BVMLT -401 Serology and Immunology

UNIT-1

INTRODUCTION TO SEROLOGY

Historical Background of Immunology Immunology is defined as the study of the molecules, cells,
organs, and systems responsible for the recognition and disposal of foreign material. Immunology began
as a branch of microbiology.

Immunology is the study of the body's immune system and its functions and disorders. Serology is the
study of blood serum (the clear fluid that separates when blood clots).

 The immunological tests used in laboratories are made by producing artificial antibodies that exactly
“match” the substance or germ in question. When these antibodies come into contact with a sample of
blood, urine or stool, they bind to the matching substance or germ if found in the sample. This reaction
shows that the germ or substance is present.

Laboratory tests

In laboratory tests, sensitive devices measure the amount of bound antibodies based on the extent of a
light or color reaction. The greater the reaction, the more of the substance or germ is present. Laboratory
tests take longer than rapid tests but they are also more accurate.

Rapid test

In rapid tests, the antibodies are usually found on paper strips (test strips), but sometimes glass is used
too. Rapid tests are easy to use and provide instant results. But they are not as sensitive as laboratory tests
and can't determine exactly how much of the substance or germ is present.

APPLICATION

Bowel cancer screening: This test looks for the blood pigment hemoglobin, a sign of blood in stool.
Blood in stool can be caused by various things, such as hemorrhoids, polyps or even bowel cancer.

 Allergy tests: to detect antibodies against allergy-triggering substances like grass pollen or

certain foods.

 Detecting germs causing an infection: If it is thought someone has bacterial tonsillitis or scarlet
fever, the test looks for Streptococcus bacteria.

 Diagnosing heart attacks and thrombosis: Shortly after a heart attack or if someone has
thrombosis , higher levels of a certain protein are found in the blood. These can be detected using
an immunological test.

 Urine test: If sugar, blood, proteins or inflammatory cells are found in urine using this rapid test,
it could be a sign of diabetes, a urinary tract infection or kidney damage.

 Pregnancy test: Women can use this rapid test to find out whether their urine contains the
"pregnancy hormone" beta-hCG.
  Rapid tests for drugs and medication: Immunological tests can also be used to look for
recreational drugs such as cannabis, ecstasy and cocaine. Medical drugs that affect the central
nervous system can also be detected in this way. These include sleeping pills (benzodiazepines),
amphetamines and morphine.

 Determining your blood group: When blood transfusions are done, the person donating the
blood and the person receiving the blood have to have the same blood group. Immunological tests
can be used to determine the blood groups before a blood transfusion.

SAMPLES FOR IMMUNOLOGY

The sample requirements are test-dependent as outlined below. Note, that with the exception of
crossmatches, none of the immunology tests are performed STAT.

PREPARATION

Serum is the preferred sample. Body fluids (e.g. peritoneal fluid) and urine should be submitted in red top
tubes for electrophoresis. CSF can be submitted as EDTA or in red top tubes for ELP. We need a
minimum of 10 ml in a red top tube for an LE prep. Please contact the Clinical pathologist on duty if you
wish to run CSF electrophoresis.

COOMBS

EDTA blood (lavender top tube) is the required sample. Samples should be run promptly to prevent false
negatives. For this reason, we run Coombs tests on the same day we receive the sample (and the request
for this test), whenever possible.
CROSSMATCH

The most important component of the crossmatch is the major crossmatch. For this we need EDTA,
citrate or citrate-phosphate-dextrose (CPD, e.g. packed red cells) from the donor (or foal or stallion for a
mare-foal or mare-stallion incompatibility test) and serum from the recipient (or mare). The minor
crossmatch is less important, but we will perform this concurrently if the samples are submitted. For this,
we need serum from the donor (foal or stallion) and red cells from the recipient (mare).

So to summarize, collect 2 tubes from both donor and recipient; i.e. 1 red top and 1 EDTA (lavender top).

ANTIGEN

Antigen is a macromolecule that causes an immune response by lymphocytes. Antigen receptor, a surface
protein located on B cells and T cells, binds to antigens and initiates acquired immune responses. The
antigen receptors on B cells are called B cell receptors (or membrane immunoglobulins) and the antigen
receptors on T cells are called T cell receptors.

Antigens may either be proteins or polysaccharides. In general, an antigen is defined as a substance that
binds to specific antibodies, which in the human body are used to find and neutralize any potentially
harmful foreign substances in the bloodstream. The specific binding between antigen and antibody is
similar to that of the lock-and-key binding model.
In human blood, the different lettering of different blood types is designated by the specific antigen
present in the individual's blood cells. While all types contain the oligosaccharide (O) antigen, the A and
B blood types are defined by having N-acetylgalactose (A) or galactose (B) monosaccharide. Likewise,
the AB blood group has both A and B antigens. Additional antigens are bound to define the positive or
negative state of the ABO blood groups. The structures of the enzymes that bind to the antigen are similar
and very slightly different, demonstrating antigen specificity.

ANTIBODY
Antibody, also called immunoglobulin, a protective protein produced by the immune system in response
to the presence of a foreign substance, called an antigen. Antibodies recognize and latch onto antigens in
order to remove them from the body. A wide range of substances are regarded by the body as antigens,
including disease-causing organisms and toxic materials such as insect venom.

When an alien substance enters the body, the immune system is able to recognize it as foreign because
molecules on the surface of the antigen differ from those found in the body. To eliminate the invader, the
immune system calls on a number of mechanisms, including one of the most important—antibody
production. Antibodies are produced by specialized white blood cells called B lymphocytes (or B cells).
When an antigen binds to the B-cell surface, it stimulates the B cell to divide and mature into a group of
identical cells called a clone. 

ANTIBODY- STRUCTURE, CLASSES AND FUNCTIONS

Functions of Antibody

1. IgG provides long term protection because it persists for months and years after the prescence of
the antigen that has triggered their production.
2. IgG protect against bacteris, viruses, neutralise bacterial toxins, trigger compliment protein
systems and bind antigens to enhance the effectiveness of phagocytosis.
3. Main function of IgA is to bind antigens on microbes before they invade tissues. It aggregates the
antigens and keeps them in the secretions so when the secretion is expelled, so is the antigen.
 4. IgA are also first defense for mucosal surfaces such as the intestines, nose, and lungs.
5. IgM is involved in the ABO blood group antigens on the surface of RBCs.

