MICROBIAL VIRULENCE

AND HOST DEFENSES

BY

D R N WA F I A I . N
(Consultant Clinical Microbiologist)
 Learning Objectives

• Koch’s postulate
• Differentiate between frank pathogen, opportunistic
pathogens and colonizers
• Virulence factors and functions
• Host defenses
• Types of immunity
• Mechanism of phagocytosis
Introduction
 Koch’s postulate

1) The microorganism must be found in diseased but not

healthy individual.

2) The microorganism must be cultured from the diseased

individual.

3) Inoculation of a healthy individual with the cultured

microorganism must recapitulated the disease.

4) The microorganism must be re-isolated from the

inoculated, diseased individual and matched to the
original microorganism.
 Definitions

• Pathogenic microorganism
- Capable of causing disease.
- Infection and disease do not always occur together.
- Some microorganism are unequivocally pathogenic
whereas some others are harmless.

• Clinical Infection
-An infection with obvious observable or detectable symptoms

• Subclinical or asymptomatic Infection

- An infection with few or no obvious symptoms
• Frank pathogens
- Causes diseases in immuno-competent and immuno-
compromised
- when isolated from a patient are considered to be a probable
agent of a disease
- Salmonella typhi
- Hepatitis A virus
- Neisseria gonorrhea

• Opportunistic pathogens
• Micro organisms that are commonly found in the host’s
environment.
• Isolated from patients whose defense mechanisms have
been compromised
 Wha
Micr is t
obia
Viru l
lence
 Definition

• Microbial virulence is a measure of the severity of the

disease caused by a microorganism
OR
• Degree of pathogenicity
 Definition……….
Pathogenicity
• Ability of the microbe to cause disease

• Dependent on ability to
• Enter the host
• Adapt and multiply in the host
• Exit from the host
• Transmit to new host

• Colonization- presence of bacteria on a body surface

without causing disease in the person.
 Virulence of organisms

• Virulence helps the organisms to

- Evade the host defenses
- Invade the tissue
- Cause disease

• Virulence involves
- Invasiveness- is the ability to enter host tissue
multiply and spread
- Toxigenicity
- Plasmid-medicated specific expression
 Adherence
• Almost all pathogens have a
means to attach to host tissue

• Prevents them from being

flushed away by mucous
secretions
- Less susceptible to the
effects of enzymes and
secretory IgA

• Examples of receptors
Adhesins
- Ligands
- Fimbriae
 Capsules

• Layer of material usually polysaccharide attached to the cell

wall or extracellular layer which lies outside the cell wall

• Function
- Capsule resist phagocytosis by WBCs
- Prevent the cell from desiccation and drying
- It protects them from mechanical and physical injury
- Capsule helps in attachment on the surface.
- Capsule prevent attachment of bacteriophage on cell
surface
 Examples of Capsulated organisms

• - Streptococcus pneumoniae/ Streptococcus mutans

- Klebsiella spp
- Haemophilus influenzae
- Pseudomonas aeruginosa
- Neisseria meninigitidis
- Cryptococcus neoformans
- Bacillus anthracis

Some killers have pretty nice capsules

 Flagella
• Whip-like appendages used primarily for locomotion
Function
- Motility
- Adhesion
• Invasion or secretion of virulence factors.
 Cell wall components

• M protein
- Found on cell surface and fimbriae of Streptococcus
pyogenes.
- Mediates attachment and helps resist phagocytosis.

• Waxes (Mycolic Acid)

- In cell wall of Mycobacterium tuberculosis
- Resistant to killing (phagocytes or antibiotics)
- Formation of biofilm
 Siderophores

They are low molecular weight compounds that chelate

iron
• Bacteria require iron for their metabolism
- Bacteria compete for Fe3+ using siderophores

• The survival of invading organism therefore depends

on its ability to scavenge iron from its environment
and deposits it within host.
• Example
• Enterochelin
 Leucocidin
• Increase the permeability of leukocytes to cations,
which can lead to rupture and prevents phagocytosis
• It ruptures and releases lysosome which contain
powerful hydrolytic enzymes causing more tissue
damage
Examples
Staphylococcus aureus
 Haemolysin
• Heamolysin causes lysis of
RBC
Alpha (α) Heamolysis
- Incomplete lysis eg
Streptococci pneumoniae
 Beta (β) Heamolysis
- Complete lysis of RBC’s
Eg Streptococci pyogenes
Gamma heamolysis
No lysis
Eg Enterococcus feacalis
• Coagulase-
• Bound coagulase (clumping factor) is bound to the
bacterial cell wall and reacts directly to activate
fibrinogen to fibrin.
- Detection in slide test
• Free coagulase involves the activation of plasma
coagulase-reacting factor. This complex in turn reacts
with fibrinogen to produce the fibrin clot.
The fibrin formed will protect bacteria from phagocytosis
eg Staphylococcus aureus
 PROTEASES

 Degradation of IgA
 Ig for protection of mucosal surfaces
- S. pyogenes

Catalase
• Breaks down hydrogen peroxide
• into water and oxygen
• Hyaluronidase- “Spreading Factor”
- Breaks down Hyaluronic acid
- Streptococci pneumonae, Staphylococci spp,
Clostridia spp

• Lecithinase
- Destroys lecithin - component of plasma membrane.
- Allowing pathogen to spread
Clostridium perfringens

• Kinases (plasminogen activating enzyme)

- Streptokinase - Streptococci pyogenes
- Staphylokinase – Staphylococci spp
 Biofilm
• Biofilm formation is a process whereby microorganisms
irreversibly attach to and grow on a surface and produce
extracellular polymer matrix.
 Toxigenicity

2 types of toxins
- Exotoxin
- Endotoxin
 Exotoxin

• A poisonous substance secreted by bacteria.

• Secreted by Gram positive and negative bacteria.
• Has two fragments
- Fragment A- Toxic unit
- Fragment B- the carrier binds to host cell receptor
and enables fragment A to enter the cell.
• Converted into toxoids - immunogens for prophylaxis
 Types of exotoxins
Cytotoxins
- Kill cells e.g. Diphtheria toxin

 Neurotoxins
- Interfere with normal nerve impulses.e.g.
Botulinum Toxin

Enterotoxins
• Affect cells lining the G.I. Tract. e.g. Cholera toxin
- Mostly causes diarrhea eg Staphlococcus spp
 Endotoxins
• Lipopolysaccharides - integral component of cell
wall of Gram negative bacteria

• Heat stable

• Most of their toxic effects are due to lipid A

component of the lipopolysaccharide.

• Physiologic effects of endotoxins include

- fever - shock - hypotