Sobel J D, Gallin J I
J Clin Invest. 1979 Apr;63(4):609-18. doi: 10.1172/JCI109343.
The elaboration of leukocyte chemotactic factors by human fibroblasts was studied. 12 lines of normal fibroblasts obtained by skin biopsy and then cultured in vitro produced chemoattractants (assessed by modified Boyden-chamber techniques) for both peripheral blood polymorphonuclear leukocytes and monocytes (obtained by Hypaque-Ficoll and dextran sedimentation). Chemotactic activity was not present performed in fibroblasts, and cycloheximide blocked its elaboration. The chemotactic activity of crude-culture supernate was heat stable (56 degrees C for 30 min), trypsin- and pronase-sensitive, and neuraminidase resistant. Characterization of the chemotactic activity by gel filtration (Sephadex G-75) showed two active fractions, one with mol wt greater than 100,000 and the other less than 10,000. In studies designed to relate these chemotactic factors to collagen, we have confirmed that type I collagen and alpha 1-chain; are chemotactically active for monocytes but not polymorphonuclear leukocytes. However, the chemotactic activity in fibroblast-culture media was media was distinct from collagen in that it attracted neutrophils, it was not precipitated by 25% ammonium sulfate, and it was resistant to collagenase treatment; ascorbic acid, in concentrations known to stimulate fibroblast collagen synthesis, had no effect on the elaboration of the chemotactic factors. Furthermore, amino acid analysis of Sephadex G-75 fractions with chemotactic activity failed to reveal amino acids such as hydroxyproline characteristic of collagen. In addition to the chemotactic factors secreted by fibroblasts, a heat-resistant factor (30 min at 56 degrees C) which generated the chemotactically active fragment of C5 (C5a) from human serum was also secreted. The elaboration of mediators of the inflammatory and immune responses by fibroblasts may initiate and(or) modulate local skin inflammatory reactions and play a protective role in vivo.
对人成纤维细胞产生白细胞趋化因子的情况进行了研究。通过皮肤活检获得12株正常成纤维细胞，然后在体外培养，这些细胞对外周血多形核白细胞和单核细胞（通过泛影葡胺 - 聚蔗糖和葡聚糖沉降法获得）产生趋化剂（通过改良的博伊登小室技术评估）。在成纤维细胞中未检测到趋化活性，放线菌酮可阻断其产生。粗培养上清液的趋化活性对热稳定（56℃ 30分钟），对胰蛋白酶和链霉蛋白酶敏感，对神经氨酸酶有抗性。通过凝胶过滤（葡聚糖凝胶G - 75）对趋化活性进行表征显示有两个活性部分，一个分子量大于100,000，另一个小于10,000。在旨在将这些趋化因子与胶原蛋白联系起来的研究中，我们证实I型胶原蛋白和α1链对单核细胞有趋化活性，但对多形核白细胞没有。然而，成纤维细胞培养基中的趋化活性与胶原蛋白不同，它能吸引中性粒细胞，不被25%硫酸铵沉淀，并对胶原酶处理有抗性；已知能刺激成纤维细胞胶原蛋白合成的浓度的抗坏血酸对趋化因子的产生没有影响。此外，对具有趋化活性的葡聚糖凝胶G - 75部分进行氨基酸分析，未发现胶原蛋白特有的氨基酸如羟脯氨酸。除了成纤维细胞分泌的趋化因子外，还分泌了一种耐热因子（56℃ 30分钟），该因子可从人血清中产生趋化活性片段C5（C5a）。成纤维细胞对炎症和免疫反应介质的产生可能启动和（或）调节局部皮肤炎症反应，并在体内发挥保护作用。
