Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum1
Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum1
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腺病毒介导的人 CD55 或 CD59 表达可保护成年猪胰岛免受人血清补体介导的细胞裂解1
DOI：
10.1097/01.tp.0000053249.39753.d6
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发表时间：
2003
期刊：
影响因子：
6.2
通讯作者：
O. Korsgren
中科院分区：
文献类型：
--
作者：
P. Schmidt;M. Goto;B. Le Mauff;I. Anegon;O. Korsgren
背景资料。在猪到人的异种胰岛移植中，对补体激活的保护可以减少急性胰岛损伤。用腺病毒转导技术在完整的成年猪胰岛(APIs)上诱导人补体调节蛋白衰变加速因子(DAF，CD55)或CD59的表达。体外检测外源基因在新鲜人血清中的功能。方法：研究方法。用腺病毒载体Ad.hDAF或Ad.hCD59或对照载体转导完整的API。3天后，胰岛被胰酶解离成单细胞悬液。将人补体活性AB血清与胰岛细胞孵育，进行细胞毒试验。用流式细胞仪和免疫组织化学方法检测转基因的表达。结果。可转导API表达hDAF或hCD59。对胰岛单个细胞的流式细胞术分析表明，只有少数细胞表达转基因基因；转导的胰岛免疫组织化学染色显示，主要位于胰岛外围的细胞表达该蛋白。未转导的胰岛细胞和表达控制蛋白的胰岛细胞对人血清裂解反应敏感(分别为66±4.0%和73±3.7%)。转导hDAF和hCD59的胰岛单个细胞对裂解有部分保护作用。表达hCD59的胰岛细胞对裂解的敏感性(33±3.3%)略低于表达hDAF的细胞(45±3.5%)。结论。这些数据表明，完整的猪胰岛可以被转导到表面表达人类补体激活调节因子，并且表达hDAF或hCD59的猪胰岛细胞对补体介导的裂解不那么敏感。
Background. Protection against complement activation may reduce acute islet damage in pig-to-human islet xenotransplantation. Expression of the human complement regulatory proteins decay-accelerating factor (DAF, CD55) or CD59 was induced on intact adult porcine islets (APIs) by adenoviral transduction. The functional capacity of the transgenes was examined in vitro after exposure to fresh human serum. Methods. Intact APIs were transduced with adenoviral vectors Ad.hDAF or Ad.hCD59 or a control vector. After 3 days, the islets were trypsin dissociated to a single-cell suspension. A cytotoxicity assay was performed in which the islet cells were incubated with human complement active AB serum. Flow cytometry and immunohistochemistry were used to evaluate transgene expression. Results. APIs could be transduced to express hDAF or hCD59. Flow cytometry analysis of islet single cells revealed that only a fraction of the cells expressed the transgene; immunohistochemical staining of transduced islets demonstrated that mainly cells located in the periphery of the islets were expressing the protein. Cells from nontransduced islets or islets expressing the control protein were sensitive to lysis in human sera (66±4.0% and 73±3.7% cytotoxicity, respectively). Single cells from islets transduced with hDAF and hCD59 were partially protected from lysis. Islet cells expressing hCD59 were slightly less sensitive to lysis (33±3.3%) than cells expressing hDAF (45±3.5%). Conclusions. These data show that intact pig islets can be transduced to express human regulators of complement activation on the surface and that pig islet cells expressing hDAF or hCD59 are less sensitive to complement-mediated lysis.
DOI：
10.4049/jimmunol.1202204
发表时间：
2013-02-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Olaru F;Wang XP;Luo W;Ge L;Miner JH;Kleinau S;Geiger XJ;Wasiluk A;Heidet L;Kitching AR;Borza DB
通讯作者：
Borza DB
负责补体介导的裂解的同源物种限制的膜因子：在 C8 和 C9 阶段起作用的除 DAF 之外的因子的证据。
DOI：
--
发表时间：
1986
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Shin,ML;Hänsch,G;Hu,VW;Nicholson-Weller,A
通讯作者：
Nicholson-Weller,A