Yohe H C, Berenson C S, Cuny C L, Ryan J L
Department of Internal Medicine, Yale University School of Medicine, West Haven, Connecticut.
Infect Immun. 1990 Sep;58(9):2888-94. doi: 10.1128/iai.58.9.2888-2894.1990.
We have analyzed both the total ganglioside composition and the surface accessibility of C3H/HeN B lymphocytes and C3H/HeJ B lymphocytes. Seventeen individual resorcinol-positive moieties were visualized by two-dimensional thin-layer chromatography of the purified gangliosides from both strains. Complete homology between strains was seen in the patterns of total gangliosides purified from the endotoxin-responsive and -hyporesponsive strains, with only minor differences in the relative concentrations of four gangliosides. In comparison, only 12 individual gangliosides were accessible to surface labeling following galactose oxidase treatment in these same strains, suggesting that some gangliosides are masked at the cell surface in both strains. However, labeling of the more polar components was greatly reduced in the endotoxin-hyporesponsive (C3H/HeJ) strain, suggesting that these gangliosides have decreased accessibility to galactose oxidase at the cell surface. Therefore, while the total ganglioside compositions of the two strains were nearly equivalent, there were dramatic differences in ganglioside surface accessibility. These findings indicate that an alteration in membrane structure that is associated with the endotoxin hyporesponsiveness observed in C3H/HeJ B lymphocytes exists.
我们分析了C3H/HeN B淋巴细胞和C3H/HeJ B淋巴细胞的总神经节苷脂组成及表面可及性。通过对两株细胞纯化的神经节苷脂进行二维薄层色谱分析,观察到17个间苯二酚阳性部分。从对内毒素有反应和反应低下的菌株中纯化得到的总神经节苷脂模式显示出菌株间的完全同源性,仅四种神经节苷脂的相对浓度存在微小差异。相比之下,在相同菌株中,经半乳糖氧化酶处理后,只有12种单个神经节苷脂可用于表面标记,这表明在两株细胞中,一些神经节苷脂在细胞表面被掩盖。然而,在内毒素反应低下的(C3H/HeJ)菌株中,极性更强的成分的标记显著减少,这表明这些神经节苷脂在细胞表面对半乳糖氧化酶的可及性降低。因此,虽然两株细胞的总神经节苷脂组成几乎相同,但神经节苷脂的表面可及性存在显著差异。这些发现表明,在C3H/HeJ B淋巴细胞中存在与内毒素反应低下相关的膜结构改变。