Human Health Sciences, Graduate School of Medicine, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Inflammation. 2012 Apr;35(2):409-19. doi: 10.1007/s10753-011-9330-8.
The immunological properties of rat S100A8 (r-S100A8) and S100A9 (r-S100A9) in immune cells are poorly understood. Enzyme-linked immunosorbent assay (ELISA) for r-S100A9 enabled us to discuss the differential functional roles of the two proteins, and their localization in the cells was observed microscopically. Recombinant human S100A8 (rh-S100A8) or S100A9 (rh-S100A9) were intravenously administrated into rats with LPS-induced liver damage. ELISA was used to measure the serum concentration of S100A9 in the rats. Western blotting and a preparative ELISA were used to prove specificity and avidity of monoclonal antibodies for r-S100A8 and r-S100A9. Immunohistochemical staining was carried out to visualize intracellular localization of the two proteins in the immune cells using the antibodies. When rh-S100A8 was intravenously injected in the rats (B group), the serum concentration of r-S100A9 apparently decreased as compared with that of the positive control rats (A group). The activities of AST, ALT, and LD in the rat sera (B group) also significantly went down in comparison with those of the rats (A group). Although both the S100A8 and S100A9 were abundantly expressed in activated immune cells, quite difference of not only their intracellular localization but also distribution of the cells expressing the two proteins was microscopically observed. In the rats (B group), less number of the immune cells or less amount of r-S100A8 and r-S100A9 in the cells than those of the rats (A group) was also seen. The r-S100A8 could serve as a regulator of acute inflammatory reaction in the rats with LPS-induced damage.
大鼠 S100A8（r-S100A8）和 S100A9（r-S100A9）在免疫细胞中的免疫特性尚未完全阐明。酶联免疫吸附试验（ELISA）用于 r-S100A9，使我们能够讨论这两种蛋白质的不同功能作用，并用显微镜观察它们在细胞中的定位。将重组人 S100A8（rh-S100A8）或 S100A9（rh-S100A9）静脉注射到 LPS 诱导的肝损伤大鼠体内。ELISA 用于测量大鼠血清中 S100A9 的浓度。Western blot 和制备型 ELISA 用于证明针对 r-S100A8 和 r-S100A9 的单克隆抗体的特异性和亲和力。免疫组织化学染色用于使用抗体可视化两种蛋白质在免疫细胞中的细胞内定位。当 rh-S100A8 静脉注射到大鼠（B 组）中时，与阳性对照大鼠（A 组）相比，r-S100A9 的血清浓度明显降低。与大鼠（A 组）相比，大鼠血清（B 组）中的 AST、ALT 和 LD 活性也显著下降。尽管 S100A8 和 S100A9 在激活的免疫细胞中均大量表达，但不仅它们的细胞内定位，而且表达两种蛋白质的细胞分布也存在明显差异。在大鼠（B 组）中，与大鼠（A 组）相比，免疫细胞的数量较少，或细胞内的 r-S100A8 和 r-S100A9 含量较少。r-S100A8 可作为 LPS 诱导损伤大鼠急性炎症反应的调节剂。
