Post on 19-Oct-2020
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
Immunology-Serology and Blood Banking
HISTORY
YEAR PERSONS INVOLVED RESEARCH/EVENT
1492 First blood transfusion recorded in history
1500 Chinese Variolation; Insufflation
- material was dried into a powder and
inhaled by non-immune person
1700 Cross-immunity (exposure to cowpox and
immunity to small pox)
1880 Louis Pasteur Attenuation*, rabies vaccine
- bacteria/viruses are weakened
1888 Phagocytosis
1901 Serum antitoxins
1903 Almoth Wright Opsonizaton
1908 Immunity
- coined the term complement
1919 Explained the complement
1930 Human blood group antigens
The Specificity of Serologic Reactions
1943 Loutit and Mollison Acid-citrate dextrose (ACD)
1957 Citrate-phosphate-dextrose (CPD)
1972 Antibody structure
1977 Radioimmunoassay
1980 George Snell, Jean
Dausset, Baruj
Beneceraf
MHC
1984 T-cell receptor gene
1987 Antibody diversity
1991 Edward Donall
Thomas, Joseph
Murray
Transplantation
Cellular immunity to TB
Paul Ehrlich Side Chain Theory
Anaphylaxis
Immunoregulation
Monoclonal antibody (Hybridoma
technology)
Precipitins
Frank Macfarlane
Burnet
Clonal Selection theory
Gel test
IMMUNITY
☻ ___________ – study of how a host’s body discriminate between self and
foreign antigens and thereby eliminating these foreign substances.
☻ __________ – state of being resistant to infections
☻ __________ – lack of immune response to self-antigens
Innate/Natural/
Nonspecific
Adaptive/Acquired/ Specific
CELLULAR Antigen-presenting cells (eg.
Dendritic cells)
Phagocytes, eosinophils,
basophils, mast cells
NK cells
T-cells (T-cytotoxic and T-
helper)
B-cells and plasma cells
HUMORAL Lysozymes, Lactoferrin
Acute phase reactants
Complement, Properdin
Pepsin, stomach acidity
Cytokines (TNF, INF, beta-lysin
Antibodies
Cytokines
RESPONSE Rapid but short (less potent) Slow but longer (more potent)
MEMORY NO YES (because of memory cells)
SPECIFICITY General Specific antigens
OTHERS *with anatomical barriers – skin (pH 6.5), mucus membranes, normal flora, tears,
cilia, saliva
INNATE IMMUNITY ______________ – process of engulfment of substances to be discriminated; done by
cells called phagocytes
(1) ________ → (2) ________ → (3) ___________ → (4) ________ → (5) ____________
____________ – process in which blood cells pass through the intact walls of
blood cells migrating to the site of injury
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
TWO FORMS OF PHAGOCYTOSIS
1. Direct Phagocytosis
2. Indirect Phagocytosis
☻ Mediated by opsonins – antibodies (IgG/IgM), CRP, complement
breakdown products
☻ _____ – most potent opsonin
_______________ – cumulative mechanism (vascular and cellular) in respose to an
injury or invasion by an infectious agent
(1) Vascular Response → (2) Cellular Response → (3) Resolution and Repair
CARDINAL SIGNS:
☻ ______ – redness
☻ ______ – heat
☻ ______ – swelling
☻ ______ – pain
☻ Functio Laesa – loss of function
_________________________ – serum proteins that significantly increase during course
of an inflammation
APR FUNCTION INCREASE RESPONSE TIME (hrs)
Opsonization 1000x 4-6
Cholesterol removal 1000x 24
Ceruloplasmin Binds copper 2x 48-72
Complement C3 Opsonization, lysis 2x 48-72
Alpha1-antitrypsin Protease inhibitor 2-5x 24
Fibrinogen Clot formation 2-5x 24
Haptoglobin Binds hemoglobin 2-10x 24
________________ – proteins that are normally present in serum that has its functions
during inflammation.
FUNCTIONS:
☻ For opsonization (C3b – most potent)
☻ For cell lysis (C5-C9 or MAC)
☻ Clearance of immune complexes
COMPLEMENT
Protein Function Disease (if deficient)
C1q, r or s C1q – binds Fc portion of IgG or IgM Lupuslike syndrome
Recurrent infections C1r – activation of C1s
C1s – cleaves C4 and C2
C4 Part of C3 convertase (C4b2a) Lupuslike syndrome
Pathogen/Microbe
Associated
Molecular Pattern
(PAMPs/MAMPs)
Primitive pattern
recognition
receptors (PPRR)
Acute infections:
PMNs
Chronic infections:
Monocytes
CLASSICAL AND LECTIN PATHWAY ALTERNATIVE PATHWAY
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
C2
(most
common def.)
