IS

IMMUNO
phagocytes: MEND (baso not phago)
APC: MBD (mono, macro, B cells, dendritic)

cytokine major producers

IFN-a: produced by null Lymphocytes / NK cells
IFN-B: fibroblast, epithelial cells
TNF-a: macrophage
TNF-B: lymphocytes (CD41 & CD8)

COMPLEMENT PATHWAYS
Classical (142356789) & Lectin
C3 convertase - C4b2a
C5 convertase - C4b2a3b
*but in Lectin walang C1, kundi MASP at MBP

Alternative/Properdin (no C1,4,2)

C3 convertase - C3bBb
C5 convertase - C3bBb3b

*Factor B - binds to C3b

*Factor D - cleaves Factor B into Bb fragment
*Factor P (Properdin) binds to C3bBb to prevent sponty decay

INHIBITORS
Factor I - cleaves C3b & C4b
Factor H - compete for Factor B (nakikipag-agawan)
S protein or vitronectin - prevent maattach si C5b67 sa cell membrane

C456789: Membrane Attack Complex

MEASUREMENT ASSAY of Complement

1. CH50 Assay
2. Radial Immunodiffusion
LYMPHOCYTE DEVELOPMENT
production - bone marrow
maturation - thymus (T) & bone marrow (B)
proliferation & differentiation - secondary lymphoid organs like spleen, lymph
nodes

T CELL DEVELOPMENT
double negative -> double positive -> mature -> activated -> sensitized

CD2 - appears first, remains present as identifying marker of T cells (loyal)

CD3 - for pre-T cell receptor
CD25 - produced by Activated T cell for IL-2 expression

B CELL DEVELOPMENT
pro-B -> pre-B -> immature -> mature -> activated -> plasma cell

Pro-B: CD19, CD45, c-Kit ligand, mu heavy chain rearrangement on

Chromosome 14
Pre-B: complete heavy chain, encoding of light chains (Kappa - Chromosome 2,
Lamba - 22)

CD25 - exhibited by Activated B cell also

plasma cells - most differentiated B cell; actual na nagpproduce ng antibodies;

Abs makikita sa cytoplasm unlike sa mature T cells na nasa surface ang Ig

MHC
– genes found on short arm of Chromosome 6 (codominant)
– taga-deliver or present ng antigen (component ng APC) sa T cells
Class 1: present in ALL nucleated cells, process intracellular or cytoplasmically
derived antigens (e.g, viruses, cancer)
Class 2: present in surface of APCs— Macrophages, monocytes, B cells,
dendritic cells, process extracellular derived antigens

*B cells are not phagocytic, but they have antibodies in surface so it it can
recognize antigens which will then be processed by MHC Class 2 and be
 presented

Class 3:
– complement C2, C4, Factor B, TNF

HLA assoc diseases

B27 = ankylosing spondylitis
DR3 = multiple sclerosis
DR2, DR3 = SLE
DR4 = RA
DR3, DR4 = TYPE 2 DM

ANTIBODIES
IgG
subclass most efficient in:
placenta transfer: 1>3>4>2
complement fixation: 3>1>2 (4 cannot)

SEROLOGY
SECONDARY IMMUNOLOGIC TESTS
UNLABELED
Precipitation
Precipitation by Passive Immunodiffusion

Pi 1. Ouidin - single diffusion, single dimension SDSD

2. Radial Immunodiffusion - single diffusion, double dimension SDDD
Ouch 3. Ouchterlony - double diffusion, double dimension DDDD
-

Agglutination
1. Direct agglutination - Ag are found naturally on surface of particles
2. Passive agglutination - Ag are artificially attached to a carrier
3. Reverse passive agglutination - Abs are artificially attached to a carrier
4. Coaggulatination - carrier bacterium
5. Agglutination Inhibition
6. Antiglobulin Technique
7. Neutralization
Complement Fixation
– positive: no agglutination
– has many components: indicator cell, hemolysin, antigen reagent,
complement

