I. Introduction II. Element of Immunity III. Immunogenetic IV. Immune Response V. Antigen & Immunogen & Vaccine VI.

Immunoglobulin VII. Complement System VIII. Cytokines

IX. Antigen-Antibody Reaction X. Immunology in Infection diseases XI. Immunoprophylaxis XII. Hypersensitivity Reaction XIII. Autoimmune Diseases XIV. Immunodeficiency

Immunology Imunis All of physiology mechanism => foreign agent => - neutralize with or - eliminate => without - metabolism tissue damage

X1st century => China XV1st => variolasi 1798 : E. Jenner : Cowpox => Smallpox 1880 : Vaccine (L. Pasteur) 1908 : => cellular (Metchnikof) => humeral (Ehrlich) >>> 1970 : molecular biology

 Genetic
Age Metabolism Environment & nutrition Anatomy

Microbe
Physiology

1. Defense 2. Homeostasis 3. Surveillance

Innate (natural) Immunity

Acquired (adaptive) Natural : all of creature (+) Such as : 1. Physical hindered 2. Cellular hindered 3. Chemical hindered

Adaptive With functional characteristic :

1. Specificity Heterogeneity 2. Differentiate : SELF & NOT SELF 3. Memory

Operator of Immunity : Limphoreticular system Phagocyte cells : MPS, Neutrophyl, Eosinophyl Lymphoid cells : B cell & T cell Mediator cells : Basophile, Mastocyt Which come from : Hematopoetic Stem Cell

(0,001 % Bone marrow)

NK cell

Consists of

: Lymphoid cells Lymphoid tissues

Lymphoid Cells
Lymphoid cell

Immunogen

Immunocyt

Specific cellular product (Ig & CMI)

BM Primary Lymph glands
Secondary Lymph gland (Prolif. & Dif)

Thymus gland
Function : Maturity T cells BM Thymus gland Cortex
Circulation

Medulla
CD4+

CD4- CD8-

CD4+CD8+ CD 8+

Circulation

Only can be found in bird family

B cell

BM

Bursa Fabricius Bone Marrow

Secondary lymphoid organ B cell

Stem cell

Secondary lymphoid organ Consists of : lien, lymph node, Payer Patch, Tonsil as antigen filter

Bone Marrow
Myeloerythroid cells
IL-2 SCF IL-7 SCF

SCF

Hematopoietic sterm cell

Lymphoid stem cell ?

B cell precursor IL-3 SCF

Virgin B lymphocyte

Secondary lymphoid organs

T h y m u s

IL-2 IL-7 SCF Thymic factors

T cell precursor

Virgin T lymphocyte

Blood Circulation

Tissues
Afferent lymph duct Lymph node Spleen

Efferent Lymph duct

Ductus Thoracicus

ANTIGEN
PULMO SKIN GIT Resp. Tract. Circulation

Peribronchial Lymphoid Tissues

Tonsil PP

Lymph node Regional

Spleen

Structure Immunology of Spleen

Periarterioler sheet Trabecular artery

Red pulp

Central arteriole

Center germinal

Immunologist Structure of Lymph node

Germinal Center
Cortex

Paracortex

Medulla

Artery Vena

Efferent Lymph canal

All of immune response processes with genetic basic. All factors which regulate Immune Response to foreign agents => hereditary Very widely of scope : HLA & Blood Group Clinical aspects : Blood grouping, tissue/organ transplantation. Autoimmune disease, producing of vaccine, etc.

MHC = HLA (man) Genetic: position: short arm of Chromosome 6 length: 3,5 x 106 bps 5 C C A T T T A A C C - - - 3 3 C C T A A A T T C C - - - 5

Class II

Class III

Class I

C4B 21A Endo 21B BF DP DQ DR TNF BC C4A C2 TNF

500 1000 1500 2000 2500 3000 3500

centromere

kilobases 0

Figure 5-1. Organization of the HLA complex on the short arm of human chromosome 6. Regions encoding the 3 classes of MHC proteins are indicated by braces. Endo denotes a cluster of genes within the class II region that encode protease components and peptide transport proteins required for processing endogenous antigens (see text). Class III proteins are unrelated to class I and II and are not involved in antigen presentation. Among proteins encoded in the class III region are tumor necrosis factors and , and complement factors C2, C4, B and F.

Extracellular region 1 2-microglobulin

CHO 86

chain 101

CLASS I HLA
In all nucleuss cells

Extracellular region membrane cytoplasm

S 2 164 S NH2 NH2 S S 203 S 3 S 259 COOH 282 306 PO4 338 COOH

Such: A, B, C => L M A
Functions: - Immune aware - Tissue rejected

Figure 5-2. Schematic representation of a class I HLA protein. The molecule consists of an MW 44,000 polymorph transmembrane polypeptide ( chain) non covalently associated with an MW 12,000 non polymorph polypeptide (2-microglobulin). The 3 extracellular domains of the chain are designated 1, 2, and 3. The binding site for immunogenic peptides (T cell determinants, is formed by the cleft between the 1 and 2 domains.

