Kadish J L, Choi C C, Czop J K
Immunol Res. 1986;5(2):129-38. doi: 10.1007/BF02917587.
Murine bone marrow cells, plated at 4 X 10(4) cells/well and cultured in 50% fibroblast CM, yielded pure populations of large, individual, adherent cells that were phagocytic and morphologically indistinguishable from macrophages. Adherent macrophages appeared in small numbers with 24 h of culture, increased to maximal cell numbers within 10 days of culture, and remained at these cell densities for at least 11 weeks in culture. The capacities of adherent macrophages to ingest unopsonized zymosan particles and EsIgG, at inputs of 1.25 X 10(7) targets, were expressed by 7 and 40% of the cells derived from 24-hour cultures, respectively, were increased at nearly identical rates to comparable maximal levels within 10-14 days of culture and were exhibited by essentially all adherent cells derived from 2-11-week cultures. The percentage of adherent macrophages from twelve 3-6-week cultures ingesting greater than or equal to 1, greater than or equal to 6 and greater than or equal to 10 zymosan particles was 89 +/- 5, 47 +/- 11 and 14 +/- 9% (mean +/- SD, n = 12), respectively, and the percentage ingesting greater than or equal to 1, greater than or equal 6 and greater than or equal to 10 EsIgG was 86 +/- 5, 49 +/- 10 and 14 +/- 8%, respectively. Incubation of adherent macrophages with mannan-free ss-glucan particles at inputs of 5 X 10(5)-5 X 10(7)/ml initiated a phagocytic response comparable to that obtained with the same doses of zymosan particles which contained mannan and beta-glucan. Preincubation of adherent macrophages with 100 micrograms/ml of a fully soluble beta-glucan, laminarin, and solubilized barley beta-glucan reduced subsequent macrophage phagocytosis of greater than or equal to 6 zymosan particles by 53 and 40%, respectively. In contrast, yeast alpha-mannan was less than 1% as active, and 10 mg/ml reduced the number of adherent macrophages ingesting greater than or equal to 1 zymosan particles by 64%. At concentrations as high as 2 mg/ml, laminarin and barley beta-glucan had no effect on Fc receptor-mediated ingestion of EsIgG, and mannan at 20 mg/ml also failed to inhibit EsIgG ingestion. Pretreatment of adherent macrophages with 20 micrograms/ml of trypsin reduced the number of cells ingesting greater than or equal to 1 zymosan particles from 89 to 10% and those ingesting greater than or equal to 6 zymosan from 43 to 0%, whereas pretreatment with as much as 100 micrograms/ml of trypsin failed to decrease macrophage ingestion of EsIgG.(ABSTRACT TRUNCATED AT 400 WORDS)
将小鼠骨髓细胞以4×10⁴个细胞/孔接种，在50%成纤维细胞条件培养基中培养，可产生大量单个贴壁细胞的纯群体，这些细胞具有吞噬作用，形态上与巨噬细胞无法区分。贴壁巨噬细胞在培养24小时时少量出现，培养10天内细胞数量增加到最大值，并在培养至少11周内保持这些细胞密度。在输入1.25×10⁷个靶标的情况下，贴壁巨噬细胞摄取未调理酵母聚糖颗粒和抗绵羊红细胞IgG（EsIgG）的能力，分别由24小时培养的细胞中的7%和40%表现出来，在培养10 - 14天内以几乎相同的速率增加到相当的最大水平，并且基本上所有来自2 - 11周培养的贴壁细胞都有此表现。来自12个3 - 6周培养物的贴壁巨噬细胞摄取大于或等于1、大于或等于6和大于或等于10个酵母聚糖颗粒的百分比分别为89±5%、47±11%和14±9%（平均值±标准差，n = 12），摄取大于或等于1、大于或等于6和大于或等于10个EsIgG的百分比分别为86±5%、49±10%和14±8%。用输入量为5×10⁵ - 5×10⁷/ml的无甘露聚糖的β - 葡聚糖颗粒孵育贴壁巨噬细胞，引发的吞噬反应与用相同剂量含甘露聚糖和β - 葡聚糖的酵母聚糖颗粒获得的反应相当。用100μg/ml完全可溶的β - 葡聚糖、海带多糖和溶解的大麦β - 葡聚糖预孵育贴壁巨噬细胞，随后巨噬细胞对大于或等于6个酵母聚糖颗粒的吞噬作用分别降低了53%和40%。相比之下，酵母α -甘露聚糖的活性不到1%，10mg/ml可使摄取大于或等于1个酵母聚糖颗粒的贴壁巨噬细胞数量减少64%。在高达2mg/ml的浓度下，海带多糖和大麦β - 葡聚糖对Fc受体介导的EsIgG摄取没有影响，20mg/ml的甘露聚糖也未能抑制EsIgG摄取。用20μg/ml胰蛋白酶预处理贴壁巨噬细胞，摄取大于或等于1个酵母聚糖颗粒的细胞数量从89%减少到10%，摄取大于或等于6个酵母聚糖颗粒的细胞数量从43%减少到0%，而用高达100μg/ml胰蛋白酶预处理未能降低巨噬细胞对EsIgG的摄取。（摘要截短至400字）
