DETECTION OF TISSUE KALLIKREIN IN THE BRONCHOALVEOLAR LAVAGE FLUID OF ASTHMATIC SUBJECTS
DETECTION OF TISSUE KALLIKREIN IN THE BRONCHOALVEOLAR LAVAGE FLUID OF ASTHMATIC SUBJECTS
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DOI：
10.1172/jci112782
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发表时间：
1987-01-01
影响因子：
15.9
通讯作者：
COCHRANE, CG
中科院分区：
文献类型：
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作者：
CHRISTIANSEN, SC;PROUD, D;COCHRANE, CG
通过放射性标记底物（121 I-高分子量激肽原[HMWK]）的裂解，检测了17名哮喘患者的24份支气管肺泡灌洗液（BAL）样本中的22份中的激肽原酶活性，这些患者对雾化变应原激发有反应或有活动性哮喘症状。相比之下，七个正常对照组中有六个缺乏酶活性。在BAL液中发现的游离免疫反应性激肽水平与激肽原酶活性的存在相关（P=0.002）。125 I-HMWK的裂解模式的BAL流体激肽原酶（占主导地位的65000-mol wt片段），和合成抑制剂的配置文件（苯-苯-精氨酸-CH 2 Cl和苯甲基磺酰氟）与组织激肽释放酶兼容。通过HPLC凝胶过滤，在20，000 - 34，000的表观分子量处洗脱出峰值激肽原酶活性。用抗人尿激肽释放酶抗体进行免疫印迹，确定了其抗原性，其活性可被该抗体抑制。在分级BAL液和纯化的HMWK（组织激肽释放酶的特征性裂解产物）孵育期间产生赖氨酰缓激肽。我们的结论是，组织激肽释放酶和激肽的量升高存在于哮喘受试者的支气管肺泡空间。激肽的产生可能直接通过水肿形成和平滑肌收缩以及通过增加预先形成的（组胺）和次级介质（如白三烯和血小板活化因子）的释放和/或产生而促进哮喘反应。
Kininogenase activity was detected by cleavage of radiolabeled substrate (121I-high molecular weight kininogen [HMWK]) in 22 of 24 bronchoalveolar lavage (BAL) fluid samples from 17 asthmatics who either responded to aerosolized allergen challenge or had symptoms of active asthma. In contrast, six of seven normal controls lacked enzymatic activity. Levels of free immunoreactive kinin found in BAL fluid correlated with the presence of kininogenase activity (P=0.002). The cleavage pattern of 125I-HMWK by the BAL fluid kininogenase (a dominant 65000-mol wt fragment), and synthetic inhibitor profile (phe-phe-arg-CH2Cl and phenylmethylsulfonyl fluoride) were compatible with a tissue kallikrein. Peak kininogenase activity eluted at an apparent molecular weight of 20,000-34,000 by HPLC gel filtration. Its antigenic identity was established by immunoblotting with anti-human urinary kallikrein antibody and its activity was inhibited by this antibody. Lysylbradykinin was generated during incubation of fractionated BAL fluid and purified HMWK, the characteristic cleavage product of the tissue kallikreins. We conclude that elevated amounts of tissue kallikrein and kinin are present in the bronchoalveolar spaces of asthmatic subjects. Kinin generation may contribute to the asthmatic response directly through edema formation and smooth muscle contraction and by augmenting release and/or production of preformed (histamine) and secondary mediators such as leukotrienes and platelet-activating factor.