-helix Peptide binding groove

2 8-strand -pleated sheet N

C 2 m C 3 Figure 5.3. Diagrammatic structure of a class I HLA molecule (side view). In this ribbon diagram of the polypeptide backbone, the polypeptides are oriented as in Fig 32, but only the extracellular region is depicted. The peptide binding site as a cleft (or groove) formed by 8 strands of pleated sheet and a pair of -helices from the 1 and 2 domains. The sheet structure forms the floor and the type helices the walls of the cleft. strands are depicted is broad arrows and helices as narrow coils.

-helix -pleated sheet forming floor of antigen-binding groove

NN

-helix

Figure 5-4. Peptide-binding site of a class I HLA molecule, viewed along an axis perpendicular to the cell surface. Eight strands of -pleated sheet contributed by the 1 and 2 domains forms the floor of the site, and 2 -helices, one from each of the 2 domains, form the walls. The groove accommodates peptides 8-9 amino acid residues long, leaving them partially accessible for interaction with the T cell antigen receptors.

 chain NH2 1 78 118 2 107 S S 163 193 214 229 COOH

CHO CHO

Extracellular region membrane cytoplasm

chain NH2 CHO 15 19 S S 1 79 Extracellular 117 region S 2 S 173 200 221 237 COOH

Class II HLA

At B cell => macrophage Functions: - T cell aware - Tissue rejected

Figure 5-5. Schematic representation of a class II HLA molecule. The molecule consists of an MW 34.000 polypeptide ( chain) noncovalently associated with an MW 29.000 polypeptide ( chain).

NH2

-helix
COOH COOH

-pleated sheet forming floor of antigen-binding groove

NH2 1

-helix
Figure 5-6. Structure of the peptide binding site of a class II HLA molecule. The binding site is similar to that of class I molecules, except that it is formed by the 1 and 1 domains of the class II molecule and is relatively open at both ends to accommodate longer peptides.

Cell surface

Peptide transporter
2m

Endogenous peptides nucleus RER

Class I MHC

To cell surfaces

Surface AgClass I MHC complex Exogenous antigen Processing

Peptides

Class II MHC

To cell surfaces

Figure 5.7. The pathway of assembly and transport for antigen-MHC complexes containing class I (top) and class II (bottom) HLA molecules. MHC polypeptide of initially expressed in the rough endoplasmic reticculum (RER). Class I proteins sequentially bind endogenous peptides and 2-microglobulin (2m) in the RER lumen and are than transported to the cell surface. Class II proteins associate with invariant chain (li) in the RER and so are prevented from binding endogenous peptides, they are translocated instead to an endosomal compartment, where li dissociates and is replaced by exogenous peptides.

Surface AgClass II MHC complex

Erythrocyte antigen:

ABO Rh

ABO Group Before 20th : transfusion 1900 Landsteiner

Sera Ery

Group

1 2 3
4 5 6

+ +
+ +

+ + + +

+ + + +

+ +
+ +

C A B
B A C

ABO

A,B

single gene ABO with 3 allele A,B,O codominant KH binding + H substrate

A anti B anti + + + Allo ab A,B anti

Genotype Phenotype Er-Ag O/O AO,AA BO,BB AB O A B AB O A B AB

B anti A anti -

Levine & Stetson (1939) => Ag + Asera from post partum mother Ag + S.I rabbit by Rhesus of erythrocyte => Rh factor => Rh. Ag. Natural antibody (-), except by immunization

Genetic of Rhesus
> 30 Ag. Rhesus type

Fisher & Race 3 gene with allele partners => 5 determinant antigen D, C, E, E, C. Wiener 1 gene locus => multiple complex allele DA Rho, rh, rh, hr, hr.

Definition: Self & not self Virgin lymphocyte (109/day), with IG & TCR => 108 antigen type Clonal restriction Clonal selection Each others cells communication.

6 Imunogen 2 1 1 1

1 5 4 3 5 4 1 3

TCR IMMUNOGEEN endosome lissome MHC II

TH cell

MACROPHAGE Antigen-presenting cell (AFC)

CD 4

Figure 3-3. Capture, processing, and presentation of antigen by an APC. The immunogen is captured by phagocytosis, receptor-mediated endocytosis, or pinocytosis and is broken down into fragments. Some fragments (antigens) become associated with class II MHC proteins and are transported to the cell surface, where they can be recognized by CD4 T cells. TCR, T cell receptor.

T cell MHC II molecules APC Autoactivation IL-1 CD4 IL-2R TH cell Activation TH cell IL-2 T cell

Processed antigens

Costimulation

Release of cytokines and other growth and differentiation factors

Figure 3-4. The cell activation. The APC presents an antigen in the context of class II MHC to the TH cell and also provides a costimulatory signal. The 2 signals lead to activation of the TH cell. The APC also releases IL-1, which acts on both the APC and the TH cell to promote activation. Activation leads to IL-2 receptor expression and IL-2 secretion by the TH cell, resulting in autocrine growth stimulation.

Helper factors CD4 TCR Ag B cell

Proliferation

B cell

Memory B cell

Progeny

IL-2R

MHC II
Ig Ag receptors

B cell differentiation

Plasma cell

Antibody

Figure 3-5. B cell activation. Antigen binding to the surface immunoglobulins, coupled with soluble or contact-mediated helper factors from an activated TH cell, lead to proliferation and differentiation. Cytokines involved in TH cell help include IL-2, IL-4 and IL-6.