PRIMARY IMMUNOLOGIC TESTS

LABELED IMMUNOASSAYS
Ligand immunoassay - use of mono or polyclonal Ab

1. Fluorescent Immunoassay or Immunofluorescent Assay (IFA)

● Direct IFA
(sample Ag + fluoresceinated Ab)

● Indirect IFA
(solid-phase Ag + sample Ab + fluoresceinated secondary Ab)

= both methods have direct relationship between fluorescence & sample conc

2. Radioimmunoassay
● Competitive RIA

⑭se
(solid-phase Ab + sample Ag + radioactive Ag)
= two Antigens will compete for the Fab site
= positive rxn is no or low radioactivity
= radioactivity is INVERSELY proportional to Ag conc
=
● Non-competitive RIA
(sample Ag + radioactive Ab)
X solid plas

di
= NO SOLID PHASE, Ag-Ab binding only
-

= positive rxn is high radioactivity

= radioactivity is DIRECTLY proportional to Ag conc

3. Enzyme Immunoassay
● Competitive/Direct ELISA
(solid-phase Ab + sample Ag + enzyme-labeled Ag)
= two Antigens will compete for the Fab site
= enzyme activity is INVERSELY proportional to Ag conc
 ● Non-competitive/Indirect ELISA
(solid-phase Ag + sample Ab + enzyme conjugated secondary antibody)
= enzyme activity is DIRECTLY proportional to Ag conc

note that Either Ag or Ab may be bound to solid phase.

● Capture/Sandwich ELISA
(solid-phase Ab + sample Ag + enzyme conjugated secondary Ab)
-
= will bind forming a sandwich, also DIRECT

[v
*Indirect IFA & Noncompetitive/Indirect ELISA - solid phase antigen Day preet
*NoncompetitiveC
RIA - no solid phase
*Competitive RIA & Competitive ELISA, Capture ELISA - solid phase antibody
= > Inverse
C
COMPETITIVE ASSAYS = activity is INVERSE to analyte conc

TERTIARY IMMUNOLOGIC TESTS ata

OTHER SEROLOGIC TESTS
FOR SYPHILIS
VDRL
– G
heated 50uL serum at 56°C for 30 mins
– principle: FLOCCULATION
–
–
S
reagents: cardiolipin, lecithin, cholesterol (CLC)
result read MICROSCOPICALLY-
– reactive = large clumps-
– false positive due to the ff: SLE, IM, Malaria, Pregnancy, RA (SLIMPR)

RPR
– unheated serum
*di na need i-heat, choline chloride is used to inactivate serum
– principle: FLOCCULATION
–
thimerosal (CLC CCT)
#
reagents: cardiolipin, lecithin, cholesterol, charcoal, choline chloride,
Luiswal Liractivate som
– result read MACROSCOPICALLY since charcoal is used for visualization
– reactive = large clumps
-
FTA-ABS
– confirmatory test for syphilis
– principle: INDIRECT IFA
– heated serum combined with Reiter strain which removes cross-reactivity
to other treponemes
– reagent: Nichol’s strain (attached & fixed to the slide)

HATTS
– principle: Hemagglutination
– -
reagent: glutaraldehyde stabilized turkey RBC coated with treponemal Ag

MHA-TP
– principle: Hemagglutination
– -
reagent: tanned formalin sheep RBC coated with treponemal Ag

TPI
– MOST SPECIFIC test for syphilis #
– reagent: live T. pallidum (which will immobilize if it reacts with patient
antibody & complement)
– positive result = >50% immobilized treponemes

TESTS FOR GROUP A STREP

1. ASO Titration
– *
principle: NEUTRALIZATION
– ASO Titer: reciprocal of highest dilution showing no hemolysis; expressed
in Todd units

