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MUST TO KNOW IN IMMUNOHEMATOLOGY (BLOOD BANKING)
ISBT 001
ABO
ISBT 002
MNS
ISBT 003
P
ISBT 004
Rh
ISBT 005
Lutheran
ISBT 006
Kell
ISBT 007
Lewis
ISBT 008
Duffy
ISBT 009
Kidd
ISBT 010
Diego
ISBT 011
Cartwright
ISBT 012
Xg
ISBT 013
Scianna
ISBT 014
Dombrock
ISBT 015
Colton
ISBT 016
Landsteiner-Weiner
ISBT 017
Chido/Rodgers
ISBT 018
H
ISBT 019
Kx
ISBT 020
Gerbich
ISBT 021
Cromer
ISBT 022
Knops
ISBT 023
Indian
Chromosome 1
Rh
Duffy
Scianna
Cromer
Knops
Chromosome 2
Gerbich
Chromosome 4
MNS
Chromosome 6
Chido/Rodgers
Chromosome 7
Cartwright
Colton
Kell
Chromosome 9
ABO
Chromosome 11
Indian
Chromosome 17
Diego
Chromosome 18
Kidd
Chromosome 19
H
Lewis
Landsteiner-Weiner
Lutheran
Chromosome 22
P
Chromosome X
Xg
Kx
Chromosome: Not known
Dombrock
Von Descatello (Decastello) AB
Sturle (Sturli)
Blood groups
(Most common) O > A > B > AB (Least common)
Page | 237

Cell typing

Uses of anti-AB
Serum typing

Gel typing

Red cell Ag-Ab reactions

Universal donor
Universal recipient
Universal donor for RBCs
Universal recipient for
RBCs
Universal donor for plasma
Universal recipient for
plasma
RCS: Red cell suspension
Genotype
Phenotype
Homozygous
Heterozygous
Dominant
Recessive
Allele
Silent/amorph
Von Dungern
Hirzfeld

Forward/direct typing
Specimen: RBCs (Ag)
Reagents:
= Anti-A (blue: trypan blue)
= Anti-B (yellow: acroflavin dye)
= Anti-AB (colorless)
Checks for the reaction of anti-A and anti-B
Detects weak subgroup of A and B because it has higher titer of anti-A & anti-B
Reverse/indirect typing
Specimen: Serum (Ab) = 3-6 months (development)
Reagents:
= A1 cells
= B cells
Major Advantage: Standardization
Medium: Dextran-acrylamide gel
0 = Unagglutinated Cells ---(media)---> Bottom of the μtube (microtube)
1+ = Agglutinated cells ---(media)---> May concentrate at the bottom of μtube
2+ = Agglutinated cells ---(media)---> Throughout the length of the μtube
3+ = Agglutinated cells ---(media)---> Concentrate near the top of the μtube
4+ = Agglutinated cells ---(media)---> Top of the media
Mixed-field:
= Agglutinated cells ---(media)---> Top
= Unagglutinated cells ---(media)---> Bottom
0 = No agglutination/hemolysis
W+ = Tiny agglutinates, turbid BG
1+ = Small agglutinates, turbid BG (25% agglutination)
2+ = Medium agglutinates, clear BG (50% agglutination)
3+ = Large agglutinate, clear BG (75% agglutination)
4+ = One solid agglutinate (100% agglutination)
Group O
Group AB
Group O (No Ag)
Group AB (No Ab)
Group AB (No Ab)
Group AB (No Ag)
BB = 2-5% RCS
Approximate = Tomato red
Exact % RCS = [Vol
...

