JP2007106691A - Method for endermic administration of antigen and kit used for the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method, a preparation and the like for simply and surely performing endermic administration of antigen without using an adjuvant. <P>SOLUTION: An antigen kit for endermic administration of antigen without using an adjuvant contains an antigen composition (A) containing an antigen but not containing the adjuvant, and a patch (B) containing none of the antigen and the adjuvant but having a water retentive agent and a dermal plaster. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a method of transdermally administering an antigen without using an adjuvant, a kit used in the method, and the like.

  Since high molecular weight protein antigens generally cannot pass through normal skin, it is considered impossible to induce an antigen-specific immune response in the living body (transdermal immunization) by applying protein antigens to normal skin. I came. However, in recent years, it was reported in experiments using mice that transdermal immunization using cholera toxin as an adjuvant is possible (Non-patent Document 1). Since many of the side reactions in vaccination and desensitization are considered to be accompanied by injection, if antigen administration by transcutaneous immunization becomes possible, it is much safer than conventional methods by injection. It is expected that improvement of immunity and reduction of injured person burden will be planned. Therefore, research has been advanced on transdermal immunization methods (Patent Documents 1 to 6, Non-Patent Documents 2 to 5 and the like).

  In these conventional methods, it is generally essential to use an adjuvant for successful transcutaneous immunization. Patent Document 1 describes an example in which no adjuvant is used, but in this example, the antigen is applied not to intact skin but to skin that has been pretreated to remove the stratum corneum in advance. Patent Document 2 describes that an antigen with high antigenicity does not require the use of an adjuvant, but does not actually describe an example of successful transcutaneous immunization without using an adjuvant. Absent.

Special table 2005-504002 Special table 2003-523937 Special table 2004-536804 Special table 2004-529906 Special table 2004-525921 Special table 2002-504522 Glenn G.M. et al., Nature, vol. 391, p. 851 (1998) Chen D.C. et al., Nature Medicine, vol. 6, no. 10, pp. 1187-90 (2000). Hammond S.A. et al., Critical Reviews in Therapeutic Drug Carrier Systems, 18 (5): 503-526 (2001). Godefroy S. et al., Vaccine, vol. 21, pp. 1665-71 (2003). Kenney R.T. et al., J. Infectious Diseases, vol. 190, pp. 774-82 (2004)

  Adjuvants are also known to cause side effects, and in some cases have been reported to enhance antigen-induced side effects. However, the conventional methods cannot avoid the disadvantages associated with the use of such an adjuvant. In addition, conventionally proposed formulations for transcutaneous immunization are mainly in the form of dry patches impregnated with an antigen and then dried. In this case, a preparation for transcutaneous immunization is produced for each antigen. It is necessary and inconvenient.

  An object of the present invention is to provide a method, a preparation, and the like for performing antigen administration easily and surely transcutaneously without using an adjuvant.

  The present inventor has found that by adopting certain conditions, an immune response can be easily induced without using an adjuvant, and the present invention has been completed.