MONOCLONAL ANTIBODIES

Monoclonal antibodies (mAb or moAb) are antibodies that are made by identical immune cells that are

all clones of a unique parent cell. Monoclonal antibodies can have monovalent affinity, in that they bind
to the same epitope (the part of an antigen that is recognized by the antibody). In contrast, polyclonal
antibodies bind to multiple epitopes and are usually made by several different plasma cell (antibody
secreting immune cell) lineages.

List and types of monoclonal antibodies (FDA approved)

Here is a list of examples some FDA-approved monoclonal antibody drugs.

 abciximab (Reopro)
 adalimumab (Humira, Amjevita)
 alefacept (Amevive)
 alemtuzumab (Campath)
 basiliximab (Simulect)
 belimumab (Benlysta)
 bezlotoxumab (Zinplava)
 canakinumab (Ilaris)
 certolizumab pegol (Cimzia)
 cetuximab (Erbitux)
 daclizumab (Zenapax, Zinbryta)
 denosumab (Prolia, Xgeva)
 efalizumab (Raptiva)

UNIT 2

SEROLOGY PRINCIPLE

IgG are immune antibodies produced as a result of exposure to the antigen and require a
special serological test (the Coomb's procedure) to detect their presence. ... Immunohematology uses
some basic serological tests to detect the presence of antibodies in an individual's serum and/or antigens
on an individual's cells.
SEROLOGY TESTS

Serologic tests are blood tests that look for antibodies in your blood. They can involve a number of
laboratory techniques. Different types of serologic tests are used to diagnose various disease conditions.

Serologic tests have one thing in common. They all focus on proteins made by your immune system. This
vital body system helps keep you healthy by destroying foreign invaders that can make you ill. The
process for having the test is the same regardless of which technique the laboratory uses during serologic
testing.

What happens during a serologic test?

A blood sample is all that the laboratory needs to conduct serologic testing.

The test will occur in your doctor’s office. Your doctor will insert a needle into your vein and collect
blood for a sample. The doctor may simply pierce the skin with a lancet if conducting serologic testing on
a young child.

The testing procedure is quick. The pain level for most people isn’t severe. Excessive bleeding and
infection may occur, but the risk of either of these is low.

TYPES OF SEROLOGY TESTS

Antibodies are diverse. So, there are various tests for detecting the presence of different types of
antibodies. These include:

 An agglutination assay shows whether antibodies exposed to certain antigens will cause particle
clumping.

 A precipitation test shows whether the antigens are similar by measuring for the presence of
antibody in body fluids.

 The Western blot test identifies the presence of antimicrobial antibodies in your blood by their
reaction with target antigens.

SYPHILIS
 Syphilis is a human infection of global importance. Its diagnosis can be challenging, requiring
construction of a serologic profile based on the results of at least two types of antibody tests: treponemal

and nontreponemal.

SYPHILIS TESTS
Syphilis is one of the most common sexually transmitted diseases (STDs). It is a bacterial infection spread
through vaginal, oral, or anal sex with an infected person. Syphilis develops in stages that can last for
weeks, months, or even years. The stages may be separated by long periods of apparent good health.
Syphilis usually starts with a small, painless sore, called a chancre, on the genitals, anus, or mouth. In the
next stage, you may have flu-like symptoms and/or a rash. Later stages of syphilis can damage the brain,
heart, spinal cord, and other organs. Syphilis tests can help diagnose syphilis in the early stages of
infection, when the disease is easiest to treat.

Other names: rapid plasma reagin (RPR), venereal disease Research laboratory (VDRL), fluorescent
treponemal antibody absorption (FTA-ABS) test, agglutination assay (TPPA), darkfield microscopy

SYPHILIS TEST USES:

Syphilis tests are used to screen for and diagnose syphilis.

Screening tests for syphilis include:

 Rapid plasma reagin (RPR), a syphilis blood test that looks for antibodies to the syphilis bacteria.
Antibodies are proteins made by the immune system to fight foreign substances, such as bacteria.
 Venereal disease research laboratory (VDRL) test, which also checks for syphilis antibodies. A VDRL
test can be done on blood or spinal fluid.
If a screening test comes back positive, you will need more testing to rule out or confirm a syphilis
diagnosis. Most of these follow up tests will also look for syphilis antibodies. Sometimes, a healthcare
provider will use a test that looks for actual syphilis bacteria, instead of the antibodies. Tests that look for
the actual bacteria are used less often because they can only be done in specialized labs by specially
trained health care professionals.

Definitions and classification of agglutination reactions

 Agglutination is defined as the formation of clumps of cells or inert particles by specific antibodies to
surface antigenic components (direct agglutination) or to antigenic components adsorbed or chemically
coupled to red cells or inert particles (passive hemagglutination and passive agglutination, respectively

C-REACTIVE PROTEIN (CRP)

A c-reactive protein test measures the level of c-reactive protein (CRP) in your blood. CRP is a protein
made by your liver. It's sent into your bloodstream in response to inflammation. Inflammation is your
body's way of protecting your tissues if you've been injured or have an infection. It can cause pain,
redness, and swelling in the injured or affected area. Some autoimmune disorders and chronic diseases
can also cause inflammation.

Normally, you have low levels of c-reactive protein in your blood. High levels may be sign of a serious
infection or other disorder.

Other names: c-reactive protein, serum

USED FOR:
A CRP test may be used to find or monitor conditions that cause inflammation. These include:

 Bacterial infections, such as sepsis, a severe and sometimes life-threatening condition

 A fungal infection
 Inflammatory bowel disease, a disorder that causes swelling and bleeding in the intestines
 An autoimmune disorder such as lupus or rheumatoid arthritis
 An infection of the bone called osteomyelitis

happens during a CRP test

A health care professional will take a blood sample from a vein in your arm, using a small needle. After
the needle is inserted, a small amount of blood will be collected into a test tube or vial. You may feel a
little sting when the needle goes in or out. This process usually takes less than five minutes.

Rheumatoid arthritis

Rheumatoid arthritis can be difficult to diagnose in its early stages because the early signs and symptoms
mimic those of many other diseases. There is no one blood test or physical finding to confirm the
diagnosis.

During the physical exam, your doctor will check your joints for swelling, redness and warmth. He or she
may also check your reflexes and muscle strength.

Blood tests
People with rheumatoid arthritis often have an elevated erythrocyte sedimentation rate (ESR, or sed rate)
or C-reactive protein (CRP), which may indicate the presence of an inflammatory process in the body.
Other common blood tests look for rheumatoid factor and anti-cyclic citrullinated peptide (anti-CCP)
antibodies.