TCR (already triggered) IL-2

IL-2R TCR Ag target cell (cell death) Tc cell MHC I

TH cell

IL-2R CD4

Autoactivation

CD8

Toxins

MHC I

Figure 3-6. To cell activation requires contact with specific antigen in the context of a class I MHC molecule on the surface of a target cell. It also requires IL-2 from a nearby activated TH cell. The activated Tc cell kills the target cell either by secreting cytotoxins (as shown) or by inducing it to commit suicide.

Definition :

1. 2. 3. 4.

Immunogen Antigen Immunogenicity Antigenicity

Classification : 1. Exogen antigen 2. Endogen antigen : - Xenogeny Ag. (Heterolog) - Autolog Ag. - Alogenic Ag.

Commonly is a macromolecule protein. 1. Molecule antigenisity 2. Molecule size 3. Complexity of Chemistry structure 4. Genetic constitution 5. Method of entry 6. Dosage 7. Digestibility

Determinant Antigenic

Hapten
IK

Hapten

carrier

Immunogen

I.K agent

Hapten

Thymus dependent Ag and Thymus independent Ag

Receptor
T B

Imunogenik

Macrophage cell
HLA DR

Plasma cell

Cross Reaction

>>> Immunogenicity antigen pathway

>>> Retention >>> Molecule size

Local stimulation

Definition :
Protein as humoral immunity effectors molecule

The function of Ig :
Binding Ag Biological activity Thus as complex molecule

Example Antibody to Viral It has particular part which could : Binding virus Be able to enter respiratory tract Not be broken by enzyme Be able to joint with leukocyte

1940 : Tiselius & Kabat Globulin - AB 1950 : Porter gave papain 3 fragments 1960 : Edelman : Multiple chains Porter : 4 chains 1969 : Edelman, AA chain.from BJ Prot > 1970 : Leder genetic

Three-dimensional structure of an immunoglobulin molecule

H3N+
H3N+ VH

Vk

Fab Ck Hinge region Fc

CH2 CH3

CH1

H chain

COOCOO-

Pepsin Papain H3 N+ H3N+

Cleavage sites

Fab

Figure 6-1. Schematic model of an IgG1 (x) human antibody Molecule showing the basic 4-chain structure and domains. Sites of enzymatic cleavage by pepsin and papain are shown

CDR I CDR 3

Figure 6-5. Three dimensional structure of a light chain, in CDR 2 this ribbon diagram tracing the polypeptide backbone, strands are shown as wide Variable domain ribbons, other region as narrow string. Each of the 2 globular domains consists of a barrel-shaped assembly of 7-9 antiparallel -strands. The three hypervariable regions Constant domain (CDR1, CDR2, & CDR3) are flexible loop that project outward from the aminoterminal end of the VL domain.

Side Chain Theory

Abundant receptors as antibody
Ag DA

Instructive Theory

DNA

globulin DA

Selective Theory
spontaneous DNA

Clonal selection Ag

Immunization

Myeloma cell culture

HPRT -IgMyeloma cells (2x107)

HPRT+Ig+ Spleen cells (10)8 Assay for antibody Clone antibody-production (positive) hybrids Tumor Induction Freeze hybridoma for future use
Monoclonal antibody

Selection of hybridcells In HAT medium

Mass culture growth

Monoclonal Antibody

Figure 12-40. Formation of hybridomas between mouse cells and myeloma cells. Mouse myeloma cells that do not produce their own immunoglobulins and lack hypoxanthine and phosphoribosyl transferase (HPRT) are fused to splenocytes From an immunized mouse with polyethylene glycol. The hybrid cells are selected in hypoxanthine-aminopterinThymidine (HAT) medium. Unfused myeloma cells are killed By HAT, and unfused splenocytes die out. The hybridomas are cloned, and antibody is produced in tissue Culture or by ascites formation. (Reproduced, with permission, From Diamond BA, Yelton DE, Scharff MD: Monoclonal Antibodies: A new technique for producing serologic reagents. N Engl J Med 1981; 304: 1344

46 + cell
V.ch N V.ch

Lysis

I
V.ch
S1

health

Lysis (+)

V.ch
S1

560C 30
V.ch

Lysis (-) Lysis

S1/SN

Complement Form & Shape

>>> Globulin : > 20 type 11 type : C1Q,R,S, C2 C9
Widely : C1

C2 C3 (5,6,7,8,9) C4 Complement : CoE

comp. Biologic function

The Complement Cascade

Classic Pathway
Ag:Ab complex C1 C4 C1 C2 C3 C3bBbP Factor D C3b C3 H2O Properdin Factor B C3(H2O)

