Immunology Review3/8/11 1 Immunology Terms Acute phase reactant Protein trauma haptoglo 3/8/11 2 glycopro Autoantibody Avidity Chemotaxin Clusters of 3/8/11 3 Antibo Strengt Chemic a partic Antige Histamine Hypersensitivity Immunity 3/8/11 Av bas Ah tiss Immunogen Re 4 An Ligand Lymphokines Lysozyme 3/8/11 A molecule configuratio Polypeptide regulate the 5 Enzyme fou . Plasma cells Postzone Prozone Seroconversion Transform Polyclonal antibody Antibody p False-nega excess False-nega 3/8/11 The chang 6 . Branches of the Immune System Branch Definition Defe Cellular 3/8/11 Cell mediated 7 Viru myco . Types of Immunity Type Natural or innate Explanation Non-specifi with which 3/8/11 8 . Adaptive Immunity Type Explanation Naturally acquires 3/8/11 An individual infected with a microorganism 9 active . Adaptive Immunity Continued Type Explanation Naturally 3/8/11 An individual is 10 acquired protected by antibo . Immunoglobulins IgG Form Monomer Ig P 3/8/11 Molecular 150000 11 9 . IgG Serum 800-1600 I 1 concentration mg/dL Antigen3/8/11 m 2 12 1 . 3/8/11 13 IgM First in an Ig at respo .IgG Other Defense against bacteria and viruses. Cells of the Immune System Cell T lymphocytes Functi Cell-m 3/8/11 14 . Cells of the Immune System Continued Cell Suppressor/cytotoxic T cells Fun Supp cells 3/8/11 15 . Cells of the Immune System Continued Cell Neutrophils Functio Principa phagocy 3/8/11 16 response . Lymphoid Organs Primary Lymphoid Organs Bone marrow Thymus 3/8/11 17 . To Obtain Lymphocytes Density gradient centrifugation Ficoll-Hypaque To Identify Lymphocytes 3/8/11 18 Laboratory Identification of Lymphocytes Direct or indirect immunofluore . Common Lymphocyte Markers T cells CD2 CD3 3/8/11 CD4 19 . Complement Definition A series o host defen are conve 3/8/11 Functions 20 Opsoniza . Immunoglobulin Structure Basic structure A fou and tw Heavy chains 3/8/11 21 γ. α. μ molecu . Immunoglobulin Structure Continued Fc fragment Fragment cr ability. heavy chains Constant region 3/8/11 The amino-t 22 the amino ac . Immunoglobulin Structure Continued Hinge region The fle and sec Joining chain 3/8/11 23 let the t A glyc . Complement Continued Definition A series of host defens are converte Functions 3/8/11 Opsonizatio 24 . Immune Phenomena Explan Primary phenomena Combin antibod 3/8/11 25 . Hypersensitivity Reactions Type I: Anaphylac Key reactant(s) 3/8/11 IgE 26 . hay fever.Hypersensitivity Reactions Continued Type I: Anaphylactic Symptoms Example 3/8/11 Immediate Anaphylaxis. 27 . Agglutination Methods Method Direct agglutination Principle The proc antigens. to form la 3/8/11 28 specific a . Agglutination Methods Continued Method Passive Principle A reaction hemagglutination are absorb are agglut 3/8/11 29 correspon . Heterophile antibodies can cause false-positive reactions in hemagglutination tests.Agglutination Methods Continued Method Principle Hemagglutination A test for Inhibition certain vir antibody. 30 3/8/11 • and hema . In • Rheumatoid factor can cause false-positive reactions in agglutination tests because it reacts with any IgG. RBCs. Precipitation Methods Method Precipitation Princip Soluble antibody complex 3/8/11 31 less sen . Precipitation Methods Continued Counter (CIE) 3/8/11 32 Antigen immunoelectrophoresis wells t charge another . Precipitation Methods Continued Method Radial immunodiffusion (RID) 3/8/11 Principle Antigen is diameter o when it dif 33 containing . Method Other Serological Methods Principle