Tarsi-Tsuk D, Levy R
Clinical Biochemistry Unit, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
J Immunol. 1990 Apr 1;144(7):2665-70.
Lipoteichoic acid (LTA) from Streptococcus faecalis stimulates the respiratory burst in peripheral blood monocytes (mon), as measured by cytochrome C reduction. The effect of LTA was time and dose dependent. LTA stimulated the respiratory burst in a biphasic manner within a range of 1 to 1000 ng/ml.10(6) mon, with maximal activity at 50 ng/ml. At this concentration LTA increased the activity from 0.97 +/- 0.2 to 4.88 +/- 0.2 nmol.10(6) mon/20 min. The role of calcium ions in the effect of LTA in stimulating respiratory burst was studied by changing the availability of calcium ions in the medium, and by measuring the effect of LTA on 45Ca2+ uptake and on intracellular Ca2+ levels. Removal of extracellular calcium ions in the presence of the calcium chelator EGTA, abolished the LTA-stimulated respiratory burst. LTA (50 ng/ml) was found to increase 45Ca2+ uptake into monocytes within seconds (from 2200 +/- 242 in the untreated cells to 4642 +/- 365 cpm/min in the LTA-treated mon). At this concentration, LTA stimulated an immediate rise in the intracellular free Ca2+ concentration to 155 +/- 15 nM as compared with 120 +/- 14 nM in the unstimulated monocytes. LTA caused a specific release of arachidonic acid indicating the involvement of phospholipase A2 in the transduction signal stimulating the respiratory burst by LTA.
粪肠球菌的脂磷壁酸(LTA)可刺激外周血单核细胞(mon)的呼吸爆发,这可通过细胞色素C还原法来测定。LTA的作用具有时间和剂量依赖性。在1至1000 ng/ml的范围内,LTA以双相方式刺激呼吸爆发。10(6)个单核细胞,在50 ng/ml时活性最高。在此浓度下,LTA使活性从0.97±0.2增加到4.88±0.2 nmol·10(6)个单核细胞/20分钟。通过改变培养基中钙离子的可用性,以及测量LTA对45Ca2+摄取和细胞内Ca2+水平的影响,研究了钙离子在LTA刺激呼吸爆发作用中的作用。在钙离子螯合剂EGTA存在的情况下去除细胞外钙离子,可消除LTA刺激的呼吸爆发。发现LTA(50 ng/ml)在数秒内可增加单核细胞对45Ca2+的摄取(从未处理细胞中的2200±242增加到LTA处理的单核细胞中的4642±365 cpm/分钟)。在此浓度下,与未刺激的单核细胞中的120±14 nM相比,LTA刺激细胞内游离Ca2+浓度立即升至155±15 nM。LTA导致花生四烯酸的特异性释放,表明磷脂酶A2参与了LTA刺激呼吸爆发的转导信号。