Imaging tests

Your doctor may recommend X-rays to help track the progression of rheumatoid arthritis in your joints
over time. MRI and ultrasound tests can help your doctor judge the severity of the disease in your body.

STREPTOCOCCI

Structure

Streptococci are Gram-positive, nonmotile, nonsporeforming, catalase-negative cocci that occur in pairs
or chains. Older cultures may lose their Gram-positive character. Most streptococci are facultative
anaerobes, and some are obligate (strict) anaerobes. Most require enriched media (blood agar). Group A
streptococci have a hyaluronic acid capsule.

Classification and Antigenic Types

Streptococci are classified on the basis of colony morphology, hemolysis, biochemical reactions, and
(most definitively) serologic specificity. They are divided into three groups by the type of hemolysis on
blood agar: β-hemolytic (clear, complete lysis of red cells), α hemolytic (incomplete, green hemolysis),
and γ hemolytic (no hemolysis). Serologic grouping is based on antigenic differences in cell wall
carbohydrates (groups A to V), in cell wall pili-associated protein, and in the polysaccharide capsule in
group B streptococci.

Hepatitis B

Hepatitis B is an infection of the liver caused by the hepatitis B virus (HBV). It can be acute and resolve
without treatment. However, some forms can be chronic, and these could lead to cirrhosis and liver
cancer.

HBV can cause infection and inflammation of the liver. A person can have HBV and transmit the virus to
others without knowing that they have it.

Some people experience no symptoms. Some only have the initial infection, which then resolves. For
others, the condition becomes chronic. In chronic cases, the virus continues to attack the liver over time
without detection, resulting in irreversible liver damage.

Symptoms

Many HBV infections occur during infancy or childhood. This is because a mother can pass HBV to her
child during childbirth. However, doctors rarely diagnose HBV in childhood, as it causes few obvious
symptoms.
Symptoms of a new HBV infection may not be apparent in children under 5 years of age or in adults with
a suppressed immune system. Among those aged 5 years and over, around 30–50%Trusted Source will
show initial signs and symptoms.

Acute symptoms appear around 60–150 days after exposure to the virus, and they can last from several
weeks to 6 months.

Early symptoms

If HBV does cause symptoms early on, they may include:

 fever

 joint pain

 fatigue

 nausea

 vomiting

 loss of appetite

 abdominal pain

 dark urine

 clay colored stools

Transmission

HBV is transmissible when blood, semen, or another bodily fluid from a person with the virus enters the
body of an individual who does not have it.

More specifically, infection can occur:

 when a woman with HBV gives birth

 during sexual activity

 as a result of sharing needles, syringes, or other drug injection devices

 as a result of practicing unsafe tattoo techniques

 by sharing personal hygiene items, such as razors and toothbrushes

Health workers may be at risk through unsafe medical practices, such as reusing medical equipment, not
using personal protection, or incorrectly disposing of sharps.

HBV cannot spread through:

  food or water

 shared eating utensils

 breastfeeding

 hugging

 kissing

 holding hands

 coughing

 sneezing

 insect bites

Risk factors

People with a high risk of HBV include:

 the infants of mothers with HBV

 the sexual partners of people with HBV

 people who engage in sexual intercourse without contraception and those who have multiple
sexual partners

 men who have sex with men

 people who inject illicit drugs

 those who share a household with a person who has a chronic HBV infection

 healthcare and public safety workers who are at risk of occupational exposure to blood or
contaminated bodily fluids

 people receiving hemodialysis, which is a type of kidney treatment

 people taking medications that suppress the immune system, such as chemotherapy for cancer

 people with HIV

 those who come from a region with a high incidence of HBV

 all women during pregnancy

Prevention

People can prevent HBV infection by:

  wearing appropriate protective equipment when working in healthcare settings or dealing with
medical emergencies

 not sharing needles

 following safe sexual practices

 cleaning any blood spills or dried blood with gloved hands using a 1:10 dilution of one part
household bleach to 10 parts water

HIV

HIV is a virus that targets and alters the immune system, increasing the risk and impact of other
infections and diseases. Without treatment, the infection might progress to an advanced disease stage
called AIDS.

Human immunodeficiency virus (HIV) is a virus that attacks immune cells called CD4 cells, which are a
type of T cell.

These are white blood cells that move around the body, detecting faults and anomalies in cells as well as
infections. When HIV targets and infiltrates these cells, it reduces the body's ability to combat other
diseases.

Causes

People transmit HIV in bodily fluids, including:

 blood

 semen

 vaginal secretions

 anal fluids

 breast milk

In the United States, the main causes of this transfer of fluids are:

 anal or vaginal intercourse with a person who has HIV while not using a condom or PrEP, a
preventive HIV medication for people at high risk of infection

 sharing equipment for injectable illicit drugs, hormones, and steroids with a person who has HIV
A woman living with HIV who is pregnant or has recently given birth might transfer the disease to her
child during pregnancy, childbirth, or breastfeeding.

The risk of HIV transmitting through blood transfusions is extremely low in countries that have effective
screening procedures in place for blood donations.

WIDAL TEST

The Widal test is one method that may be used to help make a presumptive diagnosis of enteric fever,
also known as typhoid fever. Although the test is no longer commonly performed in the United States or
other developed countries, it is still in use in many emerging nations where enteric fever is  endemic and
limited resources require the use of rapid, affordable testing alternatives. While the method is easy to
perform, concerns remain about the reliability of the Widal test. It is not specific for typhoid fever and
can be positive when a person does not have the infection.

Enteric fever is a life-threatening illness caused by infection with the  bacterium Salmonella

enterica serotype Typhi (S. typhi), usually transmitted through food and drinks contaminated with fecal
matter. It is associated with symptoms that include high fever, fatigue, headache, abdominal pain,
diarrhea or constipation, weight loss, and a rash known as "rose spots." Early diagnosis and treatment are
important because serious complications, including severe intestinal bleeding or perforation, can develop
within a few weeks.

TESTING FOR TB INFECTION

There are two types of tests for TB infection: the TB skin test and the TB blood test.   A person’s health
care provider should choose which TB test to use. Factors in selecting which test to use include the reason
for testing, test availability, and cost. Generally, it is not recommended to test a person with both a TB
skin test and a TB blood test.

The TB skin test is also called the Mantoux tuberculin skin test (TST). A TB skin test requires two visits
with a health care provider.
On the first visit the test is placed; on the second visit the health care provider reads the test.

