INTRODUCTION TO IMMUNOHEMATOLOGY  Are present on RBCs as glycolipids, proteins

(BLOOD BANKING) or glycoproteins

 Are inherited characteristics
Immunohematology  Have biological function
Combines aspects of; haematology, immunology &  Most are assigned to one of 31 blood group
genetics. o Immunologic reactions systems

Clinical Uses Blood Group Antibodies

1. Blood transfusion EXPECTED
The science of Blood Transfusion is mainly  Natural anti-A
concerned with how to provide patients with:  Natural anti-B
SAFE BLOOD (No transfusion reaction)
EFFECTIVE therapy (Haemoglobin increment) UNEXPECTED
2. Pregnancy: Prevent haemolytic disease of the  Alloimmune
new born and fetus.  Autoimmune
3. Autoimmune Haemolytic Anaemia  Passive

Review of Immunology
 The study of how the body defence itself against Blood Group Antibodies – Key Points
infections & diseases  Are stimulated by exposure to foreign
 Immune system = defence ministry antigens in the environment, or by
transfusion or pregnancy
Antigens and Antibodies  Are usually IgM and/or IgG
 BLOOD GROUP ANTIGENS: immunoglobulins
Present (predominantly) on red blood cells  Anti-A and anti-B are expected antibodies
 BLOOD GROUP ANTIBODIES (based on RBC ABO type)
Present in plasma (or serum)  All non-ABO antibodies are unexpected

Blood Group Antigens

 324 blood group antigens recognized
 33 blood group systems
 40 unassigned antigens
 Molecular biology of assigned antigens is
known Antigen-Antibody Reaction
System Notations Two Types of Tests
 Direct agglutination test for IgM antibodies
 Indirect antiglobulin test (IAT) for IgG
antibodies
Common ISBT Common Name ISBT
Name Name Name
Direct Tests – IgM Antibodies
Rh RH P P1PK ABO Typing
 Mix antibody and RBCs
Kell KEL Colton CO  Incubate (optional)
 Centrifuge (1000 x g, 15 seconds)
Duffy FY Dombrock DO  Examine
Kidd JK Cartwright YT

Lewis LE MNS MNS System Antigens Genes Phenotypes

Diego DI Lutheran LU
ABO A, B A, B, O A, B, O, AB
Antigens, Genes and Phenotypes
KEL K, k K, k K-k+; K+k+; K+k- or
K-1,2; K1,2; K-1,2
Blood Group Antigens – Key Points
FY Fya, Fyb Fya, Fyb Fy(a+b+); Fy(a+b-);
Fy(a+b-)

P P1 P1, P2 P1, P2
  Record results
dogs alive by transfusion of blood from other dogs.
Antigen-Antibody Reactions – Key Points
 Two types of tests are used to demonstrate 1667 Jean-Baptiste Denis in France and Richard
blood
group antigen-antibody reactions Lower in England separately report successful
• IgM antibodies are used (or detected) transfusions from lambs to humans. Within 10 years,
by direct agglutination tests
• IgG antibodies are used (or detected) transfusing the blood of animals to humans becomes
by indirect prohibited by law because of reactions.
antiglobulin tests
 The indirect antiglobulin test (IAT) utilizes
antihuman globulin (AHG) reagent, 1795 In Philadelphia, American physician Philip Syng
otherwise known as Coombs serum Physick, performs the first human blood transfusion,
ABO Typing although he does not publish this information.
Expected Reactions
1818 James Blundell, a British obstetrician, performs
RBCs + Plasma +
the first successful transfusion of human blood to a
Type Anti-A Anti-B A1 RBCs B RBCs patient for the treatment of postpartum hemorrhage.
Using the patient's husband as a donor, he extracts
O 0 0 + +
approximately four ounces of blood from the
A + 0 0 + husband's arm and, using a syringe, successfully

B 0 + + 0 transfuses the wife. Between 1825 and 1830, he

performs 10 transfusions, five of which prove
AB + + 0 0
beneficial to his patients, and publishes these results.
ABO and Rh Typing – Key Points
 Can be done by tube, gel and solid-phase He also devises various instruments for performing
assays transfusions and proposed rational indications.
 Two types of tests for RhD: a direct test, and
an IAT to detect weak expression of D
• Apparent D-negative donors (by 1840 At St. George's School in London, Samuel
direct tests) must be tested for weak
Armstrong Lane, aided by consultant Dr. Blundell,
D
• Apparent D-negative patients need performs the first successful whole blood transfusion
not be tested for weak of D to treat hemophilia.
 Different ethnic groups have disparate blood
group phenotype frequencies
1867 English surgeon Joseph Lister uses antiseptics to
WEEK 2 HISTORY
control infection during transfusions.
Highlights of Transfusion Medicine History

1628 English physician William Harvey discovers the 1873-1880 US physicians transfuse milk (from cows,
circulation of blood. Shortly afterward, the earliest goats, and humans).
known blood transfusion is attempted.

