Cho N, Shinomiya H, Matsumura H, Wang F Z, Nakano M
Department of Microbiology, Jichi Medical School, Tochigi, Japan.
Int Arch Allergy Appl Immunol. 1991;96(2):161-7. doi: 10.1159/000235488.
Protein antigens eliciting delayed type hypersensitivity (DTH) were analyzed and purified from the supernatants of protein-free cultures in which Salmonella typhimurium TV148 organisms had grown. DTH activity was measured by the footpad swelling test in mice immunized with living organisms of S. typhimurium TV148 or Escherichia coli K-12. DTH activity in the culture supernatant was specific to TV148-immunized mice. This activity was destroyed by pronase. DTH activity was unable to pass through an ultrafilter with an exclusion limit of 10 kD. After condensation of the supernatant and following centrifugation (100,000 g for 1 h), the DTH activities of the sediment and the supernatant were examined, and both showed DTH activity. Further analyses of DTH antigens in the supernatant by HPLC gel filtration separated the activity into three portions. The most active portion was further fractionated by hydroxyapatite HPLC, revealing the presence of two DTH antigens, with molecular weights of 65 and less than 10 kD. These results indicate that the culture supernatant of S. typhimurium TV148 organisms contains a variety of macromolecular protein DTH-eliciting antigens, and one of the antigens is 65 kD, which is dissociated partly by organic solvents into a low molecular weight (less than 10 kD) antigen.
对从鼠伤寒沙门氏菌TV148菌株生长的无蛋白培养上清液中引发迟发型超敏反应（DTH）的蛋白质抗原进行了分析和纯化。通过足垫肿胀试验测定用鼠伤寒沙门氏菌TV148或大肠杆菌K - 12活菌免疫的小鼠的DTH活性。培养上清液中的DTH活性对经TV148免疫的小鼠具有特异性。该活性被链霉蛋白酶破坏。DTH活性不能通过截留分子量为10 kD的超滤膜。将上清液浓缩并离心（100,000 g，1小时）后，检测沉淀物和上清液的DTH活性，两者均显示出DTH活性。通过高效液相色谱（HPLC）凝胶过滤对上清液中的DTH抗原进行进一步分析，将活性分为三个部分。活性最高的部分通过羟基磷灰石HPLC进一步分级分离，发现存在两种DTH抗原，分子量分别为65 kD和小于10 kD。这些结果表明，鼠伤寒沙门氏菌TV148菌株的培养上清液含有多种引发DTH的大分子蛋白质抗原，其中一种抗原分子量为65 kD，可被有机溶剂部分解离为低分子量（小于10 kD）的抗原。
