + 3.1 ± 1.1 to 3.0 ± 1.0 1.1 1.2 As shown in FIG. 6, the results of the skin prick test on milk varied in all the groups without statistical significance (p>0.05).
3. Results of Hematological Test
The respective groups subjected to the tolerance induction procedure were measured for leukocyte count, eosinophil count and the ratio of eosinophils in leukocytes. The measured results are given in Table 7 below.
Table 7:
Figure imgf000027_0001
As shown in Table 7, the number and ratio of the respective blood- corpuscles varied in all the groups without statistical significance (p>0.05).
Furthermore, the respective groups subjected to the tolerance induction procedure were measured for blood IgE concentration and the concentration of IgE specific to milk after the administration of milk. The measured results are given in Table 8 below. Table 8:
Figure imgf000028_0001
As shown in Table 8, the blood IgE concentration and the concentration of IgE specific to milk varied in all the groups without statistical significance (p>0.05).
4. Conclusion
As shown in Table 5, it can be fount that the tolerance induction method of the present invention allows tolerance to certain allergens to be effectively induced, and this effect of tolerance acquirement is a characteristic effect shown by only the tolerance induction method of the present invention, other than a residual effect of IFN-γ itself or an effect of natural tolerance acquirement caused by continuous exposure to allergy-causing foods.
Moreover, as shown in Tables 6, 7 and 8, it can be found that the acquirement of tolerance to allergens according to the tolerance induction method of the present invention is achieved by mechanisms, such as cell immunity, other that the mediation of IgE antibodies as known in the prior art.
Industrial Applicability
The tolerance induction method and kit of the present invention, and the method for acquiring tolerance using them, advantageously allow tolerance to certain allergens to be effectively induced, and since then, allow the tolerance to be maintained lastingly even upon the ordinary administration of the allergens.
The tolerance induction method and kit of the present invention, and the method for acquiring tolerance using them, will be useful in the field of the prevention and inhibition of allergy.

