TW201406786A - Novel t cell epitope peptide of japanese cypress pollen allergen - Google Patents

Abstract

Provided is a novel T cell epitope peptide of Japanese cypress pollen allergen different from known Japanese cypress pollen allergens. A peptide selected from the group consisting of amino acid sequences shown by SEQ ID NO. 3-16, or a peptide having 1-3 amino acids substituted and/or 1-10 amino acids deleted or added in this peptide, the peptide having T cell stimulatory activity.

Description

T cell epitope peptide of novel eucalyptus pollen allergen

The present invention relates to a T cell epitope peptide derived from a novel allergen protein of eucalyptus pollen, and a peptide immunotherapy composition for eucalyptus hay fever using the peptide as an active ingredient.

The amount of eucalyptus pollen scattered was only a small amount in the 1940s (1965-1974), but it increased to more than three times in the next 20 years. The cultivation of Chinese fir was concentrated in the 1920s (1945~1954), and in the second half of the 1940s (1970-1974), the pollen began to scatter. The eucalyptus was planted in the 1930s (1955-1964) after the planting of Chinese fir, and it was later than the Chinese fir. In recent years, the number of vegetation of eucalyptus is gradually equal to the number of vegetation of Chinese fir, and it is predicted that the pollen of eucalyptus will increase in the future. Therefore, the industry has pointed out the necessity of immunotherapy using the antigen specific to eucalyptus pollen.

A major problem with immunotherapy is the anaphylactic reaction. An allergic reaction is caused by a chemical medium that is released by binding of an antigen-specific IgE antibody to an antigen on an obese cell. Peptide immunotherapy using a T cell epitope is an immunotherapy that does not include a part that binds to an IgE antibody and is a T cell epitope associated with immunotolerance, and has been used clinically by cats, A peptide immunotherapy agent for T cell epitopes identified by antigens such as alfalfa, Chinese fir pollen, and ragweed pollen (Non-Patent Documents 1 and 2). Further, regarding the eucalyptus pollen, a T cell epitope peptide was also identified for Cha o1 and Cha o 2 as main antigens (Patent Document 1 and Non-Patent Documents 3 and 4).

[Previous Technical Literature] [Patent Literature]

Patent Document 1: Japanese Patent Laid-Open Publication No. 2008-195718

[Non-patent literature]

Non-Patent Document 1: Valenta R. et al., Annu. Rev. Immunol., 2010: 211 - 241, 2010

Non-Patent Document 2: Casale T. et al., J. Allergy Clin. Immunol., 127: 8 - 15, 2011

Non-Patent Document 3: Sone, T. et al., Clin. Exp. Allergy, 35: 664 - 671, 2005

Non-Patent Document 4: Sone, T. et al., Allergol. Int., 58: 237 - 245, 2009

The present invention is intended to contribute to an improvement in the effectiveness of immunotherapy for beech pollen that is thought to increase in the future, and to provide a T cell epitope peptide of a novel eucalyptus pollen allergen different from the known eucalyptus pollen allergen.

The present inventors have found a novel eucalyptus pollen allergen which is abundantly present in eucalyptus pollen and has an allergen activity equal to or higher than Cha o1 or Cha o2, and has been applied for a patent (PCT/JP2012/052110). Further, according to the case of eucalyptus pollen sensitization of cedar pollen, a T cell line specific for the eucalyptus pollen allergen is separately established, and the main T cell epitope is determined therefrom, thereby completing the present invention.

That is, the present invention relates to the following 1) to 4).

1) A peptide selected from the group consisting of amino acid sequences represented by SEQ ID NOS: 3 to 16, or 1 to 3 amino acids in the peptide, and/or 1 or 10 additions or additions Amino acid, and has T cell stimulating activity.

2) The peptide according to the above 1), wherein the peptide lacking 1 to 10 amino acids lacks the amino acid of the N-terminal side and/or the C-terminal side of the peptide.