That is, according to the present invention,
[1] An antigen kit for transdermal antigen administration without using an adjuvant, comprising A) an antigen composition containing an antigen and no adjuvant, and B) containing neither an antigen nor an adjuvant, at least water retaining A kit comprising a patch having a material and a patch;
[2] The antigen kit according to [1], which is used as a vaccine;
[3] The antigen kit according to [1], which is used as an antigen in desensitization therapy or immunotherapy;
[4] A step of applying to the skin an antigen composition containing neither an antigen nor an adjuvant, and having at least a water-retaining material and a patch, the antigen composition containing no antigen, and the skin at the application site Covering with a patch having a water-retaining material that does not contain any antigen or adjuvant and containing water, and maintaining the water-retaining material in contact with the skin of the application site for at least 2 hours. A patch used in an antigen administration method characterized in that it comprises:
[5] A patch that contains neither an antigen nor an adjuvant, and has at least a water retention material and a patch, and contains an aqueous antigen composition that contains an antigen and does not contain an adjuvant in the water retention material of the patch; And a step of applying a patch with an adhesive so that the water-retaining material is in contact with the water-retaining material, and a step of maintaining the water-retaining material in contact with the skin for at least 2 hours. Patch to be applied;
[6] An antigen composition containing an antigen and not containing an adjuvant, the step of applying this composition to the skin, and the skin at the application site containing neither antigen nor adjuvant, but water An antigen composition for use in an antigen administration method, comprising: coating with a patch having a water retaining material to be maintained; and maintaining the water retaining material in contact with the skin of the application site for at least 2 hours object;
[7] An aqueous antigen composition containing an antigen and not containing an adjuvant, wherein the aqueous composition contains at least a water retention material of a patch having a water retention material and a patch, and the skin and the water retention material. An antigen composition for use in an antigen administration method, comprising: a step of applying a patch with an adhesive so as to contact; and a step of maintaining the water-retaining material in contact with the skin for at least 2 hours;
[8] A method of using the kit according to [1], wherein the antigen composition is applied to the skin, and the skin at the application site contains neither an antigen nor an adjuvant, but water. Covering with a patch having the water retention material and maintaining for at least 2 hours in contact with the skin at the application site;
[9] A method of using the kit according to [1], wherein the patch is applied with a patch so that the skin and the water retaining material are in contact with each other, and the step of incorporating the antigen composition into the water retaining material of the patch. And maintaining the water-retaining material in contact with the skin for at least 2 hours;
[10] A step of applying an immunogen composition containing an antigen and not containing an adjuvant to the skin, and a patch having a water-retaining material containing neither the antigen nor the adjuvant and containing water, the skin at the application site. A transdermal antigen administration method that does not use an adjuvant (“the first aspect of the present invention”), comprising a coating step and a step of maintaining the water-retaining material in contact with the skin at the application site for at least 2 hours One antigen administration method ”);
[11] A step of containing an aqueous immunogenic composition containing an antigen and no adjuvant in a water retaining material of a patch having at least a water retaining material and a patch, and a patch so that the skin and the water retaining material are in contact with each other And a step of maintaining the water-retaining material in contact with the skin for at least 2 hours, wherein the method is a transdermal antigen administration method that does not use an adjuvant ("the second antigen of the present invention"). Administration method),
Is provided.

  According to the present invention, there is no fear of adverse effects such as side effects associated with the use of an adjuvant, and the method is highly safe and has a low burden on the recipient. Kits and the like are provided. The effect of antigen administration according to the present invention is comparable to conventional antigen administration by injection, and is applied to vaccination, desensitization therapy, cancer immunotherapy, etc. where antigen administration has been performed by conventional injection. Can do. In addition, as an antigen in the present invention, even if it is generally considered to have a very low antigenicity, it can be used sufficiently, so that the range of use is very wide.

  In addition, according to the present invention, immunization can be carried out so that anyone can easily and easily obtain an effect without requiring a special technique or process such as injection or removal of the stratum corneum. By obtaining and using the antigen composition, kit or the like of the present invention as a medicine prescribed by a doctor or an OTC medicine, for example, a vaccination conventionally required to be performed by a doctor or a veterinarian, Individuals can do it at home. This is particularly convenient for regularly immunizing pets and livestock.

  Furthermore, in the present invention, by changing the amount of the antigen composition used, the dose can be easily adjusted according to the animal species to be administered (human, non-human animal, non-human mammal, etc.), age, weight, condition, etc. You can choose. In addition, a special composition for each antigen as an antigen-containing patch is not required, and if an antigen composition containing an appropriate amount of antigen for each antigen is prepared, the patch is not limited to a specific antigen. Since a certain thing can be used, it is advantageous also in manufacture.

  The antigen used in the present invention is not particularly limited as long as it has antigenicity (immunogenicity). For example, proteins derived from various pathogenic organisms generally used for vaccination (tetanus toxin and its toxoid, diphtheria toxin and its toxoid, pertussis toxin and its detoxification product, influenza virus hemagglutinin, hepatitis B virus HBs antigen Etc.), various allergens (pollen such as cedar pollen, bee venom, mite antigen, house dust, etc.), cancer antigen and the like. Such an antigen may be purified to a single antigenic substance, but it is sufficient if it is purified to such an extent that it is considered necessary from the viewpoint of effectiveness and safety. In general, the required amount of antigen used for vaccination, desensitization therapy or other immunotherapy is known for each antigen. Alternatively, it can be easily determined by administering various amounts to animals and examining the presence or amount of antibodies produced in the serum.

