Chapter 1

INTRODUCTION
1. Compare prokaryotes and eukaryotes

Prokaryotes Eukaryotes
-do not have membrane bound nucleus -have distinct nucleus
hereditary material suspended in a portion of membrane-bounded nucleus
cytoplasm ( nucleoid/nuclear region )
devoid mitochondria and organelles contain mitocondria and organelles
eg: mycoplasma, ricketsia, Chlamydia, blue- eg: algae, protozoa, slime moulds, fungi
green algae

2. Define:-
a) viroids : protein free fragments of single stranded circular RNA that cause disease in
plants
b) prions : infectious proteins devoid of nucleic acid.
c) virions : complete virus particle that is composed of a nucleic acid core (DNA or
RNA) surrounded with a protein coat(capsid)

Chapter 2
BACTERIAL STRUCTURE
1) Enumerate the functions of:

a) Ribosomes

-site of protein synthesis

-selective target for antibiotic action

b) Mesosomes

-involved in cell division and sporulation

-membranous support for respiratory enzymes

c) Cytoplasmic membrane

-selective transport

-excretion of extracellular enzymes

 -respiration

-cell wall biosynthesis

-reproduction

-chemotactic system

d) Cell wall

–maintain characteristic shape of bacterium

-support against high internal osmotic pressure of protoplasm(5-25atm)

-cell division

-staining affinity of organism

e) Capsule

-protection against antibacterial agents

-protection from phagocytosis (considered as virulence factor)

-attachment to target surface to establish infection

f) Flagella

-movement of bacteria towards regions with higher concentration of nutrients and solutes or away
from disinfecting substances (negative chemotaxis)

g) Fimbriae (pili)

-adherence and attachment to host surfaces to establish infection

-conjugation

2) Compare :

a) The composition of Gram positive and Gram negative cell wall

Gram positive Gram negative

-peptidoglycan and teichoic acid -peptidoglycan,outer membrane and

periplasmic space

b) Mycoplasm and L-forms of bacteria

Mycoplasm L-form of bacteria

 -exist without cell wall -wall deficient or defective bacteria

c) Flagella and fimbriae (pili)

Flagella pili

-consist of protein called flagellin which are -composed of structural protein subunits called
highly antigenic pilins

-function: for movement of bacteria -function: adherence and conjugation

d)Short pili and sex pilus

Short pili sex pili

-enable bacteria to attach to host surfaces and -involved in transfer of DNA between bacteria
establish infection by process of conjugation

3) Give reasons for :

a) The impressive strength of cell wall

-it is formed of peptidoglycan

b) Mycoplasm do not assume a defined shape

-lack of rigid cell wall

c) Mycoplasm are resistant to penicillin

-it exists without cell wall and resistant to cell wall inhibitors as penicillin

d) The capsule is considered an important virulence factor

-protect bacterial cell from phagocytosis

e) Fimbriae are considered a virulence factor

-enable bacteria to attach to host surfaces and establish infection

f) The marked resistnce of spores

-thermal resistance is due to high content of Ca and dipicolinic acid

-impermeability of cortex and outer coat

-low content of water

 -very low metabolic and enzymatic activity

4) Comment :

a) The arrangement of cells is determined by the planes of cell divisions.

-the cocci that divide along a single plane produce diplococci or chains eg; streptococci

-the cocci that divide on many planes produce clusters eg; staphylococci

b) bacterial ribosomes are a selective target for antibiotic actions

-prokaryotic ribosomes have a sedimentation constant of 70S, smaller than the 80S

ribosomes of eukaryotes.

c) the mesosomes have a function analogous to the mitochondria in eukaryotes

-they provide a membranous support for respiratory enzymes.

d) the cause and effect of the cell wal rigidity

cause: primarily due to peptidoglycan

effect: maintains the characteristic shape of the bacterium

5) Give an account:

a- L-forms of bacteria

-they are wall defective or wall deficient bacteria

-they loose their cell wall when exposed to hydrolysis by lysozyme or by blocking peptidoglycan
biosynthesis with antibiotics, such as penicillin.

-they present in isotonic medium

-some L-forms resynthesize their walls once the inducing stimulus is removed.others, permanently
lose the capacity to produce a cell wall

-they may survive antibiotic therapy.their reversion to the walled state can produce relapses of the
overt infection.

b) Axial filaments

-composed of two groups o fibers that originate within the opposite ends of the cell and overlap in
the middle.
 -called “endoflagella” because it is structurally and chemically similar to flagella.

Eg:spirochaetes-move by means of axial filaments by rotating along its longitudinal axis,and flexes
and bends along its length.

Chapter 3
BACTERIAL GROWTH

1. Define

a) The generation time: is the time between two successive divisions.It may be as short as 13 min. in
Vibrio cholerae and may reach 24 hours in Mycobacterium tuberculosis.
b) Aerotolerant anaerobes: have an anaerobic pattern of metabolism but can tolerate the presence of
oxygen because they possess superoxide dismutase e.g: Clostridium perfringens
c) Capnophilic bacteria: microorganisms which thrive in the presence of high concentrations of carbon
dioxide (CO2) or which require the presence of carbon dioxide to survive.
d) Mesophiles: organisms that grow within a temperature range of 20-40oC.Pathogens which replicate on
or in humanbody are able to grow within this range, with an optimum temperature of 37oC which is the
the normal body temperature.
e) Psychophiles: (cold loving) are capable of growth at refrigeration temperature (0-8oC) e.g.
Flavobacterium spp.
f) Thermophilus: (heat loving) grow best at high temperature (>60oC), e.g. Bacillus stearothermophilus.
g) Growth: is an increase in the size and number of organisms.
h) Colony: macroscopic products of 20-30 cell divisions of a single bacterium on solid media.

2.Give reasons for

a) obligate anaerobes die in presence of O2 : because obligate anaerobes lack superoxide dismutase and catalase
and so they cannot grow in presence of of O2.

b)Most bacteria of medical importance are heterotrophs : because they require organic sources for carbon, as
they can not synthesize complex organic substances from simple inorganic sources.

c)Most bacteria of medical importance are mesophils : because they can grow within range of 20-40oC which
can replicate on or in human body at optimum temperature of 37oC which is the normal body temperature.

1. Structure of bacteriophage
2.Define:
a)Lysogenic bacteria:
b)Prophage
c)Lysogenic conversion
d) Eclipse phase
Chapter 4
BACTERIOPHAGES
:bacteria carrying the integrated phage genome ( the phage
DNA integrated with the bacterial chromose and divides
with it to pass into daughter cells )
:the phage DNA integrated with the bacterial chromosome and divides
with it to pass into daughter calls. ( integrated phage genome )
:acquisition of a new character coded for by a prophage DNA
:no phage components are detected inside the cells.
3.Give an account on:
a) Lytic cycle of phage replication cycle:
this cycle ends in lysis of the bacterial host cell and release of a newly formed phages
1. Adsorption: The phage attaches, by its tail, to specific receptors on the
bacterial cell. The specificity of this process determines the susceptibility of bacteria to different
phages.
2. Penetration: The tail sheath contracts and the nucleic acid is injected into the
cell. The empty head and the tail are left outside the cell.
3. Eclipse phase: in which no phage components are detected inside the cell. It
takes a short time ( minutes to hours ) during which viral nucleic acid directs
the host cell metabolism to synthesize the enzymes and proteins required for phage synthesis.
4. Intracellular synthesis: of phage nucleic acids, capsids and tails. Several
hundreds of phage components are synthesized.
5. Assembly: The phage components aggregate to form a complete phage
particles which mature into typical infectious phage.
6. Release: The bacterial cell bursts liberating a large number of phage particles
to infect new cells.

b)Outcome of the temperate phage cycle

1.the prophage may be carried inside the bacterial cell indefinitely passing to daughter cells.
2.the prophage may be induced to detach from the bacterial chromosome and start a lytic cycle. Induction
maybe spontaneous or achieved by an inducer. E.g: UV light.

c)Practical uses of bacteriophages

1. cloning vectors in recombinant DNA technology
2. phage typing.
3. research elements in some biological n genetic studies

4.Give reason: Lysogenic diphtheria bacilli are toxigenic

-only capable to produce toxic when lysogenized.
 Chapter 5
BACTERIAL GENETICS

1. Define:
a. The bacterial genome
Total set of genes present inside the bacterial cell. It comprises bacterial chromosome,plasmids,
transposable genetic elements and bacteriophage DNA (prophage).

b. Plasmid copy number

Multiple copies of the same plasmid that may exist in the same cell
c. Bacteriocins
Bactericidal substances produced by certain bacterial stains and are active against
other strains of the same closely related species.
d. Plasmid curing
The experimental kicking off of the plasmids using physical agents or chemical
agents.
e. Gene cassettes
Genes that can be clipped out of one integron and inserted into another.

2. Compare:

a.Relaxed Plasmids Stringent Plasmids

Replication Absence of protein Acquired protein synthesis
Copy number 30-50 copies/cell 1-5 copies.cell
Size Small Large

b.Conjugative Plasmids Non-conjugative Plasmids

Size Large Usually small
Copy number 1-2 >30
F factors Present Absent
Transfer among By conjugation By the help of conjugative
bacteria plasmid
Host bacteria Common in Gram-ve bacilli Common in Gram+ve cocci
Conjugation Can conjugate Cannot conjugate
 c.Insertion sequence Transposons
Simplest type of transposable elements Complex transposable elements
Encode enzyme necessary for : Encode specific genes, flanked by two IS
a) Recombination (transposition) elements.
b) The control of frequency of
transposition

3. Comment: Plasmids are dispensable.

Plasmid can be transfer from one bacteria cell to another. This indicates that most plasmids encode
properties that are essential for growth, replication or survival of the host bacterium. This is evidenced by
spontaneous loss of plasmid during cell division and plasmid curing.

4. Give a short account on:

a. Properties of plasmid
Plasmids are extra chromosomal, circular, double-stranded DNA molecules dispersed in
the cytoplasm. They are much smaller than the bacterial chromosome. They are capable
of replicating independently of the bacterial chromosome and plasmid is generally
dispensable.

b. Functions of plasmid
i. Sex pilus formation.
Some plasmid carry fertiliy (F) factors that code for the formation of sex pilus which
mediates the process of conjugation .
ii. Antibiotic resistance
Some plasmid carry genes for resistance (R-factors) to one or several microbial drugs.
They often comtrol the formation of enzymes capable of destroying the antimicrobial
Drugs
iii. Virulence plasmids
May code for exotoxins, adhesins or invasion factors.
iv. Bacteriocin production
Bactericidal substances produced by certain bacterial stains and are active against other
strains of the same closely related species.
v. Other function includes nitrogen fixation, sugar fermentation, antibiotic production,
H2S production, resistance to heavy metals and degradation of aromatic compounds.

c. Transposable genetic elements

These are non-replicating DNA segments (units) that are capable of inserting themselves
into other DNA molecules. They are also capable of mediating their own transfer from
one location to another on the same chromosome or between the chromosome and
plasmids.

d. R-factors
R-factors are usually conjugative plasmids that can be transferred among bacteria by
conjugation. This results in the rapid spread of drug-resistance among bacterial
populations and the development of multiple drug-resistant straints

e. Integrons
 They are specific class of transposons caapable of capturing and mobilizing genes
contained in mobile elements called gene cassettes.

5. Give reason for:

a. Multiple copies of the same plasmid may exist in the same cell
Plasmids are capable of replicating independently of the bacterial chromosome.

b. Plasmid may render the bacteria pathogenic.

Plasmid especially virulence plasmid may code for exotoxins , adhesions or invasion
factor.

Chapter 6

BACTERIAL VARIATIONS

1) a. Compare between phenotypic and genotypic variation.

Phenotypic variation Genotypic variation

• It occurs in respond to changes in the • It occurs as a result of a change in the

environmental condition without underlying genetic constitution
change in the genetic constitution

• Reversible • irreversible

• Not-heritable • Heritable

Example: It occurs through:

a. L-forms bacteria a. Mutation

b. Loss of flagella upon exposure to b. Gene transfer:

phenol
• Transformation

• Transduction

• Conjugation

b.Compare between generalized and specialized transduction

Generalized transduction Specialized

transduction

• Type of phage Lytic phage Temperate phage

 • Replication cycle Lytic cycle Lysogenic cycle

• The transfer Any piece of bacteria A specific piece of DNA

DNA fragment DNA(chromosomal or adjacent to the site of the
plasmid) insertion of the phage.

2) Give an account on:

a. Mutation.

• It results from a change in the nucleotide sequence of DNA that may occur either
spontaneously as replication error or induced by radiation or chemical agents

• Classified according to the nucleotide substitution, insertion or deletion into:

Single-base mutation Frame-shift mutation

• It involves in the replacement • Occurs when the nucleotide

of a single nucleotide in the is inserted into or deleted
coding sequence. fro the coding sequence.

• Eg: • Eg:

a. Same sense(Silent) a. insertion of

mutation-When its code transportable elements
for the same amino acid

b. Missense mutation-
When the mutant base
change the coding
sequence so different
amino acid is produced

b. Transformation

• Dying bacteria release DNA which can taken up by other bacteria that may be chromosomal
or plasmid in origin, and it may carry genes that transform the recipient bacteria.
 • It may become integrated with bacterial chromosome or re-established extra-
chromosomally in the recipient’s cells.

• It depend upon competence which is the ability of the recipient bacterial cell to take up DNA
and competence depends upon the presence of protein in the cell membrane that have the
ability to bind DNA and transport it into the cell

c. Transduction.

