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THE APPLICATION OF THE DENDRITIC CELL GENE THERAPY IN TRANSPLANTATION FIELD -Induction of antigen-specific T cell proliferation by Indoleamine 2,3-dioxygenase cDNA-transfected dendritic cells-

树突状细胞基因治疗在移植领域的应用-吲哚胺2,3-双加氧酶cDNA转染的树突状细胞诱导抗原特异性T细胞增殖-

基本信息

批准号：
13671250
负责人：
MIYAZAKI Kunihisa
金额：
$ 2.3万
依托单位：
Jichi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671250/
关键词：
adenovirus IDO tryptophan organ transplantation cre loxP XS106 gene therapy dendritic cell lentivirus vector apoptosis 膵頭移殖 RCAS-1 キヌレニン

项目摘要

Purpose : Indoleamine 2,3-dioxygenase (IDO) is an enzyme, involved in the catabolism of tryptophan and has been show to prevent rejection of the fetus during pregnancy, probably by inhibiting alloreactive T cells. In the present study, we investigated whether DCs that are transfected with IDO cDNA in inhibition of T cell proliferation after antigen-specific interaction. Methods : IDO was expressed with a gene delivery system using a recombinantadenoviral vector and its expression and function were confirmed by Western blot, immunology staining and kynurenine assay. The expression of the surface molecular of the IDO-transfected XS106 DC clone (derived from A/J mice) was confirmed by FACS analysis. Alloreactive T cell proliferation was assayed after culture with IDO-expressed DC. Results : A recombinant adenoviral vector expressing IDO was successfully generated and its gene expression detected by Western blot and immune staining. Its catabolic effect was confirmed by increase the kynurenine concentration. It revealed that IDO-expressing XS106 DC were no changeable for CD86, CDllc and CD69 expression. After co-cultured IDO-expressing DC with B6 allogeneic splenic T cell, the proliferation of the T cell was inhibited significantly. Conclusions : These results suggest that over expression of the IDO in the DC was effective to inhibit T cell proliferation, and may expand a new immunomodulatory strategy to prevent the allo-rejection of the organ transplantation.

目的：吲哚胺 2,3-双加氧酶 (IDO) 是一种酶，参与色氨酸的分解代谢，并已被证明可以通过抑制同种异体反应性 T 细胞来预防妊娠期间胎儿的排斥反应。在本研究中，我们研究了转染 IDO cDNA 的 DC 是否在抗原特异性相互作用后抑制 T 细胞增殖。方法：使用重组腺病毒载体通过基因递送系统表达IDO，并通过Western blot、免疫学染色和犬尿氨酸测定证实其表达和功能。通过FACS分析证实了IDO转染的XS106 DC克隆（源自A/J小鼠）的表面分子的表达。用表达IDO的DC培养后测定同种异体反应性T细胞增殖。结果：成功构建表达IDO的重组腺病毒载体，并通过Western blot和免疫染色检测其基因表达。通过增加犬尿氨酸浓度证实了其分解代谢作用。其揭示表达IDO的XS106 DC的CD86、CD11c和CD69表达没有变化。表达IDO的DC与B6同种异体脾T细胞共培养后，T细胞的增殖受到显着抑制。结论：这些结果表明，DC 中 IDO 的过度表达可有效抑制 T 细胞增殖，并可能扩展一种新的免疫调节策略，以防止器官移植的同种异体排斥。