1999 年度実績報告書

アレルギー疾患の遺伝子治療に関する基礎的研究

研究課題

研究課題/領域番号	10670419
研究機関	横浜市立大学
研究代表者	青木一郎横浜市立大学, 医学部, 教授 (00184028)
研究分担者	宮城洋平横浜市立大学, 医学部, 講師 (00254194) 石ヶ坪良明横浜市立大学, 医学部, 教授 (40137039) 奥田研爾横浜市立大学, 医学部, 教授 (40124862)
キーワード	FcR / マスト細胞 / アレルギー / 遺伝子治療 / アナフィラキシー / IgE / マウス / 発現プラスミド
研究概要	われわれはマウスIgEレセプターであるFcεRα鎖をマウス脾細胞cDNAライブラリーよりRT-PCR法にてクローニングした。そのDNAに免疫グロブリンの分泌シグナルを結合させたコンストラクトを作成しmyc-epitopeをtagとしpCAG-GSおよびpSeqTaq2Aベクター2種類の発現プラスミドに組み込んだ。 In vitroでHEK293T細胞に導入した。その培養上清の抗MYC-tag抗体を用いたウェスタンブロット法では予想される分子量のところにバンドが確認された。また、pCAGはpSeqTagに比べ約10倍の蛋白質が培養上清に分泌されていることを見出した。この培養上清中蛋白とIgEとの結合を確認するためIgEとin vitroで反応させた。これによりIgEによるラット受身皮膚アナフィラキシー(PCA)反応惹起能は培養上清蛋白の濃度依存的に減弱した。この発現プラスミドの直接投与によるPCA反応の抑制を試みた。IgE皮内投与24時間前にIgE感作部位の周囲に2ヶ所10μgのプラスミドを皮内注射したところ、ラットPCA反応の抑制に成功した。この抑制効果はプラスミド投与後96時間では消失していた。発現プラスミドのin vivo投与についてはHIVに対するDNAワクチンで投与経路やアジュバンドの利用に関して多くの知見を得たので、それらの知見を利用し、投与方法などを検討し、効果の持続性の向上をはかるとともに他のアレルギーモデルへの応用を検討したい。これらと平行してFcεRの主たる発現細胞であるマスト細胞の免疫機能を検討した。その結果、経口と経皮の同時免疫によりTh2型の免疫応答が効率的に誘導されること、その誘導にマスト細胞が深く関与していることを明らかにした。IgEを介したマスト細胞の応答がTh2型免疫の成立に重要である可能性が示唆された。今後、可溶型FcεRα鎖によりTh1/Th2免疫応答がmodulateされるかどうかを検討した。

研究成果
(42件)

すべてその他

すべて文献書誌 (42件)

