ISBB // MAVM 1

IgG IgA IgM IgD IgE

150k Da 160 to 140k Da 900k Da 180k Da 190k Da
7s 7s 19s 7s 8s
23 days 5 days 6 days 1 to 3 days 2 to 3 days
4 domains 4 domains 5 domains 4 domains 5 domains
✓ C’ fixation ✓ Alternative C’ ✓ C’ (most) ✕ ✕

Warm reacting Major in secretions Cold reacting Immunoregulation Reagenic Ab

Immunity for Serum = 1st in phylogeny, Affinity to baso +
✓ B-cell surface
newborn monomer last in senescence mast cells
2O immune Allergy +
Best in C’
response Secretory = dimer Anti-idiotypic Ab parasitic infection
✓ Precipitation ✓ Agglutination Eos (MBP)

MATURE B-CELL IgM + IgD (MD) XTRA HINGE region IgM + IgE (ME)
✓ placenta IgG (✕IgG2) ✓ complement IgG (✕IgG4)
 ISBB // MAVM 2

Type I HS Type II HS Type III HS Type IV HS

IgE IgG IgG/IgM T-cells
✕ ✓ C’ ✓ C’ ✕
Mediators from mast
Cytolysis due to Ab + C’ Ag-Ab complexes Release of cytokines
cells + basophils
Histamine; Allergens Bound Ag Unbound Ag Delayed Type HS
Anaphylaxis Transfusion reactions Serum sickness Contact dermatitis
Hay fever AIHA Arthus reaction Tuberculin test
Asthma HDN SLE Poison Ivy

PA-PA-IN PEP-SIN
Cut at the hinge region Cute below the HR
2 FAB + 1 FC F (AB) + 1 FC
2
 ISBB // MAVM 3
AUTOIMMUNE DISEASE
Thyroglobulin
HASHIMOTO’S Microsomal Ag GOODPASTURE Type IV collagen

GRAVE’S TSH receptors MYASTHENIA GRAVIS Ach receptors

CAH Smooth muscles PERNICIOUS ANEMIA Parietal cell Ag, IF
SJOGREN’S Salivary gland nucleolar PRIMARY BILLIARY
Mitochondria
SYNDROME Ag CIRRHOSIS

SYSTEMIC LUPUS Disease of CT

Butterfly rash/red wolf
ERYTHEMATOSUS (SLE)
Chronic inflammation of joints +
RHEUMATOID ARTHRITIS periarticular tissue
(+) ANA (anti-dsDNA, anti-SM)
✓ LE cell
Igs + C’
IgM against the Fc portion of IgG
(+) Anti-cyclic citrullinated peptide
 ISBB // MAVM 4
Fluorescent staining patterns
RA

HOMOGENOUS Anti-dsDNA
SLE
Sjogren’s syndrome
MCTD

PERIPHERAL Anti-dsDNA
SLE
Sjogren’s syndrome
SLE
SPECKLED Anti-Smith RA
MCTD

NUCLEOLAR Anti-RNP Scleroderma

CREST (Calcinosis, Raynaud's

DISCRETE Anti-centromere
phenomenon, Esophageal
dysmotility, Sclerodactyly, and
Telangiectasia)
 ISBB // MAVM 5
Force of attraction bet. 1 Sum of all attractive forces
AFFINITY Fab + 1 epitope
Covalent = strong bond
AVIDITY bet. Ag and Ab
Avidity = Dissociation

Contact with
LAG immunocompetent cells LOG Ab production
Ab production = Ab Ab degradation
STATIONARY degradation DECLINE Titer falls

Initial exposure Subsequent exposure

PRIMARY Longer Lag phase

Ab production SECONDARY Shorter Lag phase
Ab production

RESPONSE IgM RESPONSE IgG

 ISBB // MAVM 6
Unknown Ag + Unknown Ab +
DIRECT/FORWARD known anti-sera INDIRECT/REVERSE known Ag
Heat at 56OC for Heat at 56OC for
INACTIVATION 30 minutes REINACTIVATION 10 minutes

PRIMARY SECONDARY TERTIARY

Combination of Ag-Ab Demonstrates Ag-Ab reaction Immunologically in vivo
Non-visible reaction Visible reaction Biologic reaction is detectable

PRO-ZONE POST-ZONE FALSE (-)

Excess Ab Excess Ag Remedy with dilution
 ISBB // MAVM 7
Precipitations | soluble antigens
TURBIDIMETRY NEPHELOMETRY
Turbidity/cloudiness Light scattered LIGHT SCATERRING
Spectrophotometer Nephelometer

PASSIVE IMMUNODIFFUSION | Agarose; Longer TAT

OUIDIN TEST
Single diffusion, dimension
(+) Precipitin line
FAHEY + KCKELVEY
RADIAL IMMUNODIFFUSION OUCHTERLONY TECHNIQUE
Single diffusion, double dimension Double diffusion, dimension
(+) Precipitin ring (+) Smooth curve [Identity]
Kinetic End point
- Log. of the conc. MANCINI METHOD Diameter squared
 ISBB // MAVM 8
ELECTROPHORETIC TECHNIQUES | Separate molecules using electrical current
ROCKET IMMUNO (IEP) COUNTER (CIE)
RID + electrophoresis Double diffusion + electrophoresis Ag + Ab place on the well
peak = conc. ID of monoclonal proteins (BJP) opposite each other
One dimension electroID Grabar and Williams Albert and Johnson

