A novel method for direct measurement of complement convertases activity in human serum

被引：41

作者：

Blom, A. M. ^{[1
]}

Volokhina, E. B. ^{[3
]}

Fransson, V. ^{[1
]}

Stromberg, P. ^{[2
]}

Berghard, L. ^{[2
]}

Viktorelius, M. ^{[1
]}

Mollnes, T. E. ^{[5
,6
,7
,8
]}

Lopez-Trascasa, M. ^{[9
,10
]}

van den Heuvel, L. P. ^{[3
,4
,11
,12
]}

Goodship, T. H.

Marchbank, K. J. ^{[13
]}

Okroj, M. ^{[1
]}

机构：

[1] Lund Univ, Dept Lab Med Malmo, Malmo, Sweden

[2] Swedish Orphan Biovitrum AB, Stockholm, Sweden

[3] Radboud Univ Nijmegen, Med Ctr, Dept Pediat Nephrol, NL-6525 ED Nijmegen, Netherlands

[4] Radboud Univ Nijmegen, Med Ctr, Dept Lab Med, NL-6525 ED Nijmegen, Netherlands

[5] Univ Oslo, Rikshosp, Oslo Univ Hosp, Inst Immunol, N-0027 Oslo, Norway

[6] Univ Oslo, Oslo, Norway

[7] NLSH, Res Lab, Bodo, Norway

[8] Univ Tromso, Tromso, Norway

[9] Univ Hosp La Paz, IdiPAZ, Immunol Unit, Madrid, Spain

[10] Ctr Biomed Res Rare Dis CIBERER, Unit 754, Madrid, Spain

[11] Univ Hosp Leuven, Dept Pediat, Leuven, Belgium

[12] Newcastle Univ, Inst Human Genet, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England

[13] Newcastle Univ, Inst Cellular Med, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England

来源：

CLINICAL AND EXPERIMENTAL IMMUNOLOGY | 2014年 / 178卷 / 01期

基金：

瑞典研究理事会;

关键词：

complement system; convertase; eculizumab; OmCI; HEMOLYTIC-UREMIC SYNDROME; FACTOR-H-AUTOANTIBODIES; DECAY-ACCELERATING FACTOR; SURFACE-BOUND C3B; ALTERNATIVE PATHWAY; NEPHRITIC FACTOR; FACTOR-B; CELLS; ACTIVATION; MUTATIONS;

D O I：

10.1111/cei.12388

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

071005 [微生物学]; 100108 [医学免疫学];

摘要：

Complement convertases are enzymatic complexes that play a central role in sustaining and amplification of the complement cascade. Impairment of complement function leads directly or indirectly to pathological conditions, including higher infection rate, kidney diseases, autoimmune-or neurodegenerative diseases and ischaemia-reperfusion injury. An assay for direct measurement of activity of the convertases in patient sera is not available. Existing assays testing convertase function are based on purified complement components and, thus, convertase formation occurs under non-physiological conditions. We designed a new assay, in which C5 blocking compounds enabled separation of the complement cascade into two phases: the first ending at the stage of C5 convertases and the second ending with membrane attack complex formation. The use of rabbit erythrocytes or antibody-sensitized sheep erythrocytes as the platforms for convertase formation enabled easy readout based on measurement of haemolysis. Thus, properties of patient sera could be studied directly regarding convertase activity and membrane attack complex formation. Another advantage of this assay was the possibility to screen for host factors such as C3 nephritic factor and other anti-complement autoantibodies, or gain-of-function mutations, which prolong the half-life of complement convertases. Herein, we present proof of concept, detailed description and validation of this novel assay.

引用

页码：142 / 153

页数：12

共 41 条

[1]

Complement Factor H Autoantibodies Are Associated with Early Stage NSCLC [J].