3) The peptide according to the above 2), which is a peptide which lacks the amino acid of the N-terminal side and/or the C-terminal side of the peptide comprising the amino acid sequence of SEQ ID NO: 7 and has at least "FKEKIVYEGHW", and the deletion includes The amino acid at the N-terminal side and/or the C-terminal side of the peptide of the amino acid sequence shown in SEQ ID NO: 10 and having at least a peptide of "PYYQIIYHPAS" or a peptide comprising the amino acid sequence of SEQ ID NO: 15 The amino acid on the N-terminal side and/or the C-terminal side and having at least a peptide of "FKVITTNKQ".

4) A peptide immunotherapy agent for eucalyptus hay fever, which comprises one or more of the peptides according to any one of the above 1) to 3) as an active ingredient.

5) Use of one or more of the peptides according to the above 1) to 3) for the production of a peptide immunotherapy agent for eucalyptus.

6) One or more of the peptides of the above 1) to 3), which are used for peptide immunotherapy of eucalyptus hay fever.

7) A peptide immunotherapy of eucalyptus pollination, which comprises one or more of the peptides according to the above 1) to 3).

The peptide group of the present invention can be used as an immunotherapeutic agent against eucalyptus hay fever by using a plurality of species alone or in combination. Furthermore, by combining with the T cell epitope peptide of Cha o1 or Cha o2, which is known as a eucalyptus pollen allergen, it can be caused by eucalyptus pollen which is not effective for immunotherapy using cedar pollen extract. Peptide immunotherapy useful for sensitive cases.

Figure 1 shows the T cell stimulating activity of each overlapping peptide.

Figure 2 shows the ability of the peptide of the present invention to bind to eucalyptus pollen-specific IgE.

Figure 3 shows the T cell stimulating activity of the short peptide (peptide number: #52).

Figure 4 shows the T cell stimulating activity of the short peptide (peptide number: #24).

Figure 5 shows the T cell stimulating activity of the short peptide (peptide number: #38).

Hereinafter, the present invention will be described in detail. Furthermore, the amino acid sequence in the present specification is described in an alphabet.

The peptide of the present invention comprising the amino acid sequence of SEQ ID NOs: 3 to 16 is derived from the eucalyptus pollen allergen protein comprising the amino acid sequence of SEQ ID NO: 1, or an isoform thereof (SEQ ID NO: 1) In the amino acid sequence, the second position Pro is substituted with Thr, the 230th position Val is replaced with Ile, the 271st position Val is replaced with Ile), that is, the protein containing the amino acid sequence of SEQ ID NO: 2 In the structure, 20 amino acid residues containing a total of 528 amino acids from the N-terminal Leu to the C-terminal Leu as the amino acid of the message peptide from the N-terminus are removed (partially 18) a partial peptide of an amino acid residue). The position on the amino acid sequence shown by SEQ ID NO: 1 or 2 of each peptide of the present invention is as shown in the following Tables 1-3, that is, SEQ ID NO: 3 (peptide number: #12): 111# of SEQ ID NO: 1. 130-position, SEQ ID NO: 4 (peptide number: #13): SEQ ID NO: 1 to 121 to 140, SEQ ID NO: 5 (peptide number: #20): 191 to 210 of SEQ ID NO: 1, SEQ ID NO: 6 (peptide) No.: #21): 201 to 220 of SEQ ID NO: 1, SEQ ID NO: 7 (peptide number: #24): 231 to 250 of SEQ ID NO: 1, SEQ ID NO: 8 (peptide number: #31): SEQ ID NO: No. 301-320 of SEQ ID NO: 9 (peptide number: #37): No. 361-380 of SEQ ID NO: 1, SEQ ID NO: 10 (peptide number: #38): No. 371-390 of SEQ ID NO: 1, SEQ ID NO: 11 (peptide number: #39): 381 to 400 of SEQ ID NO: 1, SEQ ID NO: 12 (peptide number: #41): 401 to 420 of SEQ ID NO: 1, SEQ ID NO: 13 (peptide number: # 42): 411 to 430 of SEQ ID NO: 1, SEQ ID NO: 14 (peptide number: #49): 481 to 500 of SEQ ID NO: 1, SEQ ID NO: 15 (peptide number: #52): SEQ ID NO: 1 511~528, sequence number 16 (peptide number: #56): sequence 2 of 261 - 280.

