Mackin W M, Becker E L
Int J Immunopharmacol. 1983;5(5):365-75. doi: 10.1016/0192-0561(83)90011-5.
Pretreatment of rabbit peritoneal neutrophils at 37 degrees with 10-35 microM L-1-tosylamide-2-phenylethyl chloromethyl ketone (TPCK) decreases by 20-50% the detectable number of f Met-Leu-[3H]Phe binding sites. Greater TPCK concentrations, between 50 and 100 microM, cause less of a decrease or actually increase peptide binding activity to a level greater than that of untreated cells. Furthermore, Scatchard analysis indicates that the sites detected on neutrophils after TPCK treatment have 1.2-3.2 fold lower apparent Kd (higher affinity) than those detected on untreated, control cells (1.1 +/- 1.7 X 10(-8) M vs 1.7 +/- 1.5 X 10(-8) M, P less than 0.02). Thus, TPCK treatment of rabbit peritoneal neutrophils causes both a decrease in f Met-Leu-[3H]Phe receptors and increases the affinity of the remaining sites. In addition, peritoneal neutrophils incubated at 37 degrees without TPCK were found to rapidly express additional f Met-Leu-[3H]Phe receptors. These additional sites, however, were not evident on neutrophils incubated at 37 degrees with TPCK. Concomitantly with the expression of additional sites, neutrophils placed at 37 degrees were found to spontaneously release small amounts of lysozyme. However, since equivalent amounts of lysozyme were released by cells incubated with or without TPCK, we are unable to state whether expression of the additional sites is due to neutrophil degranulation. Finally, although rabbit peripheral blood neutrophils also show an increase in binding sites at 37 degrees, treatment of these cells with TPCK does not cause a decrease in their f Met-Leu-[3H]Phe binding activity.
用10 - 35微摩尔/升的L - 甲苯磺酰酰胺 - 2 - 苯乙基氯甲基酮(TPCK)在37℃预处理兔腹膜中性粒细胞,可使可检测到的f Met - Leu - [³H]Phe结合位点数量减少20% - 50%。更高浓度的TPCK(50至100微摩尔)导致的减少幅度较小,实际上反而使肽结合活性增加到高于未处理细胞的水平。此外,Scatchard分析表明,TPCK处理后的中性粒细胞上检测到的位点表观解离常数(亲和力更高)比未处理的对照细胞低1.2 - 3.2倍(分别为1.1±1.7×10⁻⁸摩尔/升和1.7±1.5×10⁻⁸摩尔/升,P<0.02)。因此,TPCK处理兔腹膜中性粒细胞会导致f Met - Leu - [³H]Phe受体数量减少,并增加剩余位点的亲和力。此外,发现未用TPCK在37℃孵育的腹膜中性粒细胞会迅速表达额外的f Met - Leu - [³H]Phe受体。然而,在用TPCK在37℃孵育的中性粒细胞上未观察到这些额外的位点。与额外位点的表达同时发生的是,置于37℃的中性粒细胞被发现会自发释放少量溶菌酶。然而,由于用TPCK孵育和未用TPCK孵育的细胞释放的溶菌酶量相当,我们无法确定额外位点的表达是否是由于中性粒细胞脱颗粒所致。最后,尽管兔外周血中性粒细胞在37℃时结合位点也会增加,但用TPCK处理这些细胞不会导致其f Met - Leu - [³H]Phe结合活性降低。