• It is transfer of DNA from one cell to another cell by bacteriophage.

Generalized transduction:

- During the lytic phage cycle, the bacterial DNA fragmented and any fragment of DNA
may be incorporated into the phage head.

The phage particle can then transfer the incorporated DNA into another bacteria.

Specialized transduction:

- Occur when phage contained lysogenized bacterial cell is induced to detach .

- Such prophage may carry with it the adjacent piece of the DNA and transfer to another
bacterial cell.

d. Conjugation.

• It is a part of mechanism of DNA transfer.

• Involves 2 cell types : donor (F+) which processed the fertility (F) factor and recipient (F-)
factor which lack the F factor.

• The F factor carries genes for the synthesis of the sex pilus which acts as conjugation tube
between the donor and recipient cells.

• The 2 DNA strands of the F factor separated, and one of them is transfer into recipient cell.

• Each strand form the complementary, thus the recipient cell acquires a copy of the F plasmid
and become F+ factor

3. Enumerate:

a) Methods of gene transfer among bacteria

• Transformation

• Conjugation
 • Transduction

a. Specialized transduction

b. Generalized transduction

b) Types of mutation.

• Single-base mutation

a. Silent mutation

b. Missense mutation

• Frame-shift mutation

4) Define

a) phenotypic variations

-changes in bacterial characters in response to changes in the environmental conditions without

change in the genetic constitution.

b) Genotypic variations

-changes in bacterial characters as a result of a change in underlying genetic constitution

c) Mutation

- change in the nucleotide sequence of DNA that may occur spontaneously or induced by chemical
agents or radiation.

d) Competence

-ability of the recipient bacterial cell to take up DNA

 Chapter 7
GENETIC RECOMBINATION

1. Define:

a)cloning vectors-vehicles used to carry and introduced foreign DNA fragments into a host cell

b)cosmids-artificially constructed cloning vectors formed of plasmoids and cohesive end (COS) of
lambda phage DNA

c)restriction endonucleases-enzymes that recognized specific nucleotide sequences within DNA

molecules and catalize the cleavage of both DNA strands at internal position within the sequence

d)genetic probes-short single stranded DNA or RNA fragments used for diagnosis of infections as well
as genetic diseases

2. Enumerate:

a)requirements for an ideal cloning vector

-as small as possible

-well characterized regarding gene location and nucleotide sequence

-possess a single cleavage site for at least 1 restriction endonuclease

-capable of autonomous replication within the host cell

-carry selective marker

 b)commonly used cloning vectors

-plasmids

-bacteriophage

-cosmides

-animal viruses

c)applications of recombinant DNA technology

-mapping of microbial genome

-production of biological product of medical importance,e.g. hormones

- production of recombinant vaccines

-preparation of genetic probes

-gene therapy

d)the properties of host organisms used for cloning properties

-strains used should be free of any restriction activity

-nucleic acid of host easily saparable from that of cloning vector

3. Give reasons for:

a)the restriction endonucleases are so-called

- activity is restricted to unmodified foreign DNA because own DNA is methylated in specific manner
to be

protected from cleavage by own enzymes produce

b)the ideal cloning vector must be small

- to contribute as little as possible to the overall size of recombinant molecule

c)the ideal cloning vector must be carry a selectable marker

- to distinguished between the host cells transformed with the vector from non-transformed cells
d)the relaxed plasmids are among the best cloning vectors

- they fulfill most of the requirements for ideal cloning vector

Chapter 8

ANTIMICROBIAL THERAPY
1. Define :

A Antimicrobial chemically synthesized substance that are used to treat

chemotherapeutic infectous disease by killing or inhibiting the growth of
agents microorganisms
B Antibiotics low molecular weight antmicobial substance that are produce
as secondary metabolites by certain groups of microorganism,
especially Streptomyces, Bacillus, and a few moulds
(Penicillium and Cephalosporium)
C Synthetic Antibiotics that are made synthetically
antibiotics
D Semisynthetic Chemical modification of certain antibiotic, to achieve the
antbitics desired properties.

E Bacterostatic an antimicrobial agent that is capable of inhibiting bacterial

agent multiplication
F Bacteriocidal an antimicrobial agent that is capable of killing bacteria
agent
G Broad spectrum antibiotics which kill or inhibit the growth of a wide range
bactercidal agent Gram-positive and Gram-negative bacteria

2. Give a short account on :

a. Selective toxicity : is the ability of an antimicrobial agent to harm a pathogen without harming
the host.It may be function of a specific receptor for the drug found in the microb but not in the
human body (eg. Peptidoglycan), or it may depend on the inhibition of a biochemical event
essential for the organism but not for the host.

b. mechanism of action of antimicrobial drugs

- inhibition of cell wall synthesis

- interference with cell membrane function

- inhibition of bacterial protein synthesis

- inhibition of bacterial nucleic acid

c. Mechanism of acquired drug resistance :

One or more of the following mechanisms used by bacteria:

a. reduction of the intracellular concerntration of the antibiotic by:

i. decrease in influx of antibiotic through reduction of permeability of the outer

membrane by modification or loss of porin

ii. efflux pumps : the antibiotic is pumped out across the cytoplasmic membrane faster
than it can diffuse in

b. Inactivation of the antibiotic, eg: production of β-lactamase leads to hydrolysis of the β-

lactam rings, thus inactivates penicillins and cephalosporins

c. Target modification : modification of the target site for the antibiotic reduces its affinity for
its receptor

d. Target elimination by developing new metabolic pathways: the bacteria createa new
metabolic pathways that bypass the original target, eg. resistance to trimethoprim

e. Target overproduction : used by S. Aureus strains with intermediate susceptibility to

vancomycin (VISA)

d. Spectrum of activity : range of action of microorganism that are affected by a certain antibiotics
is expressed as its spectrum of action. Antibiotic that kill both Gram-positive and Gram-negative
said to be broad spectrum. If effective mainly against either Gram-positive or Gram-negative
bacteria they are narrow spectrum. If effective against one of them,referred as limited spectrum.

e. Intrinsic resistance: refer the bacteria that are insensitive. It is consistent and can be expected
once the organism is known. Intrinsic resistance occurs in the follwing situations :

a. Streptomyces are protected from the antibiotics they produce.

b. Gram-negative cell membrane has pores too small to allow large antibiotic molecules,
e.g. nafcillin, to penetrate.

c. An organism lack the target or receptor for the antibiotic as in case of resistance of
enterococcus species to cephalosporin.
f. Superinfection : it occurs as a result of outgrowth of resistance members of normal flora when
the sensitve ones are eradicated during antibiotic therapy.e.g.: Pseudomembranous colitis caused
by outgrowth of the yeast Candida.

g. Empiric therapy : is a best guess. By indicating type of infection, a short list of bacteria can be
identified. Depending on the type of bacteria, there will be an antibiotic that can acts on the
bacteria, then treating the infection. Best guess treatment is not always successful as many
bacteria have unpredictable susceptibilities to antimicrobial agents. Indication : in seriously and
if no sample is available.

h. Complication of chemotherapy :

a. Toxicity

b. allergy(hypersensitivy)

c. emergence of resistant strains

d. superinfection

i. Effects of combined therapy

a. Synergistic effect(1+1=>2) : the combined action is greater than the sum of both
effect,e.g.:Vancomycin+gentamicin in treatment of methicillin-resistant staphylococci

b. Antagonistic effect (1+1<2) : the combined action is less than that of the more effective
agent when used alone,e.g. : penicillin + Chlorompenicol in treatment of meningitis

c. Indifference (1+1=1) : the combined action is no greater than that of the more effective
agent when used alone,e.g. : cefepime+ vancomycin

d. Addition (1+1=2) : the cobined action is equivalent to the sum of the action of each drug
when used alone.

j. Antimicrobial chemoprophylaxis : is the administration of an effective antimicrobial agent to

prevent rather than to treat infection with a certain microbe, thus preventing development of a
disease.e.g. : Long acting penicillin is given to rheumatic patient to prevent reinfection with
S.pyogens.
3. Comment : Bacterial ribosomes are a selective target for antimicrobial agent

Bacteria have 70S ribosomes (30S & 50S subunits) differ from mammalian cell ribosomes,
> agents acting on the 30S ribosomal subunit : eg. tetracycline
> agents acting on the 50S ribosomal subunit : eg. macrolides

4. Enumerate:
a. Methods for in vitro susceptibility testing:
• Disc Diffusion method
• Dilution method (eg. tube broth dilution)
• Gradient diffusion (E test) method
b. Possible indications for combined therapy
• severely ill patient suspected of having serious infection
• febrile neutropenia
• to delay the emergence og drug-resistant mutants
• to achieve bactericidal action through synergistic effect
• mixed infections
c. Indications for empiric therapy:
• in seriously ill patient after collecting specimens for culture
• in closed lesion (no available sample)

5. Define:
a. MIC : the lowest concerntration of a drug that prevents growth of a test organism
b. Breakpoint of antimicrobial agent: the concerntration that can be achieved in the serum with
optimal dose
-then mention their impact in determining susceptibility
Organisms with MICs at or below breakpoint are considered susceptible, while organisms with
MICs above the breakpoint are considered resistant

6. Give reason for:

a. Vancomycin could be used successfully in infections caused by β-lactam resistant bacteria

• the mechanism of resistance to β-lactam antibiotics is different from that of

glycopeptides (vancomycin). β-lactam drugs inhibit the last step of peptidoglycan
synthesis while vancomycin inhibit early steps in the biosynthesis of peptidoglycan
which occur inside the cytoplasmic membrane.

b. Antibiotics which interfere with the cytoplasmic membrane function are highly toxic
• they have narrow margin of selective toxicity

c. β-lactam antibiotics are bactericidal with minimal host tissue toxicity

 • β-lactam antibiotics inhibit the last step of pepyidoglycan synthesis by binding to
penicillin binding proteins

d. It is advisable to use as narrow antibiotic spectrum as possible

• the use of broad spectrum antibiotics is likely to induce resistance to antibiotics and
may be complicated by superinfection

e. Undue prolonged therapy should be avoided

• may result in drug toxicity and antibiotic resistance

f. Antibiotic resistance is more prolonged in hospitals

• due to the higher antibiotic use.

Chapter 9
DISINFECTION AND STERILIZATION

1- Define :-

a)Sterilization - it is complete removal, destruction or inactivation of all forms of microbial life including
bacteria, viruses,spores, and fungi.

b)Disinfection - it is the elimination of most, if not all, pathogenic microorganisms including spores.

c)Antiseptics - special types of chemical disinfectants which can be safely applied to skin and mucous
membranes but are not suitable for systemic administration.

d)Cleaning - it is a process of removal of the visible dust, dirt, organic load, and other foreign materials. The
process removes some microorganisms. It is usually done with water and soap, detergents or enzymatic
products. Cleaning must always precede disinfection and sterilization.
e)Decontamination - it is a general term that is applied to any procedure by which pathogenic microorganisms
are reduced to a level where items are safe to handle.

f)Critical items - instruments that enter sterile tissues, cavities, or vascular system.

g)Semicritical items - objects that come in contact with mucous membranes or non intact skin.

h)Non critical items - objects that contact intact skin only but not mucous membranes.

2- Enumerate

a) Main methods of disinfection

1- Boiling water : Boiling at 1000C for 20 minutes achieves high disinfection. It can be useful
in emergencies if no sterilizer is available.

2- Pasteurization: (hot water at temperatures lower than 100o C) e.g pasteurization of milk by
heating at 63oC for 30 min. or at 72oC for 20 sec., followed by rapid cooling, destroys important
pathogenic organisms e.g. Mycobacterium tuberculosis, Brucella, Salmonella, and Coxiella
burnetti.

3- Ultraviolet Radiation: Ultraviolet Radiation(UV) is a low energy, non-ionizing radiation

present in sun rays or artificially produced by mercury lamps. UV radiations have extremely
weak penetration power. Therefore UV rays can be used only for air and surface disinfection, in
operating rooms and laboratory safety cabinets.

4- Chemical disinfection

b)Main methods of sterilization

I. Heat

1.Moist heat or steam sterilization.

2.Dry heat sterilization.

II. Low temperature (“cold”) sterilization methods:

A-Chemical

1.ethylene oxide gas.

2.liquid sterilization process: commonly used liquid sterilants include:

-Glutaraldehyde
 -Liquid peracetic acid

-hydrogen peroxide 6%

B-Plasma sterilizers

III. Other sterilization methods

1.Ionizing radiation.

2.Filtration.

3.Microwaves.

c)Advantages and disadvantages of:

advantages disadvantages
Steam sterilization 1.A good ability of saturated 1.Some items cannot
steam to penetrateinto loaded withstand steam at high
porous items. temperatures.

2.Liberation of latent heat of 2.Steam sterilization is not

vaporization after cooling due suitable for sterilizing
to condensation of the steam. powders and oils.

3.Absence of toxicity

4.Low costs.

Hot air sterilization 1.The capability of being used 1.Slow and uneven
 for sterilizing powders, penetration of heat into the
waterless oils and glassware. materials to be sterilized.

2.Absence of a corroding 2.A necessity for prolonged

effect. exposure.

