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新标准化临床冷冻保存/缓慢复温方案对大鼠动脉同种异体移植物免疫原性的影响。

Influence of the new standardized clinical cryopreservation/slow thawing protocol on immunogenicity of arterial allografts in rats.

机构信息

2nd Department of Surgery-Department of Cardiovascular Surgery, 1st Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Czech Republic.

Tissue Bank, Faculty Hospital Hradec Kralove, Charles University- Faculty of Medicine in Hradec Kralove, Hradec Kralove, Czech Republic.

出版信息

PLoS One. 2020 Mar 10;15(3):e0230234. doi: 10.1371/journal.pone.0230234. eCollection 2020.

Abstract

OBJECTIVES AND DESIGN

At the present time there are two waiting list for patients with vascular prosthetic infection indicated for arterial transplantation in the Czech Republic. The inclusion of each patient for cold-stored or cryopreserved arterial transplantation is the preference of indicating surgeon. In this experimental work we studied the immunogenicity of rat aortal allografts treated by our new clinical cryopreservation/slow thawing protocol.

MATERIAL AND METHODS

Brown-Norway (BN) (N = 6, 203-217 g) or Lewis (LEW) (N = 6, 248-254 g) abdominal aortal grafts treated in accordance with our new clinical cryopreservation/slow thawing protocol were orthotopically transplanted to Lewis recipients (N = 12, 191-245 g). Aortal wall histology and infiltration by recipient immune cells, as well as donor specific anti MHC class I and II antibodies in recipient serum were studied in both isografts and allografts on day 30 postransplant. Core data of cryopreserved allografts were compared to our previous data of cold-stored aortal allografts treated in accordance with our clinical cold-storage protocol.

RESULTS

Cryopreserved allografts showed regular morphology of aortal wall with clear differentiation of all three basic anatomical layers on day 30 postransplant. Intimal layer showed no hyperplasia, luminal surface was covered by endothelial cells. No statistical difference was observed in tunica media thickness between isografts and allografts. The medial layer showed no necrosis, shrinkage or immunoglobuline G deposition in any experimental group. The adventitial infiltration by immune cells was significantly higher (P<0.05) in allografts. Cryopreserved allografts showed significant lower activation of both cell- and antibody mediated immunity compared to historical data of cold-stored allografts.

CONCLUSION

Aortal wall histology of rat allografts treated by our new standardized clinical cryopreservation/slow thawing protocol was comparable to that of the cryopreserved isografts on day 30 posttranspant. The immunogenicity of cryopreserved aortal allografts was significantly lower compared to that of cold-stored aortal allografts.

摘要

目的和设计

目前,捷克共和国有两个血管假体感染患者的等待名单,这些患者需要进行动脉移植。每位患者接受冷藏或冷冻保存的动脉移植的选择取决于指示外科医生。在这项实验工作中,我们研究了我们新的临床冷冻保存/缓慢解冻方案处理的大鼠主动脉同种异体移植物的免疫原性。

材料和方法

Brown-Norway (BN)(N = 6,203-217 g)或 Lewis (LEW)(N = 6,248-254 g)的腹部主动脉移植物按照我们新的临床冷冻保存/缓慢解冻方案进行处理,然后原位移植到 Lewis 受体(N = 12,191-245 g)。在移植后第 30 天,研究了同种异体移植物和同种异体移植物的主动脉壁组织学和受体免疫细胞浸润,以及受体血清中供体特异性抗 MHC 类 I 和 II 抗体。冷冻保存的同种异体移植物的核心数据与我们以前按照临床冷藏方案处理的冷藏同种异体移植物的数据进行了比较。

结果

冷冻保存的同种异体移植物在移植后第 30 天显示出主动脉壁的规则形态,具有明显的三个基本解剖层的分化。内膜层无增生,腔面覆盖内皮细胞。同种异体移植物和同种异体移植物之间的中膜厚度没有统计学差异。中膜层在任何实验组中均无坏死、收缩或免疫球蛋白 G 沉积。免疫细胞在 Adventitia 的浸润在同种异体移植物中明显更高(P<0.05)。冷冻保存的同种异体移植物与历史上冷藏同种异体移植物的数据相比,细胞和抗体介导的免疫激活显著降低。

结论

按照我们新的标准化临床冷冻保存/缓慢解冻方案处理的大鼠同种异体移植物的主动脉壁组织学在移植后第 30 天与冷冻保存的同种移植物相似。冷冻保存的主动脉同种异体移植物的免疫原性明显低于冷藏的主动脉同种异体移植物。

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