Holmes Beverly J, Richards Andrew C, Awwad Michel, Copeman Laura S, McLaughlin Megan L, Cozzi Emanuele, Schuurman Henk-Jan, Davies Hugh F S
Imutran Ltd, A Novartis Pharma AG Company, Cambridge, UK.
Xenotransplantation. 2002 Mar;9(2):135-47. doi: 10.1034/j.1399-3089.2002.1o056.x.
Anti-pig antibodies (APA) were analysed in serum from 28 naïve wild-caught baboons (originating from Kenya) and 31 naïve captive-bred cynomolgus monkeys (13 from the Philippines and 18 from Mauritius), using a haemolytic assay with pig erythrocytes (APA), flow cytometry on the porcine lymphoma T-cell cell line L35, and enzyme linked immunosorbent assay (ELISA) using alpha-Gal type II and type VI antigen. This was extended in baboon samples by the evaluation in two laboratories (Imutran, Cambridge, UK and Immerge, Boston, USA), and by antibody absorption using either immobilized alpha-Gal type II or alpha-Gal type VI. Anti-porcine antibodies were demonstrated in all assays with substantial variability within and between the three non-human primate groups. Immunoglobulin (Ig)M antibody levels tended to be similar to or higher than those in a pooled normal human standard serum while IgG levels tended to be lower. Highest antibody levels were recorded in Mauritius cynomolgus monkeys. There were statistically significant correlations between assays for IgM or IgG class anti-Gal antibodies using either alpha-Gal type II or alpha-Gal type VI as antigen, both for different assays and two laboratories involved. Also, significant correlations were observed between the anti-Gal and L35 binding assays. Baboon sera before and after absorption to immobilized alpha-Gal type II or type VI were analysed for anti-Gal type VI or type II antibody: levels were almost undetectable indicating that most anti-Gal antibodies react to epitopes shared between alpha-Gal type II and type VI oligosaccharides. Finally, the relation between APA and outcome of porcine heart xenotransplantation in cynomolgus monkeys and baboons showed no apparent relation between pre-transplant APA levels and the occurrence of hyperacute rejection (HAR) when compared with non-immunological cause of organ/recipient dysfunction or acute humoral xenograft rejection during the first 4 days post-transplantation or survival exceeding 4 days post-transplantation.
使用猪红细胞溶血试验（抗猪抗体，APA）、对猪淋巴瘤T细胞系L35进行流式细胞术以及使用α - Gal II型和VI型抗原的酶联免疫吸附测定（ELISA），对28只未接触过的野生捕获狒狒（原产于肯尼亚）和31只未接触过的圈养食蟹猴（13只来自菲律宾，18只来自毛里求斯）的血清中的抗猪抗体（APA）进行了分析。在狒狒样本中，通过在两个实验室（英国剑桥的Immutran和美国波士顿的Immerge）进行评估，并使用固定化的α - Gal II型或α - Gal VI型进行抗体吸收，扩展了分析。在所有检测中均证实存在抗猪抗体，三个非人灵长类动物组内部和之间存在显著差异。免疫球蛋白（Ig）M抗体水平往往与正常人类混合标准血清中的水平相似或更高，而IgG水平往往较低。毛里求斯食蟹猴的抗体水平最高。使用α - Gal II型或α - Gal VI型作为抗原，针对IgM或IgG类抗Gal抗体的检测之间，在不同检测以及涉及的两个实验室之间均存在统计学显著相关性。此外，在抗Gal与L35结合检测之间也观察到显著相关性。分析了狒狒血清在固定化α - Gal II型或VI型上吸收前后的抗Gal VI型或II型抗体：水平几乎检测不到，表明大多数抗Gal抗体与α - Gal II型和VI型寡糖共有的表位发生反应。最后，食蟹猴和狒狒中抗猪抗体与猪心脏异种移植结果之间的关系表明，与移植后前4天内器官/受体功能障碍的非免疫原因或急性体液异种移植排斥反应或移植后存活超过4天相比，移植前抗猪抗体水平与超急性排斥反应（HAR）的发生之间没有明显关系。