A
N-acetylgalactosaminyltransferase
N-acetyl-D-galactosamine
A
H
B
D-galactosyltransferase
D-galactose
B
H
AB
N-acetylgalactosaminyltransferase
N-acetyl-D-galactosamine
A
H
D-galactosyltransferase
D-galactose
B
O
---Unchanged H
Amount of H Ag
(Greatest) O > A2 > B > A2B > A1 > A1B (Least)
Bombay individual
Bhende
(-) H gene
hh or Hnull
Lack H, A and B Ag’s
Designated as Oh
w/ anti-H, anti-A and anti-B Ab’s
Mistyped as group O
Parabombay
Inherit weak H gene
w/ detectable A and B Ag’s but no detectable H Ag
Ah, Bh, ABh
Anti-H
Differentiates Group O from Oh individuals
Ulex europaeus
Lectin source of anti-H
ABO
Histoblood group = present on all tissues and organs of the body
May be expressed in secretions depending on the secretor status (SeSe or Sese)
Determination of Secretor
Specimen: Saliva
Status
Principle: Hemagglutination-inhibition
ABO antibodies
Mixture of IgM, IgG and IgA (Henry)
Anti-A
Predominantly IgM
Anti-B
React at room temp
Naturally occurring
Anti-A,B (Group O)
Predominantly IgG
React at 37’C
Immune Ab
Immune Ab’s
Production is stimulated by:
1
...
Incompatible transfusion and transplant
ABO HDN
Mother: Group O
Child: Group A or B
Group I discrepancies
Weak reacting/missing Ab’s
Newborns
Elderly patients
Hypogammaglobulinemia/agammaglobulinemia
Page | 239

Group II discrepancies

Group III discrepancies

Group IV

A3
Ax
Aend
Am
Ay
Ael
B3
Bx
Bm
Bel
Rh Nomenclatures

Immunogenicity of Rh Ag’s
R or r
1 or ‘
2 or ‘’
Z or y
G
Rh: 13, 14, 15, 16
Genetic weak D
D Mosaic (Partial D)
C Trans
f (ce)
rhi (Ce)
Hr0
Rh Ab’s

Weak reacting/missing Ag’s (Less frequent)
Leukemia
Acquired B phenomenon
Subgroups of A
Hodgkin’s disease
BGSS (Remedy: Wash RBCs)
Plasma abnormalities resulting to Rouleaux formation (Plasma factor)
 globulins: MM, WM, HL
 fibrinogen
Dextran and PVP
Wharton’s jelly (Remedy: Wash cord cells 6-8x (7x)
Miscellaneous:
Polyagglutination
Anti-I (cold agglutinin)
Cis “AB phenotype”
Unexpected ABO isoagglutinins:
= Anti-H: produced by A1 and A1B (H Ag)
= Anti-A1: produced by A2 and A2B (No A1 Ag)
A Subgroups
MF agglutination w/ anti-A & anti-A,B
Weak MF agglutination w/ anti-A& anti-A,B
MF agglutination w/ anti-A,B
(-) Agglutination, (+) AH substance in secretions
(-) Agglutination,  AH substance in secretions
(-) Agglutination, (+) H substance in secretions
B Subgroups
MF agglutination w/ anti-B & anti-A,B
MF agglutination w/ anti-A,B
(-) Agglutination, (+) BH substance in secretions
(-) Agglutination, (+) H substance in secretions
1
...
Fisher-Race (DCE) = inheritance of 3 genes (d= amorph/silent)
3
...
R0), w/c codes for an
agglutinogen (Ex
...
Rh0hr’hr’’)
(Most immunogenic) D > c > E > C > e (Least immunogenic)
D or d
Ce
cE
CE
D+C+ red cell
4 different parts of the D mosaic
D Ag’s expressed appear to be complete, but few in number
If one or more parts of D Ag is missing  weakened expression of D Ag
May produce anti-D (Ab against missing fragment)
4 fragments
D ---(trans)---> C (Ex
...
Dce/DCE]
C ---(cis)---> e [Ex
...
can be transfused
w/ Rh+ blood
Anti-LW
(Landsteiner-Weiner)
Rhnull
Rhdeleted
Lewis system
Lewis Ag’s