Binds to C4b to form C3 convertase
of classical pathway
Lupuslike syndrome
Recurrent infections
Atherosclerosis
Pivotal point Severe recurrent
infections
Glomerulonephritis
Part of Membrane Attack Complex Neisseria infections
C8 – starts pore formation
C9 – eventual cell lysis No known disease
Binds C3b to form C3 convertase
Cleaves factor B
Stabilizes C3bBb complex (C3
convertase of alternative complex)
Neisseria infections
Similar to C1q; binds to mannose Pneumococcal diseases
Sepsis
Neisseria infections
MASP-1 Similar to C1r
MASP-2 Similar to C1s Pneumococcal diseases
REGULATORY PORTEINS
Dissociates C1r and C1s from C1q
thus inhibiting it
Hereditary angioedema
Factor I Cleaves C3b and C4b Recurrent pyogenic
infections Cofactor of Factor I; inactivates C3b
C4-binding
protein
Cofactor of Factor I; inactivates C4b
Prevents the attachment of C5b67
to cell membrane
Accelerates dissociation of both C3
convertases
Paroxysmal Nocturnal
Hemoglobinuria
MIRL
ADAPTIVE IMMUNITY LYMPHOID ORGANS
PRIMARY
☻ _________
✓ Atrophies as one ages
✓ Located in the thorax
✓ Site of T-cell maturation
☻ _________
✓ Equivalent to Bursa of Fabricius in birds
✓ Site of hematopoiesis
✓ Site of B cell maturation
SECONDARY
☻ _______ – largest secondary lymphoid organ
☻ Tonsils
☻ Lymphoid tissues
☻ Peyer’s patches
☻ Appendix
☻ Mucosa-Associated Lymphoid Tissue (MALT)
LYMPHOCYTES
T CELL B CELL
Cell-mediated immunity Humoral-mediated immunity
Matures in thymus Matures in bone marrow
60-80% 20-30%
CD4+ – T helper cells
CD8+ – T cytotoxic cells
T-regulatory cells
Naive B cell
Activated B cell
Plasma cell
Rosette formation identification
CD2 – receptor for sheep RBCs
Identification by surface
immunoglobulins (IgM and IgD)
Lymphokines (products) Antibodies (products)
CD2, CD3, CD4, CD8 antigens CD19, CD20, CD21, CD40 and MHC
Class II antigens
Located in the paracortex of lymph
nodes
Located in the cortex of lymph nodes
Mitogens:
Concanavalin A
Phytohemagglutinin
Pokeweed mitogen
Mitogens:
LPS
Pokeweed mitogen
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
FORMS OF ACQUIRED IMMUNITY
1. WHAT DID YOU ACQUIRE?
☻ If antigen → Active
☻ If antibody → Passive
2. HOW DID YOU ACQUIRE?
☻ Naturally
☻ Artificially
ACTIVE PASSIVE
NATURAL Natural exposure to ANTIGEN Natural exposure to ANTIBODIES
Ex. Recovery from infections Ex. Colostrum, Transplacental
ARTIFICIAL ANTIGEN was injected ANTIBODIES were injected
Ex. Toxoids, Vaccinations Ex. RhIg, anti-rabies, Hepatitis B
Immune globulin, anti-tetanus
CYTOKINES – proteins that regulates immune system
AUTOCRINE PARACRINE
v
ENDOCRINE
ANTIGENS ☻ “antibody generators”
☻ Otherwise known as ____________
☻ Substances that may or may not elicit an immune response
FORMS:
✓ _____________ (autoantigen) – host’s OWN antigen (self-antigens)
✓ ____________ – from DIFFERENT individual of the SAME species
✓ _______________ (Heteroantigen) – from DIFFERENT species
✓ ____________ – antigens that induces an immune response thus
production of antibodies persists. The antibodies produced are specific
for these antigens
✓ ____________ – antigens that reacts with antibodies it did not induce
thus cross reaction exists.
