Hammann K P, Scheiner O, Raile A, Schulz T, Schopf R E, Peters H, Erdei A, Dierich M P
Int Arch Allergy Appl Immunol. 1983;70(4):337-45. doi: 10.1159/000233345.
Incorporation of [3H]-thymidine - [3H]-TdR - into concanavalin A (Con A) stimulated murine splenocytes and thymocytes was found to be enhanced by addition of certain concentrations of phenyl-methylsulfonylfluoride (PMSF), di-isopropylfluorophosphate (DFP), N-alpha-tosyl-L-lysyl-L-chloromethylketone (TLCK), and soybean trypsin inhibitor (SBTI). No enhancement could be observed when mononuclear cells of the peripheral blood were used, and a medium enhancement when thymocytes were applied. Furthermore, no enhancing effect of the protease inhibitors (PI) on the Con A response of murine splenocytes could be observed within the first 24 h of the culturing period. DFP, PMSF, and TLCK enhanced the Con A response to a similar degree, whereas SBTI was less effective. DFP and SBTI proved to be also effective when they were added after 15-24 h to the Con A cultures, if the cultures were harvested 48 h later. Removal of adherent and phagocytic spleen cells or reduction of the concentration of spleen cells shifted the effective DFP concentration to lower concentrations, whereas addition of adherent spleen cells caused a shift of the enhancing DFP amounts to higher concentrations. The data presented suggest that the enhancing effect of PI on the T cell response depends on the concentration of PI, the time of culturing and incubation, the PI used, the origin of the stimulated cells, and especially on the number of adherent and phagocytic cells. These findings might explain - at least in part - the different results on the effect of PI on the T cell response obtained in the past.
研究发现，添加一定浓度的苯甲基磺酰氟（PMSF）、二异丙基氟磷酸（DFP）、N-α-对甲苯磺酰-L-赖氨酰-L-氯甲基酮（TLCK）和大豆胰蛋白酶抑制剂（SBTI）可增强[3H]-胸腺嘧啶核苷 - [3H]-TdR掺入刀豆球蛋白A（Con A）刺激的小鼠脾细胞和胸腺细胞的能力。使用外周血单个核细胞时未观察到增强作用，而使用胸腺细胞时增强作用中等。此外，在培养期的最初24小时内未观察到蛋白酶抑制剂（PI）对小鼠脾细胞Con A反应的增强作用。DFP、PMSF和TLCK对Con A反应的增强程度相似，而SBTI的效果较差。如果在15 - 24小时后向Con A培养物中添加DFP和SBTI，并在48小时后收获培养物，DFP和SBTI也被证明是有效的。去除贴壁和吞噬性脾细胞或降低脾细胞浓度会使有效DFP浓度向更低浓度转移，而添加贴壁脾细胞会使增强DFP的量向更高浓度转移。所呈现的数据表明，PI对T细胞反应的增强作用取决于PI的浓度、培养和孵育时间、所用的PI、受刺激细胞的来源，尤其是贴壁和吞噬细胞的数量。这些发现可能至少部分解释了过去关于PI对T细胞反应影响的不同结果。