Genotype
ABH, lele, sese
ABH, lele, SeSe/Sese
ABH, LeLe/Lele, sese
ABH, LeLe/Lele, SeSe/Sese
Lewis Ab’s

MN Ag’s

Anti-M

(-) C’ binding = extravascular hemolysis (delayed HTR)
Mother: Rh (-)
Child: Rh (+), 2nd pregnancy
Purified anti-D
Administer w/in 72 hrs after 1st delivery
300 μg anti-D
Protect up to 30mL D+ WB or 15mL D+ RBCs
50 μg anti-D
Protect up to 5mL of D+ WB or 2
...
Abortion
Volume of FMH (mL) ÷ 30
Vol
...
No prior exposure to D Ag (males) or past childbearing age (females)
2
...
5), glucose-dependent
Page | 241

Anti-N-like Ab
SS Ag’s
Ss Ab’s
Phenotype
P1
P2
p (p null)
P 1k
P 2k
P1-like
P1 substance
Anti-P1

Anti-Tja
Anti-P

Ii Blood Group
HEMPAS
Anti-I

Anti-i

Anti-IT
K
k
Kpa
Kpb
Jsa
Jsb
Kell Ag’s

IgM
Found in renal patients dialyzed w/ formaldehyde sterilized equipment
Glycophorin B (Ss-SGP)
S = Methionine (29th)
s = Threonine (29th)
IgG
React at 37’C and AHG
Severe HTR w/ hemoglobinuria and HDN
Detectable Ag’s
Possible Ab’s
P1 and P
None
P
Anti-P1
None
Anti-P, P1, Pk (anti-Tja)
Pk and P1
Anti-P
Pk
Anti-P, anti-P1
Plasma, pigeon and turtledove droppings, turtledove eggwhite
Hydatid cyst fluid, Lumbricoides terrestris, Ascaris suum
IgM
Naturally occurring
Strong anti-P1 = Hydatid disease (E
...
sinensis and O
...
-adult) = I I = I+ii Ag in adults
Interfere w/ reverse typing (Group IV)
Benign anti-I = normal, IgM, naturally occurring, react at 4’C
Pathologic anti-I = IgM, react at 30/32’C (CAS = PAP)
Autoanti-I = M
...
monocytogenes
IgM
React at 4’C
EBV caused
Diseases of RES:
- Alcoholic cirrhosis
- Myeloid leukemia
- Reticuloses
Transition: from i  I
Yanomama Indians
Hodgkin’s lymphoma
Kell
Cellano
Penney
Rautenberg
Sutter
Matthews
Immunogenicity: 2nd to D (D>K>c>E>C>e)
Synthesized on precursor Kx
Page | 242

MacLeod phenotype
Anti-K
Fy(a+b-)
Fy(a-b-)
Fya and Fyb
Duffy gene
Anti-Fya
Anti-Fyb
Jka and Jkb
Anti-Jka
Anti-Jkb
Lua and Lub
Anti-Lua
Anti-Lub
Diego (Di)
Defect in AE molecule

Cartwright (Yt)
Xg
Scianna
Dombrock
Colton (Co)
Chido/Rodgers (Ch/Rg)

Gerbich (Ge)
Cromer (Cr)
Knops (Kn)
Indian (In)
Benneth-Goodspeed (Bg)