3.Low costs. 3.Damage to the rubber items

and some fabrics
Gas (E.O) Capability of the instrument 1.long duration of the
that cannot be subjected to the sterilization cycle
steam/hot air sterilization
without any damage 2.high costs

3.toxicity
Membrane filter 1.filter more rapidly

2.they don’t affect the filtrate

in any way

3.they adsorb very little of the

substances being filtered

4.remove microorganism from

air supplied to critical areas

3.Give an account on:

Monitoring of steam sterilizers(autoclaves)

a. Physical measurement of the tracts of the time,temperature and pressure
b. Chemical indicators:chemically impregnated strips which change their colour at certain temperature
should be used with each cycle of sterilization
c. Biological indicators:
- use paper strip containing Bacillus stearothermophilus
- the biological indicator are placed in the depth of the load to be sterilized
-after that,spore strips are incubated in a fluid medium and checked for viability
-absence of bacterial growth indicates an efficient sterilization cycle

4. Mention one method used for sterilization (disinfection) of

a. skin-chemical (biguanides-chlorhexidine)
b.air in operation room-ultraviolet radiation
c. milk-pasteurization
d. disposable plastic syringes@glove-irradiation
e. glassware-sterilization (dry heat)
f. hormones,vitamins@blood products-filtration
g. floor,walls@furnitures-chemical (quaternary ammonium compound)

5.Mention one use for each of the following methods

a. Incineration
 -For dead animal bodies, infectious hospital waste such as used surgical
dressing and needles.
b. Hot air sterilizers
-Sterilization of powders, waterless oils and glassware.
c. UV radiation
-For air and surface disinfection, in operating rooms and laboratory safety cabinets.
d. Alcohol
-Disinfection of thermometers, external surfaces of stethoscopes and as skin antiseptics
(by themselves or in mixture with iodine or chlorhexidine)
e. Iodophores
-For antiseptic purposes
f. Hydrogen peroxide
-Disinfection of endoscopes and antiseptic for open wound
g. Glutaraldehyde
-For high level disinfection or sterilization of the instruments such as endoscopes,
respiratory and anaesthesia equipments.
h. Red heat
-Sterilization of inoculating wires, loops and point of forceps.
i. Bacteria filters
-Sterilization of fluids which would not withstand heat such as antibiotic solutions, blood
products, hormones and vitamins
j. Ionizing radiation
-Sterilization of prepacked heat-sensitive power items such as bone grafts, surgical
sutures, disposable plastics syringes, gloves, catheters, plastics Petri dishes and
intravenous infusion set.
k. Chlorine-active compounds
-Decontamination of the blood splashes and laboratory working surfaces.Linen bleaching.
Disinfection of water for domestic use.

l. Phenolics
-Cleaning of floors ,walls and furnitures
m. Peracetic acid
-High level disinfection or sterilization of the instruments such as endoscopes
n. Biguanides (chlorhexidine)
-Disinfection of the skin and mucous membrane (as a mouth wash).It is often combined
with detergents for hand washing or with alcohol as a handrub.

6. Give reason.
a. Moist heat is much more efficient than dry heat as a method of sterilization.
- Because it kills microorganism by coagulating and denaturing their enzymes as
structural proteins also it is quicker in heating up the article to be sterilized.
b.Steam sterilization is the most preferred method of sterilization.
- It is suitable method for those items that can withstand high temperature and
moisture.
c. In steam sterilization, it is essential to make steam free of any residual air.
 - Because air acts as an insulator, reduces temperature and hinders penetration.

7. Give an account on plasma sterilizers.

- Plasma describes any gas that consists of electrons, ions or neutral particles. Plasma sterilizers with the use
of liquid peracetic acid or hydrogen peroxide or a mixture of both are commercially available.

Chapter 10
BACTERIAL PATHOGENESIS

1.Define:
a)saprophytic bacteria= bacteria which live freely in nature,on decaying organic matter,in soil or water.They
do not require a living host.
b)parasitic bacteria= bacteria which live on or in aliving host.They are classified according to their relation to
the host into pathogenic,non pathogenic and opportunistic pathogen.
c)opportunistic bacteria= potentially pathogenic bacteria that do not cause disease under normal conditions
but can cause disease in immunocompromised patients or when they find another site other than their normal
habitat.
d)toxoid= a preparation of the poisonous toxin that has been treated to removes its toxicity and retains its
antigenicity.Are used in vaccines.
e)virulence factors= A structure(e.g. capsule) or a product(e.g. toxins) that enables the organism to cause
disease.
2.Compare:
a)pathogenicity and virulence
pathogenicity=A qualitative description of a species of bacteria denoting ability to produce disease
virulence=A quantitative character (degree of pathogenicity) of a strain belonging toa pathogenic species
b)exotoxins and endotoxins

Exotoxins Endotoxins
source Secreted by living oraganisms bothIntegral part of the cell wall of Gram
Gram +ve(mainly) and Gram -ve –ve organisms.Liberated upon cell
disintegration
Coding genes Encoded by Encoded by genes on the
chromosomes,plasmids,bacteriophages chromosomes
or PAI
exampes C. diphteriae (phage) E. coli
Cl. tetani (plasmid) Meningococcal endotoxins
B. pertussis (chromosomes)
H. pylori (PAI)
nature protein Lipopolysaccharide (lipid A)
antigenicity Highly antigenic Poorly antigenic
Heat stability Unstable to temp. above 60 ̊c Stable to temp. above 60 ̊c
detoxification Can be converted into toxoid Can not
Specificity Every toxin has specific action Same generalized effect,all give
fever and shock (non specific action)
toxicity high low

3.Give an account on:

a)Koch’s postulates= These are criteria that were proposed by Koch in order to determine if the organism
isolated from the patient actually caused the disease.These criteria are as follows:
1)The organism must be isolated from every patient with the disease
2)The organism must be isolated free fromall other organisms and grown in pure culture in vitro
3)The pure organism must cause the disease in an healthy,susceptible animal
4)The organism must be recovered from the inoculated animal
b)Adherence factors of virulence= They enable bacteria to attach to the host surfaces.For examples:
1)The fimbriae of Neisseria gonorrhoeae and E. coli help their attachment to the urinary tract epithelium.
2)The glycocalyx of Staphylococcus epidermidis and certain viridians streptococci their adherence strongly to
the heart valve
c)Invasion factors of virulence= One of main mechanisms by which bacteria can cause disease is through
invasion of tissue followed by inflammation.This invasion is helped by:
1)Enzymes: a-collagenase:degrade collagen ------------------>allow bacteria to spread through
Hyaluronidase:degrade hyaluronic acid ----->subcutaneous tissues
 b-IgA protease:degrades IgA
c-Leukocidin:destroy both polymorphonuclear leucocytes and macrophages
d-Deoxyribonuclease:breaks down DNA
e-Lecithinase:breaks down lecithin of cell membrane
2)Antiphagocytic factors:
a-capsule:prevents phagocytes from attachment to bacteria. e.g. Strept. pneumoniae
b-cell wall proteins of Gram +ve cocci e.g: M protein(Strept. Pyogenes)
: protein A(Staph. Aureus)
c-coagulase:accelerates the formation of a fibrin clot from fibrinogen.This clot
protect bacteria from phagocytosis e.g. Staph. aureus

Chapter 11
OVERVIEW OF THE IMMUNE SYSTEM

1.Compare:
A)innate& acquired immunity
Innate Acquired

Presence Since birth Following exposure to

pathogens
Onset of action Immediately after infection Relatively delayed

Main cells Granulocytes, monocytes / B&T lymphocytes

macrophages, NK cells
Memory Absent Present

Efficiency Less efficient More efficient and improves

with each exposure
Specificity Non specific: Specific:
Present in all individuals, against all Occur in a given person,
microorganisms Against a particular pathogen
Interaction Interact with acquired immunity Interact with innate immunity
through: e.g.-antigen presentation through: e.g.- opsonization

B)T&B cell receptor

B cell receptor T cell receptor

It is an antibody molecule It is a dipeptide molecule

Has 2 antigen recognition site 1 antigen recognition site

Can be active even after detachment Cannot be active after detachment-always

need cell surface receptor
Can recognize the antigen directly Recognize the antigen only when
presented by antigen presenting cells

2.Give an account on:

A)functions of macrophages
1.phagocytosis
2.Antigen presentation: Macrophages help to show or present part of the foreign agents they have eaten to T
cells, so that the T cells can start responding to them. Thus, they are among a group of cells called antigen
presenting cells(APCs).
3.Secretion: They secrete chemical mediators called cytokines ,e. g. interleukins
4.Direct cytotoxicity: They may kill target without engulfing them. Helminthic parasite which are too large to
be engulfed can be killed by macrophages releasing their toxics contents onto them. Tumor cells can also be
killed in a similar way.

B)The clonal selection theory

-T & B lymphocytes are very specific
-they recognize the antigen by the receptor present on the surface of antigen-each naive lymphocyte have single
type of receptor of certain specificity
-only the lymphocytes which meets the antigen which their receptor recognize, will undergo activation,
proliferation and differentiation
-the result is a clone family of identical daughter cells, all having identical receptors, which can bind the same
antigens wherever they find it.
-thus, antigen specificity is maintained in the daughter cells

3.Define naive lymphocytes :

small B and T lymphocytes that have matured, but have not yet met antigen .They leave bone marrow n thymus
respectively,and circulate continually to secondary lymphoid organ such as lymph node

4. Enumerate:
a. the different lymphoid organ
-there are 2 types of lymphoid organ:
1-primary(central) lymphoid organ
-places where lymphocytes complete their maturation
-they are: =>the bone marrow - where the B cells complete their maturation
=>the thymus - where the T cells complete their maturation

2-secondary(peripheral) lymphoid organ

-places where lymphocytes meet the antigens, leading to activation of the lymphocytes
-they are:
=>the lymph node - the B lymphocytes are localized in follicles and in the cortex of the lymph node, while the
T lymphocytes are more diffusely distributed in the paracortical area
=>the spleen - the lymphocytes surrounds the arterioles entering the organ,forming the white pulp. the inner
part of this called periarteriolar lymphoid sheath, containing mainly T cells and is surrounded by a Bcells
corona
=>GALT - includes the tonsils, adenoids, appendix and Payer's Patches
=>BALT - includes the tonsils, adenoids, appendix and Payer's patches in more diffusely organized collections
of lymphocytes which protect the respirator epithelium
=>other mucosal sites

b) changes which occurs when a naive lymphocyte recognizes an antigen

-the changes which occur are:
1-activation : become lymphoblast
2-proliferation : rapid multiplication
3-differentiation : change into effector cells ~B cells changes into plasma cells, capable of secreting
antibody ~cytotoxic T cells becomes capable of killing infected cells
~helper T cells becomes capable of producing cytokines
-they leave the lymph nodes through efferent lymphatic vessels and return to the blood trough the thoracic duct.
from the blood they reach the peripheral tissue where they start functioning to eliminate the specific infection
which started their activation

Chapter 12
INNATE IMMUNITY

1. Enumerate

a) Mechanical barriers and surface secretions as a mechanism of innate immunity.

1.intact skin and mucous membrane constitute a barrier that cannot be penetrated by most
microorganisms.
2.the sticky mucous covering mucous membrane traps any foreign material.
3.cilia of respiratory tract epithelium sweep foreign material out.
4.blinking, sneezing, and coughing reflexes expel foreign particles.
5.sweat and sebaceous secretions contain substances that inhibit microorganisms.
6.saliva, tears, and mucous secretions of respiratory, alimentary and genitourinary tracts contain lysozyme
which is bactericidal.
 7.gastric and vaginal acidity inhibit growth of microorganism.
8.the flushing action of saliva, tears and urine helps in washing microbes from the body.

b) humoral defence mechanism.

1.lysozyme: enzyme that lysis bacteria by destroying the peptidoglycan of their cell wall.
2.complement: group of plasma protein that act to attack extracellular pathogen.
3.acute phase protein: present at very low levels in normal serum but rise dramatically after onset of
infection.
4.interferon: 2 types of interferons. Type 1 is innate immunity and type 2 is part of acquired
immunity

2. Give an account on:

1. Acute phase proteins  Low level in normal serum
 Rise immediately after onset of infection
 E.g endotoxins stimulate macrophage to release IL-1 & IL-6
which stimulate liver to produce large no. of APP.
 they act to limit spread of infectious agent or stimulate host
response.
 E.g fibrinogen & C-reactive protein, help in complement
activation
2. Phagocytosis a) Migration :
microorganisms & injured tissue elaborate chemotactic factors that
attract phagocytic cells to site of infection.

b) Attachment :
• Phagocytes have receptors on their surface that can
recognize non-specific molecules common to many
pathogens, allowing attachment to them.
• Some microorganisms may lack these molecules @ having
thick capsule that not recognized by any phagocyte receptor.
• Attachment may still occur if microorganism coated by
molecules which phagocyte can recognize.This is called
opsonization.
• Substance that help phagocytosis is called opsonin

c) Engulfment :
• Cytoplasmic membrane of phagocyte surround organism and
encloses it in a vacuole termed phagosome.
• Lysosomes fuse with phagosome forming phagolysosomes
• Engulf material killed & digested

d) Killing:

O2 dependent mechanism :
respiratory burst consist of steep rise in o2 consumption,
accompanied by increase activity of no. of enzymes lead to
generation reactive o2 intermediates which lethal to organism.
O2 independent mechanism :
Once ingested, microorganism digested by
lysosomal enzymes

c) Respiratory burst • one of killing mechanism in phagocytosis

• due to rise in o2 consumption
• accompanied by increase activity of enzymes & leads to
production of reactive o2 intermediates which lethal to
mocroorganism
• e.g H2O2
3.Give reason: normal body flora act as a mechanism of/play role in innate immunity.
1. they produce bacteriocins and acids that destroy microorganism.
2. they compete with pathogens for essential nutrients.

