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猪C1q与人免疫复合物的固相免疫测定

Porcine C1q and the solid-phase immunoassay of human immune complexes.

作者信息

Ziola B, Salmi A, Penttinen K

出版信息

J Immunol Methods. 1982 Mar 26;49(3):247-60. doi: 10.1016/0022-1759(82)90125-9.

DOI:10.1016/0022-1759(82)90125-9
PMID:6978362
Abstract

Experiments were undertaken to determine if porcine C1q could replace human C1q in the solid-phase immunoassay of human immune complexes (ICs). Porcine C1q was obtained by a two-cycle precipitation method involving dialysis against chelating agents in low ionic strength buffer. C1q was adsorbed to polystyrene beads and in vivo- or in vitro-formed ICs binding to the solid-phase C1q were detected with 125I-labeled or horseradish peroxidase-conjugated anti-human gamma antibodies. Unfractionated, heat-aggregated human gamma globulin (delta IgG) could be detected at 20 ng/ml when diluted in buffer only. The detection threshold changed to 40-80 ng delta IgG/ml when the assay was run with buffer containing normal human serum diluted 1: 1000 (the serum dilution used for detecting natural ICs). Analysis of systemic lupus erythematosus sera revealed that 60% contained highly significant levels of ICs (binding greater than or equal to 3 S.D. above the mean of controls). Comparison with platelet aggregation test results revealed a highly significant correlation between the two methods (P less than 0.0001), even though each assay detected ICs in several serum specimens negative in the other test. These results demonstrate that porcine C1q can functionally replace human C1q in the solid-phase immunoassay of human ICs. Since porcine blood is normally a waste product of the meat-processing industry, it is an obvious source of easily isolated C1q for use in such an assay.

摘要

开展实验以确定在人免疫复合物(ICs)的固相免疫测定中,猪C1q是否能够替代人C1q。猪C1q通过两步沉淀法获得,该方法包括在低离子强度缓冲液中用螯合剂进行透析。将C1q吸附到聚苯乙烯珠上,并用125I标记或辣根过氧化物酶偶联的抗人γ抗体检测与固相C1q结合的体内或体外形成的ICs。仅在缓冲液中稀释时,未分级的热聚集人γ球蛋白(δIgG)在20 ng/ml时即可被检测到。当用含有1:1000稀释的正常人血清的缓冲液进行检测(用于检测天然ICs的血清稀释度)时,检测阈值变为40 - 80 ng δIgG/ml。对系统性红斑狼疮血清的分析显示,60%含有高度显著水平的ICs(结合量比对照平均值高3个标准差以上)。与血小板聚集试验结果的比较显示,两种方法之间存在高度显著的相关性(P小于0.0001),尽管每种检测方法在其他检测为阴性的几个血清标本中都检测到了ICs。这些结果表明,在人ICs的固相免疫测定中,猪C1q在功能上可以替代人C1q。由于猪血通常是肉类加工业的废品,它显然是用于此类检测的易于分离的C1q的来源。

相似文献

1
Porcine C1q and the solid-phase immunoassay of human immune complexes.猪C1q与人免疫复合物的固相免疫测定
J Immunol Methods. 1982 Mar 26;49(3):247-60. doi: 10.1016/0022-1759(82)90125-9.
2
The IgG detected in the C1q solid-phase immune-complex assay is not always of immune-complex nature.在C1q固相免疫复合物测定中检测到的IgG并不总是具有免疫复合物性质。
Clin Immunol Immunopathol. 1986 Jan;38(1):120-8. doi: 10.1016/0090-1229(86)90129-7.
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[Demonstration of circulating immune complexes using a C1q-solid phase enzyme immunoassay].[使用C1q固相酶免疫测定法检测循环免疫复合物]
Biomed Biochim Acta. 1983;42(7-8):1027-35.
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Detection of C1q-bearing immune complexes by a monoclonal anti-C1q ELISA system.通过单克隆抗C1q ELISA系统检测含C1q的免疫复合物。
J Immunol Methods. 1987 Sep 24;102(2):149-56. doi: 10.1016/0022-1759(87)90071-8.
5
C1q-latex assay for immune complexes. Complexes that react with both C1q and monoclonal rheumatoid factor in lupus erythematosus and lung cancer.用于免疫复合物的C1q-乳胶检测法。在红斑狼疮和肺癌中与C1q和单克隆类风湿因子均发生反应的复合物。
J Lab Clin Med. 1982 May;99(5):678-92.
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Characterization of C1q-binding IgG complexes in systemic lupus erythematosus.系统性红斑狼疮中C1q结合IgG复合物的特征分析
Clin Immunol Immunopathol. 1984 Jan;30(1):104-16. doi: 10.1016/0090-1229(84)90011-4.
7
C1q solid-phase radioimmunoassay: binding properties of solid-phase C1q and evidence that C1q-binding IgG complexes in systemic lupus erythematosus are not bound to endogenous C1q.C1q固相放射免疫测定:固相C1q的结合特性以及系统性红斑狼疮中C1q结合IgG复合物未与内源性C1q结合的证据
J Immunol Methods. 1984 Oct 12;73(1):67-74. doi: 10.1016/0022-1759(84)90032-2.
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Evidence for the presence of autoantibodies to the collagen-like portion of C1q in systemic lupus erythematosus.系统性红斑狼疮中存在针对C1q胶原样部分自身抗体的证据。
Arthritis Rheum. 1988 Apr;31(4):457-64. doi: 10.1002/art.1780310401.
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Solid-phase Clq-binding fluorescence immunoassay for detection of circulating immune complexes.用于检测循环免疫复合物的固相Clq结合荧光免疫测定法。
J Clin Microbiol. 1982 Mar;15(3):456-64. doi: 10.1128/jcm.15.3.456-464.1982.
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[A method of isolating C1q from human serum and its use in the solid-phase immunoenzyme determination of immune complexes].[从人血清中分离C1q的方法及其在免疫复合物固相免疫酶测定中的应用]
Lab Delo. 1989(3):24-6.

引用本文的文献

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Circulating immune complexes in patients with acute measles and rubella virus infections.急性麻疹和风疹病毒感染患者的循环免疫复合物
Infect Immun. 1983 Aug;41(2):578-83. doi: 10.1128/iai.41.2.578-583.1983.
2
One-way humoral immune cross-reactivity between bovine spinal cord protein and bovine myelin basic protein.牛脊髓蛋白与牛髓鞘碱性蛋白之间的单向体液免疫交叉反应性。
Mol Immunol. 1984 Oct;21(10):999-1005. doi: 10.1016/0161-5890(84)90159-7.
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J Neurol. 1985;232(1):52-4. doi: 10.1007/BF00314042.
4
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Ann Rheum Dis. 1990 Apr;49(4):214-8. doi: 10.1136/ard.49.4.214.
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Nervous system disease, immunological features, and HLA phenotype in Sjögren's syndrome.干燥综合征中的神经系统疾病、免疫学特征及人类白细胞抗原表型
Ann Rheum Dis. 1992 Apr;51(4):506-9. doi: 10.1136/ard.51.4.506.