Ogasawara M, Granfors K, Kono D H, Hill J L, Yu D T
J Immunol. 1985 Jul;135(1):553-9.
It has been suggested that the O-side chains of the lipopolysaccharide (LPS) of serotype 0:3 strains of Yersinia enterocolitica vary quantitatively and qualitatively depending on whether they are cultured at 37 degrees C or 25 degrees C. It is uncertain whether this affects the expression of the serotype-specific antigens that are probably carried on the LPS. We studied this question with a serotype 0:3-specific monoclonal antibody, 2C1. This monoclonal antibody immunoprecipitated a 39K major protein from detergent-solubilized 125I-labeled Yersinia preparation. However, results of Western blot experiments demonstrated that the antigens reactive with 2C1 were not actually the 39K protein but the O-side chains of the LPS. Significantly, reactivity could be detected whether the bacteria were cultured at room temperature or at 37 degrees C. However, absorption experiments confirmed that there were less accessible antigenic determinants on the 37 degrees C-LPS. The LPS preparations were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. These SDS-PAGE profiles showed that less O-side chains were present in the 37 degrees C-LPS. Hence, the most likely explanation for our observation is that the 37 degrees C incubation condition induced a partial smooth to rough transition of the Yersinia LPS with a decrease in the amount of 2C1-reactive antigen.
有人提出,小肠结肠炎耶尔森氏菌0:3血清型菌株的脂多糖(LPS)的O侧链在数量和质量上会因在37℃还是25℃培养而有所不同。目前尚不确定这是否会影响可能存在于LPS上的血清型特异性抗原的表达。我们用一种0:3血清型特异性单克隆抗体2C1研究了这个问题。这种单克隆抗体从去污剂溶解的125I标记的耶尔森氏菌制剂中免疫沉淀出一种39K的主要蛋白质。然而,蛋白质印迹实验结果表明,与2C1反应的抗原实际上不是39K蛋白质,而是LPS的O侧链。值得注意的是,无论细菌是在室温还是37℃培养,都能检测到反应性。然而,吸收实验证实,37℃培养的LPS上可及的抗原决定簇较少。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和银染对LPS制剂进行了分析。这些SDS-PAGE图谱显示,37℃培养的LPS中O侧链较少。因此,对我们观察结果最可能的解释是,37℃的培养条件诱导了耶尔森氏菌LPS从部分光滑型到粗糙型的转变,同时2C1反应性抗原的量减少。