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Blood Bank Exam 1 Guide

Study Guide for Blood Bank Exam 1, contains quizzes
by

Chris Brewer

on 6 September 2013

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Transcript of Blood Bank Exam 1 Guide

Blood Bank
By: Christopher Brewer
Study Guide
The Study of the immunology of blood group antigens & antibodies
Blood Group Immunology
Definitions
*
Antigen (Ag)
: A foreign substance that can elicit an immune response
-Usually on red cell surface (includes platelets)
*
Antibody
: Product of humoral immune response
-Found in plasma/serum & some other body fluids
Immune System
*
Relies on a complex system of cells located throughout the body
*
These cells distinguish "Self" from "Nonself"
*
Ability to distinguish foreign substances from "Self" begins to develop in embryonic stage
*
Immune competency is achieved by age 4-6 months
Cells of the immune system & immune response
*
Cells of the immune system include:
Lymphs
Macrophages
Killer Cells
Natural Killer Cells
*
B Lymphocytes produce antibodies (Ab)
*Two types of immune responses
Primary immune response
Antibody produced is IgM in low levels
Detectable 3-4 weeks after antigenic stimulus
Secondary immune response
On re-exposure to an antigen
High level of antibody IgG
Antibody production begins w/in 1-2 days of re-exposure due to anamnestic response
(
remembers
)
Blood Bank
Quiz
1:
What is an Antigen?
1:
A foreign substance that can elicit an immune response
2:
2:
What is an Antibody?
Product of humoral immune response
3:
3:
The immune system relies on what?
A system of cells throughout the body
4:
4:
What cells make up the immune system?
Lymphs, Killer cells, Macrophages, Natural killer cells
5:
5:
What are the two types of immune reponses?
Primary & Secondary immune reponse
6:
6:
When does the secondary immune response start?
On re-exposure to an antigen
7:
7:
Why is the secondary response faster then primary?
Due to the anamnestic response (remembers)
Factors affecting immune response
*
Immunogenicity of antigen
(
ability to initiate immune response
)
*
Survival of antigen in circulation
*
Immune status of recipient
*
Prior exposure to antigen
*
Genetic factors
*
Others Include
-
Age
-
Nutrition
/
health status
-
Size, shapes, & ionic charge of antigen
-
Dose and addition of adjuvants
(
enhance antigen reaction
)
-
Host must perceive antigen as foreign
Blood Group Antibodies
*
Clinical significance
:
-
Can cause transfusion reactions
-
Can cause hemolytic disease of the newborn
*
Antibodies can be formed against RBCs, WBCs, and platelets
*
Major role of blood banking is to prevent this from happening or detect the antibody if already formed to assure transfusion of safe blood products to patients
Antibodies
*
Can be alloantibody
(
against foreign antigen
)
*
Can be autoantibody
(
against self antigens
)
*
Protein molecules secreted by plasma cells
*
Immunoglobulins that function as antibodies are divided into 5 classes
*
Division is based on physical, chemical, & biologic properties
*
Heavy chain determines immunoglobulin class
Blood Bank
Quiz
1:
2:
3:
4:
5:
1:
2:
3:
4:
5:
Name some factors that affect the immune response
Factors affecting immune response
*
Immunogenicity of antigen
(
ability to initiate immune response
)
*
Survival of antigen in circulation
*
Immune status of recipient
*
Prior exposure to antigen
*
Genetic factors
*
Others Include
-
Age
-
Nutrition
/
health status
-
Size, shapes, & ionic charge of antigen
-
Dose and addition of adjuvants
(
enhance antigen reaction
)
-
Host must perceive antigen as foreign
What's the clinical significance for blood group antibodies?
Can cause:
Transfusion reactions
Hemolytic of disease of the new born
What "self" cells can antibodies be formed against?
RBCs, WBCs, & platelets
What is an autoantibody?
An antibody against self antigens
What determines the immunoglobulin class?
The heavy chain
Classes of Immunoglobulins
IgG
IgM
IgA
IgD
IgE
Activates Complement:
IgG IgM
Crosses Placenta:
IgG
IgG
*
Makes up about 70-80% of normal plasma immunoglobulins
*
4 Subclasses
*
Agglutination in saline is weak/negative
*
Most clinically significant antibodies are IgG
IgM
*
5 subunits
*
Makes up about 5-10% of total plasma concentration
*
Cannot cross placenta
*
Agglutinating & hemolyzing properties can be disrupted by some chemicals
*
Strong agglutination in saline
(
unlike IgG
)

