Complement A Practical Approach

by Dodds, Alister W.; Sim, Robert B.

• ISBN13: 9780199635405
• ISBN10: 0199635404
• Format: Hardcover
• Copyright: 1997-10-02
• Publisher: Oxford University Press

Summary

Complement: A Practical Approach contains detailed step-by-step protocols for analysis of the complement system, and is aimed at new and experienced workers in complement research, as well as clinicians with an interest in immunological diseases. The areas covered include functional andimmunochemical methods for the quantitation of complement components, inhibitors, receptors, inflammatory effects of anaphylatoxin release, methods for the analysis of complement biosynthesis and for the study of the system in non-humans. Of particular interest to clinicians are chapters oncomplement and immune complexes, C2 Inh deficiency, and the genetic analysis of this highly polymorphic system.

Table of Contents

List of contributors	xvii	(2)
Abbreviations	xix
1. The complement system: an introduction	1	(18)
Robert B. Sim
Alister W. Dodds
1. What is the complement system?	1	(5)
2. The classical pathway: activation and components	6	(4)
3. The alternative pathway: activation and components	10	(3)
4. Regulation of complement activation	13	(3)
Inhibition of serine proteases	14	(1)
Control of convertases	14	(1)
Control of the membrane attack complex (MAC)	15	(1)
5. Complement receptors	16	(1)
Receptors for C3 fragments	16	(1)
Receptors for other complement proteins and fragments	16	(1)
6. The future of complement research	16	(1)
References	17	(2)
2. Haemolytic assays for whole complement activity and individual components	19	(30)
Keith Whaley
Jonathan North
1. Introduction	19	(1)
2. Collection and storage of serum samples	20	(1)
3. Preparation buffers and serum reagents	21	(1)
4. Total haemolytic complement assays	21	(9)
Preparation of erythrocytes	21	(2)
Total haemolytic complement titration (CH50)	23	(5)
Alternative pathway total haemolytic complement assays (APH50)	28	(2)
5. Assays of individual components using purified complement components	30	(13)
Preparation of haemolytic intermediates (erythrocytes bearing complement components	30	(7)
Haemolytic assays of individual components	37	(6)
6. Haemolytic assays of complement control proteins	43	(4)
References	47	(2)
3. Immunochemical methods for the detection of complement components and complement activation	49	(20)
Sven-Erik Svehag
R. Graham Q. Leslie
1. Introduction	49	(2)
2. Single radial immunodiffusion (RID) for complement components	51	(1)
3. Measurement of complement protein fragments by laser nephelometry	52	(1)
4. Electroimmunoassays for complement components and complement activation products	53	(3)
Construction and performance of assays	53	(1)
Electroimmunoassays for native complement components	54	(1)
Electroimmunoassays for complement activation products	55	(1)
5. Enzyme immunoassays (EIA) for complement activation products	56	(8)
Construction and performance of the assays	56	(3)
Enzyme immunoassays for complement activation cleavage products	59	(2)
Enzyme immunoassays for complement activation complexes	61	(2)
Enzyme immunoassays for complement regulator proteins	63	(1)
6. Radioimmunoassay for complement proteins	64	(2)
7. Conclusions	66	(1)
References	66	(3)
4. Identification, functional assessment, and quantitation of receptors and membrane regulatory proteins whose ligands are C3 and C4	69	(24)
Michael W. Nickells
John P. Atkinson
1. Introduction	69	(3)
2. Preparation of unlabelled, (125)I surface-labelled, and (35)S biosynthetically labelled detergent-solubilized extracts of cells	72	(3)
3. Antibody-based methods for the identification of cell-surface complement receptors and membrane regulatory proteins	75	(6)
4. Ligand affinity-based methods for the identification of cell-surface complement receptors and membrane regulatory proteins	81	(4)
5. Assessment of cofactor activity from detergent extracts of cells	85	(3)
6. Assessment of decay accelerating activity	88	(1)
7. Quantitation of complement receptors and membrane regulatory proteins	88	(2)
References	90	(3)
5. C1-inhibitor deficiency and angio-oedema	93	(12)
John Jackson
1. Introduction	93	(1)
2. C1-inh deficiency	93	(3)
Hereditary C1-inh deficiency	95	(1)
Acquired C1-inh deficiency	96	(1)
3. Laboratory diagnosis of C1-inh deficiency states	96	(4)
Introduction	96	(1)
Functional C1-inh assays	97	(3)
4. Assays for anti-C1-inh antibodies	100	(3)
Inhibition of C1-inh function	100	(1)
Solid phase assays	101	(2)
Correlation between the various forms of acquired C1-inh deficiency	103	(1)
5. Summary	103	(1)
References	104	(1)
6. Formation and regulation of the membrane attack complex	105	(16)
B. Paul Morgan
Carmen W. van den Berg
1. Introduction	105	(1)
2. Assembly and control of the MAC	105	(2)
