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Seminars in Immunology

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Seminars in Immunology SEMINARS IN IMMUNOLOGY SEMINARS IN IMMUNOLOGY Edited by András Kristóf Fülöp Written and translated by Éva Pállinger Edit Buzás András Falus György Nagy Marianna Csilla Holub Sára Tóth László Kőhidai Zsuzsanna Pál Scientific reviewers Edit Buzás András Falus English proofreading Adrienn Ádám Semmelweis University • Budapest, 2012 © András Kristóf Fülöp, 2012 2 Manuscript closed on 31 May 2012 ISBN 978-963-9129-82-5 Semmelweis University Responsible for publishing: Semmelweis University Editor: András Kristóf Fülöp Technical editor: Adrienn Ádám Length: 223 pages 3 CONTENTS 1. INTRODUCTION (ÉVA PÁLLINGER) ......................................................................................................... 10 1.1. Basic terms ..................................................................................................................................... 10 1.1.1. Innate and acquired immunity ......................................................................................................... 10 1.1.2. What does the immune system recognize? .................................................................................... 11 1.1.3. What types of receptors can recognize antigens? .......................................................................... 11 1.1.3.1. PRR (pattern recognition receptors, pathogen recognition receptors) ............................................ 11 1.1.3.2. T cell receptor (TCR) ....................................................................................................................... 12 1.1.3.3. B cell receptor BCR......................................................................................................................... 12 1.1.4. What are the consequences of antigen recognition? ...................................................................... 12 1.1.5. Local immune response .................................................................................................................. 13 1.1.6. Recognition is based on meeting .................................................................................................... 13 1.1.6.1. Lymphocyte recirculation ................................................................................................................ 14 1.2. Organs of the immune system ........................................................................................................ 14 1.2.1. Lymph nodes .................................................................................................................................. 15 1.2.1.1. Cell types of the lymph glands ........................................................................................................ 16 1.2.1.1.1. Sentinel lymph nodes ...................................................................................................................... 16 1.2.2. Bone marrow ................................................................................................................................... 17 1.2.3. Spleen ............................................................................................................................................. 17 1.2.4. Thymus ........................................................................................................................................... 18 1.2.5. Mucosa associated lymphoid tissues: MALT .................................................................................. 19 1.3. Immune cells ................................................................................................................................... 20 1.3.1. Laboratory evaluation of the immune system .................................................................................. 21 1.3.1.1. Peripheral blood smear and bone marrow smear ........................................................................... 21 1.3.1.2. Cytochemical reactions ................................................................................................................... 21 1.3.1.2.1. PAS reaction ................................................................................................................................... 22 1.3.1.2.2. Esterase reaction with and without NaF inhibition ........................................................................... 22 1.3.1.3. Flow cytometry ................................................................................................................................ 22 1.3.1.4. Immunohistochemistry .................................................................................................................... 23 1.3.1.5. Genetic tests ................................................................................................................................... 23 1.4. Communication between immune cells ........................................................................................... 23 2. IMMUNOASSAYS I. (Edit Buzás) ............................................................................................................... 25 2.1. Features of diagnostic antibodies ................................................................................................... 25 2.1.1. Basic terms ..................................................................................................................................... 26 2.1.2. Labeling methods of antibodies ...................................................................................................... 27 2.2. Methods .......................................................................................................................................... 28 2.2.1. Flow cytometry ................................................................................................................................ 28 2.2.2. ELISA (Enzyme Linked Immunosorbent Assay) ............................................................................. 28 2.2.2.1. Indirect ELISA ................................................................................................................................. 29 2.2.2.2. Sandwich ELISA ............................................................................................................................. 30 2.2.2.3. Competitive ELISA .......................................................................................................................... 30 2.2.2.4. What should we pay attention to during the ELISA reaction? What are the potential pitfalls? ........ 31 2.2.3. ELFA (Enzyme Linked Immunofluorescent Assay) ......................................................................... 32 2.2.4. ELISPOT ......................................................................................................................................... 33 2.2.5. Immuno blot (Western blot) ............................................................................................................. 33 2.2.6. Methods based on radioactive isotope detection ............................................................................ 35 2.2.6.1. Radioimmunoassay (RIA) ............................................................................................................... 35 2.3. Aspects of selecting an immunoassay for a given purpose ............................................................. 42 2.4. Tasks .............................................................................................................................................. 43 4 2.4.1. Lateral flow assay ........................................................................................................................... 43 2.4.2. Indirect immunocytochemical preparation ....................................................................................... 43 3. METHODS BASED ON ANTIGEN-ANTIBODY REACTION II: IMMUNSEROLOGY (ANDRÁS FALUS, GYÖRGY NAGY) .......................................................................................................................................... 45 3.1. Immune complex and precipitation.................................................................................................. 45 3.2. Two-dimensional and radial immunodiffusion ................................................................................. 49 3.3. Serum Electrophoresis .................................................................................................................... 50 3.4. Immunoelectrophoresis and immunofixation ................................................................................... 52 3.5. Rocket electrophoresis .................................................................................................................... 53 3.6. Two-dimensional immune electrophoresis ...................................................................................... 54 3.7. Turbidimetry and nephelometry ...................................................................................................... 54 3.8. Application of serological methods .................................................................................................. 55 3.9. Agglutination ................................................................................................................................... 56 3.9.1. Types of agglutination ..................................................................................................................... 57 3.10. Questions .......................................................................................................................................
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