Makioka A, Kobayashi A
J Protozool. 1986 Feb;33(1):21-6. doi: 10.1111/j.1550-7408.1986.tb05549.x.
Water-soluble and 0.6 M KCl-soluble protein fractions prepared from Tetrahymena pyriformis, when inoculated into mice, could effectively induce activated macrophages having the ability to kill Toxoplasma gondii in vitro. This effect was not induced by other proteins tested, such as bovine serum albumin, pepsin from porcine stomach mucosa and chicken egg-white lysozyme, nor by muramyl dipeptide (MDP), a potent immunoadjuvant. Five fractions obtained by DEAE-Sephadex chromatography of the water-soluble protein fraction were compared with regard to induction of toxoplasmacidal activity in macrophages. The first peak was most effective for activation of macrophages. Five fractions obtained by chromatography of the 0.6 M KCl-soluble protein fraction were also examined and it was found that the first peak had the activity. No marked difference in activity was observed between the active fractions of water-soluble and 0.6 M KCl-soluble protein fractions. For practical use, we focused on the water-soluble active fraction. The minimum effective dose of the active fraction was 100 micrograms and the fraction could activate macrophages directly in vitro. Four fractions obtained by gel filtration of the active fraction on Sephadex G-200 were compared and the first peak had the activity. The first peak contained a single protein, revealed by SDS-polyacrylamide gel electrophoresis; its apparent molecular weight was 64,000.
从梨形四膜虫制备的水溶性和0.6M KCl可溶性蛋白组分，接种到小鼠体内时，能有效诱导出具有在体外杀死刚地弓形虫能力的活化巨噬细胞。这种效应不是由所测试的其他蛋白质诱导的，如牛血清白蛋白、猪胃黏膜胃蛋白酶和鸡蛋白溶菌酶，也不是由一种有效的免疫佐剂胞壁酰二肽（MDP）诱导的。对通过DEAE-葡聚糖凝胶柱层析从水溶性蛋白组分获得的五个组分，就其在巨噬细胞中诱导杀弓形虫活性的能力进行了比较。第一个峰对巨噬细胞的活化最有效。对通过0.6M KCl可溶性蛋白组分柱层析获得的五个组分也进行了检测，发现第一个峰具有活性。水溶性和0.6M KCl可溶性蛋白组分的活性组分之间未观察到明显的活性差异。为了实际应用，我们关注水溶性活性组分。活性组分的最小有效剂量为100微克，该组分能在体外直接激活巨噬细胞。对通过在葡聚糖凝胶G-200上对活性组分进行凝胶过滤获得的四个组分进行了比较，第一个峰具有活性。通过SDS-聚丙烯酰胺凝胶电泳显示，第一个峰含有单一蛋白质；其表观分子量为64,000。
