Munzig E, Eckert K, Harrach T, Graf H, Maurer H R
Institut für Pharmazie der Freien Universität Berlin, Germany.
FEBS Lett. 1994 Sep 5;351(2):215-8. doi: 10.1016/0014-5793(94)00860-4.
The thiol protease bromelain has been shown to remove T-cell CD44 molecules from lymphocytes and to affect T-cell activation. We investigated the effect of a highly purified bromelain protease F9 (F9) on the adhesion of peripheral blood lymphocytes (PBL) to human umbilical vein endothelial cells (HUVEC). Preincubation of the lymphocytes with F9 reduced the adherence to about 20% of unstimulated and to about 30% of phorbol-dibutyrate (P(Bu)2) stimulated lymphocytes. Using flow cytometry, both crude bromelain and protease F9 reduced the expression of CD44, but not of LFA-1, on PBL. F9 was about 10 times more active than crude bromelain; at 2.5 micrograms/ml of F9 about 97% inhibition of CD44 expression was found. A mAb against CD44 was tested and found to block the F9-induced decrease in PBL-binding to HUVEC. The results indicate that F9 selectively decreases the CD44 mediated binding of PBL to HUVEC.
巯基蛋白酶菠萝蛋白酶已被证明可从淋巴细胞中去除T细胞CD44分子,并影响T细胞活化。我们研究了高度纯化的菠萝蛋白酶F9(F9)对外周血淋巴细胞(PBL)与人脐静脉内皮细胞(HUVEC)黏附的影响。用F9对淋巴细胞进行预孵育后,未刺激的淋巴细胞黏附率降低至约20%,佛波醇二丁酸酯(P(Bu)2)刺激的淋巴细胞黏附率降低至约30%。使用流式细胞术,粗制菠萝蛋白酶和蛋白酶F9均可降低PBL上CD44的表达,但不影响LFA-1的表达。F9的活性约为粗制菠萝蛋白酶的10倍;在2.5微克/毫升的F9浓度下,发现CD44表达的抑制率约为97%。对一种抗CD44单克隆抗体进行了测试,发现它可阻断F9诱导的PBL与HUVEC结合的减少。结果表明,F9可选择性降低CD44介导的PBL与HUVEC的结合。