✓ __________– antigens capable of eliciting an immune response
✓ __________– non-immunogenic by itself but can be immunogenic when
coupled with a high molecular weight substance (carrier)
☻ _________ – antigenic determinant
FACTORS AFFECTING IMMUNOGENICITY
1. Foreigness
2. Molecular size
✓ < 1kDa → non-immunogenic
✓ >1kDa but 100kDa → highly immunogenic
3. Chemical composition and complexity
Protein → Polysaccharides → Lipids &Nucleic acids (most immunogenic → least)
4. Route, dosage and timing
✓ Most effective: intraperitoneal and intravenous
5. ___________
✓ Substances that enhances the immunogenicity of an antigen
✓ Included in preparation of antigens in vaccines
✓ Ex. Freund’s complete adjuvant, Freund’s incomplete adjuvant,
Aluminum potassium sulfate and Bacterial lipopolysaccharides
“ALL immunogens are antigens but NOT ALL antigens are
immunogens”
Nearby cell
Circulation
PLEIOTROPY – single chemokine
with different actions
REDUNDANCY – different cytokines
activate same pathways
SYNERGY – acting in networks
produing effects that complement
& enhances each other
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
ANTIBODIES ✓ Otherwise known as _____________, ____________ or _______________ (because
of their electrophoretic migration)
✓ _________ – antibody determinant; binding sites
✓ May be present on the surface of B-cells or secreted by plasma cells
✓ Produced in response to immunogenic stimulation
✓ Sediment rate is expressed using ____________
✓ Can be naturally occurring or immunogenic; cold reactive or warm reactive
STRUCTURE
FUNCTION
☻ Binds extracelullar antigens that later initiates phagocytosis or complement
activation
☻ Neutralizes virus (before entry to host cells) or toxins.
MUST KNOW
☻ Location of hinge region – between CH1 and CH2
☻ Complement binding site – CH2 (IgG) and CH3 (IgM)
ISOTYPES
1. IgG
☻ Monomeric
☻ Produced during the secondary immune response
☻ Four subclasses
✓ _____ – most efficient
✓ _____ – can’t cross placenta
✓ _____ – best for complement fixation (3>1>2)
✓ _____ – short hinge; poor mediator of complement fixation
2. IgA
☻ Monomeric (IgA1 = serum) or dimeric (IgA2 = secretions)
☻ Found primarily in mucosal secretions
☻ Neutralizing antibody
☻ Aggregated form can activate alternative pathway
☻ ___________ – IgA and secretory components released to the opposite
surface of cell
3. IgM
☻ Monomeric (on surface of B-cells) or pentameric
☻ Produced during primary immune response (!Mauna)
☻ Largest and heaviest
☻ Best in agglutination!
☻ Free-state = star-like
☻ Combined = Crab-like
☻ Can be dissociated using dithiothreitol (DTT) and 2-mercaptoethanol
(2-ME)
4. IgD
☻ Present on surface of B-cells
☻ First identified in a patient with Multiple Myeloma
5. IgE
☻ “Reaginic antibody”
☻ Heat labile @ ________
☻ Responsible for allergies, helminthic infections and anaphylactic
reactions
☻ Plasma cells in lungs and skin secrete IgE
☻ 2 Fab, 1 Fc
☻ Heavy chain – 5 isotypes (γ, α, μ,
Δ, Ε); has 4-5 domains
☻ Light chain – κ (chromosome 2), λ
(chromosome 22); has 2 domains
✓ Κ : λ ratio = 2:1
☻ Joining chain: only found in IgM
and secretory IgA
☻ Variable region: located in amino
terminal end
☻ Constant region: located in the
carboxy terminal end
☻ PAPAIN cleaves Ab directly in the
hinge region = 2 Fab and 1 Fc
fragments
☻ PEPSIN cleaves Ab below the
hinge region = 1 Fc’ + F(ab)2
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
BIOLOGIC PROPERTIES OF ANTIBODIES
IgG IgM IgA IgD IgE
Molecular weight 150,000 900,000 160,000 180,000 190,000
Sedimentation
Coefficient
7S 19S 7S 7S 8S
Heavy Chain
Subclasses
4 0 2 0 0
Percent of total Ig 70-75 10 10-15
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
Immune
mediator
IgE IgG and IgM IgG and IgM T cells
Mechanism Release of
mediators
Cytolysis Deposition of
antigen-
antibody
complexes
Release of
lymphokines
Efffector cells Basophils,
mast cells
RBCs, WBCs
and platelets
Host tissue
cells
T cells
Macrophages
Complement
involvement
No Yes Yes No
Examples Anaphylaxis
Atopy
Hives
Asthma
HTR
AIHA
HDN
Goodpasture’s
syndrome
Arthus
reaction
SLE
RA
Serum sickness
TB
Sarcoid
Wegener’s
granulomatosis
Contact
dermatitis
AUTOIMMUNE DISEASES
Disease Autoantibody
Adrenocortical cells
Erythrocyte membrane proteins
Chronic active hepatitis Anti-smooth muscle antibody