In the present invention, the antigen forms a composition with at least one physiologically acceptable carrier. The carrier can be an aqueous solution, such as water, saline, isotonic solutions, buffers, and the like. When the antigen is a crudely purified product, the carrier may be one brought from the purification process. Alternatively, the antigen composition may be formulated in the form of a hydrophilic gel, powder, ointment, suspension, extract, lotion, aerosol or the like by a known method. The antigen composition may contain other additives (for example, preservatives, coloring agents, etc.) known in the field of pharmaceutical preparations in addition to the antigen and carrier. Those skilled in the art can appropriately select these various components and their addition amounts.
The antigen composition used in the present invention does not contain a known adjuvant (including a substance having an adjuvant property).

  In the first antigen administration method of the present invention, the antigen composition is applied to the skin, and then the application site is covered with a water retaining material holding a liquid containing water such as water or physiological saline. In this case, the antigen composition is preferably an aqueous liquid (for example, a solution containing water, an emulsion, a suspension, etc.), but may be a powder. In the second antigen administration method of the present invention, the antigen composition is supported on a water retaining material and brought into contact with the skin. In this case, the antigen composition is preferably an aqueous composition containing water, particularly preferably an aqueous liquid.

  The patch used in the present invention has at least a water retaining material and a patch. The water-retaining material may be water-retaining and low-irritating to the skin, and examples thereof include gauze, nonwoven fabric, absorbent cotton, and sponge. The patch has an adhesive strength that allows the patch to be applied to and held on the skin, and needs to be hypoallergenic to the skin. The patch is preferably in a form and size that can cover the contact surface of the water retention material with the outside air so as to prevent the water retention material from drying. For example, a medical adhesive tape, a bandage, etc. can be used.

Therefore, as the patch used in the present invention, a commercially available adhesive bandage with gauze, a medical patch, or the like can be used. In addition, the patch used in the first method of the present invention may be prepackaged in such a state that the water retaining material contains a liquid containing moisture in advance.
The patch used in the present invention does not contain any known antigen or adjuvant.

  In the method of the present invention, both the first method and the second method have a sufficient effect by contacting the skin and the antigen in a moist environment using a patch for 2 hours or more. can get. In general, the contact time is preferably 4 hours or longer, more preferably 8 hours or longer in order to obtain a sufficient effect even when an antigen having low antigenicity is used. The upper limit is not particularly limited as long as it does not cause undesirable effects (for example, itching due to the adhesive component of the patch) by applying the patch for a long time, and can be, for example, within 48 hours or about 24 hours. .

  By performing the antigen administration according to the present invention once, a priming effect (an effect of imparting a preparation state that causes antibody production in the second immunization to the living body) can be obtained. However, in order to increase the amount of antibody production sufficiently, it is desirable to carry out antigen administration twice or more. The administration site of the antigen is not particularly limited, but in general, administration to a site where the stratum corneum is relatively thin, for example, the inner side of the arm, the abdomen, the auricle, etc., is considered to be particularly effective. Since the priming effect obtained by the antigen administration of the present invention is not limited to the site of the first administration, the administration site for the second and subsequent times may be the same as or different from the site of the first administration.

Example 1: Induction of transdermal immune response using antigen solution application and wet patch coating Ovalbumin (OVA) is used as an antigen to induce an immune response in mice by changing the contact time between the antigen and the skin. did.

  The abdominal hair of a mouse (C57BL / 6, female, 8 weeks old, Nippon SLC Co., Ltd .; equivalent mice were used in all the following experiments) was clipped with a clipper to expose the skin. Two days later, the following immunization was performed. Ovalbumin (OVA) at a concentration of 2 mg / ml dissolved in physiological saline (Otsuka raw food injection, Otsuka Pharmaceutical Factory; the same product was used in all experiments below) on the abdominal skin surface of five anesthetized mice. The solution (Albumin from chicken egg white, Grade VI, SIGMA; the same product was used in all experiments below) was dropped by 50 μl and spread about 1 cm square. After this solution was dried, a patch in which 50 μl of physiological saline was impregnated into the site where the OVA solution was applied (protective pad for injection / Medipatch, White Cross Co., Ltd .; hereinafter, the same product was used in all experiments; The patch containing the liquid to be contained was sometimes referred to as “wet patch” in this specification) and fixed with tape. This prevented the mouse from ingesting the antigen applied to the surface of the skin orally, and kept the OVA solution application site in a moist state. After leaving the patch attached, it was left for 2 hours, 4 hours, 8 hours, or 16 hours, and then the patch was removed to thoroughly wash the application site.