4. Compare:

Innate immunity Acquired immunity

Presence Since birth Following exposure o pathogens

Onset of action Immediately after infection Relatively delayed

Main cells Granulocytes B & T lymphocytes

Monocytes/macrophage
NK cells

Memory Absent Present

Effeciency Less efficient More efficient and improves with each

exposure

Specificity Non-specific : Specific :

Present in all individuals against
 all microorganisms Occurs in given person,against a
particular pathogen

Interaction Interact with acquired immunity Interact with innate immunity through :
through : -Opsonization
- Antigen presentation

Chapter 13
ANTIGENS

1. Define:
a) Immunogen / Antigen
- substance that can stimulate the immune system to produce an immune response
- reacs specifically with the product of this response.

b) Epitopes
-small, limited parts present on antigen molecule which is recognized by immune system specifically.

c) Hapten
- low molecular weight substance
- incapable of inducing immune response alone
- act as antigen when coupled with a carrier molecule (protein)

2. Enumerate: Factors affecting immunogenicity

a) Foreigness
b) Molecular size
c) chemical nature
d) route of administration
e) dosage
f) adjuvants
g) host factors

3. Give an account on adjuvants

-non specific potentiatiors of the immune response.
-one of the commonly use adjuvants is aluminium hydroxide which is added to diphtheria and tetanus toxoids
(used for human immunization)
-aluminum hydroxyde delays the absorption of toxoids and prolongs the period of their exposure to the
immune system.

4. Give reasons:
a) Adjuvants are added to toxoids during human immunisation
- to delay the absorption of toxoids and prolongs the period of their exposure to the immune system
b) The most potent immunogens are proteins
proteins are complex molecules. the more complex the molecule, the more immunogenic it is.

Chapter 14
T-CELL MEDIATED IMMUNITY

1. Give an account on :

a) T cell surface surface molecules

Their roles are- (1)antigen recognizer,(2)interact with other cells,(3)markers in identifying T cells
and divide them into subsets. Among them are:

1)T cell receptor (TCR) -2 plypeptide chain: alpha and beta

-constant part and variable region

-all are identical and recognize the same antigen.

2)CD3 -close to TCR on all T cells

-transmitting signals from TCR inside the cells

3)CD4 and CD8 –T cells carry either of them

CD4- helper T cells (Th)….help immune system by secreting cytokines

CD8- cytotoxic T cells (Tc)…kill infected cells, tumour cells, other cells

recognized by T cells.

Both are associated with TCR…= ‘CO-RECEPTORS’

*SIGNAL DELIVERED THROUGH TCR WITH THE HELP OF THE ABOVE MENTIONED IS

THE ‘FIRST SIGNAL’ FOR ACTIVATION OF T-CELL.

4)CD28- present on all T cells

-binds to B7 on APC>>delivers second signal (for T cells activation)

>>>= ‘co-stimulatory molecule’

5)CD40L- present on activated Th cells

-involved in B cells activation by T cells

-binds to CD40
b)the professional APCs

-the only cells that capable of activating naïve T cells

-in the peripheral lymphoid tissues

1)dendritic cells…most important

So called because > have dendrites(cytoplasmic projections)

In nearly all tissues

Most efficient

Antigen presentor

2)macrophages…important phagocytic cells

Essentials for innate immune system

Contribute to acquired immune system

3)B cells…humoral immunity

Also as APCs

c)MHC restriction

-they code production of certain cell-surface gycoproteins

-another name > human leucocyte antigen (HLA)…first discovered on leucocytes

-two classes(important in T cells activation):

1) Class | (MHC |)

2) class || ( MHC ||)

-all people have both classes

2) Compare: endogenous and exogenous pathogens

Cytosolic “endogenous” Vesicular “exogenous”

 Examples All viruses,few bacteria -intracellular bacteria
-extracellular bacteria and their
products when internalized
Degraded in cytoplasm Vesicles
Peptides bind to MHC I MHC II
Presented to CD8 CD4
result Cytotoxic killing of presenting -Secretion of cytokines by CD4
cell by CD8 -help macrophages.B cells and
others

3) Give reason:

a) CD3 found close to the TCR on all T cells

-involves in transmitting signals from TCR to the inside of the cells

b) CD28 is called CD-stimulatory molecules

-its binding to B7 molecules delivers the second signals for activation of T cells.

Chapter 15
CYTOKINES

1) Define
a.cytokines
Cytokines are peptide or glycoprotein mediators that are produced by cells of the immune system and have
an effect on the behaviour and properties of many cells

b.LAK
It is the NK cells which is being activated by IL-2 to enhance their killing ability

2) Enumerate
a.General characteristis of cytokines
I. They are highly potent being acting at very low concentration
II. Not specific to antigen that produce them
 III. Act through high affinity cell surface receptor
IV. Their action is transient
V. They act in an autocrine or paracrine manner
VI. They are pleiotropic-same cytokines may have multiple effects
VII. Different cytokines may have same activity(redundancy)
VIII. They may act sequentially(network interaction),increase effect of other(synergy) or as antagonist

b. Cytokines that mediate innate immunity

a. IFN-α
b. IFN-β
c. IL-12
d. TNF- α
e. IL-1
f. IL-6
g. Chemokines
h. IL-10

c. Cytokines that mediated acquired immunity

a. TH-1 – IL-1,IFN-γ,TNF-β
b. TH-2 – IL-4,IL-5,TGF-β
c.
d. Cytokines that stimulate haematopoiesis
a. IL-3
b. IL-7
c. GM-CSF

e. Pro-inflammatory cytokines
a. TNF- α
b. IL-1
c. IL-6

5. Explain the role of:

a) IFN-α in activation of macrophages

• Promotes fusion of pagosomes containing bacteria to lysosomes containing antibacterial substances.
• Induces synthesis of nitric oxide and other bactericidal
• Induces macrophages to secrete cytokines

b) IL-4 in development of allergy (hypersensitivity)

• Promotes production of IgE
• Promotes growth and function of eosinophils
• Helps activation and growth of the B-cells

6. Give reason:
a) GM-CSF is used in treatment of leucopenia
• Promotes development of granulocytes and monocytes

b) IFN-α has shown success in treatment of viral infections

• Inhibition viral replication by causes cells to synthesize a number of enzymes that interfere with
translation of viral mRNA
• Activation NK cell
• Increased expression of MHC I molecules leads to more recognition of viral peptides and more efficient
killing of virally-infected cells by Tc cells

c) INF-α has been used in trials to treat malignancies

• Inhibition of cell proliferation

d) IL-2 is shown as autocrine growth factor of T-cells

• Produced by activated Th1 cells which then promote activation of T cells and cytokine production of T
cells.

e) IL-2 is not used in large scale as a therapeutic agent

• Due to severe toxic side effect

f) TNF α is not used on large scale to treat tumors

• Induces IL-1 production, and both have pro-inflammatory action (produces fever, promotes local
inflammation, induces synthesis of acute phase protein)
• When enters blood stream causing systemic effects such as fever, shock and even death
welcome

Chapter 16
THE HUMORAL IMMUNE RESPONSE
1.Give an account on:

(a) B cell surface molecules

B cell receptor receptor(BCR)

-This is membrane bound antibody molecule,surface immunoglobulin which act as antigen receptor
-all BCR on a single B cell are identical specificity
-then B cell then secrete antibody of the same specificity
-immature B cell express only IgM on their surface, but mature B cell bear both Igm n IgD
1) CD40: essential for the interaction between B n T cells
2) MHC II: antigen presentation to T helper cell

(b) -mature naïve B cell leaves the blood stream and enters secondary lymphoid organ
 -in the absence of it target antigen,Bcell traverses this region rapidly and enters the circulation
-in the presence of it antigen,the Bcell binds it specific antigen via the surface igs(BCR) and full activation
of B cell occur.
-to become fully activated naïve B cell must receive 2 signal:
• First signal is delivered by binding of antigen to BCR.
• Second signal is delivered by activated T helper cell.To receive this signal,Bcell engulfs the bound
antigen,degrades it into peptides and persents this peptide on the cell surface in association with
MHC II.T Helper cell will recognize the peptide –MHC class II complex. And produce CD 40
Ligand and IL-4,IL-5,IL-6,IL-10 which are B cell stimulatory cytokines.
-The B cell has received both signal,these stimulate Bcell to become a lymphoblast(activation),to divide
repeatedly(proliferation) and ti differentiate into antibody secreting plasma cell.

(c) functions (effector mechanisms) of antibodies

• Agglutination: binding of antibodies to an antigen (e.g. Bacteria) results in clumping of the pathogen
which prevents its dissemination and stimulate its removal by other mechanisms (e.g. Phagocytosis)
• Neutralization: antibodies can inhibit the infectivity of a pathogen (viruses or bacteria) or the toxicity
of a toxin molecule by binding to them, thereby preventing their attachment to their specific
receptors on their target cells.
• Opsonization: phagocytic cells have Fc receptors on their surface, that can recognize and bind Fc
portion of antibody molecules coating a pathogen. This facilitates the engulfment and subsequent
intracellular killing of the pathogen by the phagocytic cells.
• Complement activation: antibodies bound to the surface of a pathogen may activate proteins of the
complement system. Some of these complement proteins become deposited on the pathogen and also
bind to complement receptors on phagocytes, favoring the uptake and destruction of the pathogen
by the phagocyte. This is another example for opsonization. Other complement proteins may lyse the
pathogen directly by forming pores in its membrane.
• Antibody –dependent cell-mediated cytotoxicity (ADCC): it is the destruction of antibody –coated
cells by natural killer (NK) cells. NK cells possess receptors for the Fc portion of antibodies. An
antibody bound to an antigen on a target cell can also bind to the NK cell through its Fc portion
facilitating adhesion of the NK cell to the target cell and triggering its cytotoxic activity. Other cells
possessing Fc receptors, e.g. macrophages, may also exert ADCC.

d) hypervariable regions ~pg 75

- short segments in the variable domains of light(LC) & heavy chain(HC) that have variability in a.a. sequence
- not spread evenly over entire length of variable domains of LC & HC
- this regions r folded & brought together creating a single hypervariable surface or paratope
- paratope is da antigen(Ag)-binding site~complementary to & interacts with epitope of Ag

e) Immunological domain
Immunoglobulins consist of light and heavy chain and each chain is subdivided into domains which are:
a) Variable domain - wide variation of amino acid
b) Constant domain - constant amino acid
Light chain = constant of an VL and CL
Heavy chain = consist of an VH and 3 to 4 CH

(f)Sec IgA
-produced by submucosal plasma cell n found n mucosa secretions(saliva,tears,colostrums,GIT,genitourinary)
-the dimeric form of secretory is composed of two basic units, a short peptide chain that join the 2 unit together
In a secretory component
-the secretory component help the passage of IgA through the epithelial cell n protect the molecules from
Proteolytic digestion
-secretory IgA provides local immunity at the mucosal surface- its main function is neutralization as it prevent
attachment of organism to mucosal surface

(g)Immunoglobulin class switching

-class switching is mediated by a change in constatant domain of the heavy chain(CH).There is no
alteration in the Lchain or the variable domain of heavy chain(VH),so that the immunoglobulin
produced later has the same specificity as the original IGM but differs in biological characteristic.
-Class switching is dependent on cytokines release from T cell.
-IL-4 increase IgE
-IL-4,IL-5,IL-6 increase IgG
-TGF-B increase IgA
-IL-5 increase IgA

2. Enumerate: applications of monoclonal antibodies(Ab) ~pg 81

a) diagnostic applications –widely used in different kinds of serological reaction for Ag detection
i. determination of lymphocytes markers e.g CD markers
ii. detection of HLA Ag (tissue typing)
iii. detection & typing of viruses
iv. hormonal assays
v. detection of tumor Ag
b) therapeutic applications
i. antitumor therapy; da use of tumor specific monoclonal Ab linked
to cytotoxic drugs (magic bullet therapy)
ii. immunosuppressive therapy in graft rejection; da use of
monoclonal Ab against CD3 on T cells
iii. ttt of drug toxicity e.g digitalis
iv. passive immunotherapy in some viral diseases
v. prevention of Rh incompatibility; da use of monoclonal anti-Rh D
3. Define

a) Paratope:
is the antigen-binding site. It is complementary to and interact with the epitope of the antigen
b)Heterophil Ab:
because of the similarity that may be found between different antigens,antibodies produced in response to
an antigen may cross react with another one.Such cross-reacting antibodies are called heterophil antibodies.
c) monoclonal Ab
these are highly specific antibodies against a single epitope produced by a single clone of B cells. They can
be artificially produced to be used in diagnosis and therapy.
d) immunoglobulins(Igs) ~pg 73
glycoproteins that bind specifically to da Ag that induced their formation

4.Compare
 Thymus dependent Thymus independent
1)Requires second signal from t T cell 1)No need of second signal from t cell
cell 2)no t help
2)Activation of t cell
Small antigen antigen Large antigen
Bacteria pathogen
IgM then IgG production,can Ig IgM only,no class switching
change to other isotype with same
immunological specificity
Memory cell is produced Memory Memory cell absent
cell
Fast Duration Fast and earlier

IgG IgM

Single basic unit (monomer) 5 bassic unit(pentamer) held together by

disulphide bond n a single J(joining) chain

75 % circulating Igs in blood 8-10% circulating Igs in blood

Major antibody of the secondary immune Major antibody of the primary immune
respone respone

Only Igs which can cross placental barrier Cannot cross placental barrier

Biological activities: Biological activities:

 Neutralization  Agglutination
 Opsonization  Complement activation
 Complement activation  IgM found on the surface of B cell
 ADCC forming BCR

Primary response Secondary response

Induction(lag)period Long (7-10 days) Short(few hours to gew

days)

Antibody level Low High(10 times greater)

Duration Short (antibody declines Long (month)

rapidly)

Ig class Predominantly IgM Predominantly IgG

Memory cell Absent Present

5.Give reasons:

a)Most vaccines are given in more than one dose

-The antibody level is 10 times greater than during primary response.The response can be boosted to higher
levels by further exposures.For this reason,most vaccines are given more than one dose.

b) IgG is the only Ig that can pass the placenta to the foetus ~pg 77
- IgG interacts with Fc receptors in the placenta & is, therefore the only Ig that can pass the placental barrier to
the foetal circulation(placental transfer)
- provides passive protection to the newborn during the first few months of life

c) IgM is the only antibody made to ABO blood group antigens

-this is because, Ig M is the only antibody made to thymus-independent antigens

d)Presence of IgM in newly born blood indicates intrauterine infection

-IgM cannot passes placenta from the mother to the fetus. It is produced in primary immune response.
---Therefore, its presence in newborn blood indicates intrauterine infection.