Alternative Pathway

C14b
C14b(2b)2a C4a C3a

C14b(2b)2a3b
C5

or

(C3b)BbP
C6 C5b C7 C5b67 C8

C5a C9 C5b678(9)n

Terminal Components

C4a C4b

C4a C2 C4b C4b C2 C4b

C2b

C3a

C2a C2b

C2a

C2a

C3

C2aC3b

C4b

C4b

C4b

C4b

C5a C5
C2a

C5

C7

C3b C5b C5bC6 C7

C4b C4

Diagram of the complement cascade. A: The classic complement pathway. A doublet of IgG antibody molecules on a surface can bind and activate C1, a 3-part molecule composed of C1q, C1r & C1s. C1q has a core & 6 radiating arms, each of which ends in a pod. The pod recognizes & binds to the Fc fragment of the IgG. On activation the C1 binds & cleaves C4. The small fragment, C4a, is release. The large fragment, C4b, binds to the target to continue the cascade. In the presence of magnesium ion, C2 recognizes and binds to C4b. B: Once C2 is bound to C4b, it can be cleaved by C1. A small fragment C2b, is release, and the large fragment, C2a, remains bound to the C4b. This newly formed complex of 2 protein fragment can now bind and cleave C3. This molecule is, in turn, cleaved into 2 fragments, C3a & C3b. The small fragment, C3a, is release, & the large fragment, C3b, can bind covalently to a suitable acceptor, C3b molecules that bind directly to the C4b continue the cascade. C: The complex formed of C2a, C4b & C3b can bind and cleave C5. A small fragment of C5, C5a is released. The large fragment, C5b, does not bind covalently. It is stabilized by binding to C6. When C7 binds, the complex of C5b, C6 & C7 becomes hydro phonic. It is partially lipid-soluble and can insert into the lipid of the cell membrane bilayer.

C9 C9 C9

C8
C6 C5b

C7

C8
C9 C6 C7 C9 C5b C9

B
Ba

C3 C3a

C3a

B
C3b

Bb

Bb C3

BbC3b

C3b

C3b

C3b C5

D: When the C5b67 binds C8, a small channel is formed in the cell membrane. Multiple molecules of C9 can bind and markedly enlarge the channel. The channel has a hydrophobic outer surface and hydrophilic central channel that allows passage of water and ions. E: The alternative complement pathway. In the presence of magnesium ions, C3b on a surface can bind factor B, just as C4b can bind C3, factor D, a fluid-phase factor, can cleave bound factor B into 2 fragments, Ba & Bb. Ba is released. The C3bBb complex can now bind an additional molecule of C3 and cleave it, just as C4b2a can bind & cleave C3. C3a is release, & the new complex of C3bBbC3b, usually written (C3b)2Bb, can bind C5 to continue the cascade

Mechanism of complement regulation

1. Spontaneous destruction 2. Enzymatic inactivation 3. Specific bind with certain proteins

Complement Biologic Activity

Substance
C3a

Biologic activity
Smooth muscle control, capillary permeability, mastocyt degranulation Ossification PMN mobilization

C3b C3c

C4a
C52

Smooth muscle control, capillary permeability

= C3a

C5a-des-arg
BB

Chemotaxis, release En Hidrol from neutrophyl

Migration & induction inhibition, monocyt & macrophage spread

Definition: protein (peptide/glycoprotein) as product of a cell group => mediator/communicator between cells for immune system regulation. Today >>> 100 types, contain of: - lymphokine - monokine >>> local effect & very close Mechanism of action: autocrine & paracrine The most important: IL-1,-2,-3,-6,-7 TNF, IFN Synthetic cytokine: Recombinant DNA

Actions of IL-1 and TNF on hematopoietic & lymphoid tissue (A) and nonlymphoid cells & tissue (B). Activities of the two individual cytokines differ in some respects

Major properties of human interleukins and other immunoregulatory cytokines

Earlier Terms
Interleukins IL-1 and
Lymphocyteactivating factor, B cell activating factor, hematopoietin

Principal Cell Source

Macrophages, other APCs, other somatic cells

Principal Effects
Costimulation of

APCs and T cells B cell proliferation & Ig production Acute-phase response of liver Phagocyte activation Inflammation & fever hematopoiesis

Earlier Terms
IL-2
T cell growth factor

Principal Cell Source

Activated TH1 cells, TC cells, NK cells

Principal Effects
Proliferation of activated

T cells Nk and TC cell functions B cell proliferation & Ig G2 expression Growth of early hematopoietic progenitors
B cell proliferation, Ig E

IL-3

Multi-colonystimulating factor

T lymphocyte

IL-4

B cell growth factor I, B cell stimulatory factor I

TH2 cells, mast cells

expression & class II MCH expression TH2 & Tc- cell proliferation & function Eosinophil & mast cell growth & function Inhibition of monokine production

Earlier Terms
IL-5

Principal Cell Source

TH2 cells, mast cell

Principal Effects
Eosinophil growth & function
Synergistic effects with

IL-6

IFN-2, hepatocytestimulating factor, hybridoma growt factor

Activated TH2 cells, APCs, other somatic cells

IL-1 or TNF to costimulator T cell Acute-phase response of liver B-cell proliferation & Ig production Thrombopoiesis
T & B lymphopoiesis Tc cell function