Diabetes mellitus Pancreatic islet cells, insulin, glutamic acid
decarboxylase
Goodpasture syndrome Anti-glomerular basement membrane
Thyroid-stimulating hormone receptors
Anti-microsomal antibody, anti-thyroglobulin
Multiple sclerosis Anti-myelin sheath
Anti-acetylcholine receptor
Anti-parietal cell, Anti-intrinsic factor
Primary biliary cirrhosis Anti-mitochondrial
Myocardium
Rheumatoid arthritis Rheumatoid factor, Anti-CCP, Anti-DNP
Anti-dsDNA, anti-Smith
Anti-neutrophilic cytoplasmic
Anti-centromere
Idiopathic or immune
thrombocytopenic purpura
Anti-platelet
IMMUNODEFICIENCIES
Most common
Most severe (absence of T and B cells) –
“bubble boy”
Absence of B cells and all immunoglobulins
Triad: eczema, thrombocytopenia,
immunodeficiency
Uncoordinated muscle movements (ataxia)
& dilatation of blood vessels (telangiectasia);
decreased levels of IgG2, IgA and IgE
Recurrent bacterial infection & sinusitis;
selective IgG deficiency may occur
Markedly increased IgM but decreased
levels of other antibodies
Defective adhesion protein (CD18) on
surface of phagocytes
Absence of thymus gland and T cells Adapted from Immunology and Serology and Blood Banking Notes of Mr. Errol E. Coderes RMT, IMLT, MLS (ASCPi)CM
TUMOR MARKERS
Small cell carcinoma
Hepatocellular and testicular cancer
Bence-Jones Protein Multiple Myeloma
Ovarian cancer
Breast cancer
Gastric, colonic and pancreatic
adenocarcinoma
Familial medullary thyroid carcinoma
Breast, lung, colorectal and stomach cancer
Lung and breast cancer (also bladder cancer)
HCG Choriocarcinoma, testicular cancer
Urinary bladder cancer
Prostatic specific antigen (PSA) Prostate cancer Adapted from Immunology and Serology and Blood Banking Notes of Mr. Errol E. Coderes RMT, IMLT, MLS (ASCPi)CM
SEROLOGY
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
AGGLUTINATION METHODS a. ________________ – antigens are found on the surface of particle
b. ________________ – soluble antigens are attached to carrier particles (eg.
Latex, beads or charcoal). Antibody (if present) then attaches to the
particulate antigen.
c. ________________ – antigens are found on the surface of red cells
d. ________________ – antibody is the one attached to a carrier particle, thus if
an antigen is present = agglutination. (eg. CRP)
e. ________________ – the carrier attached to antibody is bacteria (usually S.
aureus)
f. ________________– positive result = lack of agglutination
g. ________________ – detects non-agglutinating antibody by means of coupling
with a second antibody
Precipitation methods
Radial
Immunodiffusion (RID)
“Fak Me, James RID”
Antibody is incorporated into the gel medium; antigen is
placed on the well, diffuses and reacts with an antibody;
diameter of precipitation is measured
• __________ Kinetic method
Measurement is done before
equivalence point
Time: 18hrs
d = log antigen conc
• __________ Endpoint method
Antigen is allowed to diffuse
complemetely before measurement
is made
Time: 24 hrs (IgG) & 50-72 hrs (IgM)
Antibody is incorporated into the gel medium; antigen is
then added, diffuses and reacts with the antibody thus
producing a precipitin band
Ouchterlony double
diffusion
Both antigen and antibody diffuses into the medium
☻ ______________ = serological identity
☻ ______________ = partial identity
☻ ______________ = non-identity
Antigen in serum is electrophoresed to separate protein
fractions. Antiserum is then added to the trough; change
in shape = abnormality
For detection of Bence-Jones protein
Radial immunodiffusion + electrophoresis = rocket band
Height is directly proportional to antigen concentration
Immunofixation
electrophoresis
Differ to immunoelectrophoresis in which the antiserum is
layered on the medium
Measures light scattered by immune complexes Adapted from Immunology and Serology and Blood Banking Notes of Mr. Errol E. Coderes RMT, IMLT, MLS (ASCPi)CM
LABELED IMMUNOASSAYS TERMS
a. Competitive – both labeled and unlabeled antigen are added
simultaneously thus competing for binding sites of antibody (inversely
proportional)
b. Noncompetitive – an excess antibody binding site is present so that all patient
analyte can be bound and measured (directly proportional)
c. _____________ – no washing (separation) step.
d. _____________ – with washing step.