Chapter 17
COMPLEMENT
1) Compare :

a) The classical and alternative complement pathways.

b) The 3 complement pathways.

Type of pathway Classical pathway Lectin pathway Alternative pathway

Type of immuniti Acquired(specific) Innate(non-specific) Innate(non-specific)
lectin
Initiation Antigen-antibody Lectin bindind to Microbial
complex pathogen surface components (e.g.
endotoxin)
Role of antibody Needed for No role No role
initiation (activation
of C1)
Role of faotor B,D No role No role Have a role
and properdin
Role of mannose- No role Has a role No role
binding lectin
The involved C1,4,2,3,5,6,7,8,9 C4,2,3,5,6,7,8,9 C3,5,6,7,8,9
component

2) Give on account on :

a) Functions of complement.
I. Direct cytolysis : insertion of MAC into the cell surface lead to killing of many cells
through osmotic lysis.
II. Opsonization : C3b become deposited on the surface of the pathogen during complement
activation.Phagocytic cells recognize C3b bound to the pathogen via their C3b
receptors.This facilitate the attachment and subsequent uptake and killing of the C3b-
coated pathogen by the phagocytes cell.
III. Immune complex clearance : C3b receptors also found on RBCs.These recognize C3b
bound to soluble immune complexes and the RBCs bound to them.RBCs transport them to
the organ that rich in fixed pgagocytes.Phagocytes removed the immune complexes from
the red cells by their own C3b and Fc receptors.This help clearance of soluble immune
complexes from the circulation and prevents the development of immune complex
diseases.
IV. Inflammatory response : by-product C3a and C5a are produced during complement
activation.The molecules are called anaphylatoxins.They have following biological
activities:
a. Degranulation of mast cells and basophils to release mediators of inflammations.
b. Recruitment of phagocytic cells to the site of inflammation (chemotaxis_and stimulate
of their phagocytes power and intracellular killing.

b) Opsonization.
C3b become deposited on the surface of the pathogen during complement
activation.Phagocytic cells recognize C3b bound to the pathogen via their C3b receptors.This
facilitate the attachment and subsequent uptake and killing of the C3b-coated pathogen by
the phagocytes cell.

c) Immune complex clearance.

C3b receptors also found on RBCs.These recognize C3b bound to soluble immune
complexes and the RBCs bound to them.RBCs transport them to the organ that rich in fixed
pgagocytes.Phagocytes removed the immune complexes from the red cells by their own C3b
and Fc receptors.This help clearance of soluble immune complexes from the circulation and
prevents the development of immune complex diseases.

3) Give reason : Host cells are protected from damage by complement.

This because the host cells are protected by series of complement regulatory proteins.e.g.C1 inhibitor
binds to and inactivates C1 preventing further cleavage of C4 and C2.
 Chapter 18
IMMUNITY TO MICROBES

1. Give an account on:

a.Immunity to extracellular bacteria

Innate immunity
Phagocytosis by neutroils, moncyte or tissue macrophage.
Activation of complement by alternative pathway:
 Complement is activated by pptidoglycan and polysaccharide of bacteria cell wall.
 Leads to production o opsonin, recruitment and activation phagocytes and lysis of
bacteria.
Specific immunity
Humoral immune response:
 Main protective against extracellular bacteria.
 Antibody function: opsonization, agglutination of bacteria(prevent spread),
neutralization of bacterial toxin (prevent it bindin to host cell), binding to pilin
protein f bacteria (prevent it adhesion to host), activation of complement through
classical pathway.
T cell response:
Extracellular bacteria are internalized by APC and peptides are presented at MHC 11.
Thus, the T- helper is stimulated. T- helper secrete cytokine that leads to:
 Stimulate antibody production.
 Induce local inflammation.
 Enhance phagocytic and microcidal activity of macrophage.

b.Immunity to intracellular bacteria.

Innate immunity: INEFFECTIVE: bacteria are resistant to degradation within phagocytes.
NK cells provide early defense, produce IFN-y, activate macrophage to kill
intracellular bacteria.
Specific immunity
Humoral immune response:
dont play a role… because inaccessible to circulating antibody.
T cells response: main protective:
 Bacteria induce production of IL-12 b macrophage and IFN-y by NK cells………
development of Th-1 ……….secrete IFN-y…………. activate macrophage to kill the
bacteria inside them.
 BUT…
• In TUBERCULOID LEPROSY…Th-1 is induced….has intracellular digestion of
bacteria and some is living In tissue. Skin and peripheral nerve is damaged
but.. disease is under control.
• In LEPROMATOUS LEPROSY…Th-2 is induced… no intracellular digestion….
The organism grow abundantly….more tissue destruction and death occur.
c.Immunity to viruses
Innate immunity:
 NK cells.. killing virally infected cell in early infection.
 Type 1 interferons(IFN-a and IFN-B)….. is produced by virally infected cell. Its function:
• Inhibit viral replication and produce anti viral state
• Activate NK cells
• Increase expressin of MHC-1…. Better presentation of viral peptide.
Specific immunity
Humoral immune response: by antibody…(before the virus enter the target cell or during viruses
released from infected cell…. Antibody function:

 Bind to virus, thus prevent their attachment or entrance to host cell.

 Opsonization to enhance phagocytosis.
 Activation of complement.. promote phagocytosis and lysis of viruses.
1-Secretory IgA… neutralize virus that enter mucosa.
2-IgM and IgG…. Neutralize virus pass in the blood stream.
3-Type 1 IFN…act early during viral infection, on viruses intracellularly,and not
specific againsts certain virus.
4-antibodies… act on late stage, against extracellular viuses and specifis in action.
T-cell response:
 Tc cell..main mechanism where the infected cell is killed.
 Th cell… contribute by secretion of cytokines:
• IL-2 promote proliferation and activation of Tc cell and also activates NK cells.
• IFN-y activate NK cells.

d.Immunity to fungi.
Innate imunity:
 Neutrofil is the most important. It liberate fungicidal substance and phagocytose
 Macrophage can also combat fungal infections.
Specific immunity:
Humoral immune response:
antibody are often produced againdt fungi…BUT it is not useful because fungi act as intracellular
bacteria.
T cell response: Major defense against fungal:
 Fungi induce production of IL-12 b macrophage and IFN-y by NK cells………development of
Th-1 ……….secrete IFN-y…………. activate macrophage to kill the fungi inside them.
 Th1 response is protective while Th2 response is harmful.

e.artificial passive immunity.

 Passive acquired immunity involves transfer of ready- made antibodies or
lymphocyte to an individual.
 It is immediate immunity but temporary.
 It is 2 type:
Humoral immunity. Transfer of antibodies (Passive immunization):
 Administration of antitoxic serum.. prophylaxis or treatment against
bactria that produced exotoxins… as in diphtheria / tetanus.
 Administration of gamma globulin to immunodeficient person.
 Administration of convalescent serum to protect from disease
(infectious hepatitis).
Cell-mediated immunity.Transfer of lymphocyte.. transfer must between
genetically identical individuals to avoid rejection reactions.
2. Give reasons for :

a) Intracellular bacteria tend to cause chronic infection

-Intracellular bacteria are resistant to degradation within phagocytes.
-Thus, innate immunity is usually ineffective in controlling infections by these organisms.
-That is why these organisms tend to cause chronic infections that last for years and may recur after apparent
cure.

b) People with neutropenia are extremely sensitive to fungal infections.

-Neutrophil is the most important cell of the innate immune system in combating fungi.
-Neutrophil liberate fungicidal substances and phagocytose fungi as well.
-Thus, with decreasing number of neutrophil cell in body, that people will be very sensitive to fungal infections.

c) In case of fungal and intracellular bacteria infections, Th1 response is protective and Th2 response is
harmful.
-Th1 cell in cell-mediated immunity is the main protective immune response against intracellular bacteria and
fungal infections.
-Th1 activates macrophages for combating these organisms.
-The result is there is intracellular digestion of the organisms that found in the tissues. Although inflammatory
responses associated with macrophage activation occur, this is still under control and patient survives
-But with Th2 cells in humoral immunity, this is useless in combating against intracellular bacteria and fungal
infections because antibodies cannot reach the organisms. Then these organisms can grow abundantly in
macrophages causing much more destruction and eventually death.

3.Compare:
a)Type 1 IFN with antibobies in immunity against viruses

Type 1 IFN Antibodies

Act early in viral infection Act at a later stage in viral infection

Again intracellular viruses Again extracellular viruses

Not specific agains certain viruses Specific in action

b)active and passive acquired immunity

Active Passive

Role of immune system Important role No role

Mechanism Stimulate B/t cell Transfer of ready made

antibobies/lymphocytes
Onset of protection Delayed immediate
 Duration of protection Longer Shorter

Memory cell yes no

Examples -Natural infection -maternally –acquired

-Acive immunization with antibodies
vaccines -passive immunization
with antitoxin serum

c)immunity to extracellular and intracellular bacteria

Immunity to Extracellular bacteria Immunity to Intracellular bacteria

Innate Process start with phagocytosis then Only phagocytosis no
immunity degradation degradation(bacterial resistance)
Activation of complement by NK cell produce IFNγ then
alternative pathway activate macrophage

Acquired Humoral immunity play role Humoral immunity no role

immunity Both Th1 and Th2 play role Only Th1 play role

Chapter 19
TUMOR IMMUNOLOGY

1.Define
-Immuno –surveillance:ability of the immune system to prevent the development of most tumors through early
recognition and destruction of tumor cell.

2. Give an account on
a) tumor Ag ~pg 92
- 2 groups of tumor Ag
i. tumor-specific Ag (TSAs)- Ag that are expressed on tumor cells but not on normal cells &
might, therefore, induce an active immune response(IR)
ii. tumor-associated Ag (TAAs)- Ag that are relatively restricted to tumor cells but may also
present on normal tissue & therefore, may not be able to stimulate an effective response
- any tumor Ag from either this group that contributes to tumor rejection is referred to as tumor associated
transplantation Ag (TATA)
- origin of tumor Ag;
i. oncofoetal Ag- present during normal foetal development
- lost during adult life
- reappear with development of tumors
- e.g. alpha foeto-protein(AFP) in hepatoma
carcinoembryonic Ag (CEA) in colon carcinomas
ii. TATA on virally-induced tumors
- virally derived peptides associated with surface MHC on tumor cells
- behv as powerful transplantation Ag
- present mainly on tumors produced by oncogenic viruses
- e.g Epstein-Barr virus (EBV) in lymphomas, hepatitis B virus (HBV) in
hepatic carcinoma
iii. tissue-specific differentiation Ag
- tumor arising from a particular tissue may express normal differentiation Ag
specific for that tissues
- e.g prostate-specific Ag(PSA)
-
(b) Tumour markers
Tumour markers are substance that present which indicating tumour.
Tumour antigens can be very useful tumour markers in the diagnosis and follow-up of various tumours.
An ideal tumour marker is:
• Release only from tumour tissue
• Specific for a given tumour type
• Detect antibodyle early upon tumour formation
• Its concentration in the blood is proportional to the tumour mass
• Present in all patients with the tumour
Most tumour marker are antigenic, circulating in the blood, and can be detected by immunoassays.
Although useful in monitoring patients for tumour recurrence after therapy, no tumour markers has definite
specificity or sensitivity for application in early diagnosis.

