Dias-Baruffi M, Cunha F Q, Ferreira S H, Roque-Barreira M C
Department of Immunology, Faculty of Medicine, Ribeirão Preto São Paulo, Brazil.
Agents Actions. 1993;38 Spec No:C54-6. doi: 10.1007/BF01991135.
We have previously reported that rat peritoneal macrophages stimulated with LPS release a factor (MNCF) which induces neutrophil migration that is not blocked by glucocorticoids. The supernatant of macrophage monolayers stimulated with LPS was submitted to affinity chromatography on immobilized sugar columns. We observed that the D-gal binding fraction retained MNCF activity. This fraction, consisting of four protein components, was submitted to chromatography on Superdex 75, yielding a homogeneous preparation of the active component. MNCF has a MW of 54 KDa (gel filtration and SDS-PAGE) and pI < 4.0 (isoelectrofocusing and chromatofocusing). D-gal did not interfere with the behaviour of known interleukins (IL-1 beta, IL-6, IL-8 TNF-alpha), but blocked MNCF activity in an in vitro migration assay. The present results reinforce our previous suggestion that MNCF may correspond to a novel monokine which induces neutrophil migration through a direct mechanism involving the D-gal binding site of the molecule.
我们之前曾报道，用脂多糖刺激的大鼠腹膜巨噬细胞会释放一种因子（MNCF），该因子可诱导中性粒细胞迁移，且这种迁移不受糖皮质激素的阻断。用脂多糖刺激的巨噬细胞单层培养上清液通过固定化糖柱进行亲和层析。我们观察到，与D - 半乳糖结合的部分保留了MNCF活性。该部分由四种蛋白质成分组成，将其在Superdex 75上进行层析，得到了活性成分的均一制剂。MNCF的分子量为54 kDa（凝胶过滤和SDS - PAGE法测定），等电点小于4.0（等电聚焦和层析聚焦法测定）。D - 半乳糖不干扰已知白细胞介素（IL - 1β、IL - 6、IL - 8、TNF - α）的活性，但在体外迁移试验中可阻断MNCF的活性。目前的结果强化了我们之前的推测，即MNCF可能对应一种新型单核因子，它通过涉及该分子D - 半乳糖结合位点的直接机制诱导中性粒细胞迁移。
