Quantitation and characterization of anti-Galα1-3Gal antibodies in sera of 200 healthy persons
Quantitation and characterization of anti-Galα1-3Gal antibodies in sera of 200 healthy persons
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DOI：
10.1034/j.1399-3089.1999.00023.x
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发表时间：
1999-08-01
影响因子：
3.9
通讯作者：
Nydegger, UE
中科院分区：
文献类型：
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作者：
Buonomano, R;Tinguely, C;Nydegger, UE
人血清中补体激活性抗Gal α 1 -3Gal抗体（抗Gal）的天然存在被认为是异种移植的主要障碍。在这项研究中，我们确定了200名健康志愿者（100名男性，100名女性）的不同年龄组的血清中的抗-半乳糖苷使用IgG和IgM-同种型特异性ELISA。此外，我们使用涉及兔红细胞（E-R）的直接血凝试验，已知兔红细胞表面大量表达Gal α 1 -3Gal抗原。抗-Gal的任意ELISA单位（U）测量范围为IgM的5 - 204 U（77 +/- 47;平均值+/- 1 SD），IgG的1 - 162 U（73 +/- 32）。抗ER血凝滴度在1：65536和1：64之间（平均1：703），所有血清样品的75%在1：8192-1：512范围内。通过琼脂糖偶联的合成Gal α 1 -3Gal抗原上的抗Gal的免疫吸收来确定测试的特异性，其将ELISA以及凝集滴度降低90%或更多。总体而言，抗Gal ELISA值和E-R凝集滴度的个体间差异超过了年龄、性别或ABO型相关变化。发现女性的抗-Gal IgM值高于男性（P < 0.005，Student's t检验），而抗-Gal IgG或ER滴度的性别差异不显著。本文还讨论了抗半乳糖酶联免疫吸附试验（ELISA）和E-R凝集试验在猪异种移植受者移植前筛选中的应用。
The natural occurrence of complement-activating anti-Gal alpha l-3Gal antibodies (anti-Gal) in human serum is considered as a major obstacle to xenotransplantation. In this study we determined anti-Gal in sera of 200 healthy volunteers (100 male, 100 female) of different age groups using an IgG- and IgM-isotype specific ELISA. In addition, we used a direct hemagglutination test involving rabbit erythrocytes (E-R), which are known for abundant surface expression of the Gal alpha l-3Gal antigen. The measured arbitrary ELISA units (U) for anti-Gal ranged from 5 to 204 U (77 +/- 47; mean +/- 1 SD) for IgM, and from 1 to 162 U(73 +/- 32) for IgG. Anti-ER hemagglutination titers were between 1: 65536 and 1: 64 (mean 1: 703), with 75% of all serum samples being in the range of 1: 8192-1: 512. Specificity of the tests was determined by immunoabsorption of anti-Gal on Sepharose-coupled synthetic Gal alpha l-3Gal antigen, which reduced ELISA as well as agglutination titers by 90% or more. Overall, inter-individual differences of both anti-Gal ELISA values and E-R agglutination titers exceeded age-, gender-, or ABO-type related changes. A tendency was found to higher anti-Gal IgM values in women than in men (P < 0.005 by Student's t-test), whereas the gender-difference for anti-Gal IgG or ER titers was not significant. Application of the anti-Gal ELISA as well as the E-R agglutination assay for pre-transplant screening of potential pig xenograft recipients is discussed.