= On WBCs: remain unconverted
...
If (-)  MacLeod phenotype
Acanthocytosis
Muscular dystrophy
IgG
React at 37’C and AHG
Chinese (90
...
vivax/P
...
9%)
Private Ag
Low-incidence Ag’s (<1%)
HTLA Ab’s
Anti-Ch
Anti-Rg
Anti-Kn
Anti-Yka
Anti-Csa
Anti-McC
Anti-JMH
Sources of Substances for Neutralization for Certain Antibodies
Anti-P1
Hydatid cyst fluid, pigeon droppings, turtledoves’ egg whites
Anti-Le
Plasma/serum, secretor saliva
Anti-Ch, Anti-Rg
Plasma/serum
Anti-Sda
Urine, guinea pig urine
Anti-I
Mother’s milk
Anti-H
Saliva
Effect of Proteolytic Enzymes on Select Antigen-Antibody Reactions
Enhanced
P1
Rh
I
Kidd
Lewis
Destroyed
MNS
Duffy
Chido
Rodgers
Cartwright
Xg
Donation Process
At least 2 persons
Bleeder
Head (BB)
Donor registration
1st step
Interview & physical exam
2nd step
Actual donor selection and
3rd step
blood collection
Collection
<15 mins (7-10mins)
>15 mins = (-) cryoprecipitate
Donor bleeding
450 angle  10-200 angle
(Venipuncture: 150 angle)
Basic Qualifications of the Potential Blood Donor
Good health
Purplish blue lesions  Kaposi’s sarcoma (HIV, HHV-8)
Age
18-65 yrs old
<18 yrs old = w/ parent’s consent
>65 yrs old = w/ physician’s consent
Body weight
Max: 10
...
of blood to be drawn
Vol
...
needed
Vol
...
to be
removed from blood bag
1:7
Temperature
Pulse
BP cuff
BP (DOH)
Min
...
5mL AC = 200mL blood
37
...
5’F)
50-100 beats/min ( athletes = acceptable)
40-60 mmHg (if used as tourniquet)
90-160 mmHg
60-100 mmHg
Allogeneic donation:
Hgb: ≥12
...
053
1 cm = distance between blood and solution
Acceptable drop of blood  sink w/in 15 secs (Hgb: ≥12
...
centrifuge (1-6’C)]
Bacterial contamination
Separate from WB w/in 6-8 hrs
Indication

Storage

Transport

1
...
expansion & 
RBC mass
2
...
 RBC mass
(Normovolemic patients)

1-6’C

1-10’C

1-6’C

1-10’C

Shelf-life
21d (ACD, CPD)
35d (CPDA-1)
42d (CPDA-2)
2d (Heparin
Open system: 24h
Closed system:
21d (ACD, CPD)
35d (CPDA-1)

Other Info
1 unit:
Hgb by 1 to
1
...
5g/dL
Hct by 3-5%

Page | 246

Blood
Component
Leukoreduced
RBCs
Washed RBCs

Frozen RBCs

Deglycerolized
RBCs
Fresh frozen
plasma

Indication

Storage

Transport

1
...
Ab’s) =
1’C in temp
...
Prevent CMV

1-6’C

1-10’C

1
...
FNHTR (Leuko
...
Prolong storage of rare
units
2
...
Centrifugation
2
...
Saline washing
Also for
polyagglutination

10 years

24h
1
...
factor def
...
Replace isolated factor
def
...
Reverse effects of
coumarin/warfarin

-18’C
-65’C

1 year (-18’C)
7 years (-65’C)

Cryoprecipitate

1
...

2
...
vWD
4
...


-18’C or
colder

1 year

Granulocyte
concentrate

1
...
Myeloid hypoplasia
1
...
factors and
complement
‡FFP: Thaw at 37’C
‡Thawed plasma:
store at 1-6’C
‡Administer w/in
24hrs once thawed
‡Contains:
-150mg fibrinogen
-80 U AHF
-vWF
-Factor XIII
‡Cryoppt
...
): store at RT’
‡Administer w/in 6
hrs
Contains 1 x 1010
granulocytes

5d (20-24’C w/
agitation)
2d (1-6’C)

1 unit will 
platelet count by
5,000- 10,000/μL
Usually RBCs and
platelets are
irradiated

