Raybourne R, Deardorff T L, Bier J W
Exp Parasitol. 1986 Aug;62(1):92-7. doi: 10.1016/0014-4894(86)90012-3.
Excretory secretory proteins produced in vitro by Anisakis simplex larvae incubated in Medium 199 or phosphate buffered saline with dextrose are similar with respect to protein content and biological activity. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis suggested that the molecular weight of the component(s) responsible for inhibition of mitogen induced lymphocyte blastogenesis is between 66,000 and 95,000. In vitro production of excretory secretory protein, approximately 1 microgram/24 hr by a single larva, was sufficient to inhibit lymphocyte blastogenesis. Serum from a human anisakiasis patient reacted with these proteins in immunoblots, indicating that, during invasion of the gastric mucosa, enough of them are produced in vivo to induce an immune response. The excretory secretory proteins significantly inhibited proliferation of transformed mammalian cell lines of lymphoid (P3/X63-Ag8) and epithelioid (HeLa) origin. As in mitogen stimulated lymphocytes, the inhibitory effect was cytostatic rather than cytotoxic. These findings suggest that, in addition to being potent immunogens, larval excretory secretory proteins are produced in sufficient quantity to modulate the host response in anisakiasis.
用199培养基或含葡萄糖的磷酸盐缓冲盐水培养的简单异尖线虫幼虫体外产生的排泄分泌蛋白，在蛋白质含量和生物活性方面相似。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明，负责抑制有丝分裂原诱导的淋巴细胞母细胞化的成分分子量在66,000至95,000之间。单个幼虫体外产生排泄分泌蛋白的量约为1微克/24小时，足以抑制淋巴细胞母细胞化。一名人类异尖线虫病患者的血清在免疫印迹中与这些蛋白质发生反应，表明在胃黏膜入侵期间，体内产生了足够量的这些蛋白质以诱导免疫反应。排泄分泌蛋白显著抑制淋巴样（P3/X63-Ag8）和上皮样（HeLa）来源的转化哺乳动物细胞系的增殖。与有丝分裂原刺激的淋巴细胞一样，抑制作用是细胞生长抑制而非细胞毒性。这些发现表明，幼虫排泄分泌蛋白除了是强效免疫原外，其产生量足以调节异尖线虫病中的宿主反应。