1884 Saline infusion replaces milk as a “blood

1665 The first recorded successful blood transfusion substitute” due to the increased frequency of adverse
occurs in England: Physician Richard Lower keeps reactions to milk.
1900 Karl Landsteiner, an Austrian physician, Dudley White, develops the Lee-White clotting time.
discovers the first three human blood groups, A, B, Adding another important discovery to the growing
and C. Blood type C was later changed to O. His body of knowledge of transfusion medicine, Lee
colleagues Alfred Decastello and Adriano Sturli add demonstrates that it is safe to give group O blood to
AB, the fourth type, in 1902. Landsteiner receives the patients of any blood group, and that blood from all
Nobel Prize for Medicine for this discovery in 1930. groups can be given to group AB patients. The terms
"universal donor" and "universal recipient" are coined.

1907 Hektoen suggests that the safety of transfusion

1914 Long-term anticoagulants, among them sodium
might be improved by crossmatching blood between
citrate, are developed, allowing longer preservation of
donors and patients to exclude incompatible mixtures.
blood.
Reuben Ottenberg performs the first blood transfusion
using blood typing and crossmatching in New York.
1915 At Mt. Sinai Hospital in New York, Richard
Ottenberg also observed the mendelian inheritance of
Lewisohn uses sodium citrate as an anticoagulant to
blood groups and recognized the “universal” utility of
transform the transfusion procedure from direct to
group O donors.
indirect. In addition, Richard Weil demonstrates the
feasibility of refrigerated storage of such
1908 French surgeon Alexis Carrel devises a way to
anticoagulated blood. Although this is a great advance
prevent clotting by sewing the vein of the recipient
in transfusion medicine, it takes 10 years for sodium
directly to the artery of the donor. This vein-to-vein or
citrate use to be accepted.
direct method, known as anastomosis, is practiced by
a number of physicians, among them J.B. Murphy in
1916 Francis Rous and J.R.Turner introduce a citrate-
Chicago and George Crile in Cleveland. The
glucose solution that permits storage of blood for
procedure proves unfeasible for blood transfusions,
several days after collection. Allowing for blood to be
but paves the way for successful organ
stored in containers for later transfusion aids the
transplantation, for which Carrel receives the Nobel
transition from the vein-to-vein method to indirect
Prize in 1912.
transfusion. This discovery also allows for the
establishment of the first blood depot by the British
1908 Moreschi describes the antiglobulin reaction.
during World War I. Oswald Robertson, an American
The antiglobulin is a direct way of visualizing an
Army officer, is credited with creating the blood
antigen-antibody reaction that has taken place but is
depots. Robertson received the AABB Landsteiner
not directly visible. The antigen and antibody react
Award in 1958 as developer of the first blood bank.
with each other, then, after washing to remove any
unbound antibody, the antiglobulin reagent is added
1927-1947 The MNSs and P systems are discovered.
and binds between the antibody molecules that are
MNSs and P are two more blood group antigen
stuck onto the antigen. This makes the complex big
systems — just as ABO is one system and Rh is
enough to see.
another.

1912 Roger Lee, a visiting physician at the

1932 The first blood bank is established in a
Massachusetts General Hospital, along with Paul
Leningrad hospital. Philadelphia, effectively treats victims of the Pearl
Harbor attack with Cohn's albumin for shock. Injected
1937 Bernard Fantus, director of therapeutics at the into the blood stream, albumin absorbs liquid from
Cook County Hospital in Chicago, establishes the first surrounding tissues, preventing blood vessels from
hospital blood bank in the United States. In creating a collapsing, a finding associated with shock.
hospital laboratory that can preserve and store donor
blood, Fantus originates the term "blood bank." 1943 The introduction by J.F. Loutit and Patrick L.
Within a few years, hospital and community blood Mollison of acid citrate dextrose (ACD) solution,
banks begin to be established across the United States. which reduces the volume of anticoagulant, permits
Some of the earliest are in San Francisco, New York, transfusions of greater volumes of blood and permits
Miami, and Cincinnati. longer term storage.