Patient Complement fixation ser incubated wit corresponding 34 and compleme 3/8/11 . If th 3/8/11 .Other Serological Method Continued Direct fluorescent antibody The specimen a glass and ove fluorescein-lab corresponding 35 antibody. If 36 corresponding an .Other Serological Method Continued Method Indirect fluorescent antibody 3/8/11 Principle Reagent antigen slide is overlaid patient serum. .g.Labeled Immunoassay Terminology Ligand Isotopic Nonisotopic 3/8/11 The substance An immunoas An immunoas 37 label. e. enz Competitive An immunoas . Labeled Immunoassay Terminology Continued Heterogeneous An immunoas sensitive than reactant from 3/8/11 38 . Comparison of Radioimmunoassay and Enzyme Immunoassay RIA Label Measurement 3/8/11 Radioisoto Counts per 39 scintillatio . Comparison of Radioimmunoassay and Enzyme Immunoassay Continued RIA Disadvantages Short shelf Radiation h 3/8/11 40 Regulation of radioiso . ↑Chemiluminescent molecules produce light energy from a chemical reaction. 3H 3/8/11 .Immunoassay Labels Radioisotopes 125 I • • • Enzymes Horseradish peroxidase β-Dgalactosidase 41 *Fluorochromes absorb energy from light source and convert itinto light of a longer wavelength and lower energy. Principles of Labeled Immunoassays Method RIA Type of Assay Competitive Heterogeneous 3/8/11 42 . Principles of Labeled Immunoassays Continued Method Immunoradiometric assay (IRMA) 3/8/11 43 Type o Non-co Isotopi Hetero . Principles of Labeled Immunoassays Continued Method Enzyme-linked immunosorbent assay (ELISA) 3/8/11 Type of As Competitiv Nonisotop Heterogen 44 . Principles of Labeled Immunoassays Continued Method Indirect or nocompetitive ELISA Type of A Noncomp Nonisoto Heteroge 3/8/11 45 . Principles of Labeled Immunoassays Continued Method Immunoenzymometric assay (IEMA) 3/8/11 46 Type Nonc Noni Heter . Principles of Labeled Immunoassays Method Sandwich enzymemultiplied 3/8/11 Continued Type of Assa Noncompetit Nonisotopic Heterogeneou 47 immunoassay . Principles of Labeled Immunoassays Continued Method Substrate-labeled fluorescent immunoassay 3/8/11 48 Type of A Competit Nonisoto Homogen . Principles of Labeled Immunoassays Continued Method Fluorescence polarization immunoassay 3/8/11 Type of Ass Competitive Nonisotopic Homogeneo 49 (FPIA) . Labeled Immunoassays Isotopic Competitive Heterogeneous Homogeneous 3/8/11 50 Radioimm . Nontreponemal Tests for Syphilis VDRL Method Detects Reagent(s) 3/8/11 51 Flocculat Reagin Cardiolip . Nontreponemal Tests for Syphilis Continued VDRL Falsepositives 3/8/11 10-30% may be e.g., infectious m 52 infectious hepat Treponemal Tests for Syphilis FTA-ABS Method Detects Reagent(s) 3/8/11 53 Indirect f Antibody Sorbent ( Treponemal Tests for Syphilis Continued FTA-AB Specimen(s) Reactivity during disease 3/8/11 54 Serum, Usually tests. S primary Treponemal Tests for Syphilis Continued FTA-ABS Other Sorbent r nonspeci 3/8/11 55 Used to c Absorbe 56 beef RBCs but not by guinea pig . 3/8/11 IM antibodies: Agglutinate sheep and horse RBCs.Test for the Diagnosis of Infectious Mononucleosis (IM) Antibodies Heterophile antibodies* ox. • weeks after infection. Undetecta 3/8/11 weeks. 