Blood Bank
Quiz
1:
2:
3:
4:
5:
6:
1:
2:
3:
4:
5:
6:
Name the classes of immunoglobulins
IgG, IgM, IgA, IgD, IgE
Which activates complement?
IgG & IgM
Which can cross the placenta?
IgG
Which has 5 subunits?
IgM
Which is the most clinically significant?
IgG
Which has a strong agglutination in saline?
IgM
IgA - IgD - IgE
*
IgA is the predominant immunoglobulin in body secretions
-
Secretory IgA usually a dimer; Plasma IgA are mostly monomers
*
IgD
:
Trace amounts in plasma
*
IgE
:
Trace amounts in plasma
-
Most bound to basophils & mast cells
-
Involved in allergic & anaphylactic reactions
Complement
*
A set of serum & cell membrane proteins
*
React in cascade format
*
Involved in immunological reactions
*
Effects of products of complement cascade include
:
-
Cell lysis & Phagocytosis
*
Some blood group antibodies activate complement
-
Detected in anti-human test
(
AHG
)
Agglutination & Complement
*
Agglutination occurs in 2 stages
1:
Sensitization
2:
Agglutination or Cell lysis
Grading Agglutination Reactions
4+
One solid aggregate: clear background
3+
Several large aggregates: clear background
2+
Medium aggregates: pink background
1+
Small aggregates: reddish background
W+
Tiny aggregates: reddish background
Neg
No aggregates, RBCs visible
1:
2:
3:
4:

IgA is predominant in what?
Body secretions
Name the 2 stages of agglutination
Sensitization, Agglutination/Cell lysis
What is a 4+ agglutination reaction?
One solid aggregate w/clear background
What is the effect of the complement cascade?
Cell lysis/phagocytosis
Blood Bank
Quiz
1:
2:
3:
4:
Complement
(
Continued
)
*
Activation to the end of the cascade
(
c9
)
results in cell lysis
*
3 Pathways to complement activation
1:
Classic
2:
Alternative
3:
Mannose-binding lectin
*Classic Pathway
-Complement is activated due to antigen/antibody reaction
-May lead to cell sensitization (stops at C3) or cell lysis (C9)
*Alternative Pathway
-Activation due to molecules on surface of microorganisms, endotoxins, foreign proteins
Factors affecting antigen/antibody reactions
*Antibody Size
*Electrical repulsion
*Antigen/antibody ratio
*pH (optimal is 6.5-7.5)
*Ionic strength of medium (use of LISS)
*Temp (Optimal temp differ)
*Incubation time (helps give a reaction
*Effect of colloids (use of enhancement medium)
*Centrifugation (Brings Ab/Ag close)
*Number & location of antigens
*Dosage effect - some antibodies give stronger reaction w/homozygous RBCs then w/heterozygous RBCs
1:
2:
3:
1:
2:
3:
What is the end result of the casade C9?
Cell lysis
Name the 3 complement pathways
Classic, Alternative, Mannose-binding lectin
Name some factors affecting antigen/antibody reactions
Blood Bank
Quiz
*Antibody Size
*Electrical repulsion
*Antigen/antibody ratio
*pH (optimal is 6.5-7.5)
*Ionic strength of medium (use of LISS)
*Temp (Optimal temp differ)
*Incubation time (helps give a reaction
*Effect of colloids (use of enhancement medium)
*Centrifugation (Brings Ab/Ag close)
*Number & location of antigens
*Dosage effect - some antibodies give stronger reaction w/homozygous RBCs then w/heterozygous RBCs
AHG Productions
*Purified human proteins (Ig & C') are injected into animals
*Animal gives immune response
1: Collection of serum or
2: Collection of spleen cells & fuse w/melanoma cell line
*Further processing of serum (item 1) gives polyclonal reagent which contains both anti-IgG & anti-complement (Anti-C3d)
*Further processing of spleen cells results in production of cloned cells. These produce a monoclonal reagent that contains either anti-IgG or anti-complement
(sorry animals)
Antiglobulin Testing
(
AHG Test
)
*
Principle
-
Animals can form Abs to human proteins
(
globulins
)
-
These anti-protein Abs will react w
/
their corresponding Ag such as immunoglobulins or complement either in serum or bound to red cells
DAT
:
D
irect
A
ntiglobulin
T
est
*
DAT: Used to demonstrate in-vivo attachment of Ab and/or complement to RBCs
Uses of DAT
1:
Investigation of autoimmune hemolytic anemias (AIHA)
2:
Drug induced hemolytic anemia
3:
Hemolytic disease of the newborn
4:
Immune reactions to recently transfused RBCs
IAT :
I
ndirect
A
ntiglobulin
T
est
*
Used to demonstrate in-vitro reactions between RBCs & "Coating" Abs
Uses of IAT
1:
Ab detection & identification
2:
Blood Grouping
3:
Compatibility Testing (Matching blood types)
4:
Ab titration
Factors Affecting AHG Test
*Washing of Cells
*Saline for washing
*Centrifugation

