Gene therapy for induction of immunological tolerance in organ allografts

诱导同种异体器官移植免疫耐受的基因治疗

• 批准号: 13671253
• 负责人: KATO Hirohisa
• 金额: $ 2.24万
• 依托单位: SHOWA University
• 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-13671253/
• 关键词: ORGAN TRANSPLANTATION; GENE TRANSFER; TOLERANCE

(Background) Gene transfer approach has the potential to introduce immunosuppressive molecules directly into the allograft, which in turn should limit systemic side effects of conventional immunosuppressive therapy to recipients. (Aim) To analyze the efficacy and putative mechanisms of adenoviral-mediated IL4 gene transfer in rat kidney transplant model. (Material ＆ Method) Renal allografts (BN rat) were perfused in-situ with Adex1CAmIL4 or AxCALacZ (RIKEN DNA Bank, respectively) incubated at 4C for 1.5, 24, 48, 72h (ex-vivo), or/and transplanted to LEW recipients. Adenovirus vector was perfused with 4C saline or UW solution, and renal grafts were preserved in 4C saline (1.5h) or UW solution (24-72h). (Results) 1. β-gal was detected after 24h preservation, but not 1.5h in ex-vivo model. After transplantation, there was mo staining of β-gal in renal allograft at day2/day7. 2. In ex-vivo preservation, IL4 was expressed in all of renal grafts (1.5h-72h) with Adex1CAmIL4 perfusion, even though it was not detected in control group perfused with cold saline in any preserved term. 3. Histological damage of renal allografts maintained for 24h after Adex1CAmIL4 gene transfer were diminished as compared with 1.5h preserved grafts at day7 after transplantation. (Conclusion) 24h preservation was prefer to induce immunosuppressive effects of Adenovirus-mediated IL4 gene therapy as compared. with 1.5h maintaining. It seems that appropriate expression of IL4 might be important at the early phase (day0-2) after transplantation. Further study will be necessary to inquire "optimal way" of gene transfer in transplant setting.

（背景）基因转移方法具有将免疫抑制分子直接引入同种异体移植物的潜力，这反过来又可以限制常规免疫抑制治疗对受者的全身副作用。(Aim)目的探讨腺病毒介导的白细胞介素4（IL 4）基因转移在大鼠肾移植模型中的作用及可能机制。（材料与方法）用Adex 1CAmIL 4或AxCALacZ（分别为RIKEN DNA Bank）原位灌注肾移植物（BN大鼠），在4 ℃下孵育1.5、24、48、72小时（离体），或/和移植到LEW受体。将腺病毒载体分别用4C生理盐水或UW液灌注，移植肾保存于4C生理盐水（1.5h）或UW液（24- 72 h）中。（结果）1.离体模型保存24 h后可检测到β-gal，而保存1.5h后未检测到。移植后第2天/第7天，移植肾组织中β-gal染色消失。2.在离体保存过程中，经Adex 1CAmIL 4灌注的移植肾在1.5h-72 h内均有IL 4的表达，而对照组在任何保存期均未检测到IL 4的表达。3.移植后第7天，Adex 1CAmIL 4基因转染后24 h的移植肾组织损伤较保存1.5h的移植肾明显减轻。（结论）与腺病毒介导的IL 4基因治疗相比，保存24 h更有利于诱导免疫抑制效应。维持1.5h。IL 4的适当表达在移植后早期（0 -2天）可能是重要的。今后的研究还需进一步探索移植背景下基因转移的“最佳途径”。