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

Primary nonfunction （PNF） limits the success of allogeneic islet transplantation。PNF是指在移植站点发生的非特定炎症物质事件，无论是在活体还是在再生胶囊下。方法：肝脏、阴道和隔离片在低温部分使用ED 1和ED 2 moAb。Islets使用collagenase digestion和不连续的dextran梯度方法来孤立。直接免疫-过氧化物酶方法被用来进行免疫病理学分析。雄性Wistar大鼠被用作捐赠者，而雌性雄性Lewis大鼠被用作受体的糖尿病。隔离片中的宏观色素的消耗用50 μ L/mL LE·C12 MDP进行了4 h共培养。Recipient Rats在150 μ L/10-g BWLE-C12 MDP或saline inraperitoneal injection约1000片在renal capsule下移植。实验动物分为四个组：组1（n=8）、4 h文化+ saline pretreatment、组2（n=8）、4 h LE-C12 MDDP co-culture + saline pretreatment;组3（n=8）、4 h文化+LE-C12 MDP pretreatment、组4（n=8）、4 h LE-C12 MDDP co-culture + LE-C12 MDP pretreatment。结果：ED 2+几乎观察到的（<1%）在islets和pancreas。ED1+were frequently observed in. LE-C12 MDP中ED 1+新孤立的孤立片、文化片和联合文化片的百分比分别为11.8%、9.2%和2.6%，相当地。LE-C12MDP i.p. and i.v. resulted in depletion of ED1+and ED2+in the liver， but ED1+remained in the pancreas. 1.4组中的PNF事件分别为62.5%、50%、50%和0%，对应。Conclusions： 1） Few tissue types of macrophage （ED2） might exist in the pancreas and islets。2）monocyte/macrophage lineage （ED 1+）可以使用purified islets和LE-C12 MDP的co-cultures，但不是由systemic LE-C12 MDP管理。（3）沉积，来自胰岛和LE-C12 MDP受体的单细胞/巨细胞线的制备在预防移植胰岛的PNF方面是有效的。

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.