57 .Test for the Diagnosis of Infectious Mononucleosis (IM) Continued Antibodies Specific viral antibodies IgM-VCA (viral capsid antigen): *Nonspecific antibodies that cross-react with antigens other than those originally responsible for their production. Rickettsia rickettssi 58 .Weil-Felix Reaction Disease Organism Rocky Mountain spotted fever* • 3/8/11 Most common rickettsial disease in U.S. Antinuclear Antibodies Antibody Against S 3/8/11 Anti-ds- Double- 59 S . Antinuclear Antibodies Continued Antibody Against Sp Anti-DNP Deoxyribonuc.Pre 3/8/11 leoprotein 60 SL . Antibody Against Spe Anti-SS3/8/11 Ribonucl61 Pres . More than on epattern will be observed. • 3/8/11 Anti- Histone 62 Presen . Seen in scleroderma and polymyositis. Fluorescence of nucleoli is due to antibody to RnA.Antinuclear Antibodies Continued Antibody Against Specif – – Multiple ANAs can be present in a specimen. Hepatitis Tests Test Hepatitis A Anti-HAV (total) Anti-HAV (IgM) 3/8/11 In P A 63 Hepatitis B . Hepatitis Tests Continued Test Hepatitis C Anti-HCV Hepatitis D 3/8/11 Indicati Acute. c Past or c 64 Anti-HD . Hepatitis Profiles Acute Hepatitis A Recovery from Hepatitis A Anti-HAV 3/8/11 Anti-HAV (total) + 65 (IgM) + . HIV Testing Test ELISA Detects Antibody to HIV Type of T Screening 3/8/11 66 . HIV Testing Continued Test Detects Ty Te Slide 3/8/11 Antibody 67 Sc agglutination to HIV . HIV Testing Continued Test Western blot Detects Antibody to HIV Type Conf 3/8/11 68 . HIV Testing Continued Test Indirect rescence 3/8/11 Detects Virus or Type o Confi immunofluo.antibody assays 69 . HIV Testing Continued Test Detects Typ Tes P24 antigen test HIV (HIV-1-Ag) 3/8/11 Scre 70 antigen . Viral Testing Test Polymerase chain reaction (PCR) 3/8/11 Detects Viral genome Ty Ad sta tes 71 . Other Serological Tests Test Common Method Rheumatoid arthritis (RA) 3/8/11 Agglutination 72 . Other Serological Tests Continued Test Cold agglutinins 3/8/11 Common Method Hemagglutination 73 . Test Common Method R Antistreptolysin O (ASO) 3/8/11 Neutralization S O R 74 . Other Serological Tests Continued Test Common Method Re C-reactive Latex protein 3/8/11 75 La agglutination par . Other Serological Tests Continued Test Common Method Reage Rubella Hemagglutin Viral ation Inhibition prepar 76 3/8/11 RBCs . Test Common Method Rea Febrile agglutinins Bacterial agglutination Salm and Bru 3/8/11 77 abor . A fourfold increase in titer from acute to convalescent specimen is diagnostic.Interpretation of Serological Tests A single titer is not usually meaningful because it can’t differentiate current infection from past infection or immunity due to immunization. IgM antibody is a sign of recent infection. 3/8/11 78 . The acute specimen should be frozen and run with the convalescent specimen to eliminate differences in titer due to technical variations. The convalescent specimen should be drawn 10-14 days after the acute specimen. Mix 2 mL of the 1:100 dilution with 8 mL of buffer. 5 mL of packed RBCs + 95 mL of buffer. How would you prepare 5 ml of a 1:10 dilution of serum? 1 =x 10 5 10x = 5 x = 0. if is undiluted? 1 x ½ x ½ x ½ = 1/8 3/8/11 79 . (2 mL + 8 mL = 2:10 = 1:5.5 mL of buffer.5 mL of serum with 4. (1 mL + 9 mL = 1:10) How would you prepare 10 mL of a 1:500 dilution from a 1:100 dilution? A 1:5 dilution of a 1:100 dilution yields a 1:500 dilution (1/5 x 1/100 = 1/500). Mix 1 mL of the 1:10 dilution with 9 mL of buffer.) What is the dilution in tube 4 of a twofold serial dilution.5 mL Mix 0. How would you prepare 10 mL of 1:100 dilution from a 1:10 dilution? A 1:10 dilution of a 1:10 dilution yields a 1:100 dilution (1/10 x 1/10 = 1/100).Serology Calculations How would you prepare a 5% suspension of human group O red blood cells? 5% = mL per mL. The End 3/8/11 80 .