[文献書誌] Sasaki S et al.: "Comparison of intranasal and intramuscular Immunization against human immuncdeficiency virus type 1 with a DNA-monophosphoryl lipid A adjuvant vaccine."Infection and Immunity. 66. 823-826 (1998)
[文献書誌] Sasaki S et al.: "Induction of systemic and mucosal immune responses to human immunodeficiency virus type 1 by a DNA vaccine formulated with QS-21 saponin adjuvant via intramuscular and intransal routes."Journal of Virology. 72. 4931-4939 (1998)
[文献書誌] Hamajima K et al.: "Intranasal administration of HIV-DNA vaccine formulated with polymer,carboxymethylcellulose,augments mucosal antibody production and cell-mediated immune response."Clinical Immunology and Immunopathology. 88. 205-210 (1998)
[文献書誌] Miyajima Y et al.: "Long-range effect of mutation of calcium binding asparates on the catalytic activity of alkaline protease from Psudomonas aeruginosa."Journal of Biochemistry. 123. 24-27 (1998)
[文献書誌] Kato Y et al.: "Stimulation of motility of human renal cell carcinoma by SPARC/Osteonectin/BM-40 associated with type IV collagen."Invasion and Metastasis. 18. 105-114 (1998)
[文献書誌] Miyata S et al.: "Stimulation of cellular growth and adhesion to fibronectin and vitronectin in culture and tumorigen*** in nude mice by overexpression of trypsinogen in human gastric cancer cells."Clinical Experimental Metastasis. 16. 613-622 (1998)
[文献書誌] Sasaki S et al.: "Adjuvant effect ubenimex on a DNA vaccine for HIV-1."Clinical Experimental Immunology. 111. 30-35 (1998)
[文献書誌] Sasaki S et al.: "The search for potent DNA vaccine againts AIDS:The enhancement of immunogenicity by chemical and genetic adjuvants."Anticancer Research. 18. 3907-3916 (1998)
[文献書誌] Aoki I et al: "In vitro Development of Th2 Cells by antigen stimulation."Yokohama Medical Bulletin. 46. 9-16 (1998)
[文献書誌] Putkonen P et al: "Immune responses but no protection against SHIV by gene-gun deliverry of HIV-1 DNA followed by recombinant subunit protein boosts."Virology. 250. 293-301 (1998)
[文献書誌] Xin K-Q et al: "Intranasal administration of human immunodeficiency virus type-1 (HIV-1) DNA vaccine with interleukin-2 expression plasmid enhances cell-mediated immunity against HIV-1."Immunology. 94. 438-444 (1998)
[文献書誌] Nagashima Y et al: "Segmental mediolytic arteritis:A case report with review of literature."Pathology Research and Practice. 194. 643-647 (1998)
[文献書誌] Ott I et al: "A Role for Tissue Factor in Cell Adhesion and Migration Mediated by Interaction with Actin-binding Protein 280."The Journal of Cell Biology. 140. 1241-1253 (1998)
[文献書誌] Hirahara F ert al: "Differential expression of trypsin in human ovarian carcinomas and low-malignant-potential tumors."Gynecologic Oncology. 68. 162-165 (1998)
[文献書誌] Zhou L et al: "Evaluation of Epstein-Barr Virus infection in hypopharyngeal carcinomas from 37 Japanese pati**"Modern Pathology. 11. 509-512 (1998)
[文献書誌] Kato Y et al: "Production of trypsins by human gastric cancer cells correlates with their malignant phenotype."European Journal of Cancer. 34. 1117-1123 (1998)
[文献書誌] Koshikawa N et al: "Expression of trypsin by epithelial cells of various tissues,leukoctes,and neurons in human and mouse."American Journal of Pathology. 153. 937-944 (1999)
[文献書誌] Okudo T et al: "Administration of an IL-12 encoding DNA plasmid prevents the development of chronic graft-versus-host disease."Journal of Immunology. 162. 4013-4017 (1999)
[文献書誌] Sasaki S et al: "Adjuvant effect of ubenimex on a DNA vaccine for HIV-1."Clinical Experimental Immunology. 111. 30-35 (1998)
[文献書誌] Sasaki S et al: "The search for a potent DNA vaccine against AIDS:The enhancement of immunogenicity by chemical and genetic adjuvants."Anticancer Research. 18. 3907-3916 (1998)
[文献書誌] Aoki I et al: "In vitro Development of Th2 Cells by antigen stimulation."Yokohama Medical Bulletin. 46. 9-16 (1998)
