Döring G, Buhl V, Høiby N, Schiøtz P O, Botzenhart K
Acta Pathol Microbiol Immunol Scand C. 1984 Oct;92(5):307-12. doi: 10.1111/j.1699-0463.1984.tb00092.x.
Sera and sputa of 12 cystic fibrosis patients suffering from chronic Pseudomonas aeruginosa lung infections were assayed for immune complexes using the Raji cell assay. Whereas all sera were negative, 33% of the sputa were positive for immune complexes. Sera and sputa of these patients were also assayed for antibodies against P.aeruginosa alkaline protease (AP) and elastase (Ela) and sputa for AP and Ela, using radioimmunoassays. All patients revealed antibody titers to the proteases in serum (1:5-1:1000) and eight patients had antibody titers to the proteases in sputum (1:5-1:100); all sputa were negative for AP and Ela. Sputum immune complexes and IgG were separated from whole sputum on a Protein A-Sepharose C1-4B column and treated with 0.75 M 2-mercaptoethanol and iodoacetamide after elution. After treatment, 58% of the sputa were positive for AP and/or Ela (5-200 ng/ml sputum). The present study shows that proteases of P.aeruginosa are bound in immune complexes after the initiation of the immune response to these antigens and it yields new insights into the role of these proteases in chronic lung infection in cystic fibrosis.
采用Raji细胞试验对12例患有慢性铜绿假单胞菌肺部感染的囊性纤维化患者的血清和痰液进行免疫复合物检测。结果显示，所有血清均为阴性，但33%的痰液免疫复合物呈阳性。采用放射免疫分析法对这些患者的血清和痰液进行抗铜绿假单胞菌碱性蛋白酶（AP）和弹性蛋白酶（Ela）抗体检测，并对痰液中的AP和Ela进行检测。所有患者血清中均显示出针对这些蛋白酶的抗体滴度（1:5 - 1:1000），8例患者痰液中针对这些蛋白酶的抗体滴度为（1:5 - 1:100）；所有痰液中AP和Ela均为阴性。通过蛋白A - 琼脂糖凝胶C1 - 4B柱从全痰中分离出痰液免疫复合物和IgG，洗脱后用0.75M 2 - 巯基乙醇和碘乙酰胺处理。处理后，58%的痰液中AP和/或Ela呈阳性（痰液中含量为5 - 200 ng/ml）。本研究表明，铜绿假单胞菌的蛋白酶在针对这些抗原的免疫反应启动后会结合在免疫复合物中，这为这些蛋白酶在囊性纤维化慢性肺部感染中的作用提供了新的见解。