1939/40 The Rh blood group system is discovered by 1943 P. Beeson publishes the classic description of
Karl Landsteiner, Alex Wiener, Philip Levine, and transfusion-transmitted hepatitis.
R.E. Stetson and is soon recognized as the cause of
the majority of transfusion reactions. Identification of 1945 Coombs, Mourant, and Race describe the use of
the Rh factor takes its place next to the discovery of antihuman globulin (later known as the “Coombs
ABO as one of the most important breakthroughs in Test”) to identify “incomplete” antibodies.
the field of blood banking.
1947 The American Association of Blood Banks
1940 Edwin Cohn, a professor of biological chemistry (AABB) is formed to promote common goals among
at Harvard Medical School, develops cold ethanol blood banking practitioners and the blood donating
fractionation, the process of breaking down plasma public.
into components and products. Albumin, a protein
with powerful osmotic properties, plus gamma 1949-1950 The US blood collection system includes
globulin and fibrinogen are isolated and become 1,500 hospital blood banks, 46 community blood
available for clinical use. John Elliott develops the centers, and 31 American Red Cross regional blood
first blood container, a vacuum bottle extensively centers.
used by the Red Cross.
1950 Audrey Smith reports the use of glycerol
1940 The United States government establishes a cryoprotectant for freezing red blood cells.
nationwide program for the collection of blood.
Charles R. Drew develops the “Plasma for Britain” 1950 In one of the single most influential technical
program — a pilot project to collect blood for developments in blood banking, Carl Walter and W.P.
shipment to the British Isles. The American Red Cross Murphy, Jr., introduce the plastic bag for blood
participates, collecting 13 million units of blood by collection. Replacing breakable glass bottles with
the end of World War II. durable plastic bags allows for the evolution of a
collection system capable of safe and easy preparation
1941 Isodor Ravdin, a prominent surgeon from of multiple blood components from a single unit of
whole blood. Development of the refrigerated recognized.
centrifuge in 1953 further expedites blood component
therapy. 1962 The first antihemophilic factor (AHF)
concentrate to treat coagulation disorders in
1953 The AABB Clearinghouse is established, hemophilia patients is developed through
providing a centralized system for exchanging blood fractionation.
among blood banks. Today, the Clearinghouse is
called the National Blood Exchange. 1962 In the US, there were 4,400 hospital blood
banks, 123 community blood centers and 55
Mid-1950s In response to the heightened demand American Red Cross blood centers, collecting a total
created by open-heart surgery and advances in trauma of five to six million units of blood per year.
care patients, blood use enters its most explosive
growth period. 1964 Plasmapheresis is introduced as a means of
collecting plasma for fractionation.
1957 The AABB forms its committee on Inspection
and Accreditation to monitor the implementation of 1965 Judith G. Pool and Angela E. Shannon report a
standards for blood banking. method for producing Cryoprecipitated AHF for
treatment of hemophilia.
1958 The AABB publishes its first edition
of Standards for a Blood Transfusion Service (now 1967 Rh immune globulin is commercially introduced
titled Standards for Blood Banks and Transfusion to prevent Rh disease in the newborns of Rh-negative
Services). women.

1959 Max Perutz of Cambridge University deciphers 1969 S. Murphy and F. Gardner demonstrate the
the molecular structure of hemoglobin, the molecule feasibility of storing Platelets at room temperature,
that transports oxygen and gives red blood cells their revolutionizing platelet transfusion therapy.
color.
1970 Blood banks move toward an all-volunteer blood
1960 The AABB begins publication donor system.
of TRANSFUSION, the first American journal wholly
devoted to the science of blood banking and 1971 Hepatitis B surface antigen (HBsAg) testing of
transfusion technology. In this same year, A. Solomon donated blood begins.
and J.L. Fahey report the first therapeutic
plasmapheresis procedure — a procedure that 1972 Apheresis is used to extract one cellular
separates whole blood into plasma and red blood cells. component, returning the rest of the blood to the
donor.
1961 The role of platelet concentrates in reducing
mortality from hemorrhage in cancer patients is 1979 A new anticoagulant preservative, CPDA-1,
extends the shelf life of whole blood and red blood
cells to 35 days, increasing the blood supply and 1996 HIV p24 antigen testing of donated blood
facilitating resource sharing among blood banks. begins. Although the test does not completely close
the HIV window, it shortens the window period.
Early 1980s With the growth of component therapy,
products for coagulation disorders, and plasma 1997 U.S. Government issues two reports suggesting
exchange for the treatment of autoimmune disorders, ways to improve blood safety, including regulatory
hospital and community blood banks enter the era of reform.
transfusion medicine, in which doctors trained National Blood Data Resource Center founded by
specifically in blood transfusion actively participate in AABB to collect, analyze and distribute data on all
patient care. aspects of blood banking and transfusion medicine.

