Effects of Kupffer cell and macrophage depletion on survival of transplanted islets

枯否细胞和巨噬细胞耗竭对移植胰岛存活的影响

• 批准号: 11671144
• 负责人: FUJIMORI Keisei
• 金额: $ 2.24万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671144/
• 关键词: islet transplantation; macrophave depletion; primary non function; acute rejection; dichloromethylene disphosphonate; macrophage subpopulation; IDDM; LE-Cl_2MDP; マクロファージ; 膵ラ島; LE-C12MDP

Primary nonfunction (PNF) limits the success of allogeneic islet transplantation. PNF is attributed to non-specific inflammatory events occurring at the transplant site, in the liver or under the renal capsule. Methods: The liver, pancreas and isolated islets were stained using ED1 and ED2 moAb on cryostat sections. Islets were isolated using collagenase digestion and the discontinuous dextran gradient method. The direct immuno-peroxydase method was performed for immunohistorologal analysis. Male Wistar rats were utilized as donors, and inbred male Lewis rats made diabetic using streptozotocin as recipients. Depletion of macrophages in isolated islets was performed by 4 h co-culture with 50 μ L/mL LE・C12MDP. Recipient rats were pretreated with 150 μ L/10-g BW LE-C12MDP or saline intraperitoneal injection About 1000 islets were transplanted under the renal capsule. Experimental animals were divided into four groups: Group 1(n=8), 4 h culture + saline pretreatment, Group 2(n=8), 4 h LE-C12MDP co-culture + saline pretreatment; Group 3(n=8), 4 h culture + LE-C12MDP pretreatment, Group 4(n=8), 4 h LE-C12MDP co-culture + LE-C12MDP pretreatment. Results: ED2+were barely observed (<1%) in islets and pancreas. ED1+were frequently observed in. Percentages of ED1+in freshly isolated islets, cultured islets and co-cultured islets with LE-C12MDP were 11.8%, 9.2%, and 2.6%, respectively. LE-C12MDP i.p. and i.v. resulted in depletion of ED1+and ED2+in the liver, but ED1+remained in the pancreas. Incidences of PNF in Groups 1.4 were 62.5%, 50%, 50% and 0%, respectively. Conclusions: 1) Few tissue types of macrophage (ED2+) might exist in the pancreas and islets. 2) The monocyte/macrophage lineage (ED1+) can be depleted using co-cultures of purified islets and LE-C12MDP, but not by systemic LE-C12MDP administration. 3) Depletion, of the monocyte/macrophage lineage from islets and LE-C12MDP pretreatment of recipients is effective in preventing PNF of transplanted islets.

原发性无功能（PNF）限制了同种异体胰岛移植的成功。PNF归因于移植部位、肝脏或肾包膜下发生的非特异性炎症事件。方法：用单克隆抗体ED 1和ED 2对肝脏、胰腺和胰岛进行染色。采用胶原酶消化和不连续葡聚糖梯度法分离胰岛。采用直接免疫过氧化物酶法进行免疫史分析。雄性Wistar大鼠作为供体，近交系雄性刘易斯大鼠作为受体。用50 μ L/mLLE·C12 MDP共培养4 h，去除分离胰岛中的巨噬细胞。将150 μ L/10-gBW的LE-C12 MDP或生理盐水腹腔注射于SD大鼠，将约1000个胰岛移植于肾包膜下。实验动物分为4组：1组（n=8），培养4 h+生理盐水预处理; 2组（n=8），LE-C12 MDP共培养4 h+生理盐水预处理; 3组（n=8），培养4 h + LE-C12 MDP预处理; 4组（n=8），LE-C12 MDP共培养4 h + LE-C12 MDP预处理。结果：胰岛和胰腺中几乎未检测到ED 2+（<1%）。ED 1+在正常人中多见。在新鲜分离的胰岛、培养的胰岛和与LE-C12 MDP共培养的胰岛中，ED 1+的回收率分别为11.8%、9.2%和2.6%。LE-C12 MDP i. p.和i. v.导致肝脏中的ED 1+和ED 2+耗尽，但ED 1+保留在胰腺中。第1.4组PNF发生率分别为62.5%、50%、50%和0%。结论：1）胰腺和胰岛中可能存在少量组织型巨噬细胞（ED 2+）。2)单核细胞/巨噬细胞谱系（ED 1+）可以使用纯化的胰岛和LE-C12 MDP的共培养物来消除，但不能通过全身性LE-C12 MDP施用来消除。3)胰岛单核/巨噬细胞系的耗竭和受体的LE-C12 MDP预处理可有效预防移植胰岛的PNF。