The above peptide of the present invention is exemplified by a sensitization of eucalyptus pollen according to Chinese fir pollinosis. A T cell line specific for the eucalyptus pollen allergen protein comprising the amino acid sequence shown in SEQ ID NO: 1 is established one by one, and the entire region covering the entire structure of the protein or its isoform is 20 The proliferative activity of 57 overlapping peptides of each residue was evaluated and identified as a T cell epitope (see Examples 2 to 4). Therefore, the peptide of the present invention has at least one T cell epitope, and the peptide of the present invention has T cell stimulating activity.

Here, the T cell stimulating activity refers to an activity of inducing a T cell response such as T cell proliferation or lymphokine secretion, and means that the immune tolerance of T cells can be induced by administration to humans or animals. The role. Here, the T cell stimulating activity can be determined, for example, by measuring the incorporation into the cell by [ ³ H]thymidine and calculating the stimulation coefficient. The stimulation factor (SI (Stimulation Index) value) of the T cell response used herein is obtained by dividing the cpm of the radioactivity of [ ³ H]thymidine taken into the cell in the presence of the peptide by the absence of the peptide ( Calculated according to the cpm of the control). The peptide system of the present invention has an average SI value of 2.0 or more.

Preferably, the "importance index" obtained by multiplying the average SI value of a certain peptide by the frequency (%) of the patient whose T cell response is expressed by the peptide is preferably at least 70 or more. No. 4 (peptide number: #13), SEQ ID NO: 5 (peptide number: #20), SEQ ID NO: 7 (peptide number: #24), SEQ ID NO: 8 (peptide number: #31), SEQ ID NO: 10 (peptide number: #38), a peptide of the amino acid sequence shown in SEQ ID NO: 11 (peptide number: #39), SEQ ID NO: 12 (peptide number: #41), and SEQ ID NO: 15 (peptide number: #52). Since the above peptide does not cause severe allergy by binding to an IgE antibody specific for an allergen of cedar pollen or eucalyptus pollen, it is particularly suitable as a T cell-inducing immune tolerance specific to a eucalyptus pollen allergen. Immunotherapeutic agents.

Further, in the present invention, in addition to the above-mentioned peptide comprising the amino acid sequence shown in SEQ ID NOs: 3 to 16, the peptide is also substituted with 1 to 3 amino acids and/or 1 or 10 amines in addition or in addition to the peptide. A peptide having a T cell stimulating activity.

Here, as a peptide in which 1 to 10 amino acids are deleted, for example, a N-terminal side and/or a deletion are included. The peptide of the amino acid at the C-terminal side may, for example, be 0 to 10 residues of the amino acid which is deleted on the N-terminal side, and 0 to 10 residues of the amino acid of the C-terminal side, and the total of 1 to 10 amines may be deleted. Acidic.

Specifically, for example, in the case of a peptide (#24) comprising the amino acid sequence shown in SEQ ID NO: 7, one to three residues of the amino acid on the N-terminal side are deleted and/or a peptide having 1 to 6 residues of the amino acid at the C-terminal side and having at least "FKEKIVYEGHW"; in the case of a peptide (#38) comprising the amino acid sequence of SEQ ID NO: 10, the N-terminal side is deleted. a peptide having 6 to 8 residues of amino acid and/or 1 amino acid of the C-terminal side and having at least "PYYQIIYHPAS"; and a peptide comprising the amino acid sequence of SEQ ID NO: 15 (#52) In the case of the above, a peptide having 1 to 2 residues of the amino acid on the N-terminal side and/or 1 to 7 residues of the amino acid on the C-terminal side and having at least "FKVITTNKQ" is deleted.