EXAMPLES OF LABELED IMMUNOASSAYS
a. Enzyme immunoassays
✓ Competitive or noncompetitive (eg. ELISA, both direct and indirect)
✓ Most common: ___________________
b. Chemiluminescence
✓ Competitive or sandwich
✓ Isoluminol or acridium esters
c. Fluorescent immunoassay
✓ Homogenous
✓ FITC, TRITC
d. Radioimunnoassay
✓ Most sensitive
✓ Most specific
✓ Competitive
✓ RIST (total IgE) & RAST (allergen specific IgE)
✓ Most common: 125I
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
SEROLOGIC TESTS OF INFECTIOUS DISEASES Certain strains of Proteus vulgaris share antigens with Rickettsial
species.
☻ P. vulgaris – OX-2 & OX-19
☻ P. mirabilis – OX-K
Organism Disease
R. prowazekii Epidemic typhus, Brill’s disease
R. ricketsii RMSF
R. tsutsugamushi Scrub typhus
R. akari Rickettsial pox
R. mooseri (R. typhi) Murine typhus
For detection of antibodies in typhoid fever
No agglutination
25% agglutination
50% agglutination
75% agglutination
100% agglutination
Tests for Syphilis Syphilis
☻ STD
☻ T. pallidum is not detected in blood products that have
been stored for more than 48 hrs @ 4°C
Syphilis
Pinta
Rabbit syphilis
Bejel or nonvenereal endemic
syphilis
Yaws
Nontreponemal Determines presence of reagin (antibody against cardiolipin -
antigen)
Based on flocculation
VDRL
☻ Principle: quantitative or qualitative slide flocculation
☻ Serum (complment) inactivaetion is required
✓ Serum is heated @56°C for 30 mins (to be used
within 4hrs)
✓ If >4 hrs = reinaactivation @56°C fir 10 mins is
required
☻ Antigen:
✓ 0.03% Cardiolipin (for reactivity)
✓ 0.9% cholesterol (to increase antigen size)
✓ 0.21% lecithin (for standard reactivity)
☻ Amount of antigen suspension:
✓ Quantitative – 100 drops
✓ Qualitative – 60 drops
☻ Centrifuged @ 180rpm for 4 mins → read microscopically
RPR
☻ Antigen similar to VDRL with addition of
✓ Charcoal (for visibility – macroscopic reading)
✓ EDTA
✓ Thimerosal (preservative)
✓ Choline chloride (stabilizes antigen & inactivates
complement)
☻ Amount – 60 drops
☻ Centrifuged @ 100rpm for 8 mins → read
macroscopically
Treponemal Detects treponemal antibodies
Treponema Immobilization (TPI) Test
☻ Reference test
☻ Interpretation
✓ Positive - > 50% of treponemes are immobilized
✓ Negative -
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
☻ Elevated
✓ > 240 Todd units (adult)
✓ > 320 Todd units (child)
✓ > 200 IU/mL (ASO Slide test)
Appears earlier in a streptococcal infection
Streptozyme
test
Slide agglutination screening test
Screening test for presence or absence of heterophil antibodies
– uses sheep RBCs (infectious mononucleosis, serum sickness,
forssman antigen)
Differentiation of heterophile antibodies
✓ IM = adsorbed only by beef erythrocytes
✓ Forssman = adsorbed only by
✓ Serum sickness adsorbed by both beef erythrocytes and
GPK
Rapid test for heterophil antibodies that uses horse RBCs
HIV Screening tests
☻ ELISA (standard)
☻ Agglutination tests
☻ Dot-blot tests
☻ Immunochromatography
Confirmatory
☻ Western blot (atleast 2
of the ff – p24, gp41
or gp120/160)
☻ Indirect
immunofluorescent
☻ Nucleic acid
amplification test
AIDS CD4 T-cell count ( 6.2
> 5.5 x 1010
20-24°C, 5 days with constant
agitation
Pooled: 4hrs
pH: > 6.2
> 3.0 x 1011
20-24°C, 5 days with constant
agitation
Should be prepared within:
6 hrs after collection for ACD
8 hrs = CPD, CP2D, CPDA-1
-18°C – 1 yr
-65°C – 7 yrs
CRYOPRECIPITATE FVIII:C = 80 IU
Fibrinogen of atleast 150
mg/unit
Frozen: -18°C, ___yr
Thawed: 20-24°C, ___hrs
Pooled: 20-24°C, ____hrs
> 1.0 x 1010 PMNs/unit 20-24°C, 24 hrs (without
agitation)
80% RBC recovery
