Gariglio M, De Andrea M, Lembo M, Ravotto M, Zappador C, Valente G, Landolfo S
Department of Medical Sciences, Medical School of Novara, Italy.
J Leukoc Biol. 1998 Nov;64(5):608-14. doi: 10.1002/jlb.64.5.608.
To examine the expression of Ifi 200 genes in vivo and add new information about their function, polyclonal monospecific rabbit antibodies, designated N-term or C-term, were raised against both the N-terminus and C-terminus of the 204 protein (p204) respectively. Western blotting analysis demonstrated that p204 and D3, another member of the Ifi 200 gene family, are constitutively expressed, though at different degrees, in bone marrow, thymus and lymph nodes, and barely detectable in the spleen. Poly rI:rC treatment did not modulate their expression. Peritoneal resident macrophages (Mphi) from untreated mice were negative, but displayed high levels of both p204 and D3 on poly rI:rC treatment. A significant increase of these proteins is also observed when Mphi are cultured overnight in vitro with IFNs or LPS. Lung, kidney and brain were negative for p204 and D3 expression. These results, together with immunohistochemical analysis, demonstrate that the 204 gene has an expression pattern restricted to cells of the myelomonocytic lineage similar to that observed for the human homolog, the myeloid nuclear differentiation antigen (MNDA) suggesting its potential involvement in the differentiation and maturation of this cell lineage.
为了检测Ifi 200基因在体内的表达并补充有关其功能的新信息,分别针对204蛋白(p204)的N端和C端制备了名为N端或C端的多克隆单特异性兔抗体。蛋白质印迹分析表明,p204和Ifi 200基因家族的另一个成员D3在骨髓、胸腺和淋巴结中组成性表达,尽管程度不同,而在脾脏中几乎检测不到。聚肌苷酸:聚胞苷酸(poly rI:rC)处理并未调节它们的表达。未经处理小鼠的腹腔常驻巨噬细胞(Mphi)为阴性,但在聚肌苷酸:聚胞苷酸处理后p204和D3均呈现高水平表达。当巨噬细胞在体外与干扰素或脂多糖一起培养过夜时,也观察到这些蛋白质显著增加。肺、肾和脑的p204和D3表达均为阴性。这些结果与免疫组织化学分析一起表明,204基因的表达模式局限于骨髓单核细胞系细胞,类似于在人类同源物髓核分化抗原(MNDA)中观察到的模式,提示其可能参与该细胞系的分化和成熟。