Examples of tumour markers are:

• Carcinoembryonic antigen (CEA): in colon carcinomas
• α-Foetoprotein (AFP): in primary hepatoma
• β-Subunit of human chorionic gonadotrophin (β-HCG): in choriocarcinoma
• prostate-specific antigen (PSA): in cancer prostate
• CA 125: in ovarian cancer
• CA 19-9: in colonic and pancreatic tumours
• CA 15-3: in breast cancer
 • Bence-Jones proteins: in myeloma
Pancarcinoma antigen (TAG-72): is used to localized occult tumour secondaries by using
radiolantibodyelled monoclonal antibody

(c) ) Effector mechanism in tumor immunology

Innate and specific (humoral and cellular) immune system can effect the growth and progression of tumor:
a- T cell =

cytotoxic T-cell : recognize the tumor antigen then kill them helper T-cell recognize sheded tumor
antigen which is internalized and presented on Antigen Presenting Cell (APC) which will secrete
cytokines that will activate Tc cells,macrophages,NK cells and B cell.
b- B cell = process and present tumour antigen to Th cells
tumour specific antigen which leads to tumour cell lysis
fix complement on tumour cell membrane,membrane attack complex leads to lysis of tumour
cell
Antibody dependent cell mediated cytotoxicity (ADCC)
c- NK cells

(d) Approaches to Tumor Immunotheraphy

1. Passive cellular immunotherapy -term used when activated cells are directly infused to a patient
a)therapy with lymphokine-activated killer(LAK) cells: the patient lymphocyte are cultured in
vitro with IL-2 then reinfused
b)therapy with T cells:lymphocytes that have infiltrated tumors are excreted from tumor
biopsies, stimulated in vitro using IL-2 n tumor antigen and then reinfused
2. Passive humoral immunotherapy-
-monoclonal antibodies directed against tumors antigens
-may be used either alone or coupled to radioisotopes, cytotoxic, drug, toxins or cytokines.
-coupling has the advantages of delivering high doses of cytotoxic drugs to the site of tumor

3.Passive cellular and humoral(combined) immunotherapy

-links 1 antibody reacting with the tumor cell to a second antibody reacting with a cytotoxic cell

4.Active specific immunotherapy

a)Allogeneic tumor cells: tumor taken from other patients, irradiated n then injected with BCG
vaccine or other adjuvants
b)Tumor antigen vaccines: cellular immunity to antigens can be induced by using short synthetic
peptide
5.Nonspecific immunotherapy
a)interferons
b)bacterial adjuvants

3.Enumerate

i.Evasion of the Immune Response

a- immunocompromised host
b- related to tumour antigens:
-tumours lack antigens that can stimulate immune response
-tumours can’t be processed and presented with MHC
 -amount too small to stimulate immune response
-sheded antigens block antibodies and T cells from reacting with tumour cell
c- tumours located at inaccessible to the immune system
d- poor in expressionof MHC 1 molecules
e- fibrin coating,masking of tumour antigens
f- tumours secrete substances that suppress the immune response

-ii.Characteristic of ideal tumor makers

1) Released only from tumor tissue
2) Specific for a given tumor type
3) Detectable early upon tumor formation
4) Its concentration in the blood is proportional to the tumor mass
5) Present in all patients with the tumor

2. Give reason:

Th cells play a role in immunity to tumors, although tumor cells express class I MHC & not class II
MHC molecules ~pg 93
- Th cells are activated by shed tumor Ag which become internalized & presented on the surface of APC
in association with MHC II
- they secrete cytokines which activate Tc cell, macrophages, NK cells & B cells
- produce TNF~directly toxic to tumor cells

Chapter 20
HYPERSENSITIVITY
1.Define:
a) hypersensitivity reactions : inappropriate immune responses to certain antigens causing tissue damage
b) desensitization : a method for reducing the effects of a known allergen by injecting, over a period,
gradually increasing doses of the allergen,until resistant is built up
2.Give an account on:
A)mechanism of hypersensitivity:

Type I
(1)Sensitization
-in normal person only low level of IgE are relesed when exposed to antigen.
-but high levels of IgE are produced in individuals having this type of hypersensivity.
-TH2 lymphocytes are responsible for production of IgE.
-TH2 cell will release IL-4 which is critical stimulus for changing production IgM into IgE.
-IgE bind to mast cells and basophils via its Fc receptor.
-Now,mast cells and basophils are sesitized during first exposure.

(2)Degranulaton of mast cell

-upon 2nd exposure to the same allergen,IgE on the mast cell cross link by the allergen.
-this induce degranulation of mast cell and release two types of mediators.
a)Preformed mediators
-histamine and PAF(platelet activating factor)
-during early phase,which occur witin 15-20 minutes after exposure.
b)Newly formed mediators
-leukotriens and prostaglandin
-during late phase,typicaly occur 5-6 hours after exposure and persist longer.
-recruitment of other effector cells e.g. TH2,eosinophils,basophils which
contribute to inflammatory respond and tissue damage.

Type 2

(1)Lysis of target cell by complemet activation.

(2)Opsonization with or without complement activation.
(3)Lysis of target cell by ADCC(antibody dependant cell mediated cytotoxicity)

Type 3
(1)Formation of immune complex(antigen with IgG/IgM)
(2)Penetration of complex to the endotelial wall and become deposited on the vascular basement membrane.
(3)Activation of complement ; C 3a and C5a(anaphylatoxin)
-react with receptor on mast cells and basophil
-release of vasoactive amines
-increase vascular permeability
-C 5a also chemotactic to neutrophils that infiltrate the area.
-neutrophils engulf this comlex and release lysosomal enzymes that destroy basement
membrane.
(4)Platelet aggregation
-release vasoactive amines
-form microthrombi that cause local ischemia and further tissue damage.
Type 4
(1)TH 1 cells recognize antigen
-release po-inflammatory cytokines.
-increase vascular permeability leads to leakage of flid ito the tissue.
-attract and activate monocytes,macrophages, and more T cells.
-inflammatory reaction leads to local tissue damage.
-T cytotoxic may also play role.

B)Atopy
-hereditary tendancy to type 1 hypersensitivity.
-produce larger amount of IgE than oter individuals in response to allergen.

C)Arthus reaction
-local immune complex disease
-due to repeated subcutaneous injection of low dose of foreign antigen
-e.g insulin and rabies vaccine
-occur at the site of inection
-immune complex deposit in the blood vessel wall,activation of comlement, and
inflammatory response occur
-resulting in local erythema,oedema,and necrosis.

D)Diagnosis & therapeutic measures of type 1 reaction

*Diagnosis
(1) -detection of antigen by skin test
-introduce various antigen to the patient skin
-Wheal flare reaction proves positive case
-it appears within 15-25 minutes at the site where the patient is allergic to certain antigen.
(2) -Measurement of total IgE levels and specific IgE for particular antigen.

*Therapeutic measures
(a)anaphylactic shock management
-an emergency case
-immediate admininstraton of adrenaline,corticosteroid,and oxygen inhalation.
(b)Atopy management
-AVOIDANCE of responsible allergen
-DESENSITIZATION by giving gradually increase dose of injection against the allergen.Its aim is at shifting
the immune response from TH2 to TH1 thus down regulate IgE
-DRUG that inhibit mediator release/couteract their effect e.g. antihistaminic,corticoseroid,antileukotriens
E)Serum sickness
-systemic immune complex disease
-due to injection of large dose of :
*foreign serum-horse antitetanic / antidiphtheritic serum
*drug-penicillin
-antigen slowly cleared form the body while antibody production starts.
-antibodies bind with remnants of antigen.
-Formed immune complex deposite at certain site cause disease manifestation.
-It leads to fever,urticaria,arthralgia,lymphadenopathy,splenomegaly(develop few days-2 weeks after time of
injection)

F)Granuloma formation
-can be seen especily in chronic infectious disease caused by intracellular bacteria (e.g. mycobacteria)
-when the intracellular bacteria resist to microbicidal effect of activated macrophages,the antigen persistently
give rise to chronic antigenic stimulus.
-Continous release of cytokines by Th 1 accumulate activated macrophages forming GRANULOMA
*granuloma:
-protecive mechanism to isolate pathogen that resist destruction
-prevent from dissemination of infection
-are fatal with its absence
-e.g in AIDS patient with mycobacterial infection since Th level is very low
-central core is infected macrophages,may include fused macrophages(multinucleated giant
cell),surronded by large macrophages (epitheloid cell)
-Th cell surrond the central core
-The cells in the center of granuloma undergo necrosis (caseation necrosis)

3.Enumerate the clinical forms (examples) of:

a)Type I Hypersensitivity : -Systemic anaphylaxis

-Localized anaphylaxis ( Atopy )
b)Type II Hypersensitivity : -Blood group incompatibility
-Autoimmune disease
-Drug interact
-Graft rejection
c)Type III Hypersensitivity : -Serum sickness
-Arthus reaction
-Post-streptococcal glomerulonephritis
-Viral infections
-Autoimmune disease
-Hypersensitivity pneumonitis
d)Type IV Hypersensitivity : -The tuberculin skin test
-Contact dermatitis
-Granuloma formation
-Autoimmune disease
-Graft rejection
 Chapter 21
TRANSPLANTATION IMMUNOLOGY
1) Classify Class I MHC and Class ll MHC

2) Enumerate.
a.Types of graft
1) Autograft - graft from one part of body to another. Not foreign and do not elicit
rejection
2) Isograft – graft between genetically individual (eg: identical twins). Do not
express antigen and not rejected.
3) Allograft – one person donates organ to genetically different individual of the
same species.This allograft express antigen which are recognized
foreign by the recipient.
4) Xenograft – represent maximal genetic disparity and rapidly reected.

MHC l MHC ll
Found on surface of all nucleated cell Found on surface of the professional
antigen presenting cell.
Composed of 2 polypeptide chain Composed of 2 glycoprotein chain ( alpha
( alpha chain and beta-2 microglobulin chain and beta chain )
chain )
Locus : HLA-A, HLA-B, HLA-C Locus : HLA-DP, HLA-DQ, HLA-DR
b. Types of rejection
1) Acute rejection – takes days or weeks to develop because of the time taken for T
cell activation
2) Hyperacute rejection – take place within minutes, caused by preformed anti-donor
antibodies
4) Chronic or late rejection – take place months or years after transplantation, depending
on genetic disparity.

c. Antigen responsible for immune rejection

1) Blood group antigen of the ABO system
2) MHC antigen
3) Minor histocompatibility complex antigens.
3. Give an account on
a. significance of MHC
1) antigen presentation
2) enables virus infected cell to be a target for Tc cell ( MHC I)
3) mismatching of donor and recipient MHC causes graft rejection
4) certain MHC alleles affect the susceptibility of an individual to certain diseases. These diseases
are often inflammatory or autoimmune in natur (e.g. ankylosing spondylitis and B27, rheumatoid
arthritis and DR4, systemic erythematosus and DR3)
5) used in forensic medicine e.g. disputed paternity

b. mechanism of graft rejection

1) acute rejection
-it is type IV hypersensitivity (cell mediated) delayed. It takes days or weeks to develop because
of the time taken for T cell activation, proliferation and migration into the donor graft.
-Tc cell causes direct destruction of transplanted cells
-Th cells release a number of cytokines which lead to activation of the immune response.
-IL-2 is required to activate Tc cells
-TNF-α and IFN-γ activates macrophage
2) hyper acute rejection
-caused by performed anti-donor antibodies binding to either ABO blood group or HLA class I
antigen on the graft. It take place within minutes of transplantation.
-The antibodies attach to the endothelial cells of the donor organ and fix complement
-Causing damage to the vascular endothelium, leading to
• haemorrhage
• platelet aggregation within the vessels
• graft thrombosis
• lytic damage to cells of transplant
• release of pro inflamatory complement compenents C3a and C5a
3) chronic or late rejection
caused by low grade cell mediated rejection or the deposition of antibodies or antigen-antibody
complexes in graft. It take place whithin months to years after transplantation
depends on
• genetic disparity between donor and recipient
• success of immunosuppressive therepy
more prominent with patient having
• chronic viral infection
• cytomegalovirus (especially)
• pre-existing autoimmune disease

c. graft versus host disease

-it occurs when the graft react against the recipient tissue not vice versa
-e.g. in bone marrow transplantation
-immunological competent T cells are transplanted into a genetically non-matching immunosuppressed
recipient.
-To reduce the risk
• careful typing
• careful use of immunosuppresive drugs
• careful removal of mature T cells from the donor stem cell
 d. prevention of rejection
1) selection of donor
• ABO matching
Perform antibodies against ABO blood group system
• Histocompatibility testing
Leucocytes carry all known HLA antigen, so it can be used for tissue typing to identify HLA
and test compatibility between donor and recipient

i. Tissue typing by lymphocytoxicity test

Purified suspension of lymphocyte + antisera + complement
If complement fixed = cell damaged n dye enters
If complement not fixed = cell remain viable, unstained
ii. Compatibilty testing by the mixed lymphocyte reaction (MLR)
Donor and recipient cells are incubated together
If recipient cell recognize foreign HLA on donor cell, it will proliferate
Proliferation is measured by the extent of radioactive thymidine uptake
iii. Molecular HLA typing
PCR
Accurate and sensitive

2) immunosupressive therapy
prevent and/or treat graft rejection n GVHD
these drugs carry risk of infection
• corticosteroids: potent anti-inflammatory
• cytokine producer inhibitor: cyclosporine n tacrolimus inhibit T cell cytokine production
mainly IL-2 and TNF-γ
• antiproliferative drugs: azathioprine and methotraxate inhibit DNA production, thus prevent
lymphocyte proliferation
• monoclonal antibodies: monoclonal antiCD# antibodies block the function of T cells

3) antigen specific induced tolerence

still under trial~

4. Give reasons:
a) cyclosporin is used to prevent graft rejection
it inhibits T cell cytokine production mainly IL-2 and TNF-γ
b) azathioprine is used to prevent graft rejection
it inhibits DNA production, thus prevent lymphocyte proliferation

Chapter 22
Tolerance and Autoimmunity
1.Define :
 A Tolerance absence of specific immune response againsts some antigen in
fully immunocompetent person

B Autoimmunity adaptive immune response to self-antigens

C Clonal elimination of immature self-reactive lymphocytes during their
deletion maturation