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
6.2)
In SDP, WBS = __ > ____ > ____
(most common to least common
blood type)
☻ Antigens are detectable 5-6
weeks in utero
☻ Full expression of antigens – 2 to
4 yrs of age
☻ Antibodies are naturally
occurring
☻ Begins to rise at 3-6 mos and
peaks bet 5-10 yrs
GENE PRODUCT IMMUNODOMINANT
SUGAR
PHENOTYPE
H L-fucosyltransferase O
A N-acetylgalactosaminyltransferase A
B D-galactosyltranferase B
A&B N-acetylgalactosaminyltransferase
D-galactosyltranferase
AB
☻ Bombay phenotype
✓ Nonsecretor, H-deficient
✓ Produces anti-A, anti-B, anti-
A,B and anti-H
✓ Positive with “O” cell
☻ Parabombay
✓ Secretor but still H-deficient
__ > __ > __ > ___ > __ > ___
(greatest to least amt of H
substance)
TYPES OF DISCREPRANCY
I Weakly reacting or missing
antibodies
☻ Causes
✓ Newborn
✓ Elderly patients
✓ Leukemia
✓ Immunodeficiencies
II Weakly reacting or missing
antigens
☻ Causes
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
✓ Acquired B phenomenon
(resolution: deacetylation)
✓ Subgroups of A and or B
III Protein or Plasma Abnormality ☻ Causes
✓ Multiple myeloma
✓ Waldenstrom’s
macroglobulinemia
✓ Plasma cell dyscrasias
✓ Plasma expanders (PVP and
Dextran)
✓ Wharton’s jelly
IV Miscellaneous ☻ Causes
✓ Polyagglutination
✓ Cold reactive antibodies
✓ Unexpected ABO
isoagglutinins
Grading Agglutination Reactions (Harmening 5th Ed)
No agglutination or hemolysis
Tiny agglutinates, turbid background
Small agglutinates, turbid background
Medium-sized agglutinates, clear background
Several large agglutinates, clear background
One solid agglutinate
002 MNS M antigen N antigen
1st position Serine Leucine
5th position Glycine Glutamic acid
S antigen s antigen
29th position Methionine Threonine
☻ Anti-M – reacts better at pH ____
☻ Anti-N – found in patients dialyzed
or in equipment sterilized with
formaldehyde
003 P ☻ _______ - neutralized by hydatid cyst
fluid
☻ _______ (Donath-Landsteiner
antibody) – assoc with PCH;
biphasic antibody – binds to P(+)
cells at lower temp. and hemolyzes
cells @ 37°C
☻ Anti-PP1Pk – formerly anti-Tja
004 Rh __ > __ > __ > __ > __
(most to least immunogenic)
☻ e – associated with warm
autoimmune hemolytic anemia
(WAIHA)
☻ antibodies are not naturally
occurring, instead are immune
antibodies
☻ cause of most severe HDN
✓ father – Rh (+)
✓ mother – Rh (-)
✓ child – Rh (+)
✓ production of anti-D
WEAK D EXPRESSIONS
1. Antigen is weaker than expected
2. When C is in trans position with D
Ex. Dce/dCe
3. Anitbodies produced are directed to
the missing parts
In transfusion:
✓ Recipient: D –
✓ Donor: D+
4. ☻ Lacks all Rh antigens
☻ Stomatocytosis
005 Lutheran ☻ Named after a donor named
Lutteran (misinterpretation)
☻ Anti-Lua – discovered in serum of a
patient with diffused lupus
erythematosus; more common
antibody
006 Kell ☻ K is strongly immunogenic (2nd to D)
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
☻ Kell antigens are synthesized on
their precursor, Kx
☻ McLeod phenotype – absence of
Kx antigen (acanthocytosis)
☻ CGD – absence of Kx antigen in
WBCs
007 Lewis ☻ Only blood group synthesized by
the RBC membrane
Le (a-b-) → Le (a+b-) → Le (a+b+)
→ Le (a-b+)
(birth → post-delivery → >10 days
→ adults)
☻ Decreased expression of antigens
in pregnancy
☻ ______ has receptors for H. pylori
008 Duffy ☻ Anti-Fya – most common Ab in the
system
☻ ________ – common in blacks;
resistant to P. vivax infection
009 Kidd ☻ NOTORIOUOS ANTIBODY
☻ Assoc with delayed HTR
☻ Jk (a-b-) – resistant to 2M urea; cell
lysis in about 30 mins
☻ Antibodies reactivity are
enhinaced by adding 4 drops of
serum instead of 2.