Irradiated blood
component

1
...
For hemophilia A

1-6’C

28 days from
irradiation or orig
...
date
whichever comes
first
Varies

1
...
Hypovolemic shock

2-10’C

5 years

Stored in
lyophilized form
‡A
...
a
...
Hypovolemic shock

Transport

2-10’C

SDP

High glycerol
(Slow freezing)
Low glycerol
(Fast freezing)
Agglomeration

Shelf-life
5 years
5 years

Methods of Freezing RBCs
Concentration
Frozen at
40% w/v glycerol -80’C

Stored at
-65’C

20% w/v glycerol

-120’C

-196’C

Other Info
80-85% Albumin
10-15% Globulin
SDP: Single donor
plasma

Equipment
Mechanical
freezer
Liquid nitrogen

Glycerol
-80’C
-65’C
Mechanical
Glucose
freezer
Fructose
EDTA
Cryoprotective agent
Prevents rupture of RBCs during freezing
Ex
...
6% NaCl -----> 0
...
9% NaCl
Cryoprecipitate
After thawing = administer w/in 6 hours
After pooling = administer w/in 4 hours
Leukapheresis
HES (Hydroxyethylstarch) = Separation bet
...
k
...
super packed platelets
From plateletpheresis
For patients who are refractory or unresponsive to RDP
Limit patient exposure to multiple donors
RDP: Random Donor Platelets
SDP: Single Donor Platelets
Preparation
From WB: Light spin  Heavy spin
Plateletpheresis
Amount
5
...
0 x 1011 platelets
Vol
...
0 (New: 6
...
0 (New: 6
...
Cesium (Ce)
b
...
NSS = the only fluid allowed to start an IV line prior to transfusion
Page | 248

b
...
Ringer’s lactate =not allowed to start (contains Ca2+  promote coagulation)
Blood warmers
37’C
Temp
...
Clot screen filter (170μm) = to remove gross clots
2
...
perfringens
V
...
pneumoniae
All produce neuraminidase
Th
E
...

Produce weaker neuraminidase
Tx
Bacterial and viral
Unknown mechanism
Tk
“CABS”
C
...
niger
B
...
marcescens
All produce endo/exogalactosidase
Altered precursor substances (Altered: ABH, Le, I, P)
Acquired B phenomenon
Bacteria: Deacetylase enzyme
N-acetylgalactosamine --(Deacetylase)--> N-acetyl + galactosamine
Galactosamine = Group B
Remedy: Add acetic anhydride
VA
Vienna, Virginia
Microbial fucosidase = fucose = H
Acquired Non-Microbially Associated Polyagglutination
Tn
(-) β-3-D-galactosyltransferase enzyme needed for the normal structure of T
receptor (Tetrasaccharide of Thomas and Winzler)
Inherited Polyagglutination
a
Sd
Cad
Adult i Ag = H Ag, Sialic acid
HEMPAS/CDA II
HbM – Hyde Park
Others
NOR = Norfolk, Virginia

Page | 249

Arachis hypogaea
Salvia sclarea
Salvia horminum
Glycine soja
Predeposit AD

Intraoperative AD
Immediate preoperative
hemodilution
Postoperative salvage
FNHTR
Allergic
Anaphylactic

Anaphylactoid
TRALI (NCPE)
Hemolytic
TA-GVHD
PTP

DHTR
TACO

Tn
+
+
+

Cad
+
+

T
+
+

Tk
+
-

Autologous Donation
Before anticipated transfusion
Requirements:
*No age limit
*No strict weight requirement (vol
...
Administration of corticosteroid
b
...
IV immunoglobulins
7 days
Immediate Nonimmune Transfusion Reactions
Administration of blood w/o equivalent blood loss
Iatrogenic: physician-caused
At risk:
a
...
Elderly patients
Page | 250

Bacterial contamination

PCITR

Iron Overload
(Hemosiderosis)