  The above immunization was performed twice at 2-week intervals, and blood was collected 2 weeks after the second immunization to separate serum. The anti-OVA-IgG antibody titer in the serum was measured by ELISA.

  The results are shown in FIG. The longer the patch application time, the more individuals were observed to produce anti-OVA-IgG antibody, and the amount of antibody produced increased. At 16 hours after patch application, significant antibody production was observed in all mice.

  Therefore, it was shown that a transdermal immune response can be induced without using an adjuvant by maintaining the contact between the antigen and the skin in a moist environment patched for at least 2 hours. In all the experiments including this example, no significant changes were observed on the local skin to which the antigen was applied.

<Comparative example>
For comparison, transdermal immunization was similarly attempted by a conventional method. Specifically, in a mouse treated in the same manner as described above, an OVA solution having a concentration of 2 mg / ml dissolved in physiological saline, or cholera toxin (CT) having an adjuvant property in an OVA solution having the same concentration (Cholera Toxin, Vibrio cholerae , Type Inaba 569B, CALBIOCHEM; the same product was used in all experiments below) to a final concentration of 2 mg / ml, 50 μl each was dropped on the surface of the abdominal skin of 5 anesthetized mice. Spread about 1cm square. After the solution was applied, the mouse was left under anesthesia without being covered with a patch for 2 hours, and then the solution application site was thoroughly washed. Another 5 mice were injected intradermally with 50 μl of OVA solution of the same concentration. The above immunization was performed 3 times at 3 week intervals. Two weeks after the third immunization, blood was collected and serum was separated. Serum anti-OVA-IgG antibody titer and anti-CT-IgG antibody titer were measured by ELISA method.

  The results are shown in FIG. 2 (panels (A) and (B)). In this method, when CT was simultaneously applied to the skin as an adjuvant (“OVA + CT”), an anti-OVA-IgG antibody with a production amount equivalent to that of the group injected intradermally was observed in the serum for all five animals. However, in the group where only OVA was applied to the skin (“OVA”), the production of anti-OVA-IgG antibody was not observed at all (FIG. 2, panel (A)). On the other hand, the production of anti-CT-IgG antibody was observed only in the group to which CT was applied (FIG. 2, panel (B)). Therefore, in this method, it was confirmed that simultaneous application of CT as an adjuvant is essential for inducing a transdermal immune response.

Example 2: Induction of transcutaneous immune response using a patch containing an antigen solution Inducing an antigen-specific immune response in a mouse by applying a patch impregnated with the antigen solution to the ear of a mouse Tried. In this method, unlike the method of administering to the abdominal skin, it is possible to avoid the possibility that the antigen is taken in from an unintended wound when using the clippers.

  A patch soaked with 50 μl of the antigen solution was applied to the pinna of 5 anesthetized mice and fixed with tape. As an antigen solution, an OVA solution having a concentration of 2 mg / ml or a solution obtained by adding CT at a final concentration of 0.2 mg / ml as an adjuvant (“OVA + CT”) was used. After applying the patch, the patch was removed at 2, 4, 8, 16, or 32 hours, and the auricle was washed. The above immunization was performed 3 times at 2-week intervals, and blood was collected 2 weeks after the third immunization to separate the serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. It was confirmed that this method can also induce a transdermal immune response without using an adjuvant. Similarly to the method described in Example 1, the amount of anti-OVA-IgG antibody produced increased as the patch application time was increased.

Example 3: Comparison of effects in dry and wet conditions Compared with the case where a patch (dry or wet) was applied to the skin surface to which the antigen solution was applied, and the case where the patch was not applied. The effect was examined.

  The abdomen of the mouse was clipped with a clipper to expose the skin. Two days later, the following immunization was performed. An ovalbumin (OVA) solution having a concentration of 2 mg / ml dissolved in physiological saline was dropped on the surface of the abdominal skin of 15 anesthetized mice in a volume of 50 μl and spread over about 1 cm square. Thereafter, the following operations were carried out on 5 mice per group: 1) left alone with the antigen application, 2) apply the patch to the antigen application site and fix it with tape (dry patch), or 3) apply to the antigen application site. Apply a patch soaked with 50 μl of physiological saline and fix it with tape (wet patch). The contact time between the antigen and the skin was 16 hours, and then the antigen application site was washed.