 The TB skin test is performed by injecting a small amount of fluid (called tuberculin) into the
skin on the lower part of the arm.
 A person given the tuberculin skin test must return within 48 to 72 hours to have a trained health
care worker look for a reaction on the arm.
 The result depends on the size of the raised, hard area or swelling

Positive skin test: This means the person’s body was infected with TB bacteria. Additional tests are
needed to determine if the person has latent TB infection or TB disease.
Negative skin test: This means the person’s body did not react to the test, and that latent TB infection or
TB disease is not likely.

There is no problem in repeating a TB skin test. If repeated, the additional test should be placed in a
different location on the body (e.g., other arm)

BETA-HCG

A simple method for the early detection of pregnancy: (A) Comparative study with radioimmunoassay of
beta-HCG: and (B) Fallacies compared to other immunological pregnancy tests.

 simple, sensitive and reliable non-radioactive method for the detection of hCG in concentrated
urine for the diagnosis of early pregnancy is reported. Twenty ml of urine were sampled, filtered
and concentrated by ultrafiltration with ultra-microporous membrane under reduced pressure
(Immersible Molecular Separator, Millipore Co) and the hCG in the concentrate was detected by
the ordinary latex agglutination inhibition method using beta-hCG antiserum to avoid cross-
reaction with high levels of hLH and hMG. Concentrated and unconcentrated urine samples taken
at different periods of amenorrhoea (1, 2 and 3 weeks) were also tested by two-slide pregnancy
tests and one-tube pregnancy test. Blood and urine samples taken at one and two weeks
amenorrhea were assayed for beta-hCG by a specific RIA.

ANTIGEN–ANTIBODY REACTION

Introduction

Although there are many different types of antigen–antibody reactions, blood bankers are often concerned
with reactions between antigens on red blood cells and antibodies in serum/plasma. These antigen–
antibody reactions can occur observably in varying proportions, with regard to volumes and strength of
reactants used. They are also reversible and are influenced by many factors. Antigen–antibody reactions
are enhanced in the laboratory to make them observable, in an effort to draw practical conclusions and
report on clinical conditions with accuracy

Common types of antigen–antibody reactions

haemagglutination

sensitization

haemolysis

neutralization

precipitation
Factors that influence antigen–antibody reactions

Use of proteolytic enzymes

Use of high molecular mass substances

Use of low ionic strength saline solution

Use of labelled antibodies

Role of complement

factors that influence the action of complement in laboratory tests

Role of antihuman globulin

application of antiglobulin tests in the laboratory

Polyclonal and monoclonal antibodies.

First and second stages of antigen–antibody reactions

When a blood sample is drawn, it is taken either into a dry test tube or into a test tube containing an
anticoagulant. Serum is the fluid part of a blood specimen taken into a dry test tube, and allowed to clot. It
has similar properties to plasma, but the coagulation factors are missing because they have been utilized
in the clotting process. The blood sample in the plain tube coagulates and in a short time the clot will
retract (shrink) sufficiently for the serum to be clearly visible.

First stage

This involves antigens and antibodies randomly bumping into each other in the test environment, and
when this occurs at the antigen site, the actual attachment of antibody to antigen takes place. It usually
happens very quickly and is affected by many variables. The reaction is not visible.

Second stage

This involves the demonstrable effect of attachment of antibody to antigen. This stage takes a longer time
to develop and may need to be enhanced in the laboratory in order for it to become observable.

The most widely used strategy in the laboratory to enhance the visibility of antigen–antibody reactions is
centrifugation. After allowing sufficient time for antibody to recognize and react with antigen, which may
be within seconds or may take much longer, up to 1 hour, tests can be centrifuged to force the cells closer
together. In this way agglutination may be enhanced, whereas cells that have not reacted with antibody
remain unagglutinated.

Sensitization
Sensitizing antibodies are IgG antibodies that are about 120 Å in length. Although they are able to
sensitize red cells with the corresponding antigens, zeta potential must be reduced or altered for the
smaller IgG antibodies to achieve haemagglutination. 

QUALITATIVE AND QUANTITATIVE AGGLUTINATION TESTS

 Qualitative tests determine the presence or absence of an antigen or antibody. Blood grouping
tests that are performed to determine the presence or absence of an antigen using an antibody of
known specificity are qualitative tests.
 For example, ABO blood grouping may be carried out by cell (or forward) grouping using
reagent anti‐A and anti‐B with red cells of unknown group, and reverse grouping of unknown
serum/plasma with reagent A and B cells.
 Quantitative tests determine the highest dilution at which an antibody is able to react with its
corresponding antigen. This endpoint is the titre of the antibody, expressed as a reciprocal (or
inverse) of the highest dilution at which agglutination was observed. For example, if the highest
dilution at which a reaction is observable is 1 in 32, then the titre is expressed as 32. Quantitative
tests may also be used to determine the international units of antibody in a serum/plasma sample.

Physical and chemical barriers

The immune system comprises both innate and adaptive immune responses. Innate immunity occurs
naturally due to genetic factors or physiology. It is not induced by infection or vaccination, but is
constantly available to reduce the workload for the adaptive immune response. The adaptive immune
response expands over time, storing information about past infections and mounting pathogen-specific
defenses.

Both the innate and adaptive levels of the immune response involve secreted proteins, receptor-mediated
signaling, and intricate cell -to-cell communication. From an historical perspective, the innate immune
system developed early in animal evolution, roughly a billion years ago, as an essential response to
infection. In the innate immune response, any pathogenic threat triggers a consistent sequence of events
that can identify the type of pathogen and either clear the infection independently or mobilize a highly-
specialized adaptive immune response.

ELISA

 An enzyme-linked immunosorbent assay (ELISA) is used to detect the presence of an antigen in a
sample. The antigen is immobilized to the well of a plate by adsorption, or captured with a bound,
antigen-specific antibody. A detection antibody is then added forming a complex with the antigen, if
present.
 Procedure

1. - Remove the coating solution and wash the plate three times by filling the wells with 100 μl PBS-
0.05%Tween20. The solutions or washes are removed by flicking the plate over a sink. The remaining
drops are removed by patting the plate on a paper towel.
2. - Block the remaining protein-binding sites in the coated wells by adding 100μl blocking buffer, 3% skim
milk in PBS per well. Incubate for 1 hour at RT with gentle shaking.
3. - Wash the plate three times with 100ul PBS-0.05% Tween 20.

4. - Add 50µl of diluted antibody to each well. Incubate the plate at 37 ℃ for an hour with gentle shaking.

5. - Wash the plate six times with 100ul PBS-0.05%Tween 20.