Further, as a peptide to which 1 to 10 amino acids are added, a peptide containing 1 to 10 amino acids is added to the N-terminal side and/or the C-terminal side, and in this case, as an additional amino acid residue Examples of the amino acid sequence shown in SEQ ID NO: 1 or 2 include 1 to 10 amino acid residues adjacent to the N-terminal side or the C-terminal side of the amino acid sequence corresponding to the peptide.

Here, the term "having T cell stimulating activity" means having a T cell stimulating activity equal to or higher than that of a peptide comprising the amino acid sequence represented by SEQ ID NOs: 3 to 16. Furthermore, the meaning of T cell stimulating activity is as described above.

The T cell epitope peptide of the present invention can be synthesized by solid phase synthesis, liquid phase synthesis, or a combination of these, which is used in the synthesis of a usual peptide. Further, in the case of synthesizing the T cell epitope peptide of the present invention by recombinant DNA technology, a host cell transformed with a nucleic acid having a sequence encoding the peptide can be cultured in a medium suitable for the cell, and used. The peptide is synthesized from the culture supernatant or from the host cell by a technique known to the practitioner. As the host, Escherichia coli, yeast, mammalian cells and the like can be used.

When the T cell epitope peptide of the present invention is administered to a human, preferably to an individual sensitized by a eucalyptus pollen allergen, the individual's allergic response to the allergen can be adjusted. Therefore, it can be used for peptide immunotherapy. Furthermore, in addition to the combination of other Elm pollen allergens, the T cell epitope peptide of Cha o1 or Cha o2, the T cell epitope peptide of Cry j1 or Cry j2, which is a cedar pollen allergen, is also used. Kaiping No. 7-118295, Japanese Patent Laid-Open No. Hei 8-47392, Sone, T. et al., J. Immunol., 161: 448-457, 1998), and can be used for the representative of Chinese fir-eucalyptus. Peptide immunotherapy for spring tree pollinosis patients.

The peptide immunotherapy agent for eucalyptus pollen of the present invention can be administered by a usual administration route such as injection (subcutaneous, intravenous injection, etc.), eye drop, nasal drops, oral, inhalation, transdermal, etc. according to the administration form. , can be prepared into tablets, capsules, granules, powders, buccal tablets, sublingual tablets, syrups, injections, suppositories, inhalants, percutaneous absorption agents, eye drops, nasal drops, nasal sprays Preparations for various dosage forms such as dressings, cataplasms, ointments, lotions, and creams.

In this case, it is preferred to prepare the T cell epitope peptide of the present invention directly or to dry it into a powder, and to add a carrier such as a diluent or an additive, such as a stabilizer or an excipient, by a usual method. A complexing agent, a dissolution aid, an opacifier, a buffer, a soothing agent, a preservative, a coloring agent, and the like.

The dosage of the peptide immunotherapy agent for the amaranth pollinosis of the present invention, for example, in the case of injection, the amount of the peptide of the present invention can be set to about 1 μg to 30 mg, preferably 20 per unit. Gg~10 mg.

Hereinafter, the present invention will be specifically described by way of examples, but the invention is not limited to the examples.

[Examples] Example 1 Purification of Alfalfa Pollen Allergen Protein

To the pollen 50 g pollen, 1000 ml of extraction buffer (25 mM Tris buffer, pH 8.0) was added and stirred for 10 minutes, followed by ultrasonic treatment for 15 minutes. The obtained suspension was centrifuged (3,000 rpm, 15 minutes), and the supernatant was obtained and filtered. This gives the extract. The extract was added to a DE52 anion exchange column equilibrated with 10 mM Tris buffer (pH 8.0) to collect non-adsorbed components. Then, it was adsorbed to a CM52 cation exchange column equilibrated with 20 mM acetate buffer (pH 5.2), and a gradient elutor was fractionated into components using a 10 mM Tris buffer (pH 7.8) containing NaCl. Recycling. A part of each component was analyzed by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and components containing a protein having a molecular weight of 66 kDa were combined.