IL-7

Thymic & marrow stromal cells Macrophages, other somatic cells

IL-8

Chemoattractant for neutrophils & T cells

Earlier Terms
IL-9 IL-10
Cytokine synthesis inhibitory factor

Principal Cell Source

Cultured T cell Activated TH2, CD8 T, & B lymphocytes, macrophages

Principal Effects
Some hematopoietic & thymopoietic effects
Inhibition of cytokine

production by TH1 cells, NK cells & APCs Promotion of B cell proliferation & antibody responses Suppression of cellular immunity Mast cell growth
Synergistic effects on

IL-11

Stromal cells

hematopoiesis & thrombopoiesis

Earlier Terms
IL-12
Cytotoxic lymphocyte maturation factor, NK cell stimulatiory factor

Principal Cell Source

B cells,
macrophages

Principal Effects
Proliferation & function

of activated Tc & NK cells IFN production TH1 cell induction, supresses TH2 cell functions Promotion of cellmediated immune responses IL-4 like effects

IL-13

TH2 cells

IL-15

Epithelial cells & Monocyte, non lymphocytic cell

Mimics IL-2 T-cell effects Mast cell NK activation

Earlier Terms

Principal Cell Source

Principal Effects

TNF

Lymphotoxin

Activated macrophages, other somatic cells

IL-1 like effect Vascular thrombosis &

tumor necrosis

INF dan Leukocyte

interferons, type I interferons

Macrophages ; Antiviral effect neutrophils, other Induction of class I somatic cells MHC on all somatic cells Activation of macrophages & NK cells

Earlier Terms
INF
Immune interferon, type II interferon

Principal Cell Source

Activated TH1 & NK cells

Principal Effects
Induction of class I MHC on all

somatic cells Induction of class II MHC on APCs & somatic cells Activation of macrophages, neutrophils & NK cells Promotion of cell-mediated immunity Induction of high endothelial venules Antiviral effect

Earlier Terms
TGF

Principal Cell Source

Activated T lymphocytes, platelets, macrophages, other somatic cells

Principal Effects
Anti-inflammatory (supression of

cytokine production & class II MHC expression Anti-proliferative for macrophages & lymphocyte Promotion of B-cell expression of Ig A Promotion of fibroblast proliferation & wound healing

Noncovalent binding: 1. Electrostatic force: - NH+ - -OOC 2. Hydrogen binding force: - OH H2N 3. Hydrophobic force: 4. Van der Waals force

Antibody affinity
AG + AB
K1 > K 2 K1 K1

AGAB
Affinity

AG AB Reaction
Primary Reaction Secondary Reaction Tertiary Reaction

Primary Reaction To look labeling: FARR Immunofluorocent RIA

ELISA

Secondary reaction Precipitate reaction Agglutinating reaction Floccules reaction Neutralisms reaction RIC

Tertiary reaction Such AG AB reaction in vivo Can be: - advantages - diseases

Antigen Antibody

Schematic figure of antigen-antibody frame work performed

Supernatant Precipitate

Free Ab

No free Ab & Ag

Free Ag

P r e c i p i t a t e d a n t i b o d y

Antigen increase

Antigen Antibody

Ab-remainder

Equivalent

Ag-remainder

Schematic figure of quantitative precipitation curve

Single radial diffusion in agar (radial immunodiffusion)

Petri dished is filled with semisolid agar solution containing antibody to antigen S. After agar hardens, the center well is filled with a precisely measured amount of material containing antigen S

Antigen S is allowed to diffuse radially from the center well for 24-48 hours

Antigen concentration (n.g/mL)

Log C = D-Do K C = Antigen concentration Do = Intercept with ordinate D = ring diameter K = Slope of line

Standard curve for single radial diffusion. Relationship between ring diameter and antigen concentration is described by the line constructed from known amounts of antigen. Equation and curve for timed interval (Fahey) method

Identity reaction

Nonidentity reaction

Partial identity reaction

A = A antigen a-A = A anti B = B antigen a-B = B anti A1 = A antigen plus a-A1 = A1 anti more determinant

Schematic figure of 3 type Ouchterlony double diffuse reaction. B, Ouchterlony double diffusion bowl shows identity reaction between 1 & 2 fraction, partially identity reaction between all of Rabbit gammaglobuline (RGG) and 2 & 3 fraction and nonidentity reaction between 1 & 3 fraction.

Single radial diffusion in agar (radial immunodiffusion)

Where antigen S meet corresponding antibody to S in the agar, precipitation results. After reaction proceeds to completion or at a timed interval, a sharp border or a ring is formed

By serial dilution of a known standard quantity of antigen S-S/1,S/2, S/4,S/8rings of progressively decreasing size are formed. The amount of antigen S is unknown specimens can be calculated and compared with standard in the timed interval (Fahey method)

Reaction of identity

Reaction of nonidentity

Reaction of partial identity

R
R S

R1
R

Reaction patterns in angular double imunodiffusion (Ouchterlony). R = antigen R, S = antigen S, R1 = antigen R1, R = antibody to R , S = antibody to S. reaction of identity: Precisely similar precipitin lines have formed in the reaction of R with R . Note that the lines intersect at a point. Reaction of nonidentity: precipitin lines completely cross owing to separate interaction of R with R and S with S when R and S are non cross reacting antigens. Reaction of partial identity: R reacts with both R and R1 but forms lines that do not form a complete cross. Antigenic determinants are partially shared between R and R1

ANTIGEN X QUANTITATION
AgX AgX/32 Antibody x AgX/16 AgX/8 AgX/4 AgX/2

ANTIBODY X QUANTITATION
X X/32

X/2
AgX

X/16 X/8

X/4

Semiquantitative analysis of antigen and antibody by double immunodiffussion. Antigen X (Ag X) is serially diluted and placed circumferentially in wells surrounding the central well containing antibody against antigen X. Precipitin lines form with decreasing thickness until no longer visible at dilution of 1:32 of antigen X. on the right, a similar pattern is generated but with serial 2-fold dilutions of antibody X (X). Formation of a single precipitin line indicates that a single antigen-antibody reaction has occurred.