2.Give an account on :
1.Mechanism of auto-tolerance :
a.central tolerance : it is the phase in the primary lymphoid organ in which B and T lymphocytes
undergo permanent inactivation after being in contact with self-antigen
b.peripheral tolerance :
i. it occurs if the elimination of self-reactive cells in primary lymphoid organ is not
complete. It is because some of the self-antigens are not expressed in them.

ii. Self-reactive B cells may mature and migrate to the secondary lymphoid organ but due
to lack of T cell help, they cannot do their function
iii. Self-reactive T cells are also unable to respond due to various mechanism

2.Aetiology of autoimmune diseases

d. Exposure of the immune system to antigens that are normallly hidden within organs

e. Structural modification or alteration of tissue proteins

f. Cross reactivity

g. Breakdown of immune network which occur due to:

i. Interference with the mechanism which normally suppress surviving self-reactive T

cells

ii. Polyclonal activation of lymphocytes

iii. Over production of IL-2 by Th1 cells

3.Diagnosis and management of autoimmune diseases

a.Diagnosis
i.Elevated level of serum immunoglobulins
ii.Detection of autoantibodies in the serum
iii.Detection of immune complexes in the serum and in tissue biopsy
iv. Decreased level of serum complement, due to uptake in immune complexes
b.Management
i.Anti-inflammatory drugs
ii.Imunosuppressive drugs
iii.Plasmapheresis
iv.Interference with cytokine network(under trial)

4.Spectrum of autoimmune diseases

 At one extreme, there are the organ-spesific diseases where the immune response is directed against
components spesfic to the organ involved, eg. Grave’s disease

At the other end, non-organ-spesific diseases. The lesion is not confined to one organ eg.SLE

In between, antibodies formed are non-organ-spesific but the lesion tends to localize to a single
organ eg. antimitochondrial antibody in 1ry billiary cirrhosis

3.Enumerate
a. Factors influencing the induction tolerance

i. high doses of antigen tolarize B-cells while minute doses given repeatedly tolarize T-cells

ii. protein antigen are more tolerogenic in soluble form than in aggregated or particulate
form

iii. to maintain acquired tolarance, the tolerogen must persist or repeatedly admnistered

iv. giving the antigen together with immunosuppresant

v. prenatal or neonatal period

b. Mechanism of tissue damage in autoimmune disorders

i. cytotoxic reactions (type II)

ii. immune complex deposition (type III)

iii. cell-mediated reactions (type IV)

iv. in Grave’s disease, anti-thyroid antibodies react with thyroid gland cells, increasing the
response of cell receptors to TSH, thus stimulates secretion of excess thyroxin, leading to
thyrotoxicosis.
 Chapter 23
IMMUNODEFICIENCY DISEASES
1. Mention the patient presentation in(or the effect of):
a) Defects of migration of phagocytic cells
o defects in migration leads to failure of neutrophils and monocytes to migrate from blood stream
to sites of infection,despite their presence in large numbers in the blood.
o patient present with infections of skin,mouth,and respiratory tract,but with little pus
formation.

b) Defects intracellular killing of phagocytic cells

o phagocytes cannot produce reactive oxygen radicals,lead to decrease ability to kill intracellular
as well as ingested bacteria
o male children present with chronic bacterial infections-some cases lead to formation of granules.

c) Defect of early component of classical pathway

o accumulation of immune complex and local tissue damage

d) Defect of early component of alternative pathway

o pyogenic infection

e) Defect of terminal complement (C5-C9)of complements(MAC)

o increased susceptibility to capsulated organism Neisseria meningitides

f) Defect of complement C1 inhibitor

uncontrolled activation of the classical pathway of complement activation
o C2 activation and generation C2a(vasoactive amine)-fluid accumulation in tissues and swelling
of epiglottis which may lead to suffocation and death.”Hereditary Angioedema”

g) X-linked agamaglobulinemia
o infants:symptomatic and natural loss of antibodies at the age 5-6 months They suffer from
chronic bacterial infection such as otitis,bronchitis, and pneumonia.

h) Transient hypogammaglobunaemiaof infancy

o infants have recurrent infections and poor response to the vaccines taken routinely at that age.

i) Deficiency of natural killer cell

o patients suffer from certain viral diseases and malignancies

j) Thymic aplasia and hypoplasia(Di George Syndrome)

o develop recurrent and chronic viral,bacterial,fungal and protozoal infections

k) Combined T and B cell deficiency(SCID)

o patients present during the first few months with failure to thrive,continuous diarrhea,as well as
viral and fungal infections.
2. Give reasons
a.Defective T-cells immunity(without affection of humoral immunity) is rare:
As the collaboration between T cells and B cells in the process of antibody formation.
Thus, production of antibodies against T-dependent antigens in particularly can be
affected in cases of T cell immunodeficiency.
In George syndrome is a severe form T cells deficiency caused by congenital aplasia or
hypoplasia of the thymus.the result is the patients develop recurrent and chronic
viral,bacteria,fungal and protozoa infection.

b.Patients suspected of suffering immunodeficiency never be given live vaccine:

the patient may die as the result of immunodeficiency due to lack of T-cells and B-cells
action which are very important in protection of the body from the foreign body(live
vaccine) organism.

3.Give an account on:

a.Leucocyte adhesion deficiency
It is defect of phagocytic function on migration that involves adhesion molecules.
Lead to failure of neutrophils and monocytes to migrate from blood stream to
sites of infection although there are in huge amount.
Patient – infections of skin, mouth, respiratory tract with little pus formations.

b.Chronic granulomatous disease

It is defect of phagocytic function on intracellular killing.
Phagocyte cant produce reactive oxygen radicals – decreased ability to kill
intracellular and ingested extracellular bacteria.
Usually an X-linked disease and male children with chronic bacterial infections
that may lead to granulomas,

c.Investigations of T cell deficiency

Quantitation of cell by fluorescein-labelled monoclonal antibodies against cell markers, e.g. with
antibodies against CD3 ( to quantitate total T cells ) and against CD4 and CD8 ( to quantitate T cell sub-
sets ).
Measure ability of T cells to proliferate in response to mitogens. ( mitogens are substances that can non-
specifically stilmulate proliferation of lymphocyte. Some stimulate T cell only, some B cell and others
stimulate both ).
Delayed type hypersensitivity test to test functional activity of T cells using extracts from organism to
which people are frequently expose, e.g. candidin test.
Assessment of cytokine production.

d.investigation of B cell deficiency

- Quantitation of cells using flurescein-labelled monoclonal antibodies against B cell markes.
- Determination of serum level of immunoglobulin classes as well as IgA in saliva.
- Immunization with commonly used vaccines such as tetanus toxoid followed by measuring the antibody
response.
 e.management of the immunodeficient patient
General measures – minimize infections by avoidance.
• Administration of suitable immunization. Patient with immunodeficiency should never be given
vaccines contain living organism since it maybe fatal.
• Prompt treatment of infections with antibiontics.
Specific measures according to defect
• Transplantation of foetal thymus or bone marrow in SCID and other deficiencies.
• Immunoglobulin therapy for cases of antibody deficiencies.
• Treatment with cytokines such as GM-CSF, IL-2, IFN-gamma

4. Enumerate

a. Causes of secondary immunodeficiency(ID)

may acquired a transient or permanent immunologic impairment later in life.
1. Malnutrition in poor and under development country
2. Human Immunodeficiency Virus (HIV) causes Acquired Immune Deficiency Syndrome
(AIDS).the virus infects CD4 Th cells
3. Other virus infection lead to transient ID eg. Measles
4. Severe bacterial infection eg. Tuberculosis
5. Parasitic manifestation eg. Schistosomiasis
6. Malignancies specially Hodgkin’s disease and leukaemias
7. Others eg. Treatment with X-rays, cytotoxic drugs, steroids,immunosuppressive drugs and
burn with severe loss of body fluids.
8. chronic debilitating disease eg. Diabetes and renal failure

b. Causes in which immunodeficiency is suspected

1. increased in frequency of infection
2. failure clearance of infection rapidly despite adequate therapy
3. dissemination of local infection to distant sites
4. occurrences of opportunistic infection
5. failure to thrive in infant and children
6. development of certain kinds of tumour
 Chapter 24
ANTIGEN-ANTIBODY REACTION

1.Give an account for each principle of Ag-Ab reaction and give one application. (Illustrate with diagram)
a.Haemagglutination inhibition
 To see the agglutination of the RBCs by antibodies, certain virus particles (influenza, mumps
viruses etc) or other substances.
 Use for diagnosis certain viral diseases because specific anti-viral antibodies bind to the viral
particles and block their ability to agglutinate RBCs.
 Eg. – a person suspected of having influenza so his serum is examined for specific antibodies.
Serum is mixed with known influenza virus and RBCs.
 If no haemagglutination occur, so the antibody is present and vice versa.

With antibodies = no agglutination No antibodies = agglutination

b.Single radial immunodiffusion

a.Used to quantitate antigens
b. The antibodies are mixed with the agar before it was poured into the plate. Then the antigen is
placed in wells punched in the agar gel
c. Precipitation will occur as a ring around the antigen well
d. The diameter of the ring is proportional to the concentration of the antigen in the well
e. Routinely used to quantitate various Ig classes in human samples eg. Quantitation of IgG by
using agar gel mixed with anti IgG antibody
 Sample 2 Sample 1 Ag standard 2 Ag standard 1

Petri dish containing antibody in agar

c. Toxin neutralization
a. Antibodies against bacterial toxin will neutralize its hazardous effects
b. Eg. In-vivo toxin neutralization test for determination of toxicity of C. diphtheriae
c. Diagram as haemagglutination test

d. Direct immunofluorescence

a. Used to detect certain antigen in tissue sections fixed in microscopic slide

b. Specific fluorescein-labelled antibody is used
c. Presence of antigen will be bonded and be visualized as a green stain specimen under the UV
light
d. For the diagnosis of rabies in brain of rabid animals

Fluorescein unknown antigen binding observed by UV microscopy

tagged antibody

e. ELISA
a. The principle is conjugation of the antibodies or antigens to an enzyme
b. Detection of the conjugation is by adding proper colorless substrate that in the presence of the
enzyme it'll be converted into a colored product
 c. Degree of color change can be measured by spectrophotometrically; considered as an indicator
of the amount of antigen or antibody in the sample
d. Examples and diagrams of technique that could be used will be referred to page 125 and 126 vol
III

f. RIA (Radioimmunoassay)
a. A radioactive assay eg. Iodine125 replaces the enzyme in ELISA technique to label antigen or
antibody
b. Ag-Ab reaction will be measured by radioactivity
c. Accompanied by hazards of radiation
d. Used to quantitate many biological substances eg. Hormones, tumour markers, drugs

g. Coombs’s Test:
Function: To detect non-agglutinating antibodies.
Direct Differences Indirect
Red cell of newborns- Sample Serum of Rh –ve mother sensitize
erythroblastosis fetalis with Rh antigen

Hemolytic anemia patients

1-Patient’s RBC washed
2-Add anti human globulin to cell
suspension
Positive result
Method
1-Serum + Rh +ve + group O red
cells => incubated
2-Washing
3-Add anti-human globulin
Agglutination Positive result

h. FlocculationFunction : Detect small insoluble particulate antigen

Example: Syphilis
Antigen: Water insoluble cardiolipin (not treponemal-difficult to obtain)
Result : Microscopical aggregates => heterophil antibody(reagin) in serum of +ve syphilis.
~in RPR-carbon particles added => result seen by naked eye.

i. Complement Fixation Test

Sensitive test.
In antigen-antibody interaction – complement is fixed(consumed)
Function : Detect and quantitate antigen-antibodies
Source of complement: Guinea pig serum-rich in complement
Method:
(a) Test system:
1-Heat the serum 56°-inactivate native complement
2-Add measured amount of antigen & complement
+ve-antigen-antibody complexes will form(cannot be seen)
(b) Indicator system:
1-Add sheep RBC+hemolysin => test presence of free complement
Result
+ve - no hemolysis
- all complement are used
-ve -hemolysis
- free complement exist => lyse the sheep RBC
Example: -Diff bacterial
-Rickettsial
-Viral
-Fungal
-Parasitic diseases

j. Viral neutralization
Cytopathogenic Effects(CPE) :Virus + app. cell culture => observable cell destruction
Function: searching virus neutralizing antibodies in a serum sample
Method:
1-Serum containing antibody + known virus suspension.
2-Inoculate susceptible cell culture into (1)
Result:
+ve CPE : virus + target cell ONLY =>no neutralizing antibodies.
-ve CPE : serum antibody + virus + target cell => antibodies present.

k. Indirect immunoflourecence
Function: Detect presence of antibody against certain organism.
Method:
1-Serum of patient (unknown antibody) + Treponema pallidum (known antigen) on slide
2-Washing
3-Overlay flourecein-labelled antihuman antibody on the slide
4-Examine with UV microscope
Result: +ve-green fluorescence => antigen antibody complexes binding.

l.Passive agglutination
Method:
Adsorbing the known reactant (soluble antigen or antibody) passively on suface of inert particle (e.g: Latex
particles or RBC )
~if RBC is used= Passive heamagglutination.
Result: +ve – agglutination occur.
Examples:
1- Rheumatoid arthritis (IgM vs IgG)
Antibody: Rheumatoid Factor(RF)
Latex coated particles + IgG = agglutination [RF present]
2- Inflammation
Antibody: C-reactive protein
Serum + latex + anti C-reactive protein = agglutination [ C-reactive ptn present]
3- Immunologic pregnancy test
Antibody: HCG
Drop of urine + latex + anti-HCG = agglutination [ +ve pregnancy test]

