Chen Z, Irani A A, Bradford T R, Craig S S, Newlands G, Miller H, Huff T, Simmons W H, Schwartz L B
Department of Medicine, Medical College of Virginia, Virginia Commonwealth University, Richmond.
J Histochem Cytochem. 1993 Jul;41(7):961-9. doi: 10.1177/41.7.7685789.
We examined the cellular distribution of rat tryptase in rat skin, lung, small intestine, and peritoneal lavage cells by immunohistochemical techniques. Tryptase purified to apparent homogeneity from rat skin was used to generate a goat polyclonal anti-rat tryptase antibody. Tryptase-containing cells were detected in lung, skin, and peritoneal lavage cells. Small intestine mucosa, on the other hand, showed few if any tryptase-positive cells. Sequential staining with Alcian blue and anti-tryptase antibody showed that tryptase is located only in mast cells. Sequential staining with safranin to identify the connective tissue type of mast cell and anti-tryptase antibody showed that tryptase resides only in this mast cell type. However, only a subpopulation of the safranin-stained mast cells contained tryptase. In lung, 53% of the mast cells stained with safranin; 94% contained tryptase. In skin, 80% stained with safranin; only 6% contained tryptase. In peritoneal cells, more than 95% of the mast cells were stained with safranin; 20% contained tryptase. In the bowel mucosa, where few cells are stained by safranin, no cells with tryptase were detected. The percentages of cells with chymase I that also contained tryptase were 80% and 84% for lung, 4% and 7% for skin, and 15% and 13% for peritoneal cells by respective simultaneous and sequential double labeling with anti-tryptase and anti-chymase I antibodies. This study suggests that the rat connective tissue type of mast cell is subdivided into two forms on the basis of the presence or absence of tryptase, whereas rat mucosal mast cells lack this enzyme. These results contrast with those in humans, in which tryptase is present in all mast cells, but are similar to mice, in which tryptase mRNA has been detected only in the connective tissue type.
我们采用免疫组化技术研究了大鼠皮肤、肺、小肠和腹腔灌洗细胞中大鼠类胰蛋白酶的细胞分布。用从大鼠皮肤中纯化至表观均一的类胰蛋白酶制备山羊抗大鼠类胰蛋白酶多克隆抗体。在肺、皮肤和腹腔灌洗细胞中检测到含类胰蛋白酶的细胞。另一方面，小肠黏膜中几乎没有类胰蛋白酶阳性细胞。用阿尔辛蓝和抗类胰蛋白酶抗体进行连续染色显示，类胰蛋白酶仅位于肥大细胞中。用番红精染色以鉴定肥大细胞的结缔组织类型并结合抗类胰蛋白酶抗体进行连续染色显示，类胰蛋白酶仅存在于这种肥大细胞类型中。然而，仅一部分经番红精染色的肥大细胞含有类胰蛋白酶。在肺中，53%的肥大细胞经番红精染色；94%含有类胰蛋白酶。在皮肤中，80%经番红精染色；仅6%含有类胰蛋白酶。在腹腔细胞中，超过95%的肥大细胞经番红精染色；20%含有类胰蛋白酶。在肠黏膜中，经番红精染色的细胞很少，未检测到含类胰蛋白酶的细胞。通过用抗类胰蛋白酶和抗糜蛋白酶I抗体分别进行同时和连续双重标记，肺中同时含有糜蛋白酶I和类胰蛋白酶的细胞百分比分别为80%和84%，皮肤中为4%和7%，腹腔细胞中为15%和13%。本研究表明，大鼠结缔组织型肥大细胞根据类胰蛋白酶的有无可分为两种形式，而大鼠黏膜肥大细胞缺乏这种酶。这些结果与人类的情况相反，人类所有肥大细胞中都存在类胰蛋白酶，但与小鼠相似，小鼠中仅在结缔组织型肥大细胞中检测到类胰蛋白酶mRNA。
