BROBERGER O, PERLMANN P
J Exp Med. 1963 May 1;117(5):705-16. doi: 10.1084/jem.117.5.705.
By means of immunofluorescent methods it has been shown that sera from children with ulcerative colitis contain antibodies which react with fetal colon cells in tissue culture. 5 out of 13 sera from patients reacted positively when tested for staining antibodies while 12 sera from healthy individuals yielded negative results. The specificity of the staining reactions was confirmed by inhibition experiments. The staining capacity of various sera was correlated to their hemagglutinating titer when tested against phenol-water extracts of human colon. The presence of blood group substances of the ABO system on fetal colon cells in tissue culture could be demonstrated by application of fluorescent H agglutinins from eel. Cross-inhibition experiments indicated that the H agglutinins stained colon antigens which were different from those reacting with the antibodies of ulcerative colitis sera. The reactivity of cultured fetal colon cells with the antibodies in ulcerative colitis sera was retained for up to 12 days, with optimal staining at 4 to 5 days. Reactivity with H agglutinins was present for a longer period, sometimes more than 20 days. Although antigen could be shown to be present on fetal colon cells in tissue culture, exposure of the culture, in the presence of fresh guinea pig serum, to sera from patients with ulcerative colitis did not lead to any visible cytotoxic damage. In order to investigate the possible cytotoxic effect of the sera with a more sensitive technique, freshly explanted fetal colon was dispersed by trypsinization and the cells labeled with (32)P-orthophosphate. Subsequently, these cells were exposed to sera, in a final concentration of 30 per cent, from patients or healthy controls in the presence of fresh guinea pig serum (final concentration 15 per cent). Approximately 20 per cent of the cellular isotope was released into the medium within 150 minutes of incubation, but the release was the same in the samples treated either with patients' sera or normal control sera. Thus, under the present conditions, the patients' sera did not exert any specific cytotoxic action on colon cells.
通过免疫荧光法已表明，溃疡性结肠炎患儿的血清中含有能与组织培养中的胎儿结肠细胞发生反应的抗体。13例患者的血清中有5例在检测染色抗体时呈阳性反应，而12例健康个体的血清检测结果为阴性。染色反应的特异性通过抑制实验得到证实。当用各种血清检测人结肠的酚-水提取物时，其染色能力与其血凝效价相关。应用来自鳗鱼的荧光H凝集素可证明组织培养中胎儿结肠细胞上存在ABO系统的血型物质。交叉抑制实验表明，H凝集素所染色的结肠抗原与溃疡性结肠炎血清抗体所反应的抗原不同。培养的胎儿结肠细胞与溃疡性结肠炎血清中的抗体的反应性可持续长达12天，在4至5天时染色最佳。与H凝集素的反应性存在的时间更长，有时超过20天。尽管在组织培养中可证明胎儿结肠细胞上存在抗原，但在新鲜豚鼠血清存在的情况下，将培养物暴露于溃疡性结肠炎患者的血清中并未导致任何可见的细胞毒性损伤。为了用更灵敏的技术研究血清可能的细胞毒性作用，将新鲜移植的胎儿结肠用胰蛋白酶消化分散，细胞用（32）P-正磷酸盐标记。随后，在新鲜豚鼠血清（终浓度15%）存在的情况下，将这些细胞暴露于终浓度为30%的患者或健康对照的血清中。在孵育150分钟内，约20%的细胞同位素释放到培养基中，但用患者血清或正常对照血清处理的样品中的释放情况相同。因此，在目前条件下，患者血清对结肠细胞未产生任何特异性细胞毒性作用。