  The above immunization was performed 3 times at 2-week intervals, and blood was collected 2 weeks after the third immunization to separate serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. In the group where only the antigen was applied, almost no antibody production was observed, whereas in the group where the patch was applied, significant anti-OVA-IgG antibody production was observed. Further, in the group to which the wet patch was applied, compared with the group to which the dry patch was applied, on the average, the amount of antibody produced was larger and antibody production with less variation was observed. In transcutaneous immunization without an adjuvant, it has been found that it is optimal to contact the antigen with the skin using a patch in a moist environment.

Example 4: Examination of antigen amount The effect of transdermal immunity depending on the amount of antigen administered was examined.
A patch soaked with 50 μl of the antigen solution was affixed to the mouse pinna and fixed with tape. Antigen solutions include OVA solutions with concentrations of 2 mg / ml, 0.2 mg / ml and 0.02 mg / ml (the dose of antigen per individual is 100 μg, 10 μg and 1 μg, respectively), and OVA at each concentration. A total of 6 types of solutions (“OVA + CT”) obtained by adding CT at a final concentration of 0.2 mg / ml as an adjuvant were used. Twenty hours after applying the patch, the patch was removed and the auricle was washed. The above immunization was performed 3 times at 2-week intervals, and blood was collected 2 weeks after the third immunization to separate the serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. In the group administered with the antigen solution to which CT was added as an adjuvant, antibody production tended to be induced by a smaller amount of antigen, but no significant difference was observed.

Example 5: Transition of antibody production The relationship between multiple administrations of antigen and the transition of antibody production was examined.
A patch soaked with 50 μl of the antigen solution was affixed to the mouse auricle and fixed with tape. As the antigen solution, an OVA solution having a concentration of 2 mg / ml, and an OVA solution having the same concentration plus CT having a final concentration of 0.2 mg / ml as an adjuvant (“OVA + CT”) were used. Twenty hours after applying the patch, the patch was removed and the auricle was washed. The above immunization was performed 3 times at 2-week intervals, and blood was collected every week until 6 weeks after the first immunization to separate serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. In both the OVA-only group (“OVA”) and the group to which CT was added (“OVA + CT”), antibody production showed a similar transition. In the group to which CT was added, the amount of antibody production was slightly higher, but it was hardly significant.

Example 6: Long-term fate of antibodies in serum The long-term fate of antigen-specific IgG antibodies induced in serum was examined.
A patch soaked with 50 μl of the antigen solution was affixed to the mouse pinna and fixed with tape. As the antigen solution, an OVA solution having a concentration of 2 mg / ml or an OVA solution having the same concentration plus CT having a final concentration of 0.2 mg / ml as an adjuvant (“OVA + CT”) was used. Twenty hours after applying the patch, the patch was removed and the auricle was washed. The above immunization operation was repeated 3 times at 2-week intervals. Blood was collected at 4 weeks, 6 weeks and 24 weeks after the first immunization to separate serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. In both the OVA-only group (“OVA”) and the CT-added group (“OVA + CT”), anti-OVA-IgG antibodies induced in the serum remained almost unchanged until 20 weeks after the final immunization. did. The group to which CT was added had a slightly higher amount of antibody production, but no significant difference was observed.

Example 7: Examination of retention period of priming effect The period during which the priming effect by the first immunization was retained was examined.
A patch soaked with 50 μl of OVA solution having a concentration of 2 mg / ml was applied to the auricle of a mouse and fixed with tape. Twenty hours after applying the patch, the patch was removed and the auricle was washed. The same immunization was repeated 2 weeks, 4 weeks, 6 weeks and 14 weeks after the above immunization. Two weeks after the second immunization, blood was collected to separate serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. Although the induction of specific antibody production by only the first immunization was hardly observed, the priming effect by the first immunization was maintained at least until 14 weeks after the immunization.

Example 8: Transmission of priming effect It was examined whether the priming effect by the first immunization was transmitted to a skin site different from the site where the antigen was administered.