6. - Add 50μl of conjugated secondary antibody, diluted at the optimal concentration (according to the
manufacturer) in blocking buffer immediately before use. Incubate at 37 ℃ for an hour.

7. - Wash the plate six times with 100ul PBS-0.05%Tween20.

8. - Prepare the substrate solution by mixing acetic acid, TMB and 0.03% H2O2 with the volume ratio of
4:1:5.

9. - Dispense 50μl of the substrate solution per well with a multichannel pipe. Incubate the plate at 37 ℃ in
dark for 15-30mins.

10. - After sufficient color development, add 100μl of stop solution to the wells (if necessary).

11. - Read the absorbance (optical density at 450nm) of each well with a plate reader.
 UNIT 3

INTRODUCTION TO IMMUNOLOGY AND ANTIGEN ANTIBODY REACTION

INTRODUCTION
Reactions of antigens and antibodies are highly specific. An antibody will react only with the antigen that
induced it or with a closely related antigen. Because of the great specificity, reactions between antigens
and antibodies are suitable for identifying one by using the other. This is the basis of serologic reactions.
However, cross-reactions between related antigens can occur, and these can limit the usefulness of the
test.

The results of many immunologic tests are expressed as a titer, which is defined as the highest dilution of
the specimen (e.g., serum) that gives a positive reaction in the test.

Physical and Chemical Barriers

Before any immune factors are triggered, the skin functions as a continuous, impassable barrier to
potentially infectious pathogens. Pathogens are killed or inactivated on the skin by desiccation (drying
out) and by the skin’s acidity. In addition, beneficial microorganisms that coexist on the skin compete
with invading pathogens, preventing infection.

Regions of the body that are not protected by skin (such as the eyes and mucus membranes) have
alternative methods of defense, such as tears and mucus secretions that trap and rinse away pathogens,
and cilia in the nasal passages and respiratory tract that push the mucus with the pathogens out of the
body. Throughout the body are other defenses, such as the low pH of the stomach (which inhibits the
growth of pathogens), blood proteins that bind and disrupt bacterial cell membranes, and the process of
urination (which flushes pathogens from the urinary tract).
Phagocytosis

Phagocytosis, process by which certain living cells called phagocytes ingest or engulf other cells or
particles. The phagocyte may be a free-living one-celled organism, such as an amoeba, or one of the body
cells, such as a white blood cell. In some forms of animal life, such as amoebas and sponges,
phagocytosis is a means of feeding. In higher animals phagocytosis is chiefly a defensive reaction against
infection and invasion of the body by foreign substances (antigens).

PHAGOCYTOSIS

Phagocytosis Definition

Phagocytosis, or “cell eating”, is the process by which a cell engulfs a particle and digests it. The word
phagocytosis comes from the Greek phago-, meaning “devouring”, and -cyte, meaning “cell”. Cells in the
immune systems of organisms use phagocytosis to devour bodily intruders such as bacteria, and they also
engulf and get rid of cell debris. Some single-celled organisms like amoebas use phagocytosis in order to
eat and acquire nutrients.

Function of Phagocytosis

The function of phagocytosis is to ingest solid particles into the cell. Phagocytosis is a type
of endocytosis, which is when cells ingest molecules via active transport as opposed to molecules
passively diffusing through a cell membrane. Only certain small molecules can pass through the cell
membrane easily; larger ones have to go through special channels in the cell or be ingested via
endocytosis. Other types of endocytosis include pinocytosis, also called “cell drinking”, and receptor-
mediated endocytosis, which is when molecules bind to specific receptors on the cell membrane that
causes the cell to engulf them.
Phagocytosis is different from pinocytosis because phagocytosis involves the ingestion of solid particles
while pinocytosis is the ingestion of liquid droplets. Phagocytosis is also used by cells to take in much
larger particles than those that are ingested through pinocytosis. Some single-celled protists, such as
amoebae, use phagocytosis to ingest food particles;

STEPS OF PHAGOCYTOSIS

Step 1:

The cell that will perform phagocytosis is activated. This can be a phagocyte, which is a cell in the
immune system that performs phagocytosis, or an organism such as an amoeba, which behaves in a
similar way to phagocytes when it carries out phagocytosis. In the case of immune cells, activation occurs
when the cells are near bacterial cells or parts of bacterial cells. Receptors on the surface of the cells bind
to these molecules and cause the cells to respond.

Step 2:

In the immune system, chemotaxis may occur. Chemotaxis is the movement of phagocytes toward a
concentration of molecules. Immune cells pick up chemical signals and migrate toward invading bacteria
or damaged cells.

Step 3:

The cell attaches to the particle that it will ingest. Attachment is necessary for ingestion to occur. Some
bacteria can resist attachment, making it harder for them to be taken into the cell and destroyed.

Step 4:

The cell ingests the particle, and the particle is enclosed in a vesicle (a sphere of cell membrane with fluid
in it) called a phagosome. The phagosome transports the particle into the cell.

Step 5:
A lysosome fuses with the phagosome and the particle is digested. Lysosomes are vesicles that contain
hydrolytic enzymes that break down molecules. A phagosome fused with a lysosome is called a
phagolysosome.

INFLAMMATION

When a wound swells up, turns red and hurts, it may be a sign of inflammation. Very generally speaking,
inflammation is the body’s immune system’s response to an irritant. The irritant might be a germ, but it
could also be a foreign object, such as a splinter in your finger.

This means that an inflammation doesn’t only start when, for instance, a wound has already been infected
by bacteria, is oozing pus or healing poorly. It already starts when the body is trying to fight against the
harmful irritant.

Causes of an inflammation

Many different things can cause inflammations. These are the most common:

 Pathogens (germs) like bacteria, viruses or fungi

 External injuries like scrapes or damage through foreign objects (for example a thorn in your
finger)

 Effects of chemicals or radiation

Diseases or medical conditions that cause inflammation often have a name ending in “-itis.” For example:

 Cystitis: an inflammation of the bladder

 Bronchitis: an inflammation of the bronchi

 Otitis media: an inflammation of the middle ear

 Dermatitis: a disease where the skin is inflamed

Signs of an inflammation

There are five symptoms that may be signs of an acute inflammation:

 Redness

 Heat

 Swelling

 Pain

 Loss of function

General responses in the body

If the inflammation is severe, it can cause general reactions in the body. These may include the following
signs and symptoms:
  Generally feeling ill, exhaustion and fever. These are signs that the immune system is very active
and needs a lot of energy, which may be lacking for other activities. If the rate of metabolism is
higher due to a fever, more antibodies and cells of the immune system can be produced.

