Immunochemistry 2 A Practical Approach

by Johnstone, Alan P.; Turner, Malcolm W.

• ISBN13: 9780199636105
• ISBN10: 0199636109
• Format: Hardcover
• Copyright: 1997-10-02
• Publisher: Oxford University Press

Summary

Immunochemistry is of immense importance to virtually all areas of modernbiology and medicine, and the space afforded by two volumes has allowed coverageof a wide range of topics. The familiar and clear Practical Approach format willmake a large body of valuable information easily accessible to a diversereadership. Topics covered in Immunochemistry 2 include: affinity and aviditymeasurements; the use of liposomes to modulate immune responses;immunfluorescence and immunoenzyme histochemistry; immunological applications ofconfocal microscopy; flow cytometry; soluble adhesion molecules; complementcomponents; and complement receptors.

Table of Contents

Contributors	xxi	(4)
Abbreviations	xxv
1. Antibody affinity measurements	1	(30)
Helene Saunal
Robert Karlsson
Marc H. V. van Regenmortel
1. Introduction	1	(1)
2. General theory	2	(1)
3. Equilibrium constant K measurement	2	(9)
Equilibrium dialysis	3	(1)
Ammonium sulfate precipitation	4	(1)
Measurement of free Ab at equilibrium by ELISA titration	4	(5)
Measurement of free Ab at equilibrium using BIAcore	9	(2)
4. Measurement of the kinetic rate constants K(a) and K(d)	11	(18)
Principles and technology of real time biosensor measurements	11	(1)
Principle of the method	12	(2)
Mass transfer during BIAcore experiments	14	(3)
More complex interactions	17	(6)
Range of kinetic rate constants measurable with the BIAcore system	23	(1)
Experimental aspects of the determination of rate and equilibrium constants	24	(1)
Example of kinetic measurements in BIAcore analyses and interpretation of experimental data	25	(4)
References	29	(2)
2. Simple solid phase assays of avidity	31	(22)
David Goldblatt
1. Introduction	31	(2)
The measurement of avidity	31	(2)
2. Competition inhibition assays	33	(2)
3. Elution assays	35	(11)
Thiocyanate elution assays	37	(5)
Urea elution assays	42	(2)
Diethylamine elution assays	44	(2)
4. Comparison of elution techniques	46	(3)
References	49	(4)
3. Liposomes	53	(18)
Eric Claassen
Freek van Iwaarden
1. Introduction	53	(1)
2. Lipids	54	(1)
3. Multilamellar vesicles	55	(3)
4. Unilamellar liposomes	58	(1)
5. Size	59	(1)
6. Content	59	(1)
7. Giant liposomes	60	(1)
8. Cytotoxic T cells	61	(1)
9. Stealth (PEG; SL) liposomes	62	(1)
10. Enhanced entrapment	63	(1)
11. Carbocyanin labelling of liposomes	64	(4)
12. Conclusions	68	(1)
References	68	(3)
4. Immunofluorescence and immunoenzyme histochemistry	71	(60)
P. Brandtzaeg
T. S. Halstensen
H. S. Huitfeldt
K. N. Valnes
1. Introduction	71	(1)
2. Immunostaining for fluorescence or light microscopy	72	(14)
Reagents	72	(8)
Immunohistochemical staining methods	80	(6)
3. Multicolour immunostaining and its evaluation	86	(24)
Primary antibodies from different species	87	(1)
Primary antibodies from the same species	87	(14)
Computerized image analysis of multicolour fluorescence	101	(9)
Confocal microscopy	110	(1)
4. Methodological considerations	110	(7)
Tissue preparation methods	110	(5)
Choice of immunostaining method	115	(2)
5. Immunohistochemical reliability criteria	117	(6)
Specificity criteria	117	(3)
Detection accuracy	120	(1)
Detection precision	121	(1)
Detection sensitivity	121	(1)
Detection efficiency	122	(1)
6. General conclusions	123	(1)
Acknowledgements	123	(1)
References	123	(8)
5. Confocal laser scanning microscopy	131	(18)
G. D. Johnson
1. Introduction	131	(2)
Principle	132	(1)
2. Practical considerations relating to the equipment	133	(3)
Microscope specification	133	(1)
Supplementary equipment	133	(1)
Operating adjustments	134	(2)