  A patch soaked with 50 μl of OVA solution having a concentration of 2 mg / ml was applied to the left auricle of the mouse and fixed with tape. Twenty hours after applying the patch, the patch was removed and the auricle was washed. Two weeks after the above immunization, the same immunization was performed on the left auricle that was the same as the first immunization or the right auricle opposite to the first immunization. As a control, an experiment using physiological saline instead of the OVA solution was performed in parallel. Two weeks after the second immunization, blood was collected to separate serum. The anti-OVA-IgG antibody titer in serum was measured by ELISA.

  The results are shown in FIG. Significant antibody production was also observed when the second immunization was performed on the pinna opposite to the first ("Left: OVA Left: OVA" and "Left: OVA Right: OVA") . Therefore, it was confirmed that the effect of transcutaneous immunity was transmitted to a skin site different from the site where the immunization was performed.

Example 9: A patch impregnated with 50 μl of a transdermal immune antigen solution using tetanus toxoid was applied to the auricle of a mouse and fixed with a tape. A tetanus toxoid (Ttd) solution (obtained from the Institute of Chemistry and Serum Therapy) with a concentration of 200 Lf / ml as an antigen solution or a Ttd solution with the same concentration plus CT with a final concentration of 0.2 mg / ml “Ttd + CT”) was used. Twenty hours after applying the patch, the patch was removed and the auricle was washed. The above immunization operation was repeated 3 times at 2-week intervals, and blood was collected at 2-week intervals until 6 weeks after the first immunization to separate serum. Anti-Ttd-IgG antibody titer in serum was measured by ELISA.

  In addition, the antibody titer of the serum 6 weeks after the first immunization was measured by the KPA method (indirect aggregation reaction using artificially synthesized particles as a carrier), which is said to have a high correlation with the toxin neutralizing activity. Furthermore, 3 weeks after the final immunization, 10 times the half-lethal dose of tetanus toxin was administered subcutaneously to the thigh, and the survival of the mice was observed 2 days later.

  The results are shown in FIG. In the measurement by ELISA (FIG. 10), although slightly inferior to the group to which CT was added as an adjuvant, significant anti-Ttd-IgG antibody production was also observed in the group immunized transdermally with Ttd alone. The antibody titer measured by the KPA method also showed a serum antibody titer about 4 times higher in the adjuvant-added group than in the adjuvant-free group (Table 1). However, 100% of mice immunized without adjuvant survived asymptomatically against tetanus toxin challenge. From this, it was confirmed that the transdermal immunization method without using the adjuvant of the present invention was sufficiently effective.

Example 10: Comparison between a method of applying a patch soaked with an antigen solution and a method of applying a wet patch to the skin after application of the antigen The hair of the abdomen of 10 mice was clipped with a clipper to expose the skin. Two days later, the mice were anesthetized and divided into two groups of 5 animals each. In one group of mice, a patch soaked with 50 μl of ovalbumin (OVA) solution at a concentration of 2.0 mg / ml was applied to the exposed normal skin of the abdomen. In the other group of mice, 50 μl of 2.0 mg / ml OVA solution was dropped on the exposed normal skin of the abdomen, spread to about 1 cm ² and dried, and then 50 μl of physiological saline was applied to the same site. A soaked patch was applied. In both groups, the patch was removed after 18 hours and the antigen administration site was thoroughly washed. The above immunization was performed 3 times at 2-week intervals. Blood was collected 2 weeks after the final immunization, and the anti-OVA-IgG antibody titer in the serum was measured by ELISA.

  The results are shown in FIG. Compared with the method of applying a patch soaked with an antigen solution, the method of applying a moist patch to the skin after application of the antigen significantly increased the amount of anti-OVA-IgG produced and was more effective. Therefore, the method of applying a patch soaked with an antigen solution can also provide a sufficient transdermal immunity effect, but the method of applying a wet patch to the skin after application of the antigen has been found to be more advantageous.