1981 First Acquired Immune Deficiency Syndrome 1998 HCV lookback campaign — a public health
(AIDS) case reported. effort to alert anyone who may have been exposed to
the hepatitis C virus (HCV) through blood
1983 Additive solutions extend the shelf life of red transfusions before July 1992 so they can receive
blood cells to 42 days. medical counseling and treatment if needed.

1984 Human Immunodeficiency Virus (HIV) 1999 Blood establishments begin using nucleic acid

identified as cause of AIDS amplification testing (NAT) under
FDA’s Investigational New Drug (IND) program;
1985 FDA approves enzyme-linked NAT employs testing technology that directly
immunosorbent assay (ELISA), first blood-screening detects genetic materials from viruses, including HCV
test to detect HIV antibodies. and HIV.

1987 Two tests that screen for indirect evidence of 2002 West Nile virus identified as transfusion
hepatitis are developed and implemented, hepatitis B transmissible.
core antibody (anti-HBc) and the alanine
aminotransferase test (ALT). 2002 Nucleic acid amplification test (NAT) for HIV
and HCV was licensed by the Food and Drug
1989 Testing of donated blood for human- Administration.
Tlymphotropic-virus-I-antibody (anti-HTLV-I)
begins.. 2003 First-ever National Blood Foundation forum
unites leaders in blood banking and transfusion
1990 Introduction of first specific test for hepatitis C, medicine
the major cause of “non-A, non-B” hepatitis.
2003 FDA issues final guidance regarding “Revised
1992 Implementation of testing donor blood for HIV- Recommendations for the Assessment of Donor
1 and HIV-2 antibodies (anti-HIV-1 and anti-HIV-2). Suitability and Blood and Blood Product Safety in
Cases of Known or Suspected West Nile Virus
 KEYPOINTS IN HISTORY:
Infection.”
1892 James Blundell of England
1901 Karl Landsteiner (ABO)
2003 First West Nile Virus-positive unit of blood 1902 Von Decastello & Sturli (AB)
1914 Hustin (Sodium Citrate)
intercepted. 1915 Lewisohn (min.amt. of Na. citrate)
1916 Rous & Turner (citrate dextrose
2003 Guidance on Implementation of New Bacteria sol’n)
1941 Dr. Charles Drew
Reduction and Detection Standard issued. 1943 Loutit & Mollison (ACD)
1957 \Gibson (CPD sol’n)
Right shift- decreases Hb affinity to oxygen, more oxide
2004 AABB receives $2.4 Million CDC grant to
release to tissue—
reduce transfusion-transmitted HIV in Africa and
Increase 2,3-bisphosphoglycerate (formerly 2,3-
South America. diphosphoglycerate/2,3- DPG) level
Left shift- increases Hb affinity to oxygen, less oxide
2005 FDA clears apheresis platelets collected with release to tissue—

certain systems for routine storage and patient Decreases 2,3-biphosphoglycerate (2,3-BPG)
level
transfusion up to 7 days when tested with a microbial
detection system release test. BLOOD BAGs
Single BB- collection, storage, transfusion of whole
blood.
2005 FDA’s Center for Biologics Evaluation and Double BB- separation of red cell & plasma.
Research publishes compliance program guidance for Triple BB- separation of red cell, platelet
concentration
inspection of human cells, tissues, and cellular and
Quadruple BB- separation of red cell, cryoprecipitate
tissue-based products (HCT/Ps). & plasma.

2005 AABB founding member Tibor Greenwalt dies.

2005 FDA approves the first West Nile virus (WNV)

blood test to screen donors of blood, organs, cells and
tissues.

