Sehgal G, Zhang K, Todd R F, Boxer L A, Petty H R
Department of Biological Sciences, Wayne State University, Detroit, MI 48202.
J Immunol. 1993 May 15;150(10):4571-80.
We have tested the role of lectin-like interactions, with particular emphasis on CR3, in insoluble immune complex (IC)-mediated activation of human polymorphonuclear leukocytes (PMN). The ability of IC to trigger intracellular Ca2+ release and O2- production by normal PMN, saccharide-treated cells, and CR3-deficient PMN (leukocyte adhesion deficiency, LAD, patients) were tested. When indo-1-labeled normal PMN were stimulated with IC in Ca(2+)-free buffer, intracellular Ca2+ rose from approximately 100 nM to approximately 230 nM. However, when LAD PMN were tested, a small rise in intracellular Ca2+ was observed. Because previous studies have shown that certain saccharides inhibit CR3-Fc gamma RIII co-capping, we tested a panel of saccharides to determine their ability to influence IC-mediated intracellular Ca2+ release. When normal PMN were exposed to 0.15 M N-acetyl-D-glucosamine (NADG), D-mannose, or alpha-methyl-mannoside, the Ca2+ response to IC was significantly reduced. However, addition of 0.15 M glucose, raffinose, sucrose, galactose, fructose, or sorbitol did not significantly affect the Ca2+ response, suggesting that the response was specific for certain saccharides. To test the physiologic consequences of these Ca2+ signals, we have examined the ability of saccharides to influence O2- production by normal PMN and the ability of LAD PMN to produce O2- upon triggering by IC. Normal PMN stimulated with IC generated 4.3 +/- 1.7 nmol/10(6) cells/30 min of O2-. In contrast, O2- production was inhibited by 0 to 20% by glucose, galactose, sucrose, sorbitol, fructose, and raffinose and > or = 50% by NADG and mannose. LAD PMN, which display diminished Ca2+ signals, were found to produce O2- at 47 +/- 6% of control levels. NADG and mannose dose-response studies indicated that they cooperatively block O2-.
我们测试了凝集素样相互作用的作用,尤其着重于CR3,在不溶性免疫复合物(IC)介导的人多形核白细胞(PMN)激活中的作用。测试了IC触发正常PMN、经糖类处理的细胞以及CR3缺陷型PMN(白细胞黏附缺陷症,LAD,患者)细胞内Ca2+释放和O2-产生的能力。当用无Ca(2+)缓冲液中的IC刺激indo-1标记的正常PMN时,细胞内Ca2+从约100 nM升至约230 nM。然而,当测试LAD PMN时,观察到细胞内Ca2+有小幅升高。因为先前的研究表明某些糖类会抑制CR3-FcγRIII共帽化,我们测试了一组糖类以确定它们影响IC介导的细胞内Ca2+释放的能力。当正常PMN暴露于0.15 M N-乙酰-D-葡萄糖胺(NADG)、D-甘露糖或α-甲基甘露糖苷时,对IC的Ca2+反应显著降低。然而,添加0.15 M葡萄糖、棉子糖、蔗糖、半乳糖、果糖或山梨醇并未显著影响Ca2+反应,表明该反应对某些糖类具有特异性。为了测试这些Ca2+信号的生理后果,我们研究了糖类影响正常PMN产生O2-的能力以及LAD PMN在IC触发时产生O(2)-的能力。用IC刺激的正常PMN产生4.3±1.7 nmol/10(6)细胞/30分钟的O2-。相比之下,葡萄糖、半乳糖、蔗糖、山梨醇、果糖和棉子糖对O2-产生的抑制率为0至20%,而NADG和甘露糖的抑制率≥50%。发现显示Ca2+信号减弱的LAD PMN产生O2-的水平为对照水平的47±6%。NADG和甘露糖的剂量反应研究表明它们协同阻断O2-。