☻ __________ – causes formation of
autoanti-Jk
010 Diego ☻ Marker of Mongolian ancestry
☻ Antigens are located on anion
exchange molecule 1 (AE-1)
011 Cartwright ☻ Antigens are located on RBC
acetylcholinesterase
012 Xg ☻ Sex-linked
☻ Xga expression: 89% (females) and
66% in males
013 Scianna ☻ Prevalence of Sc2 is higher in
Mennonite population
014 Dombrock
015 Colton ☻ Antigens are found on channel-
forming integral protein or
______________
016 Landsteiner-Weiner ☻ Agglutinates Rh+ and Rh- cells
except Rhnull cells
017 Chido-Rodgers ☻ High Titer, Low Avidity (HTLA)
☻ Associated with HLA
018 H ☻ Anti-H lectin = _______________
019 Kx ☻ Precursor of kell antigens
020 Gerbich ☻ Lecah phenotype (elliptocytosis)
021 Cromer ☻ Located on decay accelerating
factor (DAF)
022 Knops ☻ Antigens are located on
complement receptor 1 (CR1) or
CD35
023 Indian ☻ Antigens are located on CD___
027 I ☻ Product of I gene – N-
acetylglucosaminyl transferase
☻ I = adult red cells
☻ i – newborn or cord cells
☻ _________ = Mycoplasma
pneumonia (cold AIHA)
☻ __________ = infectious
mononucleosis
BLOOD DONATION
1. registration ☻ collection of patient information
☻ date of last donation
✓ 2 days or more – after plasmapheresis, plateletpheresis or
leukapheresis
✓ 4 weeks – after infrequent plasmapheresis
✓ 8 weeks – between whole blood donations
✓ 16 weeks – after 2-unit red cell collection
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
2. Health history interview DEFERRALS
Permanent Tegison medication – for severe psoriasis
Persons with HIV
Men engaged in sex with other men since 1977
Indefinite History of Chaga’s disease and Babesiosis
Lived > 3 months in UK from 1980-1996
Lived > 5 years in Europe from 1980 to present
Family history of CJD
Temporary 2 days for platelet donors Aspirin ingestion
Received attenuated vaccines for
Measles (Rubeola), Mumps, Polio (oral),
Typhoid and Yellow Fever
28 days Conclusion of pregnancy
After taking last dose of Accutane or
proscar (for benign protatic
hyperplasia)
Received live attenuated vaccine for
German measles (Rubella) or Chicken
pox (Varicella-zoster)
Avodart (teratogenic drug)
1 year History of syphilis or gonorrhoeae or
treatment
Travel to malaria endemic area
Tattoos or permanent makeup
Recently undergone surgery
Incarceration for >72 hrs
Sexual contact with an individual at
high risk for HIV
History of malaria
Lived in malaria endemic country for 5
consecutive years
Soriatane (treatment for severe
psoriasis)
3. Physical exam
GENERAL REQUIREMENTS FOR DONATION
CRITERIA ACCEPTABLE LIMIT
GENERAL APPEARANCE In good health
HEMOGLOBIN Autologous: > 11.0 g/dL or 110 g/L
Allogeneic: > 12.5 g/dL or 125 g/L
HEMATOCRIT Autologous: _____
Allogeneic: ______
BLOOD PRESSURE Systolic: _____________
Diastolic: ____________
TEMPERATURE _____°C or ____°F
No drinking of coffee or hot beverages
while waiting to donate
PULSE Between ____ and ____ bpm
Athletic donor will have a pulse 100lb (45kg) Adapted from Immunology and Serology and Blood Banking Intensive Review Notes of University of the Immaculate
Conception – Medical Laboratory Science Program
Must remember: blood collection procedure ☻ Patient identification is a MUST
☻ Blood is extracted at the antecubital fossa
☻ Tourniquet of blood pressure cuff: ____________
☻ Needle gauge: ________________
☻ During collection, instruct patient to open and close hand every
