Rosenbaum J T, Wong K, Perez H D, Raymond W, Howes E L
Invest Ophthalmol Vis Sci. 1984 Oct;25(10):1184-91.
Although a cellular exudate characterizes acute anterior uveitis, few studies have sought to identify the chemoattractant(s) contributing to this phenomenon. As a model of acute ocular inflammation, the authors have injected rabbits intravenously with endotoxin (Salmonella typhimurium LPS, 2.5 micrograms/kg). In a Boyden chamber assay, aqueous humor drawn 3 hr after LPS (post-LPS aqueous) exhibited chemotactic activity for purified rabbit granulocytes (PMN). "Checkerboard" analysis indicated that chemotaxis, rather than protein-induced chemokinesis, primarily accounted for PMN migration. Aqueous from normal rabbits demonstrated no chemotactic activity. Chemotactic activity was maximal at 3 hr post-LPS (versus 1 or 5 hr). PMN migration exhibited a direct correlation with the concentration of aqueous tested (0.5-5%). Several observations indicated that this chemotactic activity is complement (C5)-derived. It is inhibited by antibodies to C5 but not affected by antibodies to C3. Similar to rabbit C5a, chemotactic activity in post-LPS aqueous was heat stable at 56 degrees C X 30 min, attracted both human and rabbit PMN at similar concentrations and induced release of beta glucuronidase from PMN. In addition, prior incubation of rabbit PMN with partially purified C5a (densensitization) specifically inhibited chemotactic responses to both C5a and post-LPS aqueous without inhibiting responses to another chemoattractant, n-formyl-methionyl-leucyl-phenylalanine. Finally, chemotactic activity from post-LPS aqueous could be recovered from a Sephadex G75 column and eluted similarly to chemotactic activity in zymosan activated rabbit serum or 13,700 D molecular weight marker. The presence of complement-derived chemotactic activity in this model should not be construed as evidence that this activity contributes to the pathogenesis of endotoxin-induced inflammation.(ABSTRACT TRUNCATED AT 250 WORDS)
尽管细胞渗出物是急性前葡萄膜炎的特征，但很少有研究试图确定促成这种现象的趋化因子。作为急性眼部炎症的模型，作者给兔子静脉注射内毒素（鼠伤寒沙门氏菌脂多糖，2.5微克/千克）。在博伊登小室试验中，脂多糖注射后3小时抽取的房水（脂多糖后房水）对纯化的兔粒细胞（多形核白细胞）表现出趋化活性。“棋盘”分析表明，趋化作用而非蛋白质诱导的细胞运动主要导致多形核白细胞迁移。正常兔子的房水未显示趋化活性。趋化活性在脂多糖注射后3小时达到最大值（与1小时或5小时相比）。多形核白细胞迁移与所测试房水的浓度（0.5 - 5%）呈直接相关。几项观察结果表明，这种趋化活性是补体（C5）衍生的。它被抗C5抗体抑制，但不受抗C3抗体影响。与兔C5a相似，脂多糖后房水中的趋化活性在56℃×30分钟时热稳定，以相似浓度吸引人和兔的多形核白细胞，并诱导多形核白细胞释放β-葡萄糖醛酸酶。此外，用部分纯化的C5a预先孵育兔多形核白细胞（脱敏）可特异性抑制对C5a和脂多糖后房水的趋化反应，而不抑制对另一种趋化因子N-甲酰甲硫氨酰亮氨酰苯丙氨酸的反应。最后，脂多糖后房水的趋化活性可从葡聚糖G75柱上回收，洗脱方式与酵母聚糖激活的兔血清或13,700 D分子量标记物中的趋化活性相似。该模型中补体衍生趋化活性的存在不应被解释为该活性促成内毒素诱导炎症发病机制的证据。（摘要截短至250字）