Technique of immunoelectrophoresis
Semisolid agar poured onto glass slide and antigen well and antiserum trough cut out of agar

Antigen well filled with human serum

Serum separated by electrophoresis

Technique of immunoelectrophoresis
Antiserum trough filled with antiserum to whole human serum

Serum and antiserum diffuse into agar

Precipitin lines form for individual serum proteins

Comparison of patterns of zone electrophoresis and immunoelectrophoresis of normal human serum

albumin

Complement Fixation Test Indicator system +

Complement Sheep red cell coated with anti sheep red cell antibody Complement reacts with anti sheep red cell antibody and lyses cell

Positive test +
Antigen

+
Complement

+
No lysis of antibody coated red cells as complement used up

Antibody to antigen

Antibody reacts with antigen and complement combines

Complement-fixation test. The indicator system (sheep red cells coated with antibody to sheep red cells) is normally lysed in the presence of complement (fresh guinea-pig serum) top. If another antibody-antigen system is first mixed with the complement it will no longer be available to lyse the indicator system bottom.

Specificity Test
DIRECT METHOD

++
INDIRECT METHOD

Direct method +

Indirect method + +

+
Legend

Substrate Antigen Fluorescent Fluorescent Immune Unlabeled Unlabeled Fluorescent antibody antiglobulin complex antibody antiglobulin heterologous antibody
Mechanism of immunofluorescence techniques. Direct method (top): Antigen in substrate detected by direct labeling with fluorescent antibody. (bottom): Antigen-antibody (immune) complex in substrate labeled with fluorescent antiglobulin reagent. Indirect method (top): incubation of antigen in substrate with unlabeled antibody forms immune complex. Labeling performed with fluorescent antiglobulin reagent. (bottom): Immune complex in substrate reacted with unlabeled antiglobulin reagent and then stained with fluorescent antiglobulin reagent directed at unlabeled antiglobulin.

BLOCKING METHOD (Indirect method)

NEUTRALIZING METHOD

Specificity test. Direct method (Left): Substrate antigen fails to react with fluorescent antiglobulin reagent. No fluorescence results. (Right): Immune complex substrate fails to react with fluorescent antibody directed again unrelated antigen. No fluorescence results. Indirect method (Top): Unlabeled specific antiglobulin is replaced by unrelated antibody. In second step, fluorescent antiglobulin can not react directly with antigen in substrate that has not bound specific antiglobulin. No fluorescence results. (Bottom): First step performed by reacting specific antibody with substrate antigen. In second stage, the specific conjugate is replaced by unrelated fluorescent heterologous antibody. No fluorescence results. Blocking method Substrate antigen is incubated with unlabeled specific antibody prior to addition of specific fluorescent antibody . Decreased fluorescence results. Neutralizing method Substrate antigen is incubated with specific fluorescent antibody after it is absorbed with specific antigen substrate. No fluorescence results.

Infection and Infection disease Infection = microorganism invasion local / systemic alteration. Pathogenic M.O. has evasive mechanism with its photogenic factors. Balance disturbance defense <<< iatrogenic disease. Defense mechanism <<< Immune Compromised Host.

Predisposing factors Immune system effects

Immunosupression: X ray, cancer th/, alograft res. Viral Infection: Rubella, EBV Herpes, HIV Hepatitis Tumor Malnutrition CMI & humoral immunity decrease. Viral replication in limfoid cell which cause immune function disturbance. Immune cells replaced by tumor cells. Lymphoid hypolasia Lymphocyte << Phagocytes << Inflam. lung change Immune Compl. to spore Fag. Activ. <<

Infection types
Pulmonal inf., bacterium, fungal inf., UTI. Secondary bacterial inf.

Bacterium, pneumonia, UTI Measles, TB, Respiratory Tract Inf., GIT inf. COPD Allergic response. Staph. Inf., TB, Respiratory Tract Inf., bacterium.

Smoking, Dust inhalation Chronic endocrine diseases.

Primary I.D

CMI & Hum. <<

Evasi mechanism pattern:

S. aureus Streptococcus Gonococcus & Meningococcus Intracellular org. : intracellular defense Herves Vi. & EBV : complement inhibit.

: A proteine,coagulase : polisach. caps., streptolysin. : protease to IG A

factor.