Claims

What Is Claimed Is:
1. A method for acquiring tolerance to allergy, which comprises the steps of: confirmation of allergic disease, maximum avoidance of allergens, allergen challenge, minimum avoidance of allergens, tolerance induction, and tolerance maintenance.
2. The method of Claim 1, wherein the tolerance induction step comprises a first sub-step for the induction of tolerance where an IFN-γ-containing composition and allergy-causing substances (allergens) are administered to a patient with allergy against the substances; and a second sub-step for the confirmation of tolerance induction where only the substances are administered to the patient.
3. The method of Claim 2, wherein the allergens are selected from foods, including milk, eggs, soybeans, wheat flour, pork, fowl, rice, and garlic, and house dust mite, pollen, animal hair, mold, and certain chemicals.
4. The method of Claim 3, wherein each of the first sub-step for tolerance induction and the second sub-step for the confirmation of tolerance induction is carried out for 7 days.
5. The method of Claim 4, wherein the first sub-step for tolerance induction comprises administering the IFN-γ-containing composition to the patient with allergy against certain substances in a given unit per body surface area, and at the same time, administering the substances while gradually increasing the amount of the substances.
6. The method of Claim 5, wherein the first sub-step for tolerance induction comprises: administering one dose of the IFN-γ-containing composition and then immediately administering the allergens in 1/4 unit a day at days 1 and 2; administering one dose of the IFN-γ-containing composition and then immediately administering the allergens in 1/2 unit a day at days 3 and 4; administering one dose of the IFN-γ-containing composition and then immediately administering the allergens in one unit a day at days 5 and 6; and administering one dose of the IFN-γ- containing composition and then immediately administering the allergens in 1.5 units at day 7.
7. The method of Claim 6, wherein the iFN-γ-containing composition is injected subcutaneously at the amount of 3 x
Figure imgf000031_0001
of body surface area.
8. The method of any of Claims 1 to 7, wherein the second sub-step for the confirmation of tolerance induction comprises administering only the allergens during a given period while increasing the amount of the allergens as in the first sub- step for tolerance induction, without administering the IFN-γ-containing composition.
9. The method of Claim 8, wherein the second sub-step for the confirmation of tolerance induction comprises: administering the allergens in 1/4 unit a day at days
1 and 2; administering the allergens in 1/2 unit a day at days 3 and 4; administering the allergens in one unit a day at days 5 and 6; and administering the allergens in 1.5 units a day at day 7.
10. The method of Claim 9, wherein the allergens are milk.
11. The method of Claim 10, wherein one unit of milk is the amount of 200 ml per 1.71 m of body surface area.
12. A method for inducing tolerance to allergy, which comprises: a first step for the induction of tolerance where an IFN-γ-containing composition and allergy- causing substances are administered to a patient with allergy against the substances; and a second step for the confirmation of tolerance induction where only the substances are administered to the patient.
13. The method of Claim 12, wherein the allergens are selected from foods, including milk, eggs, soybeans, wheat flour, pork, fowl, rice, and garlic, and house dust mite, pollen, animal hair, mold, and certain chemicals.
14. The method of Claim 13, wherein each of the first step for tolerance induction and the second step for the confirmation of tolerance induction is carried out for 7 days.
15. The method of Claim 14, wherein the first step for tolerance induction comprises administering the IFN-γ-containmg composition to the patient with allergy against certain substances in a given unit per body surface area, and at the same time, administering the substances while gradually increasing the amount of the substances.
16. The method of Claim 15, wherein the first step for tolerance induction comprises: administering one dose of the IFN-γ-containing composition and then immediately administering the allergens in 1/4 unit a day at days 1 and 2; administering one dose of the rFN-γ-containing composition and then immediately administering the allergens in 1/2 unit a day at days 3 and 4; administering one dose of the IFN-γ-containing composition and then immediately administering the allergens in one unit a day at days 5 and 6; and administering one dose of the IFN-γ- containing composition and then immediately administering the allergens in 1.5 units at day 7.
17. The method of Claim 16, wherein the IFN-γ-containing composition is injected subcutaneously at the amount of 3 x 106 U per m2 of body surface area.
18. The method of any of Claims 12 to 17, wherein the second sub-step for the confirmation of tolerance induction comprises administering only the allergens during a given period while increasing the amount of the allergens as in the first sub- step for tolerance induction, without administering the iFN-γ-containing composition.
19. The method of Claim 18, wherein the second step for the confirmation of tolerance induction comprises: administering the allergens in 1/4 unit a day at days 1 and 2; administering the allergens in 1/2 unit a day at days 3 and 4; administering the allergens in one unit a day at days 5 and 6; and administering the allergens in 1.5 units a day at day 7.
20. The method of Claim 9, wherein the allergens are milk.
21. The method of Claim 20, wherein one unit of milk is the amount of 200 ml per 1.71 m2 of body surface area.
22. A kit for inducing tolerance to allergy, which comprises: an INF-γ-containing composition of a quantity to be administered in the first step for the induction of tolerance, allergy-causing substances of a quantity to be administered together with the INF-γ-containing composition in the first step for tolerance induction, and allergy-causing substances of a quantity to be administered in a second step for the confirmation of tolerance induction.
23. The kit of Claim 22, wherein the allergens are selected from foods, including milk, eggs, soybeans, wheat flour, pork, fowl, rice, and garlic, and house dust mite, pollen, animal hair, mold, and certain chemicals.
24. The kit of Claim 23, which comprises a first container unit containing the
IFN-γ-containing composition, and a second container unit containing the allergy- causing substances.
25. The kit of Claim 23, which comprises a first container unit containing the IFN-γ-containing composition to be used in the first step for tolerance induction, and a second container unit containing the allergy-causing substances to be used in the second step for the confirmation of tolerance induction.
26. The kit of any of Claims 22 to 25, wherein each of the INF-γ-containing composition and the allergy-causing substances is contained in one container or several different small containers.
27. The kit of Claim 26, which comprises the INF-γ-containing composition of a quantity to be administered in the first step for the induction of tolerance for 7 days, the allergy-causing substances of a quantity to be administered together with the INF-γ-containing composition in the first step for tolerance induction for 7 days, and the allergy-causing substances of a quantity to be administered in a second step for the confirmation of tolerance induction for 7 days.
28. The kit of Claim 27, which comprises the first container unit containing seven doses of the IFN-γ-containing composition, and a second container unit containing 10 units of the allergy-causing substances.
29. The kit of Claim 28, wherein the first container unit consists of one small container containing seven doses of the IFN-γ-containing composition, or seven small containers each containing one dose of the composition.
30. The kit of Claim 29, wherein the second container unit consists of one small container containing 10 units of the allergy-causing substances, or consists of 4 containers each containing 1/4 unit of the substances, 4 containers each containing 1/2 unit of the substance, 4 containers each containing one unit of the substances, and 2 containers each containing 1.5 units of the substances.
31. The kit of Claim 27, which comprises the first container unit containing seven doses of the IFN-γ-containing composition and 5 units of the allergy-causing substances, and the second container unit containing 5 units of the allergy-causing substances.
32. The kit of Claim 31, wherein the first container unit consists of one small container containing seven doses of the IFN-γ-containing composition, and one small container containing 5 units of the allergy-causing substances, or consists of 7 small containers each containing one dose of the composition, and 2 containers each containing 1/4 unit of the substances, 2 containers each containing 1/2 unit of the substance, 2 containers each containing one unit of the substances, and one container containing 1.5 units of the substances.
33. The kit of Claim 32, wherein the second container unit consists of one small container containing 5 units of the allergy-causing substances, or consists of 2 containers each containing 1/4 unit of the substances, 2 containers each containing 1/2 unit of the substance, 2 containers each containing one unit of the substances, and one container containing 1.5 units of the substances.
34. The kit of Claim 30 or 33, wherein the IFN-γ-containing composition is injected subcutaneously at the amount of 3 x 106 U per m2 of body surface area.
35. The kit of Claim 34, wherein the allergens are milk.
36. The kit of Claim 35, wherein one unit of milk is the amount of 200 ml per 1.71 m2 of body surface area.
PCT/KR2003/001270 2002-06-28 2003-06-27 Method and kit for the tolerance induction to allergy-causing substances, and the tolerance achievment against the substances thereby WO2004002520A1 (en)

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