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)

• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671246/
• 关键词: Lymphoid Irradiation; Donor Bone Marrow Cells; FK506; chimerism; kidney Transplantation; Lymphoid Irradiation; 骨髄chimerism; 腎移植; Iymphoid lrradiation; Chimerism; PCR SSCP; donor骨髄移植; Presenvation

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

我们已经开发出一种安全的方案，可以在组织不相容的比格犬之间产生嵌合和对肾脏同种异体移植的耐受性。基本方案包括术前FLI、肾移植时输注dbmc （KGT）和FK术后IM。方法;根据MLR （S.I.=10）选择狗，通过PCR-SSCP确认性别错配、无亲缘关系和匹配等级。分别给予FLI （150rad/d）治疗3天，pha -无反应dbmc输注（2 × 10 μ d/ kg）和FK （0.08mg/kg/d）治疗90天。监测方面采用mlr阻断试验；制备针对(D)或固定第三方(T)细胞作为刺激细胞的应答细胞（术前冷冻储存的细胞），共培养细胞如下；R外周淋巴细胞、R淋巴结细胞和R骨髓细胞；核型嵌合分析；lymphocytotoxicity测试;肾(D)和肾(T)植皮（SGT）。结果;他们被分为5组，如下；A组)未治疗（n=9）；B)颗(n = 6);C)FLI，DBMC，…More (n=3)；D) FLI颗(n = 8);E)FLI、DBMC和FK（n=l3）。A)、B)、C)组，MSD；11、5、7.3天。在D组中，7/8的所有kg在90天内（MSD； 53.2天）被拒绝。第118天拒收1 KG， (D)和(T)的SGs均未延长。在E组（MSD， 178.8天），持续嵌合的Rs（5/8）存活了70天，其中1例完全接受或仅来自肾脏的SGs (D)。同样，3个同性组合中的2个只接受来自肾脏的SGs存活了200天。在(D)和(T)中，具有3/8暂时嵌合的Rs在137天内均被排斥，而不延长SGs。在2例长期嵌合存活者（E组）中，在mlr阻断研究中观察到再生R的外周淋巴细胞、淋巴结细胞和bmc的特异性抑制。相反，在嵌合消失而被排斥的(R)s中，观察到任何淋巴样细胞不抑制(D)特异性淋巴细胞毒性抗体升高。结论;FLI、dbmc输注和Fk联合治疗可有效延长KGs，嵌合骨髓中的移植物促进因子特异性阻断R抗(D)活性。少

项目成果

K.Watanabe et al: "Donor Bone Marrow Cell(OBMC)-infusion is effective in inducing tolerance in dogs treated with fractionated lymphoid ivoadiation(FLI) and FK506"6th Congress of the Asion Society of Transplantion. Abstract Book. 316 (1999)

K.Watanabe 等人：“供体骨髓细胞 (OBMC) 输注可有效诱导接受分次淋巴碘辐射 (FLI) 和 FK506 治疗的狗的耐受性”Asion 移植学会第六届大会。

K.Watanabe et al.: "prolongation effect of FK506 on the survival of 3-day preserved kidney allografts in dogs" Transplantation Proceedings. 30(7). 3603-3605 (1998)

K.Watanabe 等人：“FK506 对狗中保存 3 天的同种异体肾移植物的存活的延长作用”移植论文集。

Koji Watanabe et al: "Prologation effect of FK506 on the survival of 3-day preserved kidney allografts in dogs"Transplantation Proceedings. 30. 3603-3605 (1998)

Koji Watanabe 等人：“FK506 对狗 3 天保存肾同种异体移植物存活的延长作用”移植论文集。

Koji Watanabe, et al.: "Induction of immunological tolerance in adult dogs."Annual Report of the Research on Immunology, Allergy, and Organ Transplantation (Health Sciences Research Grants from Ministry of Health and Welfare, Japan). 18-23 (1998)

Koji Watanabe 等人：“成年犬免疫耐受的诱导”。免疫学、过敏和器官移植研究年度报告（日本卫生福利部健康科学研究资助）。

渡部浩二他: "死体腎移植における術前リンパ組織照射をFK506投与による免疫寛容誘導法"第97回日本外科学会総会臨時増刊号. 98. 405 (1997)

Koji Watanabe 等人：“使用 FK506 给药进行尸体肾移植术前淋巴组织照射的免疫耐受诱导方法”，第 97 届日本外科学会年会，特刊 98. 405 (1997)。