Asaoka H, Nishinaka S, Wakamiya N, Matsuda H, Murata M
Department of Microbiology and Hygiene, Faculty of Applied Biological Science, Hiroshima University, Japan.
Immunol Lett. 1992 Mar;32(1):91-6. doi: 10.1016/0165-2478(92)90205-3.
Two chicken monoclonal antibodies (MAbs), HU/Ch2-7 and HU/Ch6-1, against heterophil Hanganutziu-Deicher (HD) antigens with N-glycolylneuraminic acid (NeuGc) at a terminal carbohydrate were established by cell fusions using chicken B cell lines lacking thymidine kinase and spleen cells from chickens immunized with II3NeuGc alpha-LacCer (HD3). The reactivities of these MAbs against several gangliosides including NeuGc-containing glycosphingolipids were examined by a thin-layer chromatography/immunostaining method. MAb HU/Ch2-7 (IgG) reacted strongly with HD3 and IV3NeuGc alpha-nLc4Cer (HD5) and weakly with VI3NeuGc alpha-nLc6Cer (HD7) and 4-O-acetyl-HD3. HU/Ch6-1 (IgG) reacted with HD3 and HD5, but did not react with the other HD antigens. The reactivities of these MAbs against HD antigen were greatly reduced by pre-treatment of the antigen with neuraminidase. These MAbs did not react with N-acetylneuraminic acid-containing gangliosides (GM1 and GM3). These results indicate that these two chicken MAbs are directed toward the antigenic epitope containing the NeuGc.
通过使用缺乏胸苷激酶的鸡B细胞系与用II3NeuGcα-LacCer（HD3）免疫的鸡脾细胞进行细胞融合，建立了两种针对带有末端碳水化合物N-糖基神经氨酸（NeuGc）的异嗜性汉氏-戴氏（HD）抗原的鸡单克隆抗体（MAb），即HU/Ch2-7和HU/Ch6-1。通过薄层色谱/免疫染色法检测了这些单克隆抗体对包括含NeuGc的糖鞘脂在内的几种神经节苷脂的反应性。单克隆抗体HU/Ch2-7（IgG）与HD3和IV3NeuGcα-nLc4Cer（HD5）反应强烈，与VI3NeuGcα-nLc6Cer（HD7）和4-O-乙酰基-HD3反应较弱。HU/Ch6-1（IgG）与HD3和HD5反应，但不与其他HD抗原反应。用神经氨酸酶预处理抗原后，这些单克隆抗体对HD抗原的反应性大大降低。这些单克隆抗体不与含N-乙酰神经氨酸的神经节苷脂（GM1和GM3）反应。这些结果表明，这两种鸡单克隆抗体针对的是含有NeuGc的抗原表位。
