A soluble form of Fc gamma RIII is present in human serum and other body fluids and is elevated at sites of inflammation.
A soluble form of Fc gamma RIII is present in human serum and other body fluids and is elevated at sites of inflammation.
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DOI：
10.1182/blood.v79.10.2721.bloodjournal79102721
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发表时间：
1992
期刊：
影响因子：
20.3
通讯作者：
H. Fleit;C. Kobasiuk;C. Daly;R. Furie;PC Levy;RO Webster
中科院分区：
文献类型：
--
作者：
H. Fleit;C. Kobasiuk;C. Daly;R. Furie;PC Levy;RO Webster
我们开发了一种高灵敏度和特异性的三明治酶联免疫吸附试验（ELISA）来测量血清和其他体液中Fc γ RIII的浓度。该ELISA基于Fc γ RIII单克隆抗体（MoAb） （3G8）和Fc γ RIII兔抗血清。该酶联免疫吸附法的检测下限为1.5 nmol/L。正常血清中可溶性Fc γⅲ的浓度范围为7.3 ~ 75.9 nmol/L。可溶性Fc γⅲ也存在于其他正常生物体液中，如唾液、尿液和精液，但浓度远低于血清。兔抗fc γⅲ免疫印迹多肽用MoAb 3G8免疫沉淀。从中性粒细胞裂解液中免疫沉淀的Fc γ RIII从40 Kd迁移到76 Kd，而从同一供者血清中免疫沉淀的Fc γ RIII从40 Kd迁移到66 Kd。可溶性Fc γ RIII明显与血清IgG结合，因为可溶性Fc γ RIII被MoAb 3G8共沉淀多肽免疫沉淀，这些多肽被山羊抗人IgG鉴定。中性粒细胞体外4℃和37℃孵育显示，37℃孵育30分钟后Fc γ RIII释放，以确定生物体液中可溶性Fc γ RIII浓度与炎症性疾病之间是否存在相关性。我们测量了成人呼吸窘迫综合征（ARDS）患者支气管肺泡灌洗液和各种关节炎患者滑液中Fc γⅲ的浓度。在ARDS中，我们发现可溶性Fc γⅲ的浓度比健康成人支气管肺泡灌洗液中的浓度高5至7倍。类风湿性关节炎患者滑液中可溶性Fc γⅲ的浓度范围为10 nmol/L至28 nmol/L。这些结果表明，活化的中性粒细胞，如炎症部位的中性粒细胞，可能释放Fc γ RIII。
We have developed a highly sensitive and specific sandwich enzyme-linked immunosorbent assay (ELISA) to measure the concentration of Fc gamma RIII in serum and other body fluids. This ELISA is based on the use of monoclonal antibody (MoAb) (3G8) to Fc gamma RIII and a rabbit antiserum against Fc gamma RIII. The lower limit of detection of this ELISA was 1.5 nmol/L. The concentration of soluble Fc gamma RIII in normal serum ranged from 7.3 to 75.9 nmol/L. Soluble Fc gamma RIII was also present in other normal biologic fluids such as saliva, urine, and seminal fluid, but at much lower concentrations than that found in serum. Rabbit anti-Fc gamma RIII immunoblotted polypeptides immunoprecipitated with MoAb 3G8. Fc gamma RIII immunoprecipitated from a neutrophil lysate migrated from 40 to 76 Kd, whereas Fc gamma RIII immunoprecipitated from serum from the same donor migrated from 40 to 66 Kd. The soluble form of Fc gamma RIII apparently was bound to serum IgG, because immunoprecipitation of soluble Fc gamma RIII by MoAb 3G8 coprecipitated polypeptides that were identified by goat antihuman IgG. Incubation of neutrophils in vitro at 4 degrees C and 37 degrees C showed that Fc gamma RIII was released after 30 minutes of incubation at 37 degrees C. To determine whether there was a correlation between the concentration of soluble Fc gamma RIII in biologic fluids and inflammatory diseases, we measured the concentration of Fc gamma RIII in the bronchoalveolar lavage fluid from patients with adult respiratory distress syndrome (ARDS) and in the synovial fluid from patients with various forms of arthritis. In ARDS, we found concentrations of soluble Fc gamma RIII that were five to seven times higher than that found in the bronchoalveolar lavage fluids from healthy adults. The concentration of soluble Fc gamma RIII in the synovial fluid from patients with rheumatoid arthritis ranged from 10 nmol/L to 28 mumol/L. These results suggest that activated neutrophils, such as those at sites of inflammation, may release Fc gamma RIII.