Photocontrolled Immune Responses using Photochromic Antigen

使用光致变色抗原的光控免疫反应

• 批准号: 05558089
• 负责人: NAKANISHI Mamoru
• 金额: $ 9.92万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05558089/
• 关键词: photochromic antigen; antigen-antibody reaction; photocontrol; T cell; immune response; antigen-presenting cell; cis-trans isomerization; calcium ion; 抗原抗体反応光制御; フォトクロミック分子; T細胞レセプター; アゾベンゼン; 抗体; 免疫リポソーム; 細胞内カルシウムイオン

One of the major characteristics of antigen-antibody reactions is their precise molecular recognition that can distinguish even a small change in the antigen structure. This precise molecular recognition is useful for many applications in technology as well as in medical sciences. Monoclonal antibodies against a tetrapeptide carrying a photochromic azobenzene moiety (Glu-azoAla-Gly-Gly, azoAla=L-p-phenylazophenylalanine) were prepared. The antibodies bind the hapten peptide when the azobenzen moiety is in the trans form, but release the peptide in the cis form. The mechanism of photoreversible binding and release was studied using a pulse laser light. Photoisomerization of the hapten peptide was found to occur inside the binding site, indicating thatthe latter is flexible enough to allow the trans-cis photoisomerization wthin a few ten picoseconds. Further, we prepared T-cell hybrodomas against photochromic antigen. Using these photochromic T-cell hybridomas we studied the interaction between T cell and antigen-presenting cells by confocal fluorescence microscopy. It was found that antigen presentation caused activation-driven cell death (apoptosis) in T-cell hybridomas. These results indicated that apoptosis in T cells are able to be photocontrolled by using these T-cell hybridoma systems.

抗原抗体反应的主要特征之一是它们的精确分子识别，即使可以区分抗原结构的微小变化。这种精确的分子识别对于技术和医学科学的许多应用都是有用的。制备了针对携带光致变色偶氮苯一部分的四肽的单克隆抗体（Glu-azoala-gly-gly，Azoala = L-P-P-苯基唑酚丙氨酸）。抗体结合偶然肽时，当偶氮芬部分处于反式形式时，但以顺式形式释放肽。使用脉搏激光光研究了光疗结合和释放的机制。在结合位点发现了触觉肽的光异构化，这表明后者足够灵活，可以允许几十个picseconds的反式传播光异构化。此外，我们制备了针对光致变色抗原的T细胞杂种。使用这些光致变色T细胞杂交瘤，我们通过共聚焦荧光显微镜研究了T细胞和抗原呈递细胞之间的相互作用。发现抗原表现引起了T细胞杂交瘤中激活驱动的细胞死亡（凋亡）。这些结果表明，使用这些T细胞杂交瘤系统可以光控制T细胞的凋亡。

项目成果

R.Teshima: "Herbimycin A and ST637 inhibit receptor-activated phosphoinositide breakdown and Ca^<2+> signals in rat basophilic leukemia cells" Biochim.Biophys.Acta. (in press). (1994)

R.Teshima：“Herbimycin A 和 ST637 抑制大鼠嗜碱性白血病细胞中受体激活的磷酸肌醇分解和 Ca 2+ 信号”Biochim.Biophys.Acta。

中西,守: "免疫応答とイメージング" 生物物理. 34. 1-5 (1994)

Mamoru Nakanishi：“免疫反应和成像”生物物理学 34. 1-5 (1994)。

T.Iwaki: "Antibodies for fluorescent molecular rotors" Biochemistry. 32. 7589-7592 (1993)

T.Iwaki：“荧光分子转子的抗体”生物化学。

M.Harada: "Photocontrolled uptake and release of photochromic haptens by monoclonal antibodies" Bull.Chem.Soc.Jpn.67. 1380-1385 (1994)

M.Harada：“单克隆抗体对光致变色半抗原的光控摄取和释放”Bull.Chem.Soc.Jpn.67。

中西,守: "生体超分子システム(免疫系と超分子システム)" 共立出版, 340(9) (1993)

Mamoru Nakanishi：“生物超分子系统（免疫系统和超分子系统）”Kyoritsu Shuppan，340（9）（1993）