2006 AABB starts collaborating with Centers for

Disease Control and Prevention to create CDC
PRESERVATIVE SOLUTION
National Healthcare Safety Network Hemovigilance
Module. ACRONYM Blood Chrom. Blood grp Chrom.
Acid citrate ACD-A 21 days grp location location
2014 FDA
dextrose approves first U.S. pathogen ABO 9 Cartwright 7
inactivation systems
(formula A*) for platelets and plasma.
MNS 4 Scianna 1
Citrate- CPD 21 days P 22 Colton 7
2017 FDA
phosphate- approves first two chimeric antigen
receptor (CAR) T cell therapies to treat cancer. Rh 1 Landsteiner- 19
dextrose Weiner
Citrate- CP2D 21 days Luthera 19 Chido/Rodgers 6
2018 FDA grants emergency use authorization
phosphate- n
(EUA) enabling U.S. military to use freeze-dried
double- Kell 7 H 19
plasma to treat hemorrhage in combat settings.
dextrose Lewis 19 Gerbich 2
Citrate- CPDA-1 35 days Duffy 1 Cromer 1
phosphate-
dextrose-
adenine
 Kidd 18 Knops 1 meiosis.
Diego 17 Indian 11

Additive Abbreviation Days

Sol’n
Adsol AS-1 42
(Baxter (mannitol)
Health Care)
Nutricel AS-3 (citrate 42
(Pall & phosphate)
Corporation
Optisol AS-5 42
(Terumo (mannitol)
Corporation WEEK3- REVIEW IN IMMUNOLOGY
) ANTIGENS
An antigen is a substance (molecule) that, when
Introduced into a human(or animal) who lacks that
DIFFERENCE OF GENOTYPE BETWEEN PHENOTYPE substance, triggers the production of antibody by
the body’s immune system. The antibody thus produced
Genotype actual genetic make up of individual
will rect specially with the antigen in an observable way.
Phenotype observable expression of inherited traits
ANTIBODIES
An antibody, also known as an immunoglobulin, is a
protein produced by the immune system following
exposure to foreign antigen. Antibodies, usually found
in plasma, react with cells carrying the foreign antigen
in a very specific manner.
foreign substance exposure
“Immunoglobulins”
Type O- Universal Donor (contains antigens)
Type AB- Universal Recipient

Immune Response- reaction of body to substance that

are foreign.
Immune Tolerance- tolerates transfused cells.

IMMUNE SYSTEM
Mendelial Principle INNATE/ ADAPTIVE/
Law of Independent Assortment Natural immunity Acquired immunity
The law of independent assortment says that genes (Primary): (Secondary):
for different traits segregate independently of each  non specific  specific
other. It means that separate traits are separately  fast  takes time
inherited. This is because during meiosis the  more effective
chromosomes line up randomly before the cell
divides, allowing for gamete formation. - first line of defence o The second line of
o fast, non specific defence
Law of Dominance o examples: oprotect the body against a
The law of dominance says that there are a. physical repeated attack by the same
dominant and recessive traits. Dominant traits are barriers, agent ( eg. Vaccination)
defined as whichever phenotype is expressed in an b. biochemical o specific, takes time but more
organism that is heterozygous for the trait. effectors effective
(complement) o Eg: lymphocytes
Law of Segregation
The law of segregation says that everyone has two and antibody formation
versions (called alleles) for each trait—one from
each parent—and that these alleles segregate
randomly (see independent assortment) during
 NON-SPECIFIC DEFENSES SPECIFIC PASS Natur Transfer x / Short
DEFENSES IVE al in
INNATE ADAPTIVE vivo/colo
1ST LINE 2ND LINE 3rd LINE strum
Skin Phagocytic Lymphocytes Artifi Infusion x / short
leukocyte cial of
Mucous Antimicrobial Antibodies serum/pl
membrane proteins asma
Secretion Inflammatory Memory cells injection
of skin & response
mucous
membrane
fever
Double (-) Stage- lacks CD4 & CD8
1ST LINE Double (+) Stage- expresses CD8 & CD4 antigens
Toll-like receptor- main feature of innate; pattern
recognition that recognize pathogen associated ANTIGENS
molecular patterns; to signal immune system that Factors affecting immunogenicity macromolecular
invaders are already present! size
Cytokines- plays an important role in innate chemical composition
immunity. molecular complexity
ability to processed & presented w/ MHC molecules
2nd LINE
EPITOPE- combined with antibody HAPTEN
phagocytosis- produces cytokines
molecules carrier molecules complete antigen
Complement system- classical, alternative, lymph.
pathway, cell lysis
RELATIONSHIP OF ANTIGEN TO HOST
*Inflammatory Response- for tissue damage. Increases
Autoantigens- produced by Ag of self
blood flow, blood vessel permeability
- react with Ag on pt’s own cells
 RUBOR- redness
(autoimmune reaction) & with same Ag
 CALOR-heat
on cells of other individuals
 TUMOR- swelling
 DOLOR- pain
Alloantigens- produced as result of immune stimulation
 FUNCTIO LAESA- loss of function
with Ag of the same species (human to human)
eg; transfusion or pregnancy
COMPONENTS OF IMMUNE RESPONSE
React with foreign Ag not present on pt’s own RBC
CELLULAR HUMORAL
IMMUNITY IMMUNITY
Heteroantibodies:
T lymphocyte B lymphocyte Ab produced from Ag of other species eg; vaccination
Produce cytokines Produce antibody
Acts against Acts against MAJOR HISTOCOMPATIBILITY COMPLEX
intracellular extracellular -Mount an immune response is linked to a group of
pathogen pathogens molecules originally referred to as HUMAN
LEUKOCYTE ANTIGEN.
2 TYPES OF ADAPTIVE IMMUNITY -The French scientist Dausset, gave them this , first
Mode of Antib Imme Durat defined by discovering an antibody response to
Acquisiti ody diate ion of circulating WBC.
on prod Respo Imm -These antigens “MHC” determined whether
uce nse une transplanted tissue is histocompatible & thus
by Resp accepted/recognized as foreign & rejected.
Host onse -MHC molecules are found on all nucleated cells in the
ACTI Natur Inf. / x long body.
VE al -Gene coding for MHC molecules in humans are found
Artifi Vacc. / x long in SHORT ARM OF CHROMOSOME 6.
cial
 CLASS I CLASS II CLASS activation complement complement
III if conditions activator
Gene Loci HLA-A, HLA-DP, C2, are optimal
HLA-B, HLA-DQ, C4A, Placental optimum 37̊C 4̊C
HLA-C HLA-DR C4B temperature
Chain α chain Α chain Number of antigen 2 10
structure β chain β chain site
Cell All nucleate APC & B Not
distributio cell cell expressed
n
Function Presentation Presentation Code for
of antigen of antigen C4,
to CD8 + T to CD4 + T Factor B,
cell cell TNF
IgM: first Ig to appear
ANTIBODIES LAG no antibody detected
is a specifically reactive immunoglobulin (Ig) produced LOG antibody titer increased arithmetically
in response to immunogenic stimulus (foreign) PLATEU stabilize
DECLINE PHASE antibody catabolize
Immuno: because of their function Ig- 15 days before antibody appears in serum
Globin: because of their nature (composition)

