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OVERVIEWOF IMMUNOLOGYHj.NOORMARTANY. dr, SpPK (K), M.SiCLINICAL PATHOLOGY OFMEDICAL FACULTY OF MEDICINEPADJADJARAN UNIVERSITY1REFERENCESImmunologgy Ivan Roitt, Jonathan .B, David.M.Essentials of Immunology and SerologyJ. StanleyBasic & Clinical Immunology Daniel P.S, Abba I.T, Tristram G.P.Immunology: an introductionTizardBasic Immunology Abbas A.K & Lichtman A H6. Imunologi : Diagnosis & Prosedur LaboratoriumSiti Budina Kresno 2InnateImmune systemAdaptiveNo MemoryNo SpecificityMemorySpecificityImmune system3Host Defenses In Innate ImmunityCells Antigen Presenting Cells Basophils Eosinophils Mast cells NK cells PhagocytesHumoral Factors Complement proteins Lactoferin Lysozyme Pepsin Stomach acidity Anatomical Barriers Cilia Mucus Skin Resident Flora Mainly nonpathogenic bacteria4Antigen Presenting Cells Present antigen to T cells

Basophils & Mast cells Increase Vascular Permeability

Eosinophils Destroy helminths NK cells Destroy infected cells (virally invected cells) Phagocytes Engulf antigen extracellular in to vacuole and destroyed Cells of Innate Immunity and theirrole in host defense5Host Defenses In Adaptive ImmunityCells B cells Plasma cells T cytotoxic cells T helper cellsHumoral Factors Antibodies Cytokines6THE ELEMENTS IN IMMUNOLOGYIMMUNE RESPONSE MANIFESTATIONCLINICAL IMMUNOLOGYIMMUNNOLOGICALDIAGNOSTICSLABORATORYIMMUNOLOGICAL TECHNIQUES7THE ELEMENTSINIMMUNOLOGYANTIGENS ANTIBODIES COMPLEMENTSLYMPHORETICULERSYSTEMMHC /HLASURFACE LEUCOCYTEMOLECULES8ANTIGENSA MOLECULE WHICHINDUCES THE FORMATIONOF ANTIBODYANTIBODYSECRETED IMMUNOGLOBULIN PROTEINSPARTICULARLY THOSE THAT CAN BIND SPECIFICALLYTO A GIVEN ANTIGEN9Antibodi

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Polypeptide Chain Structure Four Human Ig G Subclasses Structur of IgGANTIBODIES 11

Structure of Ig EStructure of Ig D12

Structure of Ig MStructure of IgA1Structure of sIgA13

COMPLEMENTS14

COMPLEMENTS15

LYMPHORETICULERSYSTEM16MHC /HLA

HLA CLASS IHLA CLASS II17SURFACE LEUCOCYTEMOLECULES

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Origin of Cells Involved In The Immune Response19MANIFESTATIONOFIMMUNE RESPONSESHUMORALIMMUNE RESPONSESCELLULARIMMUNE RESPONSESHYPERSENSITIVITYREACTIONSINFECTIONSIMMUNE RESPONSES20

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Some non-Specific Mechanisms Involved In Immunity22CLINICAL IMUNNOLOGY1.IMMUNODEFICIENCY2.LYMPHOPROLIVERATIVEDISORDERS3.HYPERSENSITIVITY4.AUTOIMMUN DISEASE5.IMMUNE COMPLEX23IMMUNODEFICIENCYI. ANTIBODY (B CELL) IMMUNODEFICIENCY DISORDERSII. T CELL IMMUNODEFICIENCY DISORDERSIII. COMBINED ANTIBODY (B CELL) AND CELLULAR (T CELL) IMMUNODEFICIENCY DISORDERSIV. PHAGOCYTIC DYSFUNCTION DISEASES V.COMPLEMENT DEFICIENCIES24LYMPHOPROLIVERATIVEDISORDERSACUTE LYMPHOCYTIC LEUKEMIACHRONIC LYPHOCYTIC LEUKEMIAPLASM CELL NEOPLASMS & DYSCRASIAS * MULTIPLE MYELOMA * WALDENSTROMS MACROGLOBULINEMIA * LYMPHOMA * MONOCLONAL GAMMOPATHY UNDETERMINED SIGNIFICANCE (MGUS) * HEAVY CHAIN DISEASE25HYPERSENSITIVITY* TYPE I - ASTHMA - RHINITIS - URTICARIA - ECZEMA - HAY FEVE* TYPE II(ANTIBODY DEPENDENT CYTOTOXIC)- TRANSFUSION REACTIONS- HAEMOLYTIC DESEASE OF THE NEWBORN- AUTOIMUNE HAEMOLYTIC ANEMIA- MYASTHENIA GRAVIS* TYPE III - AUTO IMMUNE COMPLEXES DISEASE - THE ARTHUS REACTION* TYPE IV - DELAYED TYPE OF HYPERSENSITIVITY - JONES MOTE HYPERSENSITIVITY - DERMATITIS CONTACT - TUBERCULIN-TYPE HYPERSENSITIVITY - GRANULOMATOUS HYPERSENSITIVITY26