3. MAC assembly, isolation, and analysis	107	(6)
Production of depleted sera and erythrocyte intermediates	107	(3)
Isolation of terminal complement complexes (TCCs)	110	(2)
Measurement of TCCs	112	(1)
4. Isolation and characterization of MAC inhibitors	113	(5)
Isolation of fluid-phase inhibitors	113	(2)
Isolation of CD59	115	(3)
Assays for MAC inhibitors	118	(1)
References	118	(3)
7. Purification of complement components	121	(14)
Mohamed R. Daha
1. Introduction	121	(1)
General references for complement protein purification	121	(1)
Purchase of purified complement components	122	(1)
2. Methods	122	(10)
Purification of C1	122	(1)
Purification of C1q	123	(1)
Purification of C1r and C1s	124	(1)
Purification of C4	124	(1)
Purification of C2	125	(1)
Purification of C3 and C5	125	(1)
Purification of C6	126	(1)
Purification of C7	127	(1)
Purification of C8	127	(1)
Purification of C9	128	(1)
Purification of factor D	128	(1)
Purification of factor B	129	(1)
Purification of properdin	129	(1)
Purification of C1 inhibitor	130	(1)
Purification of factor I	130	(1)
Purification of factor H	131	(1)
References	132	(3)
8. The anaphylatoxins	135	(30)
Jorg Kohl
1. Introduction	135	(3)
2. Generation and purification of anaphylatoxins	138	(9)
Generation and purification of C3a and C5a from plasma	138	(3)
Recombinant anaphylatoxins from E. coli	141	(6)
3. Assays of biological activity	147	(11)
Heterologous assays	147	(5)
Homologous assays	152	(6)
4. Measurement of anaphylatoxins in inflammation	158	(2)
ABICAP C3a/C3adesArg and C5a/C5adesArg assay	160	(1)
Acknowledgements	160	(1)
References	160	(5)
9. Complement genetics	165	(34)
Peter M. Schneider
Christian Rittner
1. Introduction	165	(1)
2. Complement polymorphism	165	(23)
Chromosomal location of complement components and related proteins	165	(2)
The components--polymorphic systems and typing methodology	167	(18)
Population genetics	185	(3)
3. Inherited complement deficiencies	188	(5)
Detection	188	(1)
Disease manifestations	189	(4)
Genetic basis of deficiency	193	(1)
Acknowledgements	193	(1)
References	194	(5)
10. Biosynthesis of complement components	199	(24)
Antonella Circolo
Thomas B. Nutter III
Robert C. Strunk
1. Introduction	199	(2)
2. Cell culture techniques	201	(6)
General considerations for fibroblasts	201	(2)
Culture of human fibroblasts	203	(1)
Culture of human blood monocytes	204	(1)
Culture of alveolar macrophages	205	(1)
Determination of number of cells adherent to tissue culture surface	206	(1)
3. Meditators for stimulation of complement synthesis and conditions for stimulation	207	(1)
Mediators	207	(1)
Stimulation conditions	207	(1)
4. Analysis of complement protein synthesis using biosynthetic radiolabelling with [(35)S] methionine plus [(35)S] cysteine	208	(5)
Biosynthetic radiolabelling of cells	208	(1)
Pulse times for labelling	208	(2)
Immunoprecipitation of radiolabelled proteins	210	(1)
SDS-PAGE detection of complement proteins	211	(1)
Quantitation of proteins in SDS-PAGE	211	(2)
5. Analysis of complement protein synthesis by direct measurement of complement protein levels in tissue culture media	213	(3)
ELISA	213	(2)
Measurement of complement proteins by haemolytic assay	215	(1)
6. Analysis of steady-state levels of complement protein-specific mRNA	216	(4)
Guanidine/CsCI method for isolation of RNA from cells cultured in vitro	216	(2)
Isolation of RNA from tissue	218	(1)
RNA agarose gel separation and Northern blot analysis	218	(2)
References	220	(3)
11. Complement and immune complexes	223	(24)
Peter J. Spath
Jurg A. Schifferli
1. Introduction	223	(3)
2. Immune complexes in plasma and their detection using complement-related methods	226	(8)
The extended C1q-binding assay (eC1qBA)	226	(5)
Other methods for detecting immune complexes in body fluids	231	(1)
Commercially available test kits for detection of C1q-binding or C3-split products bearing immune complexes: a comparison	231	(1)
Cryoglobulins	232	(2)
3. Autoantibodies against complement	234	(4)
Anti-C1q autoantibodies	235	(1)
Nephritic factor (Nef)	236	(2)
4. Complement deposition in tissues	238	(4)
5. Complement synthesis in tissues	242	(2)
6. Usefulness of the detection of complement activation and IC in diseases	244	(1)
Acknowledgements	244	(1)
References	244	(3)
12. Non-human complement	247	(18)
Masaru Nonaka
1. Introduction	247	(1)
2. Functional approach	248	(4)
Haemolytic assay	248	(3)
Phagocytosis assay	251	(1)
3. Immunological approach	252	(2)
Antigenic cross-reactivity with human components	252	(1)
Production of antibody against C3 using zymosan	253	(1)
4. Protein structural approach	254	(1)
5. Approach at the nucleic acid level	254	(9)
Cross-hybridization with human sequences	254	(5)
Reverse transcriptase-polymerase chain reaction (RT-PCR)	259	(4)
References	263	(2)
A1. List of suppliers	265	(5)
Index	270