Tolerance promoting cells were derived from infused donor bone marrow cell (DBMC) in dogs treated with fractionated lymphoid irradiation (FLI) and FK506 (FK)

耐受促进细胞源自经分次淋巴照射 (FLI) 和 FK506 (FK) 治疗的犬体内输注的供体骨髓细胞 (DBMC)

基本信息

批准号：
09671246
负责人：
WATANABE Koji
金额：
$ 1.22万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

We have developed safe regimen that can produce chimerism and tolerance to kidney allografts between histoincompatible beagles. The basic regimen included preoperative FLI, DBMC-infusion at kidney grafting (KGT) and postoperative IM of FK. Methods; Dogs were selected on the basis of MLR (S.I.=10), sex-mismatched, unrelated pairs and matching grade was confirmed by PCR-SSCP. They were treated with FLI (150rad/day) for 3 days, PHA-nonresponding DBMC-infusion(2x10ィイD17ィエD1/kg) and FK (0.08mg/kg/d for 90 days). As for monitoring, MLR-blocking test was performed; Responder cells(preoperatively frozen stored cells) against (D) or fixed third-party(T) cells as stimulator cells were prepared, and cocultured cells were as follows; R peripheral lymphocytes, R lymphnode cells, and R BMCs; chimerism analysis by karyotyping; lymphocytotoxicity test; and skingrafting(SGT) from kidney (D) and (T). Results; They were divided into 5 groups, as follows; group A) non-treated(n=9); B)FK,(n=6); C)FLI,DBMC, … More (n=3); D)FLI, FK(n=8); E)FLI, DBMC, and FK(n=l3). In group A), B), C), MSD;11, 5, 7.3 days. In group D), all KGs in 7/8 were rejected within 90 days(MSD; 53.2 days). One KG rejected on 118th days without prolongation of SGs from both (D) and (T). In group E) (MSD; 178.8 days), Rs with persistent chimerism(5/8), survived >70 days, one of which had complete acceptance or SGs only from kidney (D)'s. And also, two of three with same sex-comination survived >200 days with acceptance of SGs only from kidney (D)s. Rs with temporary chimerism in 3/8 were rejected by 137 days without prolongation of SGs from both (D) and (T). In 2 long survivors with chimerism in group E), specific suppression by repopulated R's peripheral lymphocyte, lymphnode cells and BMCs was observed in MLR-blocking study. On the contrary, in (R )s which were rejected with disappearance of chimerism, nonsuppression by any lymphoid cells observed with elevation of (D) specific lymphocytotoxic antibody. Conclusion; Combined treatment with FLI, DBMC-infusion, and Fk was effective in prolongation of KGs. Graft promoting factors derived from chimerric bone marrow specifically blocked R anti-(D) activity. Less

我们开发了安全的方案，可以在组织不相容的比格犬之间产生嵌合和对同种异体肾移植的耐受性。基本方案包括术前FLI、肾移植时DBMC输注和术后IM。 (S.I.=10)，性别不匹配、不相关的配对和匹配等级通过 PCR-SSCP 得到确认，他们接受 FLI (150rad/天) 治疗。 3天，PHA无反应DBMC输注(2x10D17D1/kg)和FK(0.08mg/kg/d，持续90天)，针对(术前冷冻保存的细胞)进行MLR阻断测试； d)制备或固定第三方(T)细胞作为刺激细胞，共培养细胞如下：R外周淋巴细胞、R淋巴结；细胞和R BMC；通过核型分析；和来自肾脏的皮肤移植(SGT)；结果将它们分为5组，如下：A)组； 9); B)FK,(n=6); C)FLI,DBMC, … 更多 (n=3); E)FLI, DBMC，和FK(n=13)，MSD；11、5、7.3天在D)组中，7/8中的所有KG在90天内被拒绝(MSD；53.2)。天）。第 118 天拒绝一公斤，而 E 组的 SG 没有延长（MSD；178.8）。天），具有持续嵌合状态的 Rs（5/8），存活 >70 天，其中一个完全接受或仅来自肾脏（D）的 SG，而且，三分之二的同性组合存活 >200 天。仅接受来自肾脏 (D) 的 SG，其中 3/8 的患者在 137 天时被拒绝，而来自 (D) 和 (T) 的 SG 均没有延长。在E组中，在MLR阻断研究中观察到重新增殖的R的外周淋巴细胞、淋巴结细胞和BMC的特异性抑制，相反，在因嵌合体消失而被排斥的(R)组中，观察到任何淋巴细胞的非抑制升高。 (D) 特异性淋巴细胞毒性抗体的结论；FLI、DBMC 输注和 Fk 的联合治疗可有效延长源自嵌合体的移植物促进因子。骨髓特异性阻断 R 抗 (D) 活性较少。