Disease transmission
In utero
Neonatal period
Treatment

Full X-match
Abbreviated X-match
Electronic X-match

c
...
Therapeutic phlebotomy
b
...
O2 therapy
Cause: Endotoxin production by psychrophilic/cryophilic bacteria
Y
...
coli
P
...
During phlebotomy
b
...
During thawing
Prevention:
a
...
Visual inspection of unit
→ Blood unit = Brown, purple, hemolysis, clot
→ Plasma = Murky (dark brown) purple, red
Causes:
- Small bore needle
- Warming blood above 50’C
- Freezing blood w/o cryoprotective agent
- Citrate toxicity
Delayed Nonimmune Transfusion Reactions
Patients w/ normovolemic anemia
Transfusion-dependend patients:
- Aplastic anemia
- Congenital hemolytic anemia
Prevention:
a
...
Neocytes = young RBCs, has longer lifespan
HBV, HCV, HDV, CMV, EBV, HTLV-I and II, HIV, T
...
, B
...
cruzi, T
...
Intrauterine transfusion
- In utero
- Corrects anemia
- X-match: Mother’s serum
2
...
weak Ab’s = 133:1
pH 7
...
4
30-120 mins (majority: 30 mins)
1
...
PEG
3
...
enterocolitica
Density
Requirements for blood
products to be transfused
to infants
Allogeneic donor blood
from outside sources
w/in 30 mins
Procedure when HTR
occurs
Specimens for investigation
of HTR

LISS
ZZAP
True chimerism
Artificial chimerism

AB cis genotype
Acquired A antigen

Collect new specimen every 3 days in a series of transfusion
Keep for 7 days after transfusion (at 1-6’C)
Series of tests that ensure safety of transfusion to recipient and donor
1
...
ABO and Rh
3
...
Cross matching
a
...
Minor X-match = PC/DS
Occur 3 to 7 days after transfusion
Heating in a liquid state w/ LMW stabilizers
Heating in lyophilized state
UV irradiation
 Bacterial contamination of blood
Separates neocytes from mature RBCs
1
...
<7 days
3
...
(-) CMV
Repeat ABO and Rh typing
Time limit when a unit of blood is removed from 2-8’C and returned back to the
refrigerator
1
...
Keep IV line open
3
...
Patient pre-transfusion blood sample
2
...
2 g/L or 20 mg/dL
= Red: Hgb >1 g/L or 100 mg/dL
3
...
Blood bag
= GS/CS
Glycine/glucose + saline
5-15 mins incubation
Cysteine-activated papain
Mixture of DTT and papain
Presence of 2 cell population
Ex
...
Tn activation
2
...
mirabilis
Page | 253

Acquired B antigen

HAT medium
Hybridoma cells
Polyspecific AHG reagent
Monospecific AHG reagent
Type 1 precursor substance
Type 2 precursor substance
Agglutination in Gel
technology
Affinity column technology
(Gamma ReACT)
Wash RBCs
# cryobags
# units to test
Enzyme technique

Elution
Adsorption
Biphasic hemolysin
Perfluorocarbons

“EPIC”
1
...
coli O86
2
...
vulgaris
3
...
Carcinoma of the colon and rectum
Hypoxanthine, Aminopterin, Thymidine medium
Culture for hybridoma cells
Plasma cells (mouse) + myeloma cells + PEG
Anti-IgG and anti-C3d
Prepared by conventional technique (rabbits)
Anti-IgG or anti-C3d
Prepared by monoclonal Ab production (mice)
β 1 3
ABH substance in secretions (glycoproteins)
HAB, Se, Le genes
β 14
ABH on RBC (glycoproteins)
HAB genes
Incompatible
Affinity adherence of IgG-sensitized RBCs to immunologically active matrix
2-3x to remove unbound globulins
[Desired level of Factor VIII x volume of plasma (mL)] ÷ 80
# units required ÷ frequency of donors (-) for relevant Ag
1
...
Bromelin = pineapple
3
...
Trypsin = pig’s stomach
Breaking of bonds between Ag and Ab by:
a
...
Chemical: ether, acid
Removal of Ab in serum using appropriate RBC
Cold = attaches to RBCs
Warm = RBC lysis
O2 carrying capacity
RBC substitute

Page | 254



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