3. Application of CLSM in immunological studies	136	(9)
Improved definition	136	(1)
Image intensity enhancement	137	(1)
Optical sectioning and 3D reconstruction	137	(2)
Multichannel analysis	139	(3)
Quantitative image analysis	142	(3)
4. Summary	145	(3)
Future prospects for CLSM in immunology	146	(2)
References	148	(1)
6. Flow cytofluorimetry	149	(32)
G. Damgaard
C. H. Nielsen
R. G. Q. Leslie
1. Introduction	149	(1)
2. Applications of flow cytofluorimetry	150	(2)
3. Investigation of membrane glycoprotein expression and molecular interactions at the cell surface	152	(7)
Quenching and pH dependence	152	(1)
Calibration	153	(1)
Measurement of a membrane glycoprotein on blood leucocytes	154	(5)
Investigation of molecular interactions at the cell surface	159	(1)
4. Measurement of intracellular proteins and mRNA	159	(5)
Detection of intracellular proteins	160	(1)
Measurement of intracellular enzymatic activity	161	(1)
Measurement of mRNA	162	(1)
General remarks regarding the investigation of intracellular components	162	(2)
5. Investigation of cellular activation	164	(1)
Measurement of intracellular calcium ions	164	(1)
Measurement of surface activation markers	164	(1)
6. Investigation of cellular replication	165	(3)
PCNA and BrdU approaches	166	(1)
The macromolecular dilution approach	166	(2)
7. Measurement of cellular uptake and ingestion of soluble and particulate materials	168	(4)
Binding of opsonized immune complexes to whole blood cells	169	(2)
Measurement of binding and ingestion by phagocytes	171	(1)
8. Measurement of phagocytic oxidative processes	172	(1)
9. Measurement of cell-mediated cytotoxicity and apoptosis	173	(5)
Cell-mediated cytotoxicity	173	(4)
Measurement of apoptosis	177	(1)
References	178	(3)
7. Analysis of soluble adhesion molecules	181	(16)
M. G. Bouma
M. P. Laan
M. A. Dentener
W. A. Buurman
1. Introduction	181	(1)
2. E-selectin	182	(1)
3. ICAM-1	183	(1)
4. VCAM-1	184	(1)
5. Potential roles of soluble adhesion molecules in health and disease	185	(1)
6. ELISA for sICAM-1	185	(8)
7. Potential pitfalls	193	(1)
Acknowledgements	193	(1)
References	193	(4)
8. Immunochemical assays for complement components	197	(28)
R. Wurzner
T. E. Mollnes
B. P. Morgan
1. Introduction	197	(4)
The complement system	197	(1)
Clinical importance of the complement system	197	(3)
Assays for the assessment of complement activation	200	(1)
2. Collection and preservation of samples	201	(2)
3. EIAs for the assessment of complement activation	203	(4)
4. Native-restricted and neoepitope-specific mAbs	207	(8)
The classical pathway	207	(2)
C3 and the alternative pathway	209	(2)
The terminal pathway	211	(4)
5. mAbs directed against complement control proteins	215	(3)
6. Allospecific mAbs	218	(2)
References	220	(5)
9. Assays for complement receptors	225	(32)
Istvan Bartok
Mark J. Walport
1. Introduction	225	(2)
2. Identification of complement receptors using fluorescence-labelled antibodies	227	(3)
Isolation of human neutrophils	228	(1)
Assay of cell surface receptors	229	(1)
3. Radioligand binding assay using C3b dimers to enumerate CR1	230	(7)
Binding of radiolabelled ligand to CR1	231	(2)
Calculation of data	233	(1)
Scatchard plot analysis	234	(3)
4. Complement receptor production and surface expression	237	(5)
Immunoprecipitation of surface CR1	237	(2)
Biosynthetic labelling of CR1	239	(3)
5. Rosette formation assays	242	(11)
Generation of fixed fragments on sheep erythrocytes	242	(3)
Enumeration of complement fragments on sheep erythrocytes	245	(3)
Rosette formation	248	(5)
6. Complement receptor-dependent phagocytosis	253	(2)
Preparing C3bi-coated particles	253	(1)
Phagocytosis of C3bi-coated fluorescent microspheres	254	(1)
7. Conclusion	255	(1)
References	255	(2)
A1 List of suppliers	257	(8)
Index	265