 Multiple defenses

Characteristic : Natural immunity Adaptive immunity

Skin as first line defense Pathogen bacteria PMN come from blood vascular

Tissue macrophage

Schematic form of phagocytes by poly morphonuclear leukocyte (PMN) and tissue macrophage after penetrating skin and the pathogen bacteria entry to the deeper part of the tissue. PMN are more efficient in phagocyting than macrophage. Attention to PMN which are mobilized to the tissue and vascular in inflammatory response

Natural Immunity
Preventing of entry Intact skin Mucous membrane normal flora Defense for attacking Humoral mechanism Cellular mechanism as phagocytes, killing microorganism with : Oxidize intra cellular ADCC Cytokine

 The last mechanism is very various depend on the

etiology Consist of : Immunity to bacterial infection : toxin extra cell intracellular Immunity to viral infection Immunity to fungal infection Immunity to bacterial toxin
ec : C. tetani, V. Cholera, The most responsible is IgG

C. Diphtheria

Plasma cell

Activating lymphocyte Organism Toxin Antitoxin Ab-Ag complex

Toxin degradation PMN Macrophage

Schematic form of immunology mechanism in neutralizing toxin by antibody. Toxin-antitoxin complex, which is neutralized, is showed being ingested and destroyed in two type of phagocyte cells

Immunity to Extra cellular Infection

Through specific immunoglobulin :
IgG & IgM : Opsonisasi IgA for bacterial inside the lumen IgM and Ig (1,2,4) through C lyses IgG & IgM : agglutination phagocytes Inhibit Fe uptake by bacteria IgE at mastocyt cell histamine Bacterial motility <<<

Immunity to Intracellular infection

Phagocytes & humoral immunity is not effective CMI with APC pathway CD4 Cytokine Activation of CD8 & CD4 as cytolysis cell Another cytolytic cell is NK cell

Immunity to Viral Infection

Vi is non cellular-organism, always intrasel with way :
Immune system Infection Various of membrane Antigen Moving antigen

Interferon AMI CMI

Function : Delayed viral replication (type I = & ) Activation immunity system (Type II = )

Working Mechanism of Interferon

Virus

dsRNA

Host Cell
mRNA

(A)
Interferon receptor

Gene activation

NK activation, macrophage activation, increase expression of MHC molecules

IFN SYNTHESIS

IFN

IFN

IFN

(B)
Protein kinase dsRNA Phosfhorylated eiF2

mRNA Protein synthesis 2,5 A Synthesize 2,5 A Activated endonuclease

Inhibit protein synthesis

FIGURE

FIGURE

Figure :
Proposed mechanisms of induction of interferon synthesis and production of resistance to virus infection. Cell (A) is induced to produce interferon (IFN) by the presence of double stranded RNA (dsRNA).

The interferon ( or depending on

the type of cell) is released and binds to receptors on other cells.

The interferon ( or depending on the type of cell) is released and binds to receptors on other cells. This interaction can cause activation of host effectors functions and induce an intiviral state in neighboring cells (B). mRNA = messenger RNA; 2, 5-A = 2, 5 oligoadenylate.

Working Mechanism :
(2 5 oligoadenylate synthetase-inactive) + ds RNA 2 5 oligoadenylate synthetase Active

(endonucleaseinactive)

endonuclease

Active

RNA degradation

Protein Kinase + ds RNA

elF 2 active (elF 2 INACTIVE) Impaired protein synthesis

Mx protein (and its analogues in other species) Specific influenza virus inhibition in mice

Specific Antibody : Delayed mix with receptor Making immune complex Stimulating viral coagulation

AB was not effective for intracellular viral Could changed membrane cell Antigen Example : Oncogenic vi., Vaccinia vi., Influenza vi. Paramyxo vi., Toga virus, Papova, Rubella, Rabies. Form main defense on viral infection. The effectors is Tc (CD 8 & cd 4).

Immunity for Fungal Infection

Manifestation of Fungal Infection :

Superficial mycosis /cutaneus Subcutaneous mycosis Systemic mycosis

Defense Mechanism ? AMI or CMI ? Cutaneus as DTH Subcutaneous and systemic depend on activity from neutrophyl, macrophage, lymphocyte, NK cell ?

Although we have immunity to fungal infection :

Patient with neutropenia easy to get infection as: Candidiasis, Aspergilosis, Zigomycosis. Patient with CMI disturbance easy to get infection as : Cryptococcosis, Histoplasmosis, Coccidiomycosis

Basic of Immunoprophylactic Knowledge of the immune system Immune Response Defense mechanism like AMI & CMI R.I have response of memory. Process : Immunization Active immunization Passive immunization

Active Immunization
Immunity gets actively.

Requirement :

Immune System must be normal. Booster

The immunogen : vaccine consist of :

Conventional vaccine : Toxoid Killed Vaccine Subunit Vaccine Attenuated Living Vac.

Genetic device vaccine Ex. : Hepatitis B

Vaccine Preparation :
Bacteria cell : Pertusis, Typhoid, BCG Toxoid : Tetanus, Difteri Virus : Poliomyelitis, Morbilli, Rubella,Mumps Polysaccharide capsule : Pneumococcus, Meningococcus, H. Influenza type B

The successful of immunization depend on : Kind of Vaccine Booster Infection before How to give Immunization target in Indonesia : Neonatal until child with school age

Kind of Immunization : Obliged : Diphtheria, Pertusis, Tetanus (DPT) Tuberculosis (BCG) Polio (Sabin) Measles

Immunization Procedure
Each country is different.

