Gillery P, Serpier H, Polette M, Bellon G, Clavel C, Wegrowski Y, Birembaut P, Kalis B, Cariou R, Maquart F X
Laboratory of Biochemistry, CNRS URA 610, Faculté de Médecine, Reims/France.
Eur J Cell Biol. 1992 Apr;57(2):244-53.
Three-dimensional collagen lattice cultures of fibroblasts mimic the in vivo situation better than monolayer cultures. Here, skin fibroblasts from scleroderma patients and healthy controls were cultivated in collagen lattices, and the effects of recombinant human gamma-interferon (IFN-gamma) on these cultures investigated. IFN-gamma inhibited collagen lattice retraction in a dose-dependent way at concentrations ranging from 10 to 10,000 U/ml. This effect was independent of any alteration to the cell proliferation within the lattices. The inhibition was of the same order of magnitude in normal and pathological fibroblasts. The synthesis of collagen and non-collagen proteins, particularly fibronectin, was increased in scleroderma cultures. It was inhibited in both normal and scleroderma fibroblasts by IFN-gamma, with a maximal effect at the concentration 1000 U/ml, but the inhibition of protein synthesis was far more intense in scleroderma than in normal cells. In situ hybridization, Northern blot and dot blot analyses showed that mRNA coding for pro alpha 1(I) collagen was decreased in IFN-gamma-treated cells, indicating an effect at the pretranslational level. IFN-gamma also inhibited glycosaminoglycan synthesis, but in scleroderma cells only. This study shows that IFN-gamma regulates cell behavior in three-dimensional collagen matrices: (i) it decreases protein and specifically glycosaminoglycan synthesis in scleroderma fibroblasts, (ii) it modulates the interactions between cells and matrix that lead to the retraction of the lattice. Whereas collagen synthesis is largely decreased in lattice cultures like in vivo, it remains increased in the case of scleroderma compared to normal fibroblasts and may be down-regulated by IFN-gamma. Similar conclusions may be drawn for fibronectin and glycosaminoglycans. The inhibitory effect of IFN-gamma on the retraction capacity of fibroblasts and on their ability to synthesize increased amounts of extracellular matrix macromolecules may be of potential interest for therapeutic use of IFN-gamma in scleroderma patients.
成纤维细胞的三维胶原晶格培养比单层培养能更好地模拟体内情况。在此,将硬皮病患者和健康对照者的皮肤成纤维细胞培养于胶原晶格中,并研究重组人γ干扰素(IFN-γ)对这些培养物的影响。IFN-γ在10至10000 U/ml的浓度范围内以剂量依赖方式抑制胶原晶格收缩。该效应与晶格内细胞增殖的任何改变无关。正常和病理成纤维细胞中的抑制作用程度相同。硬皮病培养物中胶原和非胶原蛋白质,特别是纤连蛋白的合成增加。IFN-γ在正常和硬皮病成纤维细胞中均抑制其合成,在浓度为1000 U/ml时作用最大,但硬皮病细胞中蛋白质合成的抑制远比正常细胞强烈。原位杂交、Northern印迹和斑点印迹分析表明,经IFN-γ处理的细胞中编码原α1(I)胶原的mRNA减少,表明在翻译前水平有作用。IFN-γ也抑制糖胺聚糖合成,但仅在硬皮病细胞中。本研究表明,IFN-γ调节三维胶原基质中的细胞行为:(i)它减少硬皮病成纤维细胞中的蛋白质尤其是糖胺聚糖合成,(ii)它调节导致晶格收缩的细胞与基质之间的相互作用。与体内情况一样,晶格培养中胶原合成大幅减少,但与正常成纤维细胞相比,硬皮病时胶原合成仍增加,且可能被IFN-γ下调。对于纤连蛋白和糖胺聚糖也可得出类似结论。IFN-γ对成纤维细胞收缩能力及其合成大量细胞外基质大分子能力的抑制作用可能对硬皮病患者使用IFN-γ进行治疗具有潜在意义。