IG -humoral branch of immune response. IgG: secondary response; predominant Ig.

-end product of B lymphocyte (plasma cells) stimulation -lasts longer
COMPOSITION: 82%-96% polypeptide
2%-14% carbohydrates CLASSES OF IG
ELECTROPHORESIS: ph 8.6 gamma(Y)bands IgA- main IG responsible for mucosal immunity.
MAJOR CLASSES: IgG, IgM, IgA, IgD, IgE  Found in secretion (milk, saliva, tears, etc.)
---BASIC STRUCTURE OF IG---  Secretory IgA.
Light (L) Chain: kappa& lambda  Composed of 2 H2L2 subunits 7 one molecule
-both L chain types occur in all IG classes but  J chain
only contains 1 type of L chain.  Secretory component: protein derived from
Heavy (H) Chain- mu, gamma, alpha, delta, epsilon cleavage of poly-IG receptor; exists in serum as
---IG VARIABILITY--- monomer; secretions as dimer
Isotypic Variation- type of heavy chain that is unique to  TWO SUBCLASSES: IgA1, IgA
each IG class.
Allotypic Variation- genetic variation in the constant IgD- acts as antigen receptor when present in cells
regions of IG molecule. surface of B lymphocyte.
Idiotypic Variation- diversity at the binding site & in  -in serum, present in trace amounts
particular relates to hypervariable segments of antibody  -function is UNCLEAR
combining site (paratope)  (HALF LIFE: 2-3 DAYS)

COMPARISON OF IgG & IgM IgE- triggers allergic responses of immediate type (Type
CHARACTERSTIC IgG IgM I HSR) through release of chemical mediators.
Clinical Significance Causes in Usually does  -responsible for mounting immune response
vivo not cause in against parasitic infections. (mast cells)
hemolysis vivo
due to hemolysis
antibody IgG vs. IgM Antibodies
coating RBC
Size Monomer Pentamer Protein IgG IgM
Serum conc. Found in Found
largest conc. relatively Characteristic Immune Natural
Of all Ig in small
plasma amounts Stimulus Protein Carbohydrate
Complement Can activate Very good
 150
Blood Group weight 900 kDa
kDa
Complement binding Rarely Yes
Antigen binding
2 10
sites
Placental Transfer Yes No