Conditions to give immunization :

Less protection The disperse specific antigen rate is highest. The biggest risk.

Immunization Program in Indonesia

Variety of vaccine Immunization Count Time Interval (Weeks)
4 8 6 8 -

Age (months)

Basic immunization
BCG DPT Polio Measles

1x 3x 4x 1x

0 11 2 11 2 11 9 15 Table

Variety of vaccine

Immunization Count

Time Interval (Weeks) -

Age (months)

Booster :
DPT Polio 1 x 1 x 1,5 2 1,5 2

DT Td

1 x 1 x

4 6 12 14
(every 10 yrs)

Suggestion Immunization
MMR
Hepatitis B

1 x
3 x

>1 years
Anytime
(every 5 yrs)

Immunization Program at Posyandu/ Puskesmas

Variety of Immunization
BCG, DPT I, Polio I HB I, DPT II, Polio II HB II, DPT III, Polio III HB III, Measles, Polio IV

Age
2 months 3 months 4 months 9 months

Direct giving AB which needed :

Homolog AB Heterolog AB Autolog AB

Specific Antibody

Maternal Gamma Globulin Heterolog Antibody

N Immunity

Ag

Tolerance

Hypersensitive

Allergen

Allergy

Coombs & Gell :

I, II, III IV V : A.M. Hypersensitivity : DTH + C.M.Hypersensitivity : Stimulatory

Type I = Anaphylactic
Ag Ag

IgE
Vasoactive amin

Degranulation Bronchus autonomy muscle Capiler activation

Amine vasoactive substances : histamines slow-reacting substance of anaphylaxis (SRS-A) ECF-A serotonine

Substances effects to arachidonic acid metabolism : leukotriens (LTC4 & LTD4) prostaglandin tramboxan

People with possibility to hypersensitive reaction : Atopi

Type II reaction : cytotoxic

Binding between Ig G & Ig M (FAB) with cell antigen : phagocytes cytotoxic lysis e.c. : isoimmune reaction autoimmune reaction drugs reaction

Type III reaction : Immune Complex

Antigen-antibody complex abundant elimination not perfect precipitate in tissue and vascular blood Antigen can produce from ; Pathogen persistent infection Inhalant antigen Autoimmune disease Anomaly because immune complex depends on : Absolute rate antigen-antibody complex Antigen-antibody proportion Antibody >>> local Antigen >>> systemic

Type IV reaction : CMI : DTH

cell

Tissue damage
e.c. : Contact dermatitis Tuberculin test Tissue rejected

Type V reaction
Antibody CTC induce self tissue, e.c. thyroid tissue

Secretion

XIII.Autoimmunity and autoimmune disease

Body complement Self Immune system Antibody (Autoantibody) Autoimmunity Autoimmune disease

Not Self

Pathogenesis
1. Forbidden-clone theory 2. Sequestered-antigen theory 3. Immunodeficiency

4. Go away from T cell tolerance

5. <<< T suppressor cell function

Forbidden-Clone theory

Positive mutant as antigen Normal lymphocyte Destroyed by normal lymphocyte

Normal lymphocyte

Negative mutant as antigenic (Forbidden-Clone)

Survive, become sensitive and attack target tissue

Immune Deficiency Theory

Positive mutant as antigenic

Immunoglobulin deficiency lymphocyte Positive mutant as antigenic Position & negative mutant attack tissue target

Microbe pathogen survive

Antigen change Or unknown antigen release

+ Antibody

Cellular injury (Type IV) Or + Complement Injury mediated by antigen Injury immune complex (Type III)

THYROID EMBRYONIC EMBRYONIC LYMPHOID CELL

Antigen surfaces

Unknown antigen

Unknown antigen that affected by before

ADULT THYROID

Sensitized lymphocyte

Activated autoimmune process

Spectrum of Autoimmune Disease

Variety of Diseases Hashimotos Thyroid Primary Miksedema Graves disease Antigen Tyro globulin Surface cell TSH receptor HLA Link DR 5 DR 3 DR 5 Relative Risk 3,2 5,7 3,7

Diabetic autoimmune

Islet cells

DR 5, DR 4, DR 3/4

5,0 - 6,0 14,3

Variety of Diseases

Antigen

HLA Link

Relative Risk

Goodpasture Syndrome
Primary Cirrhosis Billiary Colitis ulcerative

Glomerulus Basal membrane of lung Mitochondria

DR 2

13,1

Colon lypopolisacharida

Rheumatoid arthritis
S.L.E.

Ig G
nucleoprotein

DR 4
DR 3

4,2
5,8

Pioneer :
Bruton found the 8 yrs child who has hypogamaglobulinemia. As clinical squelae S.I. Disturbance.

Genetic Metabolic & Biochemistry deficiency Vitamins & mineral deficiency Disturbance Embryogenesis Autoimmune diseases Acquired immunodeficiency.

B Cell Immunodeficiency T Cell Immunodeficiency B Cell & T Cell Immunodeficiency (combined) Phagocytic Dysfunction