Fundamentals of

Immunology
Prepared by:
Winona Mei A. Reyes, RMT
 Roles of the Immune System
✔ Protection from pathogens and foreign substances
✔ Removal of abnormal and damaged host cells
Mechanism of the Immune System

• Recognition of self and nonself

cells, tissues, organisms
• Removal of unwanted cells, tissues,
organisms
• Repair of damaged host tissues
Branches of IS
 2 Arms of Acquired IS

2. Cellular: 2. Humoral:
T cells and lymphokines B cells and Ab production
 Acquired Immunity in
Transfusion Medicine
• Antibodies DO NOT always remain in plasma at levels
observable with serologic testing, and if antigen-positive
RBC units are transfused in a sensitized patient, the
second antibody response against the transfused cell
antigens can be more vigorous, resulting in intravascular
RBC hemolysis :(
SENSITIVITY LEVEL

“Antibodies DO NOT
VALUES LESS always remain in plasma at
THAN THE
levels observable with
SENSITIVITY LEVEL
serologic testing.”
What occurs in
the primary
response?
What occurs in the primary response?
What occurs the next time the immune system
encounters the same antigen?
What occurs
the next time
the immune
system
encounters
the same
antigen?
What occurs
the next time
the immune
system
encounters
the same
antigen?
Primary Antibody Response VS Anamnestic Antibody Response
 Cell Lineages and Markers
• Pluripotent hematopoietic stem cell progenitors (CD 34+)
• Myeloid or the lymphoid lineage
• Granulocytes: all have 3 receptors (1) Fc portion of IgG (2)
C5a (3)CR3
– Neutrophils (purple color)
– Eosinophils (reddish orange color)
– Basophils (bluish black color): receptor for Fc portion of IgE
• NK cells (Large granular lymphocytes): CD16, CD56, CD94
antibody-dependent cell mediated cytotoxicity
Cell Lineages and
Markers

• Pluripotent
hematopoietic
stem cell
progenitors
(CD 34+)
Cell Lineages
and Markers

• Myeloid or the
lymphoid
lineage
 Cell Lineages
and Markers
• Granulocytes:
All have 3 receptors (1) Fc
portion of IgG (2) C5a
(3)CR3
– Neutrophils (purple color)
– Eosinophils (reddish
orange color)
– Basophils (bluish black
color): receptor for Fc
portion of IgE
Wright-Giemsa Stain and Granulocytes
 Cell Lineages
and Markers
• NK cells
(Large granular
lymphocytes):
CD16, CD56, CD94
antibody-dependent
cell mediated
cytotoxicity
Lymphocytes
h
t
t
. Properties B-Cells T-Cells . Properties B-Cells T-Cells
p
s
:
/
/
m 1 Name B lymphocytes T lymphocytes 9 Secretion They secrete antibodies They secrete
i
c
r
Lymphokines
o 2 Origin Bone Marrow Thymus
b
i
o
10 Function В-cells form humoral or T-cells form
l 3 Position Outside Lymph Node Interior of Lymph Node
o
g
antibody-mediated cell-mediated immune
y
i
4 Membrane BCR (= immunoglobulin) TCR for antigen immune system (AMI). system (CMI).
n
f
o
.
receptor for antigen 11 Blood 20% of lymphocytes 80% of lymphocytes;
c
o
m CD4 > CD8
/
d
i
f
5 Connections B-cells can connect to T-cells can only connect 12 Formation They form plasma cells They form killer, helper
f
e antigens right on the to virus antigens on the and memory cells. and suppressor cells.
r
e
n
surface of the invading outside of infected cells.
c
e virus or bacteria. 13 Movement to Plasma cells do not move Lymphoblasts move to
s
-
b
Infection Site to the site of infection. the site of infection.
e
t
6 Tissue Germinal centres of Parafollicular areas of
w
e Distribution lymph nodes, spleen, cortex in 14 Function Plasma cells do not react Killer cells react against
e
n
-
gut, respiratory tract; nodes, periarteriolar in against transplants and transplants and cancer
b
- also subcapsular and spleen cancer cells. cells.
c
e
l medullary cords of lymph
l 15 Function Plasma cells have no Suppressor cells inhibit
s nodes
-
a inhibitory effect on immune system.
n
d
-
7 Life Span Life span is short Life span is long immune system.
t
-
c
e 8 Surface Surface Antibodies Absence of surface 16 Function They defend against They defend against
l
l
s
Antibodies present antibodies viruses and bacteria that pathogens including
/
enter the blood and protists and fungi that
9 Secretion They secrete antibodies They secrete lymph. enter the cells.
 Lymphocytes
B cell T cell
• Plasma cell • MHC class II
• B memory cell molecules and APCs
• Nonreactive, • Th cell
opsonization, • Tsup cell
complement activated • Tc cell
antigen-antibody complex • T memory cell
Lymphocytes
Immune system organs
MHC / HLA
MHC / HLA • HLA-DR, HLA-DQ, HLA-DC
• APCs
 APCs
• B cells
• Neutrophils
• Macrophages
– Langerhans cells
(Skin)
– Glial cells
(Nervous tissue)
– Kupffer cells
(Liver)
– Osteoclasts (Bone)
 Cytokines

