Wershil B K, Mekori Y A, Murakami T, Galli S J
Department of Pathology, Beth Israel Hospital, Boston, MA 02215.
J Immunol. 1987 Oct 15;139(8):2605-14.
We investigated the clotting associated with IgE-dependent immediate hypersensitivity reactions in the mouse by injecting monoclonal mouse anti-dintrophenyl IgE antibodies i.d. and, the next day, administering 125I-guinea pig fibrinogen i.v. 10 to 30 min before i.v. antigen (2,4-dinitrophenylated human serum albumin) challenge. In normal mice, 2-hr passive cutaneous anaphylaxis (PCA) reactions were associated with substantial leakage of 125I-fibrinogen and deposition of 125I-fibrin. Thus, ears injected with IgE contained up to six times the total cpm of 125I and up to 30 times the cross-linked 125I-fibrin-associated cpm of 125I than did control ears. Several lines of evidence indicated that the 125I-fibrin deposition associated with the PCA reactions was dependent on the activity of mast cells: 1) Mast cell degranulation occurred at sites of PCA reactions. 2) Antigen-induced influx of 125I-fibrinogen and deposition of 125I-fibrin were virtually abolished by heating the IgE (56 degrees C, 1 hr) before i.d. injection. 3) Little or no IgE-dependent 125I-fibrinogen influx or 125I-fibrin deposition occurred in mast cell-deficient WBB6F1-W/Wv or WCB6F1-S1/S1d mice X 4) Adoptive transfer of cutaneous mast cell populations into WBB6F1-W/Wv mice (by each of three approaches: i.v. transplantation of normal bone marrow cells or local i.d. injection of cultured, growth factor-dependent mast cells 2 days or 9 to 10 wk before antigen challenge) conferred on the recipients the ability to express the 125I-fibrinogen influx and 125I-fibrin deposition associated with PCA reactions. These data demonstrate that 125I-fibrinogen influx and 125I-fibrin deposition occurs in association with PCA reactions in the mouse, and that the reaction is largely or entirely dependent on the function of cutaneous mast cells. The experiments also demonstrate the utility of a novel model system for the analysis of mast cell function in vivo: WBB6F1-W/Wv mice locally reconstituted with mast cells by the injection of mast cell populations generated in vitro.
我们通过皮下注射单克隆小鼠抗二硝基苯基IgE抗体，然后在静脉注射抗原（2,4 - 二硝基苯基化人血清白蛋白）激发前10至30分钟静脉注射125I - 豚鼠纤维蛋白原，研究了小鼠中与IgE依赖性速发型超敏反应相关的凝血情况。在正常小鼠中，2小时的被动皮肤过敏反应（PCA）与125I - 纤维蛋白原的大量渗漏和125I - 纤维蛋白的沉积有关。因此，注射了IgE的耳朵中125I的总计数每分钟衰变数（cpm）比对照耳朵多高达6倍，交联的125I - 纤维蛋白相关的125I - cpm比对照耳朵多高达30倍。几条证据表明，与PCA反应相关的125I - 纤维蛋白沉积依赖于肥大细胞的活性：1）肥大细胞脱颗粒发生在PCA反应部位。2）在皮下注射前将IgE加热（56℃，1小时），抗原诱导的125I - 纤维蛋白原内流和125I - 纤维蛋白沉积几乎被消除。3）在肥大细胞缺陷的WBB6F1 - W/Wv或WCB6F1 - S1/S1d小鼠中，很少或没有IgE依赖性的125I - 纤维蛋白原内流或125I - 纤维蛋白沉积。4）通过三种方法中的任何一种（在抗原激发前2天静脉移植正常骨髓细胞或在局部皮下注射培养的、生长因子依赖性肥大细胞9至10周）将皮肤肥大细胞群体过继转移到WBB6F1 - W/Wv小鼠中，使受体具有表达与PCA反应相关的125I - 纤维蛋白原内流和125I - 纤维蛋白沉积的能力。这些数据表明，125I - 纤维蛋白原内流和125I - 纤维蛋白沉积与小鼠的PCA反应相关，并且该反应在很大程度上或完全依赖于皮肤肥大细胞的功能。这些实验还证明了一种新型模型系统在体内分析肥大细胞功能的实用性：通过注射体外产生的肥大细胞群体对WBB6F1 - W/Wv小鼠进行局部肥大细胞重建。