--Following only affect IAT--
*Ratio of plasma to cells
*Reaction temperature
*Incubation time
*Reaction medium

AHG Testing :
False Positives
*
Polyagglutinable cells
*
Bacterial contamination of cells or saline
*
Cells that are DAT positive used in IAT procedure
*
Abs to reagents
(
LISS
)
*
Overcentrifugation
*
Poor reading technique
False
Negatives
*
Improper washing
*
Inactive AHG reagent
*
Failure to add AHG reagent
*
Failure to add plasma
(
IAT only
)
*
Undercentrifugation
*
Under incubation
*
Delays in testing
*
Sample deterioration
*
Cell suspension too weak
/
heavy
*
Poor reading technique
1:

2:

3:




4:

1:

2:

3:




4:

what is DAT used for?
Blood Bank
Quiz
1: Investigation of autoimmune hemolytic anemias (AIHA)
2: Drug induced hemolytic anemia
3: Hemolytic disease of the newborn
4: Immune reactions to recently transfused RBCs
What is IAT used for?
1: Ab detection & identification
2: Blood Grouping
3: Compatibility Testing (Matching blood types)
4: Ab titration
Factors affecting AHG test?
*Washing of Cells
*Saline for washing
*Centrifugation
--Following only affect IAT--
*Ratio of plasma to cells
*Reaction temperature
*Incubation time
*Reaction medium
Name some false positives
*Polyagglutinable cells
*Bacterial contamination of cells or saline
*Cells that are DAT positive used in IAT procedure
*Abs to reagents (LISS)
*Overcentrifugation
*Poor reading technique
Blood Bank reagents
*
The majority of blood bank testing falls into 2 categorie
1:
Use of known Ab to detect unknown Ag
(
plasma
/
serum
)
2:
Use of known Ag to detect unknown Ab
Sources of Antigen & Antibody
Known Source
Unknown Source
Antigen
Antibody
Reagent RBCs
Patient RBCs
Commercial Antisera
Patient Serum/Plasma
Routine Testing Procedures in Immunohematology
*
ABO & Rh typing
*
Ab screen
*
Ab identification
*
Crossmatch
(
compatibility testing
)
Categories of Reagents
*
Commercial antisera & reagent red cells are licensed by the FDA
-
Minimum standards for specificity & potency must be met
1:
Reagent red cells
(
contain known RBC antigens, usual concentration 2-5%
)
a:
A1 and B cells
(
ABO serum testing
)
b:
Screening cells
(
Antibody detection
)
c:
Panel cells
(
Antibody identification
)
-
panel & screen cells are always group O
2:
Antisera
(
contain known RBC antibodies
)
a:
Antisera for ABO typing
b:
Antisera for Rh typing
c:
Antisera for other blood groups
d:
Plant lectins
(
plant extracts that contain antibodies
)
3:
AHG Reagents
a:
AHG serum
/
reagent
-Monospecific
-Polyspecific
b:
Coombs control cells
(
Check cells
)
-
used as control for negative reactions
4:
Potentiators or Enhancementmedia adjust to in-vitro test environment to promote agglutination

-
LISS
(
Low ionic strength saline
)
-
PEG
(
Polyethylene glycol
)
-
BSA
(
Bovine serum albumin
)
-
Proteolytic enzymes
(
Ficin, papain
)
*
enzymes may enhance, inhibit, or have no effect on antigen
/
antibody reactions
Other Testing Methods in Blood Banking
*
Tube testing is most often used for routine blood bank testing.
Other methods in use include:
-
Gel technology
(
eliminates the wash step of AHG testing
)
*
Uses 0.8% RBC suspension
-
Microplate testing
:
uses microtiter plates; used in large blood centers
-
Solid phase adherence
:
Similar to microplate testing
Blood Bank
Quiz
1:
2:
3:

4:
5:

1:
2:
3:

4:
5:

Name the known source for RBCs
Reagent RBCs
What is the unknown source for RBCs?
The patient's RBCs
What is a routine test for Immunohematology?
ABO/Rh typing
Ab screen
Ab Identification
Compatibility testing (Crossmatch)
What does antisera contain?
Known RBC antibodies
What does gel technology eliminate?
The wash step of AHG testing
All credit
for the information contained in this prezi/study guide go to

Angela Njoku
Created by:
Christopher Brewer
Extra Credit Pending
Created on: 9-5-13
Full transcript