Direct agglutinin Rarely Yes

Example Anti-Rh Anti-A, -B

COMPLEMENT PATHWAY
 20 proteins WEEK4- REVIEW IN SEROLOGY
 present in BLOOD in an inactive form Blood group serology
(proenzymes)
 can be activated by multiple pathways  the study of antigenic molecules present on
 when activated undergo a serious of reactions
various cellular & soluble components of whole
activation of C3 protein = results in cell lysis
blood, together with Abs and lectins recognize

them and their interactions

 In practice, this term is restricted to RBC surface

Ag and their interaction with specific Abs

Factors:
1. Intermolecular Binding Forces- responsible
for non-covalent bond.
2. Antibody Properties
FUNCTIONS: Affinity: strength of interaction between
antigen and antibody’s binding site at one
1. Enhances phagotcytosis
individual site.
2. Antibody function
Avidity: used to express binding strength of a
3. Clears immune complexes in blood
multivalent antigen with antisera produced in
4. Kills microbes by cell lysis
5. Causes the increased in vascular immunized individual.
permeability
Monovalent- antibody binds & complex is
6. Enhances platelet aggregation
7. Enhance smooth muscle contraction formed known as EPITOPE.
Multivalent- binding of one or more antigen-
8. Aids in viral neutralization
binding site on antibody molecule to more
CLASSICAL PATHWAY- activated by IgM/IgA. than one antigenic determinant.
ALTERNATIVE PATHWAY- activated by microbe
surface molecules. *Antigenic determinant of stimulating antigen-strong
LECTIN PATHWAY- activated by mannose/other
affinity.
sugar on microbe surface.
(Initiation, amplification, membrane attack) *Cross-reacting antigen-affinity not as strong.
 3. Host factors- includes nutritional status, (special type of agglutination), precipitation,
hormones, genetic inheritance, age, race, sex, agglutination inhibition and hemolysis.
physical activity level, environmental exposure -In vitro reaction in an artificial environment
and occurrence of disease or injury. such as in a test tube, microplate, or column.

4. Tolerance- exposure to an antigen during fetal AGGLUTINATION

life usually produces tolerance to that antigen. -involves particulate antigens (cells) which
-The induction of tolerance is used to prevent D- aggregate to form larger complexes when
negative mothers from developing anti-D specific antibodies are present.
antibodies after delivering Rh (+) infants.
*disease is called HEMOLYTIC DISEASE OF STAGES:
A NEWBORN. 1st Sensitization: occur when antigen-antibody
*mother should receive Rh immune globulin. become closely associated with antigenic
determinants or epitopes of the antigen.
DETECTION OF RBC ANTIGEN-ANTIBODY
REACTION

2nd Lattice formation: occurs when antibodies

on coated cells form cross-linkages between
cells resulting in visible clumping.

5. Centrifugation- mechanical way to bring

antigens and antibodies closer to each other,
increases the changes of agglutination.
-effective way to enhance a agglutination
reaction because it decreases the reaction time
by increasing gravitational forces on the
reactants and bringing reactants closer together.
-In vitro testing for the detection of antigens or
antibodies may be accomplished by commonly
used technique, including hemagglutination
Antigen-Antibody Ratio
Zone of equivalence- number of the binding
sites of multivalent antigen and antibody are
approximately equal.

Prozone effect- excess antibody causing false (-)

reaction.

Postzone effect- excess antigen causing false (-)

reaction.

Dosage effect- antibody reacts more strongly with

red blood cell carrying a double dose that a single
dose of an antigen.

DOSAGE EFFECT
 Fy (a+b-) exhibits a stronger reaction with anti- combining sites and a lack of agglutination is an
Fya antiserum than a person whose phenotype is indicator of (+) reaction.
Fy (a+b+).
Kidds and Duffy the Monkey (Rh) eat lots of -Involves HAPTEN that are complexed to
M&Ns. proteins. The hapten-protein conjugate is then
Kidds- Jka, Jkb attached to a carrier particle.
Duffy- Fya, Fyb ---COAGGLUTINATION---
Rh- C, c, E, e  name given to systems using bacteria as inert
MNSs- M, N, S, s particles to which antibody is attached.

