Patella V, Marinò I, Lampärter B, Arbustini E, Adt M, Marone G
Department of Medicine, University of Naples Federico II, School of Medicine, Italy.
J Immunol. 1995 Mar 15;154(6):2855-65.
We have isolated, partially purified, and characterized the mast cells from human heart tissue. The histamine content of left and right ventricles and septum of hearts obtained from 25 patients undergoing heart transplantation was 5.4 +/- 0.6, 5.3 +/- 0.5, and 5.6 +/- 0.5 micrograms/g of wet tissue, respectively. Ultrastructural study of cardiac mast cells revealed scroll, crystal, and mixed granules, homogeneously dense granules, and lipid bodies in the cytoplasm. A mild collagenase digestion was used to disperse the heart mast cells; the average yield was 3.2 +/- 0.6% (range: 0.8 to 13.6%). The average histamine and tryptase content/heart mast cells was 3.3 +/- 0.2 pg (n = 25) and 24.2 +/- 4.3 micrograms/10(6) cells (n = 11), respectively. Survival of cardiac mast cells after overnight culture was 71.9 +/- 5.4% (n = 23). The purification of human heart mast cells can be brought from less than 0.1 to 12% by a combination of low-speed centrifugation over albumin (2%) solution and Percoll gradient. Viability as shown by trypan blue exclusion was greater than 90%. Heart mast cells released histamine in response to immunologic (anti-IgE, anti-Fc epsilon RI, and C5a) and nonimmunologic stimuli (recombinant human stem cell factor, A23187, and compound 48/80) but did not respond to substance P, FMLP, 12-O-tetradecanoylphorbol-13-acetate, or acetylcholine. There was a linear correlation between the percentage of release caused by anti-IgE and anti-Fc epsilon RI, whereas there was no correlation between the release caused by C5a and anti-IgE-mediated stimuli. Cross-linking with anti-IgE of IgE on heart mast cells induced the release of tryptase (10.1 +/- 2.1 micrograms/10(7) cells; n = 10) and the de novo synthesis of PGD2 (17.3 +/- 4.3 ng/10(6) cells; n = 10) and of leukotriene C4 (19.1 +/- 4.5 ng/10(6) cells; n = 10). There was a linear correlation between the percentage of histamine secretion and tryptase release (r = 0.67; p < 0.001) induced by cross-linking of Fc epsilon RI. similarly, there was a significant correlation between percentage of histamine secretion and PGD2 (r = 0.63; p < 0.001) and LTC4 (r = 0.64; p < 0.001) release. Immunoelectron microscopy demonstrated the presence of chymase in cardiac mast cells. Mast cells isolated from human heart can be a useful model with which to study the role of these cells and their mediators in cardiac anaphylaxis and cardiovascular diseases.
我们已从人心脏组织中分离、部分纯化并鉴定了肥大细胞。对25例接受心脏移植患者的心脏左心室、右心室和室间隔的组胺含量进行检测，结果分别为5.4±0.6、5.3±0.5和5.6±0.5微克/克湿组织。对心脏肥大细胞的超微结构研究显示，细胞质中有卷轴状、结晶状和混合颗粒、均匀致密颗粒以及脂滴。采用温和的胶原酶消化法来分散心脏肥大细胞；平均产量为3.2±0.6%（范围：0.8%至13.6%）。每个心脏肥大细胞的组胺和类胰蛋白酶平均含量分别为3.3±0.2皮克（n = 25）和24.2±4.3微克/10⁶个细胞（n = 11）。过夜培养后心脏肥大细胞的存活率为71.9±5.4%（n = 23）。通过在2%白蛋白溶液上进行低速离心和Percoll梯度离心相结合的方法，可将人心脏肥大细胞的纯度从低于0.1%提高到12%。经台盼蓝排斥试验显示，细胞活力大于90%。心脏肥大细胞可对免疫刺激（抗IgE、抗FcεRI和C5a）和非免疫刺激（重组人干细胞因子、A23187和化合物48/80）产生组胺释放反应，但对P物质、FMLP、12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯或乙酰胆碱无反应。抗IgE和抗FcεRI引起的组胺释放百分比之间存在线性相关性，而C5a和抗IgE介导的刺激引起的组胺释放之间无相关性。用抗IgE交联心脏肥大细胞上的IgE可诱导类胰蛋白酶释放（10.1±2.1微克/10⁷个细胞；n = 10）以及PGD₂（17.3±4.3纳克/10⁶个细胞；n = 10）和白三烯C₄（19.1±4.5纳克/10⁶个细胞；n = 10）的从头合成。FcεRI交联诱导的组胺分泌百分比与类胰蛋白酶释放之间存在线性相关性（r = 0.67；p < 0.001）。同样，组胺分泌百分比与PGD₂（r = 0.63；p < 0.001）和LTC₄（r = 0.64；p < 0.001）释放之间存在显著相关性。免疫电子显微镜显示心脏肥大细胞中存在糜酶。从人心脏分离出的肥大细胞可成为一个有用的模型，用于研究这些细胞及其介质在心脏过敏反应和心血管疾病中的作用。