 Changes in the blood, such as an increased number of immune system cells.

Acute inflammation

An acute inflammation is one that starts rapidly and becomes severe in a short space of time. Signs and
symptoms are normally only present for a few days but may persist for a few weeks in some cases.

Examples of diseases, conditions, and situations that can result in acute inflammation include:

 acute bronchitis

 infected ingrown toenail

 a sore throat from a cold or flu

 a scratch or cut on the skin

 high-intensity exercise

 acute appendicitis

 dermatitis

 tonsillitis

 infective meningitis

 sinusitis

 a physical trauma

Chronic or acute inflammation

The following table shows the key differences between acute and chronic inflammation:

Acute Chronic

Caused Harmful bacteria or tissue Pathogens that the body cannot break down, including
by injury some types of virus, foreign bodies that remain in the
 system, or overactive immune responses

Onset Rapid Slow

Duration A few days From months to years

Inflammation improves, turns

Tissue death and the thickening and scarring of
Outcomes into an abscess, or becomes
connective tissue
chronic

CHRONIC INFLAMMATION

This refers to long-term inflammation and can last for several months and even years. It can result from:

 failure to eliminate whatever was causing an acute inflammation

 an autoimmune disorder that attacks normal healthy tissue, mistaking it for a pathogen that causes
disease

 exposure to a low level of a particular irritant, such as an industrial chemical, over a long period

Examples of diseases and conditions that include chronic inflammation:

 asthma

 chronic peptic ulcer

 tuberculosis

 rheumatoid arthritis

 periodontitis

 ulcerative colitis and Crohn's disease

 sinusitis

 active hepatitis

A high body temperature, or fever, is one of the ways our immune system attempts to combat an
infection. Usually, the rise in body temperature helps the individual resolve an infection. However,
sometimes it may rise too high, in which case, the fever can be serious and lead to complications.
Doctors say that as long as the fever is mild, there is no need to bring it down - if the fever is not severe, it
is probably helping to neutralize the bacterium or virus that is causing the infection. Medications to bring
down a fever are called antipyretics. If the fever is causing undue discomfort, an antipyretic may be
recommended.

Symptoms

 feeling cold when nobody else does

 shivering

 lack of appetite

 dehydration — preventable if the person drinks plenty of fluids

 depression

 hyperalgesia, or increased sensitivity to pain

 lethargy

 problems concentrating

 sleepiness

 sweating

Factors influencing Immunogencity

Nature of the Immunogencity
1. Foreignness
2. Molecular size
3. Chemical composition and heterogeneity
4. Ability to be  processed and presented with an MHC molecule on the surface of Antigen
Presenting Cells (APCs) or altered self-cell
Biological system that the antigen encounters
1. Genotype of the recipient animal
2. Dosage and route of administration

Factors influencing Immunogenicity

Ⅰ. Foreignness
Antigens must be recognized as non-self by the biological system
Degree of immunogenicity depends on the degree of foreignness    i.e. The greater the phylogenetic
distances between two species, the greater the structural (and therefore the antigenic) disparity between
them.

e.g. If Bovine serum albumin (BSA) is injected in Cow, Rabbit and Chicken, the order of
Immunogenicity will be:
EPITOPE 
An epitope is the part of an antigen that interacts with the antigen-specific receptor or antibody. Strictly
speaking, an epitope is determined by the specificity of the clonally distributed receptor to which an
antigen binds.
However, the part of an antigen that can reproducibly elicit B cell or T cell responses is often referred to
as an epitope without reference to the specificity defined by the particular receptor. Epitopes, which are
often used interchangeably with antigenic sites or antigenic determinants, can be classified as B cell
epitopes or T cell epitopes on the basis of the types of cellular responses they elicit.

PRINCIPLE:

Immunoelectrophoresis is powerful technique to characterized antibodies. This technique is based on

the principle of electrophoresis of antigen for immunodiffusion with a poly specific antiserum to form
precipitin bands.

CFT

Combating the Financing of Terrorism (CFT) involves investigating, analyzing, deterring, and preventing
sources of funding for activities intended to achieve political, religious, or ideological goals. CFT is
achieved through violence and the threat of violence against civilians.

WESTERN BLOTTING
Western blotting (or immunoblotting) is a widely used method for protein detection, using antibody-based
probes to obtain specific information about target proteins from complex samples. It is a routine method
in molecular biology, biochemistry, and cell biology fields with a multitude of applications.
This method can be used to obtain information about quantity, molecular weight, and  post-translational
modifications of proteins. Due to high affinities of antibody toward their epitopes and amplificatory
nature of Western blotting, it is a very sensitive method and even picogram quantities of target proteins
can be detected.
UNIT-4

DIFFERENCE BETWEEN HUMORAL AND CELL-MEDIATED IMMUNITY

The primary difference between them is the mechanism of immunity, where the Humoral

immunity produces antibodies against the antigens which are present outside the infected cells or free
circulating in the blood. Cell-mediated immunity works inside the infected cells, where it destroyed the
pathogens or microorganisms by the process of lysis by the releasing cytokines.

Humoral immunity shows quick response against the pathogens, while cell-mediated immunity is slow
in action. Both the type are part of the adaptive immune system. Our immune system provides the
protection and resistance against the infectious disease, which is offered by the host cell present in the
body.

The immune system has complex networks of the molecules, cells and their interactions are designed to
eradicate the infectious organisms from the body. Immunity or immune system is divided into two types –
innate (non-specific) and acquired or adaptive (specific) immunity.

IMMUNOGLOBULINS - STRUCTURE AND FUNCTION  

I. DEFINITION 

Immunoglobulin (Ig)

Immunoglobulin’s are glycoprotein molecules that are produced by plasma cells in response to an
immunogen and which function as antibodies. The immunoglobulins derive their name from the finding
that they migrate with globular proteins when antibody-containing serum is placed in an electrical field
(Figure 1).

II. GENERAL FUNCTIONS OF IMMUNOGLOBULINS

A. Antigen binding
Immunoglobulins bind specifically to one or a few closely related antigens. Each immunoglobulin
actually binds to a specific antigenic determinant. Antigen binding by antibodies is the primary function
of antibodies and can result in protection of the host. The valency of antibody refers to the number of
antigenic determinants that an individual antibody molecule can bind. The valency of all antibodies is at
least two and in some instances more.

