Immune Complexome Analysis of Serum and Its Application in Screening for Immune Complex Antigens in Rheumatoid Arthritis
Immune Complexome Analysis of Serum and Its Application in Screening for Immune Complex Antigens in Rheumatoid Arthritis
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DOI：
10.1373/clinchem.2010.157776
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发表时间：
2011-06-01
影响因子：
9.3
通讯作者：
Kuroda, Naotaka
中科院分区：
文献类型：
--
作者：
Ohyama, Kaname;Ueki, Yukitaka;Kuroda, Naotaka
背景：对免疫反应过程中产生的循环免疫复合体(CIC)的分析可能有助于阐明这一过程的某些方面。CICs中结合的抗原的鉴定提供了可能有助于制定自身免疫性疾病、感染、癌症和移植治疗的诊断和治疗策略的信息，这些信息可能比游离抗原的信息更相关。由于CICs可能含有多种抗原，因此对这些抗原的全面鉴定和图谱分析比免疫印迹检测更有效。方法：我们开发了一种新的蛋白质组学策略(免疫复合体分析)，即从血清中分离免疫复合物(IC)，直接用胰酶消化，然后进行纳米液相色谱-串联质谱分析来鉴定和图谱CICs中的抗原。我们将这一策略应用于21例类风湿性关节炎(RA)患者的CICS分析。结果：81%和52%的RA患者血清中含有TSP-1和52%的血小板因子4(PF4)，而对照组中未发现。结论：大多数RA患者血清中的ICs含有TSP-1或PF4，这些ICs可能是潜在的替代生物标志物。我们的免疫复合体分析技术使用常规的临床样本、简单的程序和广泛可用的设备。这种方法可能普遍适用于研究CICs与某些与动物和人类免疫反应相关的疾病之间的关系。
BACKGROUND: Analysis of circulating immune complexes (CICs) produced during an immune response may be useful in elucidating some aspects of this process. Identification of antigens incorporated into CICs provides information that may be helpful in developing diagnostic and treatment strategies for autoimmune diseases, infection, cancer, and transplantation therapy, and such information might be more relevant than information on free antigens. Because CICs may contain many antigens, comprehensive identification and profiling of such antigens is more effective than immunoblotting detection.METHODS: We developed a novel proteomic strategy (immune complexome analysis) in which immune complexes (ICs) are separated from serum, digested directly with trypsin, and then subjected to nano-liquid chromatography-tandem mass spectrometry for identifying and profiling antigens in CICs. We applied this strategy to the analysis of CICs in 21 rheumatoid arthritis (RA) patients. Serum samples from 13 healthy donors and 8 osteoarthritis patients were used as controls.RESULTS: CICs containing thrombospondin-1 (TSP-1) and platelet factor 4 (PF4) were found in the serum of 81% and 52% of RA patients, respectively, and in none of the controls.CONCLUSIONS: The ICs in the serum of a majority of the RA patients contained TSP-1 or PF4, and these ICs may have potential as alternative biomarkers. Our technique for immune complexome analysis uses routine clinical samples, simple protocols, and widely available equipment. This method may be generally applicable to the study of the relationship between CICs and certain diseases associated with the immune response in animals and humans.