2. Rapid Latex/ASO Latex Agglutination

– =
principle: PASSIVE AGGLUTINATION
– significant titer: >200IU/mL

E C
*most reliable of recent GAS infection, & for acute rheumatic fever &

-
glomerulonephritis - AntiDNASE B (neutralization)
#
FEBRILE AGGLUTININS
– O
for Typhoid fever, Typhus fever, Brucellosis, Tularemia (TTBT)
 1. Widal Test S .

typri
– for typhoid fever by Salmonella typhi (widal-sal)
– principle: DIRECT agglutination

2. Weil-Felix Frickettsial
– nonspecific test that antibodies against Rickettsial diseases (that cross-
reacts with Proteus Ags)
– principle: DIRECT agglutination
=
– uses Proteus antigens
– OX-19 & OX-2 of P. vulgaris (tunay teen ay vulgar = 2,19, vulgaris) emst
–
>
-

OX-K of P. mirabilis (kilometer mabilis)

– RMSF*
++++ with OX-19
-

types
– Scrub typhus-
++++ with OX-K
(kuya Weil gumamit ng Proteus para madetect ang Rickettsial diseases)
-

3. Typhus
4. Brucellosis
– agglutination test

5. Lyme
– confirmatory: Western blot
– other tests: IFA, EIA, PCR

6. Primary Atypical Pneumonia

PAP-cold agent of
– Rapid Screening test for Cold Agglutinins
–
-

principle: hemagglutination
– antigen: human Group O cell
– antibody: cold agglutinins in PX serum
– incubate tubes at 4C for 18-24 hrs
-

VIRAL
HEPATITIS
Detected in feces & Self-limiting = Hepa A, E
Sexual transmission = Hepa B, D
Blood/Parenteral transmission = Hepa B,C,D
HEPATITIS A (infectious/shorter incubation hepatitis)
F
IgM anti-HAV - onset
IgG anti-HAV - recovery & immunity

HEPATITIS B (serum hepatitis)

HBsAg - best indicator for early infection, Australian B11
- -

HBcAg - found in hepatocytes (wala sa serum)

HBeAg - highO
enfectivity
Anti-HBc
●=
IgM - indicator of current infection (useful for detecting infection during
window period)
●-
IgG - lifelong marker
Anti-HBe - recovery/convalescent
-

Anti-HBs - immunity or viral clearance of HBV

-

Anti
order: HBsAg → HBeAg → Anti-HBc → Anti-HBe → Anti-HBs seaces
Hepatitis C (posttransfusion hepatitis)
gold std test: RT-PCR

INFECTIOUS MONONUCLEOSIS (di ata lalabas sa compre)

Heterophile Antibodies
1. Paul-Bunnel Test
– HEMAGGLUTINATION

2. Davidson Differential Test

– ABSORPTION-HEMAGGLUTINATION
Heterophile Abs
● in Serum Sickness: sheep, beef, horse, guinea pig (SBHG) cells
● in IM: reacts with SBH (no absorption/high agglutination with Guinea
pig RBC)
● in Forssman: SHG (no absorption:high agglutination with beef RBC)

3. Monospot
– ABSORPTION-HEMAGGLUTINATION
 HIV
HIV markers
C -
p24 = core coat or major capsid protein; most abundant viral antigen detected
by ELISA; 1st
-
antibody produced is against p24
gp41 = transmembrane protein (gp120 kumakabit sakanya to form HIV
-

envelope)
gp120 =[
binds to CD4 on T cells (helper T cells) Not alls
gp160 = facilitates virus entry

HIV Screening: ELISA

1st gen detects Anti-HIV1 only
2nd gen to 4th gen detects Anti-HIV1&2

HIV confirmatory: Western Blot (positive result if 2/3 bands present)

ELISA at 6 weeks & 3 months after exposure

Western blot at 5 months after exposure

complement fixn - no hemolysis

aso, antidnase - neutralization (no agglu)
hepa c - PCR
rubella - ELISA
toxoplasma - EIA

ccr5
antimicrosomal antibodies - chronic hepatitis
antibodies to smooth muscle - autoimmune hepatitis

monocyte - precursor of dendritic