B. Effector Functions
Frequently the binding of an antibody to an antigen has no direct biological effect. Rather, the significant
biological effects are a consequence of secondary "effector functions" of antibodies. The
immunoglobulins mediate a variety of these effector functions. Usually the ability to carry out a particular
effector function requires that the antibody bind to its antigen. Not every immunoglobulin will mediate all
effector functions. Such effector functions include:

1. Fixation of complement - This results in lysis of cells and release of biologically active molecules (see
chapter two)

2. Binding to various cell types - Phagocytic cells, lymphocytes, platelets, mast cells, and basophils have
receptors that bind immunoglobulins. This binding can activate the cells to perform some function. Some
immunoglobulins also bind to receptors on placental trophoblasts, which results in transfer of the
immunoglobulin across the placenta. As a result, the transferred maternal antibodies provide immunity to
the fetus and newborn
 

III. BASIC STRUCTURE OF IMMUNOGLOBULINS

The basic structure of the immunoglobulins is illustrated in figure 2. Although different immunoglobulins
can differ structurally, they all are built from the same basic units.

A. Heavy and Light Chains

All immunoglobulins have a four chain structure as their basic unit. They are composed of two identical
light chains (23kD) and two identical heavy chains (50-70kD)

B. Disulfide bonds

1. Inter-chain disulfide bonds - The heavy and light chains and the two heavy chains are held together
by inter-chain disulfide bonds and by non-covalent interactions The number of inter-chain disulfide bonds
varies among different immunoglobulin molecules.

2. Intra-chain disulfide binds - Within each of the polypeptide chains there are also intra-chain disulfide
bonds.

C. Variable (V) and Constant (C) Regions

When the amino acid sequences of many different heavy chains and light chains were compared, it
became clear that both the heavy and light chain could be divided into two regions based on variability in
the amino acid sequences. These are the:

Factors Affecting Antibody Production

Immunogenicity: Only a few red cell antigens are very immunogenic (e.g., ABH in the ABO system, D
in the Rh system, and Kk in the Kell system). Other Rh antigens (CcEe) and antigens in the Kidd and
Duffy systems are not nearly as immunogenic. The strong immunogenicity for the D antigen is shown by
the fact that approximately 70% of Rh(D) negative persons produce anti-D upon exposure to the D
antigen. After the D antigen, the K antigen appears to be the most immunogenic.
Antigen Volume: Exposure to a very small volume of antigen may result in immune tolerance. Similarly,
exposure to a large volume of foreign antigens simultaneously may produce immune paralysis. In general,
the greater the volume of foreign antigen, the more likely it is that antibody will be produced.

The mechanism of cell mediated immunity includes:

(1) Activation of antigen-specific cytotoxic T-lymphocytes that is able to induce apoptosis in body cells
displaying epitopes of foreign antigen on their surface such as cells with intracellular bacteria, virus-
infected cells and cancer cells displaying tumor antigens.

(2) Activating macrophages and natural killer cells, enabling them to destroy intracellular pathogens.

(3) Stimulating cells to secrete a variety of cytokines that influence the function of other cells involved in
immune responses.

Irrespective of the type of cells involved in CMI reaction, the complete mechanism of cell mediated
immunity proceeds in three phases.

(a) Binding of the cytotoxic cells to the target cells (b) Release of cytokines (c) Lysis of
the target cell.

T CELLLS

T cells (also called T lymphocytes) are one of the main components of the adaptive immune system.
They are vital in hosting an immune response against pathogens.

T cells play a major role in defence against intracellular pathogens such as viruses, protozoa and
intracellular bacteria, and in immunity to extracellular pathogens by providing help for the antibody
response.

This article shall discuss the production of T cells, the different types present in the immune system and
relevant clinical conditions.

Types of T Cells
There are 4 main types of T cells.

CD4+ Helper Cells

CD4+ helper cells help in the maturation of B cells into plasma cells and memory B cells. They also help
activate cytotoxic T cells and macrophages.

They become activated when they are presented with peptide antigens by MHC Class II molecules,
which are expressed on the surface of antigen presenting cells (APCs).

Once activated, they divide rapidly and secrete small proteins called cytokines that regulate or assist in
the active immune response. There are various subtypes present within the immune system that are able to
secrete different cytokines depending on the immune response occurring, however these are beyond the
scope of this article.

CD8+ Cytotoxic Cells

CD8+ cytotoxic cells cause lysis of virus-infected and tumour cells. They are also involved in transplant
rejection. These cells recognize their targets by binding to antigen associated with  MHC Class I
molecules which are present on the surface of all nucleated cells.

Memory T Cells

Once they come into contact with an antigen naive T cells differentiate into effector cells (CD4+ and
CD8+ cells) and memory T cells. Memory T cells are long-lived and can quickly expand to large numbers
of effector T cells upon re-exposure to the antigen.

They provide the immune system with “memory” against previously encountered pathogens. Memory T
cells may be either CD4+ or CD8+.

Natural Killer T Cells

They bridge the adaptive immune system with the innate immune system. Whilst most T cells function
based on recognition of MHC class molecules, natural killer T cells are able to recognise other antigen
classes. Once activated they are also able to perform the same functions as CD4+ and CD8+ cells.

complement system

The complement system is a part of the immune system, consists of a series of proteins that interact with
one another in a highly regulated manner, in order to eliminate pathogens. It helps antibodies and
phagocytic cells to clear pathogens and damaged cells; promote inflammation and attack pathogen’s
plasma membrane. Proteins that take part in the complement system are called complements that
collectively work as a biological cascade; 

Complement Activation and cell lysis

The complement activation occurs via three pathways; which are:

1. Classical pathway
2. Alternative pathway
3. Lectin pathway (or mannose binding lectin pathway)

4. Difference Between T Cells and B Cells

5. Definition
6. T Cells: T cells are a type of lymphocyte, which develops in the thymus, circulates in
the blood and lymph and
7. mediates the immune response against malignant or infected cells in the body by the secretion of
lymphokines or by
8. direct contact.
 9. B Cells: B cells are a type of lymphocyte, which develops in the bone marrow, circulates in the
blood and lymph, and
10. upon recognizing a particular pathogen, differentiates into a plasma cell clone, secreting
specific antibodies and a
11. memory cell clone, for the subsequent encountering of the same pathogen
DIFFERENCE BETWEEN T CELLS AND B CELLS

T cells and B cells differ in their functions, like T cells are known to develop various immune
response such as invading bacteria from body’s immune system, virus attacks, not supporting the organ
transplant, etc., while B cells produce antibodies against the antigen. Despite showing variance in their
working, T and B cells struggle with the same aim of destroying the invader or foreign particles which are
harmful to the body.