4.Mention the type of Ag-Ab reaction in each of the following tests

1. Blood grouping – Slide agglutination

2. Elek's test – Precipitation
3. Antistreptolysin O – Passive agglutination
4. Diagnosis of rabies in brain of rabid animals – Complement fixation
5. Diagnosis of newborns with erythroblastosis fetalis – Coomb's (antiglobulin) test
6. Immunologic pregnancy test – Passive agglutination
7. VDRL test for syphilis – Flocculation
8. Quantitation of hormones – Radioimmunoassay
9. Detection of viral infections – Haemagglutination inhibition test
10. Detection of anti-Rh in the mother's serum – indirect Coomb's (antiglobulin) test
EXAM : GENERAL BACTERIOLOGY

1) Compare :
a)compare gram +ve and gram –ve cell wall

Gram +ve Gram –ve

As many as 40 sheets Thin, 1-2 sheets
Peptidoglycan
Comprising up to 50% of cell Comprising 5-10% of cell
layer
wall material wall material
Teichoic acid and cell wall
Teichoic acid associated protein are major Absent
surface antigen
Outer surface is composed
of molecules of
lipopolysaccharide (LPS).
Outer
Absent Lipid A of LPS forms the
membrane
endotoxin, while
polysaccharides are major
surface antigen
Space between cytoplasmic
and outer membrane
Periplasmic
Absent containing peptidoglycan
space
layer and gel-like solution of
protein

b)compare obligate aerobes and obligate anaerobes

Obligate Aerobes Obligate Anaerobes

Oxygen Require complete absence of
Require oxygen for growth
requirement oxygen
Example Pseudomonas aeruginosa Bacteroides fragilis

c)compare conjugative and non-conjugative plasmids

Conjugative Non-Conjugative
Size Large Usually small
Copy number 1-2 (stringent) >30 (relaxed)
F factors Present Absent
Sex pilus formation Yes No
By the help of conjugative
Transfer among bacteria By conjugation
plasmid
Host bacteria Common gram –ve bacilli Common in gram +ve cocci

d)exotoxins and endotoxins

Exotoxins Endotoxins
Secreted by living organism Integral part of cell wall of
Source both garm +ve( mainly ) and – gram –ve organisms.liberated
ve upon cell disintegration
Encoded by
Coding Encoded by genes on the
chromosomes,plamids,bacterio
genes chromosome
phages and PAI
c.diptheriae (phage)
cl.tetani (plasmid) e.coli and meningococcal
Examples
b,pertussis (chromosome) endotoxins
h.pylori (PAI)
Nature Protein Lipopolysaccharide (lipid A)

Antigenicity Higly antigenic Poorly antigenic

Heat Stable temp. >60 for several
Unstable to temperature >60
stability hours
Detoxificati
Can be converted into toxoid Can not
on
Same generalized effect ( non
Specificity Every toxin has specific action specific action),all give fever
and shock.
Toxicity high Low

2) Give an account on :
a) Outcome of temperate phage cycle

1. The prophage may be carried inside the bacterial cell indefinitely passing to
daughter cells.
2. The prophage may be induced to detach from the bacterial chromosome and start a
lytic cycle. Induction may be spontaneously or achieved by an inducer, e.g U.V light
3. During the process of induction, the prophage may carry with it few genes of the
bacterial chromosome. When it infectes another bacterium, it passes this fragment
to it giving it new characters. This is known as “specialized transduction”.

b) Complication of chemotherapy
1. Toxicity-may be dose dependent or independent
 Tetracycline may cause staining of teeth in infants
 Streptomycin may affect the 8th cranial nerve leading to vestibular
dysfunction.
  Aminoglycosides may cause nephrotoxicity
 Chloramphenicol can cause bone marrow depression

2. Allergy (Hypersensitivity): usually not dose dependent

• Penicillins may cause urticaria, anaphylactic shock or serum sickness.
• Local application of sulphonamides may result in contact dermatitis.

3. Emergence of resistant strains:

• The abuse of antibiotics ( low dosage, interrupted course, no real indication
and improper choice) encourages the emergence of resistant mutants. These
mutants will overgrow and replace the originally susceptible bacteria. It is
recommended that in vitro susceptibility testing should be performed to
guide the selection of antibacterial drugs.

4. Superinfection
• It occurs as a result of outgrowth of resistant members of normal flora when
the sensitive ones are eradicated during antibiotic therapy: eg:-
pseudomembranous colitis caused by outgrowth of clostridium difficile.
• -oral thrush caused by overgrowth of the yeast candida

3) Give reason for :

a) Mycoplasma are resistant to penicillin

Mycoplasma is the only group of bacteria that exist naturally without cell wall so they are
naturally resistant to cell wall inhibitor like penicillin.

b) Plasmid may render the bacteria pathogenic

Plasmid may render the bacteria pathogenic as it has fuction as virulence plasmid that
may code for exotoxins, adhesions or invasion factors.

c) The ideal cloning vector should carry a selectable marker

It must carry a selectable marker so that the host cells transformed with the vector can be
distinguished from non transformed cells.

4) Define:

a) Prophage- integrated phage genome.

b) Bacterial genome- total set of genes present inside the bacterial cell.
c) Competence- the ability of the recipient bacterial cell to take up DNA.
d) Opportunistic pathogens- These are potentially pathogenic bacteria that do not cause
disease under normal condition but can cause disease in immunocompromised patients or
when they find their way to another site other than their normal habitat.
e) Disinfection- It is the elimination of most, if not all, pathogenic microorganism excluding
spores.
 f) Selective toxicity- is the ability of an antimicrobial agent to harm a pathogen without
harming the host.

CH 14-T cell mediated immunity

1) Compare :
a) Effector and naïve T cells

Effector T cells Naïve T cells

Ability to respond
quickly and
efficiently when Yes No
encounter antigen an
target cells

Can be trigger to act

without need co-
stimulation Yes No

b) Ordinary antigen and superantigen

Ordinary Antigen Superantigen

Processing inside Yes No

APCs

Presentation by MHC Yes No

molecules

Site of binding to Peptide-binding cleft Outside peptide-binding

MHC molecule cleft

Binding to TCR Variable portion of α Variable portion of β

and β chains chain
Specificity of TCR to Very specific Not very specific
it

Acquired immune Stimulated Supressed

response

Development of Yes No
memory

Result of T cell Usually beneficial to Usually harmful to host

stimulation host

c) NK cells and Tc cells

Tc cells NK cells

MHC restriction Resticted to MHC I No restriction

Antigen specificity Specific to certain Not specific

antigen

Mechanism of killing Direct killing

Direct killing Antigen dependant

cellular cytotoxity
(ADDC)

2. Give an account on

a) Sequence of events in activation of naïve T cells

• Any cytosolic peptide is delivered on a MHC1 molecule to the surface of the APC. A CD8
(cytotoxic) T cell with TCRs specifics for that peptide binds he MHC1 peptide complex.
• Similarly, any vesicular peptide is delivered on an MHC 2 molecule to the surface of the
APC. A CD4 (helper) T cell with receptor specific for that peptide binds the MHC 2 peptide
complex.
 • In either case, this leads to delivery of first signal required for T cell activation.
• The second signal is delivered by binding of CD28 on the T ell to B7 on the APC(co-
stimulation).This signal is very important and without it the T cell ‘shuts down’ and
becomes non-responsive, a state called anergy.
• During this events, certain molecules called adhesion molecules. Present on the T cell and
APC help to hold the two cells together.
• The T cell become a lymphoblast (activation), divides repeatedly (proliferation or clonal
expansion) and its progeny (daughters) finally become interleukin-2 (IL2), a cytokine
secrete by the T cell itself.

b) Superantigens

• Superantigen is certain protein that secreted by some pathogens do not act by ordinary
antigens
• They are not processed and presented to T cells like ordinary antigens, but have the
ability to bind directly to the MHC 2 molecules on the surface of the APC without entering
the cell, and at the same time to the variable portion of the β chain of the TCR, acting as a
clamp between both molecules
• The type of binding to TCR is not very specific as in ordinary activation a T cells and,
consenquently very large numbers of Th cells can be activated by one kind of
suprantigen.
• The result is release of huge amounts of cytokines, which not beneficial to the host and
even causes systemic toxicity.
• There is supprson of the normal acquired immune response and no memory cells are
produced
• Superantigens are produced by any different bacteria and viruses and are afctive at very
low concentratins
• Examples of superantigens are staphylococcal enerotoxins and toxic shock syndrome-
toxin.
• Overproduction of cytokines in response to these toxins accounts for many of their toxic
effects on the body

c) Functions of effector CD4 T cells (helper Tcells)

(or mechanismof activation of macrophages by Th1)

The main function of of effector CD4 T cells is to secrete cytokines

Classified to Th1 or Th2 cells according to cytokines they secrete

1.Th1 cells

Produced cytokines which predominantly help macrophages and promote inflammation.

Mechanism of activation of macrophages by Th1 cells

• An infected macrophage displays a peptide MHC 2 complex to an effector T cell

• If the TH1 cell is specific for that peptide MHC2complex, it is induced to secrete
cytokines,most important of which is interferon-γ (IFN- γ).This causes activation of the
macrophages,making more capable of killing the bacteria it harbous:
- Ther is more efficient fusion between the phagosomes containing the bacteria and
the lysosomes containing the acterial enzmes

-There is also increased producton of oygen radicals, nitric oxide and antibacterial
enzymes bymacriphage.

Activation of Th1 cells and consequent macrophage activation can sometimes cause significant
tissue damage. However, its absence can lead to serious consequences of disseminated
infection. This is typically seen in AIDS patients with microbacterial infection, since the number of
Th cells in this patients is very low.

2.Th2 cells

Produced cytokines which predominantly help B cells and promote humoral immunity.

d)Functions of effector CD8 Tcels (cytotoxic T cells)

(or mechanism of killing by Tc cells)

The main function of effector CD8 T cells is to eliminate abnormal cells. Such as virus-infected
cells or tumor cells, which could be dangerous to the body as a whole.

• The target cell,such as avirus-infected cell,displays on its surface viral peptide in

conjugation in an MHC 1 molecule.
• An effector T cell with TCR specific for that peptide MHC2 complex recognizes it and
delivers a lethal hit, leading to death of target cell.
• After inducing death of target cell, the Tc cell detaches and search for other target cells to
kill.

Mechanism of killing by Tc cells

Two mechanisms:

1.Induction of apoptosis
 The T cell induces the target cell to undergo suicide. This process of suicide is called

Apoptosis. It may occur one or two mechanisms

• release of cytoplasmic granules. The cytotoxic cell releases two kinds of graules
Called perforins and granzymes. The perforins create perforations( holes) in the

Cytoplasmic membrane of the target cells. Through those holes granzymes enter

The target cell and cause activation of enzymes naturally present in it, leading to

degradation of cells’s DNA.

• Interaction of cell surface molecule. A receptor called FAS, pesent normally on many cells,
binds to a molecule called FAS-ligand, present on cytotoxic cells, giving a signal for
apoptosis in the target cell.

Apoptosis is a clean death in which the cell destroys itself from within, shrinking and degrading
itself until little is left. The enzymes which are activated to destroy cellular DNA can also degrade
viral DNA, thus preventing release of viral particles to infect other cells.

2.Osmotic lysis

The pores produced in the target cell membrane by the perforins may allow entrance of fluid into
cell, leading to death of the target cell by osmotic lysis.

Death apoptosis is faster than osmotic lysis and is probably the main mechanism involved n
killing of target cells by Tc cells.

3) Give reason :

a) superantigens cause systemic toxicity

Superantigens are non specific protein that attaches to the MHC II and TCR without the need of
presentation by APC. Being non-specific, they activate a very large number of Th cells leading to
release of huge amount of cytokines, which cause systemic toxicity.

b)apoptosis is a suitable mechanism of killing virally infected cells

Through apoptosis, the cell dies by destroying itself from within, shrinking and degrading itself.
The enzymes which are activated to destroy cellular DNA can also degrade viral DNA, thus
preventing release of viral particles to infect other cells.

4) Explain :

a) significance of 2nd signal of activation of naïve T-cells

Absence of the 2nd signal of activation will cause T-cells to become non-responsive, a state called
T-cell anergy.

b) the lifestyle of the pathogen decides the type of immune response

if it is extracellular pathogen, such as bacteria and its toxin, it will stimulate humoral immune
response. Thus the elimination occurs through antibodies present in plasma and extracellular
fluid.
But if it is intracellular such as viruses and some bacteria, it will stimulate cell-mediated immune
response (antibodies are not accessible to the pathogens). The destruction of these pathogens is
the function of T lymphocytes.

c) the significance of the present of MHC I on all nucleated cells of the body
any cell on the body is liable to be infected by a virus, so all cell must possess MHC I molecules
in order to be able to show viral peptides to the Tc cells so that it can kill that infected cell.
Otherwise it would escape killing. The presence of MHC I molecules on all body cells is necessary
so that these cell can be targets for the Tc cells if the need arise.

d) the significance of present of MHC II on only a few cells of the immune system
presentation of antigen to Th cells result in production in cytokines. These cause activation of
many cells of the immune system,in many consequences.there is no need for the present of MHC
II on all cells of the body. In fact this would lead to over stimulation of Th cells and over
production of cytokines, which would be dangerous to body.

e)CD8 T cells are MHC I restricted

CD8 T cells recognize peptides bound to MHC I molecule only

f) CD4 T cells are MHC II restricted

CD4 T cells recognize peptides bound to MHC II molecule only