HYPERSENSITIVITY27SPECTRUM OF AUTOIMMUNE DISEASEORGAN SPESIFIC

NON ORGAN SPESIFICHashimotos thyroiditisPrimary myxoedemaThyrotoxicosisPernicious anemiaAutoimmune atrophic gastritisAddisons diseasePremature menopause (few cases)Insulin dependent Diabetes mellitusGoodpastures syndromeMyasthenia gravisMale infertility (few cases)Pemphigus vulgarisPemphigoidSymphatetic ophthalmiaPhagogenic UveitisMultiple Sclerosis (?)Autoimmune Haemolytic anaemiaIdiophatic Thrombocytopenic purpuraIdiophatic leucopeniaPrimay biliary cirrhosisActive chronic hepatitis (HBsAg - )Cryptogenic cirrhosis (some cases)Ulcerative CollitisSjogrens SyndromeRhematoid arthritisDermatomyositisSclerodermaMixed Connective tissue diseaseDiscoid lupus erythematosusSysthemic lupus erythematosus 28IMMUNE COMPLEXIMMUNE COMPLEX ALLERGIC DISEASE * THE ARTHUS REACTION * SERUM SICKNESSIMMUNE COMPLEX GLOMERULONEPHRITISIMMUNE COMPLEX SYSTEMIC LUPUSERYTHEMATOSUS IMMUNE COMPLEX HEMOLYTIC ANEMIA 29

The Arthus Reaction The Site of Complex Deposition In The Kidney Deposition of Immune Complexes in Blood Vessel Wall Type 1&230ImmunodiagnosticsImmunologicCompetenceTestImmuno SerologicTestTumor MarkerTestImmuno diagnostics31ImmunologicCompetenceTestNon SpecificImmuneResponseTestLeukocyteFunctionTestChemotactic FsTestIngestion FsTestIntracellularKilling Fs TestSpecificImmuneResponseTestQuantitativeLymphocyteT & B countLymphocyte TSubset countQualitative*Lym.Transfor T.*DNA Syn.rate T.*Mixed cult. reac.*Cytotoxicity.T*Lym. Produc.T.32LeukocyteFunctionTestIngestion andmetabolism Function TestTo analyses Reduction NBTTo count bacteriain leukocyte withFlowcytometryTo analyses Reactive oxygen species with:Spectrofotometry,Flourometry orChemiluminescenSyntesis andSecretion cytokineWith RIA or ELISA33ImmunoserologicTestTyphoid StreptococcusGonococcusSyphilisHepatitis etcTumor Marker Test CEA AFP PSA Ca 19-9 Ca 125 MCA Cyfra 21 HCG etc34THANK YOU

3536IMMUNOLOGICAL TECHNIQUES *ANTIGEN-ANTIBODY INTERACTIONS*ISOLATION OF PURE ANTIBODIES*ASSAY FOR COMPLEMENT*ISOLATION OF LYMPHOCYTE POPULATIONS*EFFECTOR CELL ASSAYSFLOWCYTOMETRY AND CELL SORTING(FLOURECENCE ACTIVATED CELL SORTER)STRUCTUR ANALYSIS AND EXPRESION GEN37ANTIGEN-ANTIBODY INTERACTIONS A. Precipitation Reaction in Gels a. Immuno-double diffusion b. Immunoelectrophoresis c. Single Radialimmunodiffusion d. Countercurrent electrophoresis e. Rocket electrophoresis B. Haemaglutination C. Complement Fixation D. Immunofluorescence a. Direct b. Indirect c. Indirect Complement Amplified 138 E. Radioimmunoassay (RIA)

F. Enzyme-Linked Immunosorbent Assay (ELISA)

G. Radioallergosorbent test (RAST)

H. Radioimmunosorbent test (RIST)

I. Immunoblotting

39ISOLATION OF PURE ANTIBODIESASSAY FOR COMPLEMENT 2340ISOLATION OFLYMPHOCYTE POPULATIONS4 * Lymphocyte Separation on Ficoll Isopaque * Isolation of Lymphocyte Sub-populations-rosetting * Isolation of Lymphocyte Sub-population-panning

415EFFECTOR CELLASSAY * Plaque forming cell assay-I

* Plaque forming cell assay-II a. Indirect plaques b. Direct plaques c. Reversed plaques

* The lymphocyte stimulation test

* Cytotoxicity assay by chromium release.

42FLOWCYTOMETRYAND CELL SORTING6STRUCTUR ANALYSISAND EXPRESION GEN743A. Precipitation Reaction in Gels

44Immuno - double diffusion - II

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46Single Radial Immunodiffusion

47Countercurrent electrophoresisandRocket Electrophoresis

48HAEMAGLUTINATION

49COMPLEMENT FIXATION TEST

50DIRECT AND INDIRECT IMMUNOFLUORESCENCE

51ENZYME-LINKED IMMUNOSORBENT ASSAY

52RADIOIMMUNOASSAY (RIA)

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RADIOALLERGOSORBENT TEST (RAST)54

RADIOIMMUNOSORBENT TEST (RIST)55

IMMUNOBLOTTING56

ISOLATION OF PURE ANTIBODIES ( Affinity Chromatography)57

MONOCLONAL ANTIBODY PRODUCTION58

ASSAY FOR COMPLEMENT COMPONENT59

LYMPHOCYTE SEPARATION ON FICOLL ISOPAQUE

ISOLATION OF LYMPHOCYTE SUBPOPULATIONS ROSETTING60

ISOLATION OF LYMPHOCYTE SUBPOPULATIONS PANNING61

T CELL LINES62

PLAQUE FORMING CELL ASSAY. I63

PLAQUE FORMING CELL ASSAY. II64LYMPHOCYTE STIMULATION TESTCYTOTOXICITY ASSAY BY CHROMIUM RELEASE

65ImmunodiagnosticsImmunologic competence test IMUNOSEROLOGIC TUMORMARKER* CEA * AFP * HCG * PSA * TPS* Ca 19-9 * Ca 125 * MCA * Cyfra 21.1Typhoid Streptococcus Syphilis GonococcusHepatitis A, B, C, D, E, GNon specific immune response testSpecific immune response testLeukocyte function testSwallowing fn testChemotactic fn testIntracellular killing fn testQuantitativeQualitativeLymph. T &B countLymph. T subsetcountLymphocyte transformation testDNA synth. rate testMixed culture reactionCytotoxicity testLymphocyte production test66