Our body’s immune system is supported by many essential cells, among them, lymphocytes are one of
them. As these are such white blood cells which are produced in bone marrow and further become
specialized in two main parts which are T cells and B cells. When the body is attacked by the virus or
bacteria or any parasites, suddenly the immune alarm activates and starts with the chain of reactions of
cellular activity in the body’s immune system.

The term "cytokine" is derived from a combination of two Greek words - "cyto" meaning cell and "kinos"
meaning movement. Cytokines are cell signalling molecules that aid cell to cell communication in
immune responses and stimulate the movement of cells towards sites of inflammation, infection and
trauma.

ytokine function / Function of cytokine

Cytokines are a large group of proteins, peptides or glycoproteins that are secreted by specific cells of
immune system. Cytokines are a category of signaling molecules that mediate and regulate immunity,
inflammation and hematopoiesis.

LYMPHOKINES

Lymphokines are a subset of cytokines that are produced by a type of immune cell known as a
lymphocyte. They are protein mediators typically produced by T cells to direct the immune system
response by signaling between its cells.

This response, if successful, leads to the elimination of the foreign substance and thus removes the
associated pathogen from the body. The immune response consists of the expansion and differentiation of
cells bearing clonally expressed surface receptors capable of binding the foreign substance. This leads to
the secretion of specific antibodies and the production of a set of potent polypeptides that regulate the
activation, growth, and differentiation of the cells of the immune system. These polypeptides also mediate
the inflammatory responses that are elicited as a result of interactions among immunocompetent cells

DIFFERENCE BETWEEN T CELLS AND B CELLS

Definition

T Cells: T cells are a type of lymphocyte, which develops in the thymus, circulates in the blood
and lymph and

mediates the immune response against malignant or infected cells in the body by the secretion of
lymphokines or by

direct contact.

B Cells: B cells are a type of lymphocyte, which develops in the bone marrow, circulates in the blood and
lymph, and

upon recognizing a particular pathogen, differentiates into a plasma cell clone, secreting specific
antibodies and a

memory cell clone, for the subsequent encountering of the same pathogen.

Origin

T Cells: T cells originate in the bone marrow and mature in the thymus.

B Cells: B cells originate and mature in the bone marrow.

Complement System 

The defense against pathogens such as viruses and bacteria are mediated by the immune system which is
in principle divided in two parts. The adaptive or acquired immune system is evolving throughout life and
where one of the key elements is the development of specific antibodies. The other part is the innate
immune system which is already in place at birth. The innate system is a non-specific first line of defense
comprised of cells and mechanism to defend against infection caused by other organisms. The
complement system plays an important part of the innate immune system.

The complement system has four major functions, including:

 Lysis of infectious organisms - rupturing membranes of foreign cells

 Activation of inflammation
 Opsonization - enhancing phagocytosis of antigens
 Immune clearance
What are T Cells
T cells are a type of lymphocytes that develop in the thymus. They are also called T lymphocytes. These
cells are
primarily produced in the bone marrow and migrate to the thymus for maturation. The immature T cells
differentiate
into three types of T Cells: helper T cell, cytotoxic T cells, and suppressor T cells. The helper T
cells primarily
recognize antigens and activate both cytotoxic T cells and B cells. The B cells secrete antibodies and
cytotoxic T cells
destroy the infected cells by apoptosis. The suppressor T cells modulate the immune system in such a way
to tolerate
 the self-antigens, preventing autoimmune diseases
What are B Cells
B cells are the other type of lymphocytes produced and develop in the bone marrow. B cells are
also called B
lymphocytes. They mediate the humoral or the antibody-mediated immunity (AMI). That means B
cells produce
antigen-specific immunoglobulin (Ig) or antibodies, which are directed against the invaded pathogens.
The naïve B
cells can bind to antigens on the circulation through B cell receptors (BCR) present on the
surface. This binding
promotes the differentiation of the naïve B cells into antibody-producing plasma cells and
memory cells. Some
antigen types require the participation of T helper cells with the plasma cells to produce
antibodies. These type of
antigens are called T-dependent antigens. But, some antigens are T-independent antigens. When a plasma
cell binds
to a T-dependent antigen, the helper T cells, which contain CD4 coreceptors, stimulate the production
of antibodies.
The T-dependent antigens produce antibodies with high affinity. In contrast, the T-independent
antigens trigger the
production of low-affinity antibodies.
MONOCLONAL ANTIBODIES

Pregnancy test kits use monoclonal antibodies. These have been designed to bind with a hormone called
HCG which is found only in the urine of pregnant women. Monoclonal antibodies are attached to the end
of a pregnancy test stick onto which a woman urinates. If she is pregnant, HCG will be present in her
urine and will bind to the monoclonal antibodies on the test stick. This will cause a change in colour or
pattern which will indicate pregnancy. These specific monoclonal antibodies in the pregnancy test will
only bind with HCG.

Cancer diagnosis and treatment

Cancerous cells have antigens. Monoclonal antibodies can be designed to bind specifically with these
antigens. When injected into a person's body, the monoclonal antibodies will bind with these cancer cells
and clump them together. This makes it easier to identify a cancerous tumour, which can then be treated
or removed.

Monoclonal antibodies have also been designed to treat cancer by:

 carrying drugs that have been attached to them, to the tumour

 encouraging your immune system to attack the cancer cells directly

Difference Between T Cells and B Cells

Definition
T Cells: T cells are a type of lymphocyte, which develops in the thymus, circulates in the blood
and lymph and
mediates the immune response against malignant or infected cells in the body by the secretion of
lymphokines or by
direct contact.
B Cells: B cells are a type of lymphocyte, which develops in the bone marrow, circulates in the blood and
lymph, and
upon recognizing a particular pathogen, differentiates into a plasma cell clone, secreting specific
antibodies and a
memory cell clone, for the subsequent encountering of the same pathogen

Difference Between T Cells and B Cells

Definition
T Cells: T cells are a type of lymphocyte, which develops in the thymus, circulates in the blood
and lymph and
mediates the immune response against malignant or infected cells in the body by the secretion of
lymphokines or by
direct contact.
B Cells: B cells are a type of lymphocyte, which develops in the bone marrow, circulates in the blood and
lymph, and
upon recognizing a particular pathogen, differentiates into a plasma cell clone, secreting specific
antibodies and a
memory cell clone, for the subsequent encountering of the same pathogen