[文献書誌] Putkonen P et al: "Immune responses but no protection against SHIV by gene-gun deliverry of HIV-1 DNA followed by recombinant subunit protein boosts."Virology. 250. 293-301 (1998)
[文献書誌] Xin K-Q et al: "Intranasal administration of human immunodeficiency virus type-1 (HIV-1) DNA vaccine with interleukin-2 expression plasmid enhances cell-mediated immunity against HIV-1."Immunology. 94. 438-444 (1998)
[文献書誌] Nagsdhima Y et al: "Segmental mediolytic arteritis:A case report with review of literature."Pathology Research and Practice. 194. 643-647 (1998)
[文献書誌] Ott I et al: "A Role for Tissue Factor in Cell Adhesion and Migration Mediated by Interaction with Actin-binding Protein 280."The Journal of Cell Biology. 140. 1241-1253 (1998)
[文献書誌] Hirahara F ert al: "Differential expression of trypsin in human ovarian carcinomas and low-malignant-potential tumors."Gynecologic Oncology. 68. 162-165 (1998)
[文献書誌] Zhou L et al: "Evaluation of Epstein-Barr Virus infection in hypopharyngeal carcinomas from 37 Japanese pati**"Modern Pathology. 11. 509-512 (1998)
[文献書誌] Kato Y et al: "Production of trypsins by human gastric cancer cells correlates with their malignant phenotype."European Journal of Cancer. 34. 1117-1123 (1998)
[文献書誌] Koshikawa N et al: "Expression of trypsin by epithelial cells of various tissues,leukoctes,and neurons in human and mouse."American Journal of Pathology. 153. 937-944 (1999)
[文献書誌] Okudo T et al: "Administration of an IL-12 encoding DNA plasmid prevents the development of chronic graft-versus-host disease."Journal of Immunology. 162. 4013-4017 (1999)
[文献書誌] Xin K-Q et al: "IL-15 Expression Plasmid Enhances Cell-Mediated Immunity Induced by an HIV-1 DNA Vaccine."Vaccine. 17. 858-866 (1999)
[文献書誌] Asakura Y et al: "DNA-plasmids of HIV-1 induce systemic and mucosal immune responses."Biological Chemistry. 380. 375-379 (1999)
[文献書誌] Ihata A et al: "Immunomodulatory effect of a plasmid expressing CD40 ligand on DNA vaccination against human immunodeficiency virus type-1."Immunology. 98. 436-442 (1999)
[文献書誌] Xin K-Q et al: "Evidence of HIV Type 1 Glycoprotein 120 Binding to Recombiant N-Methyl-D-Aspartate Receptor Subunits Expressed in a Baculovirus System."AIDS Research and Human Retroviruses. 15. 1461-1467 (1999)
[文献書誌] Lu Y et al: "Macrophage inflammatory prorein-1 α (MIP-1 α) expression plasmid enhances DNA vaccine-induced immune response against HIV-1."Clinical Experimental Immunology. 115. 335-341 (1999)
[文献書誌] Aoki I et al: "Contrition of mast cells to the T helper 2 response induced by simultaneous subcutaneous and oral immunization."Immunology. 98. 519-524 (1999)
[文献書誌] Sasaki S et al: "Cenes that induce immunity-DNA vaccines."Microbiology Immunology. 43. 191-200 (1999)
[文献書誌] Kawano N et al: "Desmoplastic small round-cell tumor of the partesticular region:report of an adult case with demonstration of EWS and WT1 gene fusion using paraffin-embedded tissue."Modern Pathology. 12. 729-734 (1999)
[文献書誌] Miyata S et al: "Expression of trypsin in human cancer cell lines and cancer tissues and its tight binding to soluble fprm of alzheimer amyloid precursor protein in culture."Journal of Biochemistry. 125. 1067-1076 (1999)
[文献書誌] Fischer EG et al: "Tumor cell adhesion and migration supported by interaction of a receptor-protease complex with its inhibitor."Journal of Clinical Investigation. 104. 1213-1221 (1999)
[文献書誌] Kato H et al: "Rectal and vaginal immunization with a macromolecular multicomponent peptide vaccine candidate for HIV-1 infection induces HIV-specific protective immune reponses."Vaccine. 18. 1151-1160 (2000)
[文献書誌] Kusakabe K et al: "The timing of GM-CSF expression plasmid administration influences the Th1/Th2 response induced by an HIV-1specific DNA vaccine."Journal of Immunology. (In press).