AGGLUTINATIONS | PARTICULATE antigens

DIRECT agglutination Ag on the surface

PASSIVE agglutination Ag on the carrier

REVERSE PASSIVE agglutination Ab on the carrier

COAGGLUTINATION Bacterium as carrier

AGGLUTINATION INHIBITION
NEUTRALIZATION Ag activity is stopped by specific Ab
 ISBB // MAVM 9
SYPHILIS
PRIMARY SECONDARY LATENT TERTIARY
Hard chancre (painless) Condylomata Asymptomatic Gummas
Darkfield Mx Neurosyphilis
Darkfield Mx (+) Serologic test
Serologic tests Congenital

Hutchinson’s teeth ✓ Congenital syphilis

HUTCHINSON TRIAD Interstitial keratitis
Nerve deafness
Other S/S: fissuring (mouth, anus),
saddle nose, skeletal lesions

TREPONEMAL
FTA-ABS Fluorescent
Treponemal Ab Absorption Test
HATTS Hemeagglutination
Treponemal Test for Syphilis
TPI T. pallidum Immobilization Test
Indirect Fluorescent
Immunoassay
Hemagglutination
Ab against T. pallidum + C’ =
immobilize treponemes
Nichol’s strain (slide)
Reiter treponemes (absorbent)
Sheep RBC
(+) >50% immobilized
treponemes
 ISBB // MAVM 10
NON-TREPONEMAL
VDRL RPR
Heated serum, microscopically Unheated serum, macroscopically
Flocculation Flocculation
Cardiolipin + cholesterol + lecithin Cardiolipin + Lecithin + Charcoal + Choline chloride
+ Thimerosal

QUALI 180 RPM / 4 MINS. 18 G 60 DROPS

VDRL

SERUM
QUANTI 180 RPM / 4 MINS. 19 G 75 DROPS
CSF 180 RPM / 8 MINS. 21/22 G 100 DROPS
RPR 100 RPM / 8 MINS. 20 G 60 DROPS
 ISBB // MAVM 11
GROUP A STREPTOCOCCAL INFECTIONS
ü Streptococcus pyogenes (beta-hemolytic)
ü Acute Glomerulonephritis, Rheumatic fever
ü STREPTOLYSIN O: subsurface hemolysis, higly immunogenic, O2 labile

ASO TITRATION Neutralization

(+) Highest dilution with no hemolysis
Children: <125 Todd units
Adults: <166 Todd units

RAPID LATEX AGGTN Passive agglutination >200 IU/mL

 ISBB // MAVM 12
FEBRILE AGGLUTININS
Typhoid fever
WIDAL TEST Slide: Direct agglutination
Tube: High dilutions showing 2+
>80 Todd units

TYPIDOT IgG/IgM to Salmonella typhi

IgG: past; IgM: current
IgG + IgM: reinfection

WEIL FELIX Abs against rickettsial diseases

Direct agglutination
RMSF: (+) OX-19, OX-2 (P.V)
Scrub: (+) OX-K (P.M.)

BRUCELLOSIS Agglutination Cross react with F. tularensis

LYME DISEASE Erythema chronicum migrans

Bull’s eye rash
Borrelia burgdorferi (Ixodes)

1O ATYPICAL PNEUMONIA Hemeagglutination

IgM Abs target I Ag on RBC
Cold agglutinins
M. pneumoniae (Eaton’s)
 ISBB // MAVM 13
HEPATITIS
A Infectious (Picornaviridae), short incubation
Most common type; Serum: Anti-HAV; RIA/ELISA: HAV
Fecal-oral

B 1st: Ouchterlony; 2nd: CEP, Rheophoresis, C’

3rd: Reversed Passive Latex/Hemeagglutination, ELISA, Radioimmunoassay
Parenteral,
vertical, sexual

C Surrogate: ALT + anti-HBc

ELISA/RIA: Anti-HCV; RIBA: HCV Ab; RT-PCR: HCV RNA
Blood transfusion
Co-infection and superinfection with HBV
D Co-infection: (+) Anti-HBc IgM; Superinfection: HBV HDV
Parenteral,
sexual

E Water borne Hepatitis

Self-limiting
Fecal-oral

Best indicator
High degree of
HBsAg of early acute HBeAg infectivity HBcAg Hepatocytes
infection
1st serologic IgM: window
Immunity;
Anti-HBs Viral clearance Anti-HBe evidence of Anti-HBc IgG: lifelong
convalescent marker
 ISBB // MAVM 14
INFECTIOUS MONONUCLEOSIS | Kissing disease / Glandular fever
ü Epstein-Barr Virus
ü Targets B-lymphocytes (CD21)
ü (+) Downey cells

PAUL BUNNEL Screening

Hemagglutination
2% sheep RBCs

Davidson Absorption-Hemagglutination
Indicator: sRBCs
Guinea pig cells + beef RBCs

MONOSPOT Absorption-Hemagglutination
Indicator: hRBCs
More senstive

IM FORSSMAN SERUM SICKNESS

ADSORPTION (+) Beef RBCs (+) Guinea Pig Cells (+) Beef RBCs, Guinea Pig Cells

AGGLUTINATION (+) GPK (+) Beef cells (+) GPK

BI // GF // SS
 ISBB // MAVM 16
CD2, CD3 CD19
T-CELLS CELL MEDIATED B-CELLS HUMORAL IMMUNITY

Antigen Antibody
ACTIVE Long term + slow response PASSIVE Short term + immediate response

T-helper T-suppressor CD56, CD16

CD4 MHC II CD8 MHC I NK LGL