FIG. 1 is a graph showing individual anti-OVA-IgG antibody titers in mice in which mice were immunized transcutaneously at various times by applying a wet patch after applying an antigen solution. FIG. 2 shows anti-OVA-IgG (panel A) and anti-CT-IgG (panel B) according to individual of mice immunized transcutaneously without applying a patch after applying the antigen solution and mice immunized by intradermal injection. It is a figure which shows an antibody titer. “OVA” = ovalbumin, “OVA + CT” = ovalbumin + cholera toxin (hereinafter the same). FIG. 3 is a view showing the anti-OVA-IgG antibody titer of mice subjected to transcutaneous immunization by applying a patch soaked with an antigen solution. FIG. 4 is a diagram showing the effect of using a patch after application of an antigen solution according to the anti-OVA-IgG antibody titer for each mouse. FIG. 5 is a graph showing the anti-OVA-IgG antibody titers of individual mice subjected to transcutaneous immunization by applying patches impregnated with solutions containing various amounts of antigen. FIG. 6 is a diagram showing the relationship between multiple antigen administrations and the transition of antibody production. Arrows indicate the time of antigen administration. FIG. 7 is a diagram showing the relationship between antigen administration over a longer period and the transition of antibody production. Arrows indicate the time of antigen administration. FIG. 8 is a diagram showing retention of the priming effect by the first immunization for each mouse. FIG. 9 is a diagram illustrating transmission of a priming effect. FIG. 10 is a diagram showing the effect of transcutaneous immunity when tetanus toxoid is used as an antigen. FIG. 11 is a diagram comparing the effects of applying a patch soaked with an antigen solution and applying a wet patch after applying the antigen solution.

Claims (11)

An antigen kit for transdermal antigen administration without using an adjuvant,
A) a kit containing an antigen containing an antigen and no adjuvant, and B) a patch containing neither an antigen nor an adjuvant and having at least a water retaining material and a patch.   The antigen kit according to claim 1, which is used as a vaccine.   The antigen kit according to claim 1, which is used as an antigen in desensitization therapy or immunotherapy.   A step of applying to the skin an antigen composition containing neither an antigen nor an adjuvant, and having at least a water retaining material and a patch, the antigen containing and not containing the adjuvant; And a step of covering with a patch having a water-retaining material containing neither water nor an adjuvant, and maintaining the water-retaining material in contact with the skin of the application site for at least 2 hours. Patches used in the featured antigen administration method.   A patch that contains neither an antigen nor an adjuvant, and has at least a water-retaining material and a patch, and an aqueous antigen composition that contains an antigen and does not contain an adjuvant in the water-retaining material of the patch; A patch for use in an antigen administration method, comprising: a step of applying a patch with an adhesive so as to contact the material; and a step of maintaining the water-retaining material in contact with the skin for at least 2 hours .   An antigen composition that contains an antigen and does not contain an adjuvant, the step of applying the composition to the skin, and the water retaining material that does not contain either the antigen or the adjuvant, but contains water. An antigen composition for use in an antigen administration method, comprising: a step of coating with a patch comprising: a step of maintaining the water-retaining material in contact with the skin of the application site for at least 2 hours.   An aqueous antigen composition containing an antigen and not containing an adjuvant, wherein the aqueous composition is contained in a water retention material of a patch having at least a water retention material and a patch, and the skin and the water retention material are in contact with each other. An antigen composition for use in an antigen administration method, comprising: a step of applying a patch to the skin with a patch; and a step of maintaining the water retaining material in contact with the skin for at least 2 hours.   The method of using a kit according to claim 1, wherein the step of applying the antigen composition to the skin, and the skin of the application site, containing neither the antigen nor the adjuvant, the water retaining material containing water. Coating with a patch having and maintaining for at least 2 hours in contact with the skin at the application site.   The method of using the kit according to claim 1, wherein the antigen composition is contained in the water retention material of the patch, and the patch is affixed with a patch so that the skin and the water retention material are in contact with each other. Maintaining the water-retaining material in contact with the skin for at least 2 hours.   A step of applying an immunogen composition containing an antigen and not containing an adjuvant to the skin; and a step of coating the skin of the application site with a patch having a water-retaining material containing neither an antigen nor an adjuvant and containing water. And a step of maintaining the water-retaining material in contact with the skin at the application site for at least 2 hours, and a method for transdermal antigen administration without using an adjuvant.   A step of containing an aqueous immunogenic composition containing an antigen and no adjuvant in a water retention material of a patch having at least a water retention material and a patch, and affixing the patch with the patch so that the skin and the water retention material are in contact with each other A transdermal antigen administration method without using an adjuvant, comprising: a step of maintaining the water-retaining material in contact with the skin for at least 2 hours.

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