Example 2 Synthesis of overlapping peptides

The overlapping peptides were synthesized by the PepTrack Peptide Library service (JPT Peptide Technologies). Based on the first-order structure of the eucalyptus pollen allergen protein (hereinafter referred to as "protein X") (SEQ ID NO: 1) obtained in Example 1, the synthesis consists of Leu starting from the first position of the N-terminus to the C-terminus. 52 (#1~#52) overlapping residues of 20 residues (#52 (SEQ ID NO: 52) are 18 residues) in the overlapping portion of 10 residues (Tables 1-2). Further, in the amino acid sequence represented by SEQ ID NO: 1, the Pro at the second position is replaced with Thr, and the Val at the 230th position is replaced with Ile and Val at position 271. Five (#53 to #57) overlapping peptides (Table 3) were prepared in the same manner for the protein of Ile (SEQ ID NO: 2).

Example 3 Establishment of a novel eucalyptus pollen-specific T cell line

In the peripheral blood of 20 patients sensitized by eucalyptus pollen from cedar pollen, the lymphocytes were isolated from the peripheral blood using the commonly used Ficoll-paque gravity centrifugation method, and a part was stored in liquid nitrogen. The peripheral blood lymphocytes (1.2×10 ⁷ ) were suspended in RPMI-1640 supplemented with 6 ml of autologous plasma, and cultured with 7-well culture plates on the 24-well culture plate. day. At a time when it was confirmed under the microscope that T cells activated by antigen stimulation were added, 20 Unit/ml of IL-2 was added and cultured overnight. From the next day, the culture medium is changed for 10 to 12 days by using 20 units/ml of IL-2 and RPMI-1640 supplemented with 10% of autologous plasma, and the culture is determined by T cell antigen. A T cell line for identification.

Example 4 Identification of peptides containing T cell epitopes

The T-cell line established from 20 patients with hay fever was cultured together with the separately synthesized overlapping peptides to identify protein X-specific T cell epitope peptides. That is, the T cell line as an antigen-presenting cell is melted with a peripheral blood mononuclear ball from a frozen reservoir of the same patient, and then irradiated with X-rays (36 Gy) and suspended in RPMI-1640 supplemented with 10% autologous plasma. in. After the antigen-presenting cells were seeded on a 96-well flat-bottomed plate (1 × 10 ⁵ cells/well), protein X (final concentration: 10 μg/ml) or each peptide (final concentration: 2 μM) was added to each well. Then, the T cell line (2×10 ⁴ cells/well) suspended in RPMI-1640 supplemented with 10% autologous plasma was inoculated into each well, and cultured at 37° C. under 5% CO ₂ for 48 hours. 1 μCi of [ ³ H] thymidine was added to the wells and cultured for further 16 hours. The cells were first captured on a glass filter using a harvester and dried, and the radioactivity (cpm) of [ ³ H]thymidine taken into the cells was measured using a microplate luminescence analyzer (manufactured by PerkinElmer). As an indicator of the proliferative activity, the value obtained by dividing the radioactivity generated by the peptide stimulation by the T cell line by the radioactivity of the unadded peptide (control) is calculated as the SI value, and the peptide value of the SI value of 2 or more is identified. It is a T cell epitope peptide. Figure 1 shows the "average SI value" of all overlapping peptides (the average of the SI values of 20), the frequency of occurrence (%) (the ratio of the SI value of 20 to 2 or more) and the "importance index" ( The value obtained by multiplying the average SI value by the frequency of occurrence).