5. ph- most antibodies bind red blood cell at Staphylococcus aureus is the most frequently used
optimum pH 6.5-7.5 because it has a protein on its outer surface called
-acidic pH enhances reaction with anti-D, anti- PROTEIN A.
M, and anti-Pr.
 ABSORBS FRAGMENT
6. Temperature- warm reactive antibodies are  CRYSTALLIZABLE PORTION OF ANTIBODY
reactive at 37̊C- IgG  MOLECULES LIKE IgG.
-cold reactive antibodies react at room *PROTEIN A absorbs Fc portion of the
temperature IgM. antibody.

DIRECT AGGLUTINATIOn AHG- Mediated Agglutination

 occur when antigen are found naturally on a “Coomb’s Test”
particle.
 typically, patient serum is diluted into a series of  Detects non-agglutinating antibody by means of
tubes or wells on a slide and reacted with coupling with second antibody .
bacterial antigens specific for the suspected  Key component test is antibody to human globulin
disease. that is made in animals or by means of
*If the agglutination reaction involves red blood cells, it HYBRIDOMA TECHNIQUE.
is called HEMAGGLUTINATION

 The Coomb’s test can be divided into two

HEMAGGLUTINATION. different types:
-interpretation of test is done on the basis of the cell
DIRECT- which detects in vivo
sedimentation pattern.
Test tubes also can be centrifuged and then shaken to see
if the cell can be evenly suspended. The degree of
agglutination is then graded.

---AGGLUTINATION INHIBITION---
-It is based on competition between particulate
and soluble antigens for limited antibody-
INDIRECT- in vitro sensitization (SERUM as the O A B AB
sample)
Asian 40% 28% 25% 7%
Black 51% 26% 19% 4%
Caucasian 45% 40% 11% 4%
Hispanic 57% 31% 10% 2%

ABO Genotypes and Phenotypes

INHERITANCE OF THE A,B. A AND H

WEEK 5 ABO BLOOD GROUPS ANTIGENS
ABO BLOOD GROUP SYSTEM
● An individual inherits one ABO gene
The ABO system is the most important of all blood
from each parent and that these two genes
groups in transfusion practice.
determine which ABO antigens are
present on the RBC membrane.
▸ Only blood group system in which people have
antibodies in their serum to antigens that are ● ABO genes: found in the long arm of
absent from their RBCs. chromosome 9
▸ Due to these antibodies, transfusion of
incompatible ABO type may result in immediate ● H genes: found in the chromosome 19
lysis of donor RBCs. (secretor of Locus) Lu-Lutheran, Le-
Lewis genes
DISCOVERY, LAWS, PROPERTIES INHERITANCE ● O genes: silent genes, called amorph, it
Landsteiner’s rule: rule stating that normal, is a gene that is unable to formed in
healthy individuals possess ABO antibodies to the detectable antigens.
ABO blood group antigens absent from their red
cells.
The inheritance of ABO genes follows simple
Mendelian genetics. OCCURRENCE and LOCATION of the ABO antigens -
O genes: silent genes, called amorph, it is a gene influenced by three genetically independent loci:
that is unable to formed in detectable antigens.  ABO, H, and Se.
Common structure: Oligosaccharide chain
ABO PHENOTYPE FREQUENCIES attached to either a protein of a lipid carrier
ABO group frequencies vary according to ethnicity. This will molecule
be an important fact to consider when working through ABO • Terminal sugars:
discrepancy resolution. The table below delineates frequencies
• D-galactose
of ABO occurrences according to ethnic groups.
• N-acetylglucosamine
 Type 1: body fluids
Type2 : glycolipids and glycoprotein in red cell VARIATION OF H-ANTIGEN CONCENTRATIONS
membrane IN ABO PHENOTYPES
Se Gene
▸ Se gene is responsible for the secretion of A,
DEVELOPMENT OF H ANTIGEN B, H antigens in the body fluids.
• H antigen is the only antigen in the H blood
group system.
• Chromosome 19 (closely linked with Se locus) • ▹ 80% of the population are secretors; 20% are
Produced by persons with the HH or Hh gene • H non-secretors
gene is present in >99.99% of the population ▹ Associated with saliva, tears, urine, digestive
• L-fucose added to the terminal galactose of the juices, bile, milk, amniotic fluid, serous fluids
type 1 and type 2 chain is called the and ovarian cyst.
immunodominant sugar for H antigens.
• The precursor of A and B antigens is the H
antigen.

The specificity of A and B antigen is defined by

immunodominant sugars:
• N-acetylgalactosamine (A antigen)
• D-galactose (B antigen)

COMPARISON OF ABH ANTIGENS ON RBCS

WITH A, B, AND H SOLUBLE SUBSTANCES

Formation of ABH Antigens