液氮冻存人同种心脏瓣膜、大动脉免疫原性的对比研究

Comparative study on the immunogenicity of human homologous cardiac valve and great artery homograft cryopreserved with liquid nitrogen

目的进行液氮冻存人同种带瓣主、肺动脉（CVH）瓣叶、动脉壁和瓣下心肌组织的免疫原性对比研究。方法随机解冻13例液氮冻存人同种带瓣管道标本，获取瓣叶、动脉壁及瓣下心肌组织，制成单细胞悬液，其中7例获得CVH瓣叶（实验组A组）、10例获得CVH动脉壁（实验组B组）活的单细胞悬液；采用CVH瓣叶、动脉壁的单细胞悬液分别与相同血型受体的淋巴细胞混合培养法，检测混合培养后不同时点CD25、HLA—DR的表达；随机选取13例中的10例行单纯淋巴细胞培养（对照组C组），将实验组间、实验组与对照组进行对比研究。结果13例均未获得CVH瓣下心肌细胞悬液。获得的CVH瓣叶、动脉壁和受体淋巴细胞在培养前和培养后各时点行锥虫蓝染色，活细胞均〉96．2％；随着培养期的延长，各组培养细胞CD25、HLA—DR的表达均呈下降趋势；CVH瓣叶、动脉壁与受体淋巴细胞混合培养液CD25、HLA—DR水平显著高于对照组，说明CVH瓣叶及动脉壁具有一定免疫原性，且CVH动脉壁的免疫原性较其瓣叶强，表现为CD25、HLA—DR高水平表达；实验中未能获得CVH瓣下心肌的细胞悬液。结论液氮冻存人CVH动脉壁的免疫原性较其瓣叶的免疫原性强。

Objective To study the differences on the immunogenicity of the leaflets, arterial wall and myoeardium of conduit valved homografi （ CVH ） eryopreserved with liquid nitrogen. Methods Monocell suspension of leaflets cells and arterial cells of CVH were respectively co-cultured with human lymphatic cells whose blood groups were the same with that of CVH donors. Expressive levels of CD25 and HLA-DR of these lymphatic cells were detected by flow cytometry in the different cultural duration and compared with that of lymph cells alone cultured （ comparative group）. Results The immunogenicity of CVH artery walls was more severe than that of CVH leaflets, and expressive level of whose CD25 and HLA-DR was higher. The immunogenicity of CVH myocardium was not studied because the myocardial cell suspension were not be acquired in this study. Conclusion It is proved that in vitro experimental study that the immunogenicity of arterial walls of eryopreserved CVH is more severe than that of leaflets.