{"title":"Monocyte chemoattractant activity associated with human colon mucosal cells.","authors":"W Beeken, J Bigelow, J Fabian","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":23085,"journal":{"name":"The Journal of laboratory and clinical medicine","volume":"126 4","pages":"358-64"},"PeriodicalIF":0.0000,"publicationDate":"1995-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Journal of laboratory and clinical medicine","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
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.
从人结肠切除的18个无病段和7个细胞系制备的粘膜上皮细胞进行了化学引诱特性的检测。粘膜细胞在乙二胺四乙酸中连续孵育,从固有层分离,以1.5 x 10(6)个细胞/ml的速度收获,并裂解。测定裂解物对单核细胞和中性粒细胞的化学引诱活性。通过凝胶过滤层析进一步纯化化学引诱剂制剂,并对选定的化学引诱剂组分进行氨基酸分析。正常肠粘膜对单核细胞表现出显著的化学引诱特性,比中性粒细胞强15倍。棋盘分析显示趋化性而非趋化运动。细胞培养物和固有层细胞裂解物都没有表现出统计学上显著的化学吸引力,尽管活性在某些分离细胞培养物的制备中是明显的。人粘膜化学引诱剂制剂的色谱一致给出活性峰在2000道尔顿范围内。这些结果产生了一致的氨基酸谱,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。这种肽明显不同于其他已知的趋化剂,可能在单核吞噬细胞募集到人结肠粘膜中发挥作用。