Example 5 Confirmation of binding ability to eucalyptus pollen-specific IgE

The sera of five eucalyptus hay fever patients were used to confirm the ability to bind to eucalyptus pollen-specific IgE. Preferred T cell epitope peptides of the invention (Importance Index is 100 The above or the eucalyptus pollen allergen was added to Maxisorp 96-well culture plate (Nunc) at a rate of 1 μg/50 μL/well, and then allowed to stand at 4 ° C overnight to solidify. After the culture plate was washed the next day, PBS (Phosphate Buffered Saline, phosphate buffered saline) containing 1% bovine serum albumin was added thereto, and allowed to stand at room temperature for 1 hour, thereby blocking. After washing the plate, the patient serum or the negative control serum diluted 5 times was added and allowed to stand at room temperature for 2 hours. The negative control used a healthy donor serum (Radioallergosorbent Test score = 0). After washing the plate, a secondary antibody (HRP (Horseradish Peroxidase) labeled goat anti-human IgE antibody, Novus) diluted 7000 times with PBS containing 1% bovine serum albumin was added and allowed to stand at room temperature. 40 minutes. After the culture plate was washed, TMB (tetramethylbenzidine, tetramethylbenzidine, TMS Substrate Reagent Set) (BD Biosciences) was allowed to stand at room temperature for 20 minutes to develop TMB. After completion, color development was stopped by adding 1 N sulfuric acid, and the absorbance at a wavelength of 450 nm was measured using a Microplate Reader. This result is shown in FIG. Further, in Fig. 2, V012, V014, V031, V047, and V051 respectively indicate the IDs of five beech pollen patients.

It was confirmed that the T cell epitope peptide of the present invention did not bind to eucalyptus pollen-specific IgE.

Example 6 Establishment of a 20 mer epitope-specific T cell line

From the cedar pollinosis, the peripheral blood of the patient sensitized by eucalyptus pollen is separated from the lymphocytes by the Ficoll-paque specific gravity centrifugation method usually used, and a part is stored in liquid nitrogen. The peripheral blood lymphocytes were suspended in RPMI-1640 supplemented with autologous plasma, and cultured in a 96-well culture plate (1.5×10 ⁵ cells/well) together with a final concentration of 1 μM of 20 mer epitope peptide. 7~8 days. At the same time, on the 6th to 7th day of culture, antigen-presenting cells required for subculture were prepared as described below. That is, the peripheral blood mononuclear ball from the same patient's frozen reserve is awakened and suspended in RPMI-1640 medium supplemented with 10% autologous plasma, followed by the addition of 20 mer epitope peptide at a final concentration of 10 μM. The cells were cultured overnight on a 24-well culture plate. The next day, after X-rays (45 Gy) were irradiated to the cells, the 20 mer epitope peptide in the medium was washed and removed, and the obtained cells were used as surviving antigen-presenting cells. On the 7th to 8th day of lymphocyte culture, 1/3 of each well in the lymphocytes inoculated on a 96-well culture plate and human IL-2 with a final concentration of 100 Unit/ml, human IL with a final concentration of 50 ng/ml. -4 and submerged antigen-presenting cells suspended in RPMI-1640 medium supplemented with 10% autologous plasma were cultured for 7 days in a 96-well culture plate. For the remaining 2/3 of the lymphocytes of each well, they were used for evaluation of antigen specificity. All the lymphocytes that were determined to be 20-mer epitope-specific T-cell-positive lymphocytes were collected from IL-2, IL-4, and subcultured antigen-presenting cells for 7 days, and the final concentration was determined. 100 units/ml of human IL-2, human IL-4 with a final concentration of 50 ng/ml, and subcultured antigen in RPMI-1640 medium supplemented with 10% autologous plasma and prepared by the above method The culture was expanded for 7 days on a 24-well culture plate. Thereafter, the cells were further expanded by using the antigen-presenting cells every 7 days, and the cell population showing the 20 mer epitope-specific cell proliferation was designated as a 20 mer epitope-specific T cell line.