_____________
☻ Mixing of blood: 1-2 times/min or 45 seconds
☻ Donation time for a unit of whole blood: 8-12 mins, if >15 mins = platelets, FFP
and cryoprecitated AHF can’t be prepared
☻ Blood should be processed within ______________
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
Testing
Compatibility TESTING ☻ Aka serologic crossmatch test
☻ Test to detect unexpected antibodies
☻ Ensures to produce a blood product safe for transfusion
______________________
☻ Tests for alloantibodies in patient’s plasma against donor cells
☻ Donor’s cell + Patient’s plasma
_______________________
☻ Tests for alloantibodies in donor’s plasma against patient’s cells
_______________________
☻ aka enhancement media (increases reactivity of IgG antibodies)
☻ Low ionic saline solution
✓ Action: decreases zeta potential
✓ 10-15 mins incubation time
☻ Polyethylene glycol (PEG)
✓ Action: removes water
☻ 22% albumin
✓ Action: zeta potential is decreased through buffering
Antihuman globulin test PRINCIPLE:
OR
_______________ ☻ Detection of sensitization of red cells in vivo
☻ Examples: AIHA, HDN, HTR
_______________ ☻ Detection of sensitization of red cells in vitro
☻ Sensitization occurred during incubation period
☻ Examples: Weak D testing, AHG-phase compatibility testing, antibody screen
and identification
TRANSFUSION
REACTION
INFORMATION PREVENTION/TREATMENT
1°C increase in temperature
Cause: Anti-leukocyte antibodies
(patient)
Leukoreduced RBCs
ALLERGIC Cause: donor plasma with foreign
proteins
Washed RBCs
Patient has IgA deficiency w/ anti-
IgA antibodies
Patient is afebrile (no fever)
Washed RBCs
IgA-deficient donor
(rare)
Patient has normal IgA with anti-IgA
antibodies
Patient is afebrile
Washed RBCs
IgA-deficient donor
(rare)
MONOCLONAL ANTIBODIES POLYCLONAL ANTIBODIES
AH
G (from immunized animal)
Human IG
PROPERTY OF MEDTECH REVIEW NOTES
DO NOT DISTRIBUTE
Signs & symptoms resemble
respiratory distress
Cause: Anti-leukocyte antibodies
(donor)
Leukoreduced RBCs
Cause: T lymphocyte proliferation
(donor)
Irradiated RBCs
PTP Cause: anti-platelet antibodies
(HPA-1a negative platelets)
Corticosteroid
Exchange transfusion
IV immunoglobulins
Cause: rapid infusion of large vol. of
blood products (administration of
blood w/o equivalent blood loss)
Iatrogenic
Therapeutic
phlebotomy
IV diuretics
Oxygen therapy
BACTERIAL
CONTAMINATION
Cause: endotoxin production by
psycrophilic bacteria (Y.
enterocolitica)
Sterile technique
Visual inspection of unit
Cause: small bore needle, warming
blood above 50°C, freezing blood
without cryoprotective agent,
citrate toxicity
At risk: transfusion-dependent
patients (aplastic anemia,
thalassemia)
Iron-chelating agent
(deferroxamine)
Transfusion of neocytes Adapted from Immunology and Serology and Blood Banking Notes of Mr. Errol E. Coderes RMT, IMLT, MLS (ASCPi)CM
REFERENCES
Clinical Immunology and Serology: A Laboratory Perspective by Christne Dorresteyn
Stevens
Essentials of Immunology and Serology by Jacqueline Stanley
Modern Blood Banking and Transfusion Practices by Denise Harmening
Fundamentals of Immunohematology by Mary Louise Turgeon
Immunology-Serology and Blood Banking by Rodolfo Rabor
Immunology and Serology and Blood Banking Notes of Mr. Errol E. Coderes RMT, IMLT,
MLS (ASCPi)CM
Blood Banking Checkpoint notes of Ms. Judea Marie Policarpio, RMT
Intensive Review Notes of University of the Immaculate Conception – Medical
Laboratory Science Program