Watt K W, Brightman I L, Goetzl E J
Immunology. 1983 Jan;48(1):79-86.
Human leucocyte lysosomal polypeptides of mol. wt 4000-5000, which constitute the neutrophil-immobolizing factor (NIF), were isolated from the 22,000 g supernate of sonicates of human neutrophils by filtration on Sephadex G-75. The larger (NIF-1) and smaller (NIF-2) of the polypeptides were resolved by filtration on Bio-Gel P6 and purified to homogeneity by sequential reverse-phase high performance liquid chromatography and paper electrophoresis. The results of analyses of amino acid composition indicated that NIF-1 and NIF-2 are distinct polypeptides composed of an apparent total of 41 and 38 amino acids, respectively. Both NIF polypeptides contain one cysteine and one methionine, lack isoleucine, tyrosine and phenylalanine, and are rich in histidine and proline. The sequence of 20 of the amino-terminal amino acids of both NIF polypeptides is identical, but NIF-2 possesses an additional alanine at the amino-terminus. Highly purified NIF-1 and NIF-2 inhibited human neutrophil random migration and chemotaxis to diverse stimuli in a concentration-dependent manner, with 50% inhibition of chemotaxis by 0.31-1 x 10(-8) M NIF-1 and 1-3 x 10(-7) M NIF-2. Neither NIF polypeptide was cytotoxic for neutrophils, altered neutrophil phagocytosis or release of lysosomal enzymes, or inhibited mononuclear leucocyte chemotaxis. The leucocyte and functional specificity of the NIF polypeptides and the quantitites released upon stimulation of the human leucocytes suggest that the transition to a mononuclear leucocyte population in chronic inflammation may be attributable in part to the NIF derived from the leucocyte infiltrates of acute responses.
从人中性粒细胞超声裂解物的22,000g上清液中，通过Sephadex G - 75过滤分离出分子量为4000 - 5000的人白细胞溶酶体多肽，其构成中性粒细胞固定因子（NIF）。通过Bio - Gel P6过滤分离出较大的（NIF - 1）和较小的（NIF - 2）多肽，并通过连续反相高效液相色谱和纸电泳纯化至同质。氨基酸组成分析结果表明，NIF - 1和NIF - 2是不同的多肽，分别由明显总共41个和38个氨基酸组成。两种NIF多肽都含有一个半胱氨酸和一个甲硫氨酸，缺乏异亮氨酸、酪氨酸和苯丙氨酸，并且富含组氨酸和脯氨酸。两种NIF多肽的20个氨基末端氨基酸序列相同，但NIF - 2在氨基末端有一个额外的丙氨酸。高度纯化的NIF - 1和NIF - 2以浓度依赖性方式抑制人中性粒细胞的随机迁移和对各种刺激的趋化性，0.31 - 1×10⁻⁸M的NIF - 1和1 - 3×10⁻⁷M的NIF - 2对趋化性有50%的抑制作用。两种NIF多肽对中性粒细胞均无细胞毒性，不改变中性粒细胞的吞噬作用或溶酶体酶的释放，也不抑制单核白细胞趋化性。NIF多肽的白细胞和功能特异性以及人白细胞刺激后释放的量表明，慢性炎症中向单核白细胞群体的转变可能部分归因于急性反应中白细胞浸润产生的NIF。