• Lymphokines
• Monokines
• Chemokines
• Interleukins
• Interferons
• TNF
IMMUNOGLOBULINS
IMMUNOGLOBULINS
IMMUNOGLOBULINS
 Antibody
• Naturally occurring or isoagglutinins
– ABH, Hh, Ii, Lewis, MN, and P
• Immune
– Rh, Kell, Duffy, Kidd, and Ss
• Unexpected antibodies:
all other antibodies that react with RBC antigens
• Alloantibody:
produced after exposure to nonself antigens (RBC, WBC,
platelets)
• Autoantibody:
warm or cold; DAT or direct Coomb’s test;
special adsorption and elution techniques
 Immunoglobulins in
Transfusion Medicine
• Antibody screening
• Rh typing, passive anti-D antibody
• Anti-IgA antibody: can increase effect of IgG induced RBC
hemolysis
• IgE: histamine and urticaria
• Sulfhydryl reducing reagents such as
2-mercaptoethanol (2-ME) or ß-dithiothreitol (DTT)
 Complement
system
1. Final lysis of abnormal or
pathogenic cells via binding
of antibody
2. Opsonization and
phagocytosis
3. Mediation of Inflammation
 Complement System
▪ 3 Pathways for Complement Activation:

a) CLASSICAL PATHWAY
b) ALTERNATIVE PATHWAY
c) LECTIN PATHWAY

• 3 main stages of activation:

1st: RECOGNITION UNIT
2nd: ACTIVATION UNIT
3rd: MEMBRANE ATTACK COMPLEX
 THE CLASSICAL PATHWAY
• Ab-directed mechanism
• IgM, IgG1, IgG2, IgG3
• Others:
– CRP, Viruses, Mycoplasmas,
Gram-neg (e.g. E. coli)
• Recognition unit: C1 (qrs)
• Activation unit: C2, C3, C4
– End prod.: C5 convertase
(C4b2a3b)
• MAC: C5-C9

RECOGNITION UNIT
THE CLASSICAL PATHWAY

• MAC mechanisms:
– Channel formation
– Binding of
phospholipids to
decrease membrane
integrity
THE CLASSICAL PATHWAY
 THE ALTERNATIVE PATHWAY
Triggered by bacterial cell walls (lipopolysaccharide), fungal cell
walls, yeast, viruses, virally-infected cells, tumor cells,
Trypanosomes.
Unique constituents: PROPERDIN, Factor B, Factor D
 THE LECTIN PATHWAY
By the use of MANNOSE/MANNAN-BINDING LECTIN that binds to
mannose or related sugars found in glycoproteins or CHO of wide
variety of microorganisms.
Unique constituents: MASP-1, MASP-2, MASP-3
 Complement system in
Transfusion Medicine
• Evaluation of C3b and C3d complement components in
transfusion medicine testing procedures is useful in the
investigation of hemolytic transfusion reactions and
autoimmune hemolytic anemias.
• Intravascular vs Extravascular RBC hemolysis
Intravascular VS Extravascular
Hemolysis
Intravascular VS Extravascular
Hemolysis
 Factors that Influence
Agglutination Reactions
• Centrifugation
• Antigen-antibody ratio: there should be a loss of
equivalence
– Prozone
– Postzone
• pH effect: pH 6.5 to 7.5 (Why?)
– Acidification of serum: anti-M, anti-Pr
• Temperature
– 22ᵒC, IS phase
– 37ᵒC, AHG phase
 Factors that Influence
Agglutination Reactions
• Immunoglobulin type
• Enhancement media: decreases zeta potential
– LISS: 0.2% NaCl
– Protein media: PEG, polybrene
• Proteolytic enzymes
– Ficin, papain, trypsin, bromelin
– ENHANCE: Rh, Kidd, P, Lewis, I
– DESTROYS: Fya, Fyb, M, N, S
Why are RBC antigen either destroyed or
enhanced by enzymes?
 Factors that Influence
Agglutination Reactions
• AHG reagent: designed to determine if RBCs are coated
with Ab or complement or both; no agglutination but only
sensitization, AHG will allow cross-linking of Abs on the
RBCs
– Polyspecific: IgG, C3b/C3d
– Monospecific: IgG or complement
• Chemical Reduction of IgG and IgM molecules
– IgM: Dithiothreitol (DTT), B-2-mercaptoethanol (2-ME)
– IgG: ZZAP
 Traditional Laboratory Testing Methods

• Hemagglutination
1. Sensitization:
Antigen-antibody binding
2. Lattice structure : Multiple
antigen-antibody bridges
form
• Agglutination inhibition
• Hemolysis
 Nontraditional Laboratory Methods

• Flow Cytometry:
– Fluorescent-labeled antibody emit fluorescent color of a
specific wavelength
– Quanti and quali data and sort cell populations
– Direct procedure (1ᵒ Ab), Indirect procedure (2ᵒ Ab)
– Fetalmaternal hemorrhage, identify transfused cells,
cell-bound IgG, homozygous and heterozygous expression
of blood group antigens
 Nontraditional Laboratory Methods
• Flow Cytometry:
1. Scattering of light as cells are bathed in a fluid stream through
which a laser beam enters
2. Cells move into a chamber one at a time and absorb light from
laser
3. Distinction between different wavelengths of light
 Blood Product Transfusions and
the Immune System
• Transfusion-related immunomodulation (TRIM):
depression of immune system
• Clonal deletion: lymphocytes are inactivated or removed
• Immune suppression: cytokines
• Leukoreduction Filters