Castells M C, Irani A M, Schwartz L B
J Immunol. 1987 Apr 1;138(7):2184-9.
Murine monoclonal and goat polyclonal antibodies against tryptase, the dominant neutral protease and protein component in secretory granules of human mast cells, were used to assess the presence of tryptase in peripheral leukocytes. Carnoy's fluid-fixed cytocentrifuge preparations of enriched populations of lymphocytes, monocytes, eosinophils, and neutrophils showed no reactivity with anti-tryptase antibodies by a sensitive indirect immunoperoxidase procedure. Dispersed human lung mast cells showed strong granular cytoplasmic staining with both antibodies, whereas only approximately 50% of the peripheral blood basophils detectable with Wright's stain were detected with anti-tryptase antibodies, and these showed a staining pattern that was faint, granular, and cytoplasmic at high concentrations of antibody. At lower antibody concentrations mast cell staining was still intense, whereas basophils were not stained. Extracts of neutrophils and lymphocytes of up to 90% purity had undetectable amounts of tryptase by an ELISA sandwich immunoassay, as well as undetectable enzymatic activity with tosyl-L-gly-pro-lys-p-nitroanilide (a sensitive substrate for tryptase) in the presence of soybean trypsin inhibitor. Extracts of basophil-enriched (6 to 50% purity) preparations contained 0.046 +/- 0.013 pg of tryptase per basophil by the immunoassay along with 2 X 10(-9) +/- 0.8 X 10(-9) U of tryptase-like enzyme activity per basophil, compared with corresponding values of 12 pg, 480 X 10(-9) U of tryptase per human lung mast cell. Thus very small amounts of tryptase are present in human basophils (approximately 0.4% of that found in mast cells), but not in other peripheral leukocytes.
针对类胰蛋白酶（人类肥大细胞分泌颗粒中的主要中性蛋白酶和蛋白质成分）的鼠单克隆抗体和山羊多克隆抗体，被用于评估外周血白细胞中类胰蛋白酶的存在情况。通过灵敏的间接免疫过氧化物酶法，对经卡诺氏液固定的淋巴细胞、单核细胞、嗜酸性粒细胞和中性粒细胞富集群体的细胞离心涂片进行检测，结果显示这些细胞与抗类胰蛋白酶抗体无反应。分散的人肺肥大细胞与两种抗体均呈现强烈的颗粒状胞质染色，而用瑞氏染色可检测到的外周血嗜碱性粒细胞中，只有约50%能用抗类胰蛋白酶抗体检测到，且这些细胞在高浓度抗体下呈现出微弱、颗粒状的胞质染色模式。在较低抗体浓度下，肥大细胞染色仍然强烈，而嗜碱性粒细胞未被染色。通过ELISA夹心免疫测定法，纯度高达90%的中性粒细胞和淋巴细胞提取物中未检测到类胰蛋白酶，并且在大豆胰蛋白酶抑制剂存在的情况下，用甲苯磺酰-L-甘-脯-赖-对硝基苯胺（类胰蛋白酶的灵敏底物）检测不到酶活性。通过免疫测定法，嗜碱性粒细胞富集（纯度为6%至50%）制剂的提取物中，每个嗜碱性粒细胞含有0.046±0.013 pg类胰蛋白酶，同时每个嗜碱性粒细胞具有2×10⁻⁹±0.8×10⁻⁹ U类胰蛋白酶样酶活性，相比之下，人肺肥大细胞中相应的值为12 pg、480×10⁻⁹ U类胰蛋白酶。因此，人类嗜碱性粒细胞中存在极少量的类胰蛋白酶（约为肥大细胞中发现量的0.4%），而其他外周血白细胞中则没有。
