Kirshenbaum A S, Goff J P, Albert J P, Kessler S W, Metcalfe D D
Allergic Diseases Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Md 20892.
Int Arch Allergy Immunol. 1994 Dec;105(4):374-80. doi: 10.1159/000236786.
To determine the fate of Fc epsilon RI+ cells on fibroblasts in vitro, human bone marrow derived CD34+ cells were cultured in the presence of recombinant human interleukin 3 and recombinant human hematopoietic stem cell factor for 3 weeks, and Fc epsilon RI+ cells were purified by immunomagnetic selection. This enriched Fc epsilon RI+ cell population consisted of 92-94% basophils and 3-5% mast cells as determined by morphologic, immunohistochemical, and ultrastructural criteria. The Fc epsilon RI+ cells were then cocultured with 3T3 fibroblasts. Basophils decreased markedly by 1 week and were absent from cocultures by 2-3 weeks, while the mast cell numbers on the fibroblast monolayers remained constant. Ultrastructural examination of cocultures at 2 days demonstrated phagocytosis of basophils by fibroblasts. By 1 week, phagocytosed basophil membranes and granules gave fibroblasts the superficial appearance of mast cells by toluidine blue staining. Mast cells surviving in cocultures could be distinguished from granule-containing fibroblasts by IgE surface labeling and by ultrastructural demonstration of tryptase-positive granules. Thus, while mast cells remain viable in coculture with 3T3 fibroblasts, basophils do not survive and are internalized and degraded by the fibroblast monolayer.
为了在体外确定成纤维细胞上FcεRI⁺细胞的命运，将人骨髓来源的CD34⁺细胞在重组人白细胞介素3和重组人造血干细胞因子存在的情况下培养3周，然后通过免疫磁珠分选纯化FcεRI⁺细胞。根据形态学、免疫组织化学和超微结构标准确定，这种富集的FcεRI⁺细胞群体由92 - 94%的嗜碱性粒细胞和3 - 5%的肥大细胞组成。然后将FcεRI⁺细胞与3T3成纤维细胞共培养。嗜碱性粒细胞在1周时显著减少，在共培养2 - 3周时消失，而成纤维细胞单层上的肥大细胞数量保持恒定。共培养2天时的超微结构检查显示成纤维细胞对嗜碱性粒细胞的吞噬作用。到1周时，被吞噬的嗜碱性粒细胞膜和颗粒通过甲苯胺蓝染色使成纤维细胞表面呈现肥大细胞的外观。通过IgE表面标记以及通过对类胰蛋白酶阳性颗粒的超微结构显示，可以将共培养中存活的肥大细胞与含颗粒的成纤维细胞区分开来。因此，虽然肥大细胞在与3T3成纤维细胞共培养时仍能存活，但嗜碱性粒细胞不能存活，会被成纤维细胞单层内化并降解。
