Ahlstedt S, Birgegård G, Hammarström I, Into-Malmberg M B, Jontell M, Denburg J, Akerblom E
Immunology. 1986 Dec;59(4):549-55.
The development of FcR epsilon-bearing histamine-containing basophilic cells was studied in cultures of peripheral blood leucocytes and bone marrow leucocytes from normal individuals. The determination of basophilic cells was performed blindly. Before cultivation there were fairly similar numbers of basophilic cells in the samples from the three different sources (1.6 +/- 1.6 n = 13; 1.4 +/- 2.0 n = 21; 4.3 +/- 4.7 n = 8, respectively). During cultivation spontaneously appearing large blast-like basophilic cells were seen in good correspondence with a formation of histamine in the cultures (Spearman rank correlation coefficient = 0.9135, n = 13, P less than 0.001). This was more accentuated with bone marrow cells than with peripheral blood and cord blood cells. Conditioned medium (CM) was prepared from cells isolated from various tissues and stimulated by different means, e.g. peripheral blood from atopic individuals stimulated with allergen and unstimulated tonsil cells. Addition of the CM resulted in increased development of histamine-containing basophilic cells. Optimum stimulation was achieved with 10% CM. The basophilic stimulation by CM, as assessed as indices vs. unstimulated cultures, was more accentuated in cultures of peripheral blood cells than of bone marrow and cord blood cells (2.8 +/- 1.2; 1.8 +/- 0.5; 2.0 +/- 0.4, respectively). In contrast, the histamine formation was particularly evident in stimulated cultures of bone marrow cells, where more than four-fold increases of histamine were found. In bone marrow cells the histamine levels per basophilic cell also increased, whereas this was not the case in cord blood cells. A pronounced development of basophilic cells was achieved when using leukoagglutinin, provided the mitogen in the CM was eliminated. The formation of basophilic cells was prevented with mitomycin c and cycloheximide. In conclusion, the system described may provide important information on the development of histamine-containing basophilic cells at various maturation stages from different compartments, and mechanisms in a developing atopic disease.
在来自正常个体的外周血白细胞和骨髓白细胞培养物中,研究了含FcRε的嗜碱性粒细胞的发育情况。嗜碱性粒细胞的测定是在不知情的情况下进行的。培养前,来自三种不同来源的样本中嗜碱性粒细胞数量相当相似(分别为1.6±1.6,n = 13;1.4±2.0,n = 21;4.3±4.7,n = 8)。培养过程中,自发出现的大的母细胞样嗜碱性粒细胞与培养物中组胺的形成密切相关(斯皮尔曼等级相关系数 = 0.9135,n = 13,P < 0.001)。与外周血和脐血细胞相比,骨髓细胞的这种情况更为明显。条件培养基(CM)由从各种组织分离并通过不同方式刺激的细胞制备,例如用变应原刺激的特应性个体的外周血和未刺激的扁桃体细胞。添加CM导致含组胺嗜碱性粒细胞的发育增加。用10% CM可实现最佳刺激。与未刺激的培养物相比,以指数评估的CM对嗜碱性粒细胞的刺激在外周血细胞培养物中比在骨髓和脐血细胞培养物中更明显(分别为2.8±1.2;1.8±0.5;2.0±0.4)。相比之下,组胺形成在骨髓细胞的刺激培养物中尤为明显,其中组胺增加了四倍以上。在骨髓细胞中,每个嗜碱性粒细胞的组胺水平也增加,而在脐血细胞中则不然。如果消除CM中的促细胞分裂剂,使用白细胞凝集素可实现嗜碱性粒细胞的显著发育。丝裂霉素c和环己酰亚胺可阻止嗜碱性粒细胞的形成。总之,所描述的系统可能为不同隔室中不同成熟阶段的含组胺嗜碱性粒细胞的发育以及特应性疾病发展中的机制提供重要信息。