APT070 (mirococept), a membrane-localizing C3 convertase inhibitor, attenuates early human islet allograft damage in vitro and in vivo in a humanized mouse model.
APT070 (mirococept), a membrane-localizing C3 convertase inhibitor, attenuates early human islet allograft damage in vitro and in vivo in a humanized mouse model.
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DOI：
10.1111/bph.13388
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发表时间：
2016-02
影响因子：
7.3
通讯作者：
Lombardi G
中科院分区：
文献类型：
--
作者：
Xiao F;Ma L;Zhao M;Smith RA;Huang G;Jones PM;Persaud S;Pingitore A;Dorling A;Lechler R;Lombardi G
胰岛细胞移植的一个主要障碍是由于即时血液介导的炎症反应（IBMIR）导致移植胰岛的早期损失。补体途径的激活在 IBMIR 中发挥着核心作用。本研究的目的是测试用 APT070（一种膜定位 C3 转化酶抑制剂）“绘制”人类胰岛对体外暴露于人血清以及在人源化糖尿病小鼠模型中体内移植引起的炎症的抑制作用。 在体外，将与 APT070 预孵育的人胰岛暴露于同种异体全血。在体内，将经过类似处理的胰岛移植到链脲佐菌素诱导的糖尿病 NOD-SCID IL2rγ−/− 小鼠的肾囊下方，这些小鼠已用人 CD34+ 干细胞重建。使用酶联免疫吸附测定法测定补体激活和胰岛激素含量。使用细胞计数珠阵列测定上清液和血清的细胞因子。使用免疫荧光染色评估体外和移植肾中与人血清一起孵育的胰岛的形态。与 APT070 预孵育可减少体外 C 肽释放和 iC3b 产生，同时减少 C4d 和 C5b-9 在嵌入血栓中的胰岛中的沉积。在体内，经过 APT070 处理的胰岛保持了完整的结构，并且比未经处理的胰岛显示出更少的炎症细胞浸润。预处理还显着减少了上清液和血清中的促炎细胞因子。用 APT070 预处理胰岛可以减少胰岛内炎症，同时保留 β 细胞的胰岛素分泌。 APT070可以作为胰岛移植的潜在治疗工具。
A major obstacle to islet cell transplantation is the early loss of transplanted islets resulting from the instant blood‐mediated inflammation reaction (IBMIR). The activation of complement pathways plays a central role in IBMIR. The aim of this study was to test the inhibitory effect of “painting” human islets with APT070, a membrane‐localizing C3 convertase inhibitor, on inflammation evoked by exposure to human serum in vitro and by transplantation in vivo in a humanized diabetic mouse model. In vitro, human islets pre‐incubated with APT070 were exposed to allogeneic whole blood. In vivo, similarly treated islets were transplanted underneath the kidney capsule of streptozotocin‐induced diabetic NOD‐SCID IL2rγ−/− mice that had been reconstituted with human CD34+ stem cells. Complement activation and islet hormone content were assayed using enzyme‐linked immunosorbent assays. Supernatants and sera were assayed for cytokines using cytometric beads array. Morphology of the islets incubated with human serum in vitro and in graft‐bearing kidney were evaluated using immunofluorescence staining. Pre‐incubation with APT070 decreased C‐peptide release and iC3b production in vitro, with diminished deposition of C4d and C5b‐9 in islets embedded in blood clots. In vivo, the APT070‐treated islets maintained intact structure and showed less infiltration of inflammatory cells than untreated islets. The pretreatments also significantly reduced pro‐inflammatory cytokines in supernatants and sera. Pre‐treatment of islets with APT070 could reduce intra‐islet inflammation with accompanying preservation of insulin secretion by beta cells. APT070 could be as a potential therapeutic tool in islet transplantation.