1999 年度 実績報告書

アレルギー疾患の遺伝子治療に関する基礎的研究

研究代表者

青木 一郎 横浜市立大学, 医学部, 教授 (00184028)

研究成果

[文献書誌] Sasaki S et al.: "Comparison of intranasal and intramuscular Immunization against human immuncdeficiency virus type 1 with a DNA-monophosphoryl lipid A adjuvant vaccine."Infection and Immunity. 66. 823-826 (1998)

[文献書誌] Sasaki S et al.: "Induction of systemic and mucosal immune responses to human immunodeficiency virus type 1 by a DNA vaccine formulated with QS-21 saponin adjuvant via intramuscular and intransal routes."Journal of Virology. 72. 4931-4939 (1998)

[文献書誌] Hamajima K et al.: "Intranasal administration of HIV-DNA vaccine formulated with polymer,carboxymethylcellulose,augments mucosal antibody production and cell-mediated immune response."Clinical Immunology and Immunopathology. 88. 205-210 (1998)

[文献書誌] Miyajima Y et al.: "Long-range effect of mutation of calcium binding asparates on the catalytic activity of alkaline protease from Psudomonas aeruginosa."Journal of Biochemistry. 123. 24-27 (1998)

[文献書誌] Kato Y et al.: "Stimulation of motility of human renal cell carcinoma by SPARC/Osteonectin/BM-40 associated with type IV collagen."Invasion and Metastasis. 18. 105-114 (1998)

[文献書誌] Miyata S et al.: "Stimulation of cellular growth and adhesion to fibronectin and vitronectin in culture and tumorigen*** in nude mice by overexpression of trypsinogen in human gastric cancer cells."Clinical Experimental Metastasis. 16. 613-622 (1998)

[文献書誌] Sasaki S et al.: "Adjuvant effect ubenimex on a DNA vaccine for HIV-1."Clinical Experimental Immunology. 111. 30-35 (1998)

[文献書誌] Sasaki S et al.: "The search for potent DNA vaccine againts AIDS:The enhancement of immunogenicity by chemical and genetic adjuvants."Anticancer Research. 18. 3907-3916 (1998)

[文献書誌] Aoki I et al: "In vitro Development of Th2 Cells by antigen stimulation."Yokohama Medical Bulletin. 46. 9-16 (1998)

[文献書誌] Putkonen P et al: "Immune responses but no protection against SHIV by gene-gun deliverry of HIV-1 DNA followed by recombinant subunit protein boosts."Virology. 250. 293-301 (1998)

[文献書誌] Xin K-Q et al: "Intranasal administration of human immunodeficiency virus type-1 (HIV-1) DNA vaccine with interleukin-2 expression plasmid enhances cell-mediated immunity against HIV-1."Immunology. 94. 438-444 (1998)

[文献書誌] Nagashima Y et al: "Segmental mediolytic arteritis:A case report with review of literature."Pathology Research and Practice. 194. 643-647 (1998)

[文献書誌] Ott I et al: "A Role for Tissue Factor in Cell Adhesion and Migration Mediated by Interaction with Actin-binding Protein 280."The Journal of Cell Biology. 140. 1241-1253 (1998)

[文献書誌] Hirahara F ert al: "Differential expression of trypsin in human ovarian carcinomas and low-malignant-potential tumors."Gynecologic Oncology. 68. 162-165 (1998)

[文献書誌] Zhou L et al: "Evaluation of Epstein-Barr Virus infection in hypopharyngeal carcinomas from 37 Japanese pati**"Modern Pathology. 11. 509-512 (1998)

[文献書誌] Kato Y et al: "Production of trypsins by human gastric cancer cells correlates with their malignant phenotype."European Journal of Cancer. 34. 1117-1123 (1998)

[文献書誌] Koshikawa N et al: "Expression of trypsin by epithelial cells of various tissues,leukoctes,and neurons in human and mouse."American Journal of Pathology. 153. 937-944 (1999)

[文献書誌] Okudo T et al: "Administration of an IL-12 encoding DNA plasmid prevents the development of chronic graft-versus-host disease."Journal of Immunology. 162. 4013-4017 (1999)

[文献書誌] Sasaki S et al: "Adjuvant effect of ubenimex on a DNA vaccine for HIV-1."Clinical Experimental Immunology. 111. 30-35 (1998)

[文献書誌] Sasaki S et al: "The search for a potent DNA vaccine against AIDS:The enhancement of immunogenicity by chemical and genetic adjuvants."Anticancer Research. 18. 3907-3916 (1998)

[文献書誌] Aoki I et al: "In vitro Development of Th2 Cells by antigen stimulation."Yokohama Medical Bulletin. 46. 9-16 (1998)

[文献書誌] Putkonen P et al: "Immune responses but no protection against SHIV by gene-gun deliverry of HIV-1 DNA followed by recombinant subunit protein boosts."Virology. 250. 293-301 (1998)

[文献書誌] Xin K-Q et al: "Intranasal administration of human immunodeficiency virus type-1 (HIV-1) DNA vaccine with interleukin-2 expression plasmid enhances cell-mediated immunity against HIV-1."Immunology. 94. 438-444 (1998)

