Induction of tolerance in the pig-to-human Xenotransplantation by using Xenoreactive antibody producing B cells and Xenoantigen.

实验研究1为了研究产生异种反应性国家抗体（XNA）的B细胞在人体内的分布，我们从脾脏扁桃体腺和外周血中获得淋巴细胞。为了对产生xna的细胞进行免疫选择，我们使用磁珠包被表达gal - 1,3 - gal末端残基的猪甲状腺球蛋白（PTg）。FACS分析证实，PTg珠选择的几乎所有细胞都显示CD20的存在。在将ebv转化为PTg-bead选择的B细胞和正常B细胞的实验中，虽然只有10%的正常B细胞显示XNA，但PTg-bead选择的B细胞中有44%的孔显示XNA。CD5在产生XNA的B细胞上的总表达率与未分离的B细胞无显著差异。sIgM和sIgD在产生xna的B细胞上的表达率明显高于未分离的B细胞。实验研究2将仓鼠心脏移植到Lewis大鼠和裸鼠2个群体。各组均给予CyA，并于移植当日行脾切除术（Spx）。Spx和CyA联合处理可显著提高移植物存活时间。在CyA处理的Lewis大鼠和裸鼠中，Spx将Xab的产生降低到检测不到的水平。

Experimental study 1To examine the distribution of the xeno-reactive national antibody (XNA)-producing B cells in humans, we obtain the lymphocytes from the spleen tonsilla gland and peripheral blood. For the immunoselection of XNA-producing cells, we used magnetic beads coated with porcine thyroglobulin (PTg) which expressed Gal-1, 3-Gal terminal residues. It was confirmed by FACS analysis that almost all cells selected by PTg beads showed the presence of the CD20. On an experiment of transforming EBVto PTg-beads -selected B cells and normal B cells, although only 10% of the normal B cells showed XNA, 44% of the wells showed XNA in PTg-bead selected B cells. The overall expression rate of CD5 on XNA producing B cells was not significantly different to that n non-separated B cells. The expression rate of sIgM and sIgD on XNA-producing B cells was significantly higher than that on non-separated B cells.Experimental study 2Hamster hearts were transplanted to 2 colonies of Lewis rats and nude rats. CyA was given and splenectomy (Spx) was performed on the day of transplantation in same groups. The combined treatment of Spx and CyA showed a significant increase in graft survival time. Spx reduced Xab production to undetectable level in CyA treated Lewis rats and nude rats.