Example 7 Presumption of the epitope sequence core sequence

The following peptides were synthesized: peptides obtained by deleting one amino acid residue of the amino acid from the C-terminus of the epitope 20 mer (#52-1~#52-8, #24-1~#24-8 #38-1~#24-3), a peptide belonging to each amino acid residue of the amino acid is deleted from the N-terminus (#52-9~#52-13, #24-9~#24- 13), and a peptide lacking 6 to 9 amino acid residues from the N-terminus (#38-4~#24-7). The epitope determinant core sequence was estimated by culturing the established 20 mer epitope peptide-specific T cell line together with the short peptide described above. That is, the T cell line as an antigen-presenting cell is melted with a peripheral blood mononuclear ball from a frozen reservoir of the same patient, and then irradiated with X-rays (45 Gy) and suspended in RPMI-1640 supplemented with 10% autologous plasma. in. After the antigen-presenting cells were seeded on a 96-well flat-bottomed plate (1 × 10 ⁵ cells/well), 20 mer peptide or each short peptide (final concentration 2 μM) was added to each well. Then, the T cell line (2×10 ⁴ cells/well) suspended in RPMI-1640 supplemented with 10% autologous plasma was inoculated into each well, and cultured at 37° C. under 5% CO ₂ for 48 hours. 1 μCi of [ ³ H] thymidine was added to the wells and cultured for further 16 hours. The cells were first captured on a glass filter using a harvester and dried, and the radioactivity (cpm) of [ ³ H]thymidine taken into the cells was measured using a microplate luminescence analyzer (manufactured by PerkinElmer). As an index of the proliferative activity, the value obtained by dividing the radioactivity generated by the peptide stimulation by the T cell line by the radioactivity (control) of the unadded peptide was calculated as the SI value. One of the results is illustrated in FIGS. 3 to 5.

The terminal sequence was determined to be the core sequence based on the smallest sequence (S.I. value of 2 or more) which exhibits T cell proliferation activity by using the short N-terminal and C-terminal short peptides. The core sequences on #24 epitope peptide, #38 epitope peptide and #52 epitope peptide successfully predicted by the above method are as follows: #24 epitope peptide (SEQ ID NO: 7) is position 4-14 "FKEKIVYEGHW", #38 epitope peptide (SEQ ID NO: 10) is "PYYQIIYHPAS" at positions 9 to 19, and #52 epitope peptide (SEQ ID NO: 15) is "FKVITTNKQ" at positions 3 to 11.

<110> Rishang Dapeng Pharmaceutical Industry Co., Ltd.

<120> T cell epitope peptide of novel eucalyptus pollen allergen

<130> TH0083

<150> JP 2012-166053

<151> 2012-7-26

<160> 59

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<210> 38

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 38

<210> 39

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 39

<210> 40

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 40

<210> 41

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 41

<210> 42

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 42

<210> 43

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 43

<210> 44

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 44

<210> 45

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 45

<210> 46

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 46

<210> 47

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 47

<210> 48

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 48

<210> 49

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 49

<210> 50

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 50

<210> 51

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 51

<210> 52

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 52

<210> 53

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 53

<210> 54

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 54

<210> 55

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 55

<210> 56

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 56

<210> 57

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 57

<210> 58

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 58

<210> 59

<211> 20

<212> PRT

<213> Labor

<220>

<223> Synthetic peptide

<400> 59

Claims (6)

A peptide selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 3 to 16, or 1 to 3 amino acids in the peptide, and/or 1 or 10 amino groups in addition or in addition. It is acidic and has T cell stimulating activity. The peptide of claim 1, wherein the peptide lacking 1 to 10 amino acids lacks the amino acid of the N-terminal side and/or the C-terminal side of the peptide. The peptide of claim 2, which is a peptide lacking the amino acid of the N-terminal side and/or the C-terminal side of the peptide comprising the amino acid sequence of SEQ ID NO: 7 and having at least "FKEKIVYEGHW", the deletion comprising the sequence number The amino acid at the N-terminal side and/or the C-terminal side of the peptide of the amino acid sequence shown by 10 and having at least a peptide of "PYYQIIYHPAS" or a peptide lacking the peptide comprising the amino acid sequence of SEQ ID NO: 15 Amino acid on the terminal side and/or the C-terminal side and having at least a peptide of "FKVITTNKQ". A peptide immunotherapy agent for eucalyptus pollen containing one or more of the peptides according to any one of claims 1 to 3 as an active ingredient. A use of one or more of the peptides according to any one of claims 1 to 3 for the manufacture of a peptide immunotherapy agent for eucalyptus. One or more of the peptides according to any one of claims 1 to 3, which are used for peptide immunotherapy of alder pollinosis.