[文献書誌] Nagsdhima Y et al: "Segmental mediolytic arteritis:A case report with review of literature."Pathology Research and Practice. 194. 643-647 (1998)

[文献書誌] Ott I et al: "A Role for Tissue Factor in Cell Adhesion and Migration Mediated by Interaction with Actin-binding Protein 280."The Journal of Cell Biology. 140. 1241-1253 (1998)

[文献書誌] Hirahara F ert al: "Differential expression of trypsin in human ovarian carcinomas and low-malignant-potential tumors."Gynecologic Oncology. 68. 162-165 (1998)

[文献書誌] Zhou L et al: "Evaluation of Epstein-Barr Virus infection in hypopharyngeal carcinomas from 37 Japanese pati**"Modern Pathology. 11. 509-512 (1998)

[文献書誌] Kato Y et al: "Production of trypsins by human gastric cancer cells correlates with their malignant phenotype."European Journal of Cancer. 34. 1117-1123 (1998)

[文献書誌] Koshikawa N et al: "Expression of trypsin by epithelial cells of various tissues,leukoctes,and neurons in human and mouse."American Journal of Pathology. 153. 937-944 (1999)

[文献書誌] Okudo T et al: "Administration of an IL-12 encoding DNA plasmid prevents the development of chronic graft-versus-host disease."Journal of Immunology. 162. 4013-4017 (1999)

[文献書誌] Xin K-Q et al: "IL-15 Expression Plasmid Enhances Cell-Mediated Immunity Induced by an HIV-1 DNA Vaccine."Vaccine. 17. 858-866 (1999)

[文献書誌] Asakura Y et al: "DNA-plasmids of HIV-1 induce systemic and mucosal immune responses."Biological Chemistry. 380. 375-379 (1999)

[文献書誌] Ihata A et al: "Immunomodulatory effect of a plasmid expressing CD40 ligand on DNA vaccination against human immunodeficiency virus type-1."Immunology. 98. 436-442 (1999)

[文献書誌] Xin K-Q et al: "Evidence of HIV Type 1 Glycoprotein 120 Binding to Recombiant N-Methyl-D-Aspartate Receptor Subunits Expressed in a Baculovirus System."AIDS Research and Human Retroviruses. 15. 1461-1467 (1999)

[文献書誌] Lu Y et al: "Macrophage inflammatory prorein-1 α (MIP-1 α) expression plasmid enhances DNA vaccine-induced immune response against HIV-1."Clinical Experimental Immunology. 115. 335-341 (1999)

[文献書誌] Aoki I et al: "Contrition of mast cells to the T helper 2 response induced by simultaneous subcutaneous and oral immunization."Immunology. 98. 519-524 (1999)

[文献書誌] Sasaki S et al: "Cenes that induce immunity-DNA vaccines."Microbiology Immunology. 43. 191-200 (1999)

[文献書誌] Kawano N et al: "Desmoplastic small round-cell tumor of the partesticular region:report of an adult case with demonstration of EWS and WT1 gene fusion using paraffin-embedded tissue."Modern Pathology. 12. 729-734 (1999)

[文献書誌] Miyata S et al: "Expression of trypsin in human cancer cell lines and cancer tissues and its tight binding to soluble fprm of alzheimer amyloid precursor protein in culture."Journal of Biochemistry. 125. 1067-1076 (1999)

[文献書誌] Fischer EG et al: "Tumor cell adhesion and migration supported by interaction of a receptor-protease complex with its inhibitor."Journal of Clinical Investigation. 104. 1213-1221 (1999)

[文献書誌] Kato H et al: "Rectal and vaginal immunization with a macromolecular multicomponent peptide vaccine candidate for HIV-1 infection induces HIV-specific protective immune reponses."Vaccine. 18. 1151-1160 (2000)

[文献書誌] Kusakabe K et al: "The timing of GM-CSF expression plasmid administration influences the Th1/Th2 response induced by an HIV-1specific DNA vaccine."Journal of Immunology. (In press).

1999 年度実績報告書

青木一郎横浜市立大学, 医学部, 教授 (00184028)