Beeken W, Bigelow J, Fabian J
Department of Medicine, University of Vermont, Burlington 05405, USA.
J Lab Clin Med. 1995 Oct;126(4):358-64.
Mucosal epithelial cell preparations from 18 disease-free segments of human colon resections and 7 cell lines were examined for chemoattractant properties. Mucosal cells were dissociated from lamina propria by sequential incubations in ethylenediaminetetraacetic acid, harvested at 1.5 x 10(6) cells/ml, and lysed. Lysates were tested for chemoattractant activity for monocytes and neutrophils. Chemoattractant preparations were further purified by gel filtration chromatography, and amino acid analysis was performed on selected chemoattractant fractions. Mucosa from normal bowel exhibited significant chemoattractant properties for monocytes, up to 15 times greater than for neutrophils. Checkerboard analysis indicated chemotaxis rather than chemokinesis. Neither cell culture nor lamina propria cell lysates exhibited statistically significant chemoattraction, although activity was evident in certain preparations of isolated cell cultures. Chromatography of human mucosal chemoattractant preparations consistently gave peaks of activity in the 2000 dalton range. These yielded consistent amino acid profiles, with aspartic acid, glutamic acid, glycine, alanine, and lysine being dominant in all preparations. This peptide is apparently different from other known chemotactic agents and could play a role in recruitment of mononuclear phagocytes to the mucosa of the human colon.
对取自18段无病变的人类结肠切除组织以及7种细胞系的黏膜上皮细胞制剂进行趋化特性检测。通过在乙二胺四乙酸中连续孵育,使黏膜细胞与固有层分离,收获细胞并调整至1.5×10⁶个细胞/毫升,然后进行裂解。检测裂解物对单核细胞和中性粒细胞的趋化活性。趋化制剂通过凝胶过滤色谱法进一步纯化,并对选定的趋化活性组分进行氨基酸分析。正常肠黏膜对单核细胞表现出显著的趋化特性,其趋化能力比中性粒细胞高15倍。棋盘分析表明是趋化作用而非化学增活作用。细胞培养物和固有层细胞裂解物均未表现出具有统计学意义的趋化活性,尽管在某些分离的细胞培养制剂中可见活性。对人类黏膜趋化制剂进行色谱分析,始终在2000道尔顿范围内出现活性峰。这些活性峰呈现出一致的氨基酸谱,天冬氨酸、谷氨酸、甘氨酸、丙氨酸和赖氨酸在所有制剂中占主导地位。这种肽显然不同于其他已知的趋化剂,可能在单核吞噬细胞向人类结肠黏膜的募集过程中发挥作用。