Decellularization reduces the immune response to aortic valve allografts in the rat
Decellularization reduces the immune response to aortic valve allografts in the rat
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DOI：
10.1016/j.jtcvs.2005.03.021
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发表时间：
2005-08-01
影响因子：
6
通讯作者：
Ross, DB
中科院分区：
文献类型：
--
作者：
Meyer, SR;Nagendran, J;Ross, DB
目的：在先天性心脏病手术中使用的冷冻保存的同种异体瓣膜移植物与显著的细胞和体液免疫应答相关。这可以通过去除抗原性细胞成分（脱细胞化）来减少。本研究的目的是确定大鼠同种异体瓣膜模型中脱细胞的免疫学效果。方法：布朗挪威和刘易斯大鼠主动脉瓣脱细胞与一系列的低渗和高渗缓冲液，蛋白酶抑制剂，温和的洗涤剂（Triton X-100），和磷酸盐缓冲盐水。将瓣膜以同基因和同种异体组合植入刘易斯大鼠体内。蜂窝用形态计量学分析评估CD 3和CD 8浸润，用流式细胞术评估体液反应。形态学分析表明，CD 3（+）细胞浸润显著减少，（每平方毫米小叶组织细胞数）与非脱细胞同种异体移植物相比，（79 +/- 29 vs 3310 +/- 223，P < .001），2（26 +/- 11 vs 109 +/- 20，P = .004），4周（283 122 vs 984 +/- 145，P < .001）。抗-CD 8染色证实大多数浸润是细胞毒性T细胞。流式细胞仪平均通道荧光强度鉴定为负移（废除抗体形成）与非脱细胞同种异体移植物相比，（19 +/- 1 vs 27 +/- 3，P = 0.033），4（35 +/- 2 vs 133 +/- 29，P = 0.001），16周（28 2 vs 166 54，P = 0.017）。结论：脱细胞显著降低了对同种异体组织的细胞和体液免疫反应。这可以延长同种异体瓣膜移植物的耐久性，并可能防止同种异体移植物受体的免疫致敏。
Objectives: Cryopreserved valve allografts used in congenital cardiac surgery are associated with a significant cellular and humoral immune response. This might be reduced by removal of antigenic cellular elements (decellularization). The aim of this study was to determine the immunologic effect of decellularization in a rat allograft valve model.Methods: Brown Norway and Lewis rat aortic valves were decellularized with a series of hypotonic and hypertonic buffers, protease inhibitors, gentle detergents (Triton X-100), and phosphate-buffered saline. Valves were implanted into Lewis rats in syngeneic and allogeneic combinations. Cellular (CD3 and CD8) infiltrates were assessed with morphometric analysis, and the humoral response was assessed with flow cytometry.Results: Morphometric analysis identified a significant reduction in CD3(+) cell infiltrates (cells per square millimeter of leaflet tissue) in decellularized allografts compared with that seen in nondecellularized allografts at 1 (79 +/- 29 vs 3310 +/- 223, P < .001), 2 (26 +/- 11 vs 109 +/- 20, P = .004), and 4 weeks (283 122 vs 984 +/- 145, P < .001). Anti-CD8 staining confirmed the majority of infiltrates were cytotoxic T cells. Flow cytometric mean channel fluorescence intensity identified a negative shift (abrogated antibody formation) for decellularized allografts compared with nondecellularized allografts at 2 (19 +/- 1 vs 27 +/- 3, P = .033), 4 (35 +/- 2 vs 133 +/- 29, P = .001), and 16 weeks (28 2 vs 166 54, P =.017).Conclusions: Decellularization significantly reduces the cellular and humoral immune response to allograft tissue. This could prolong the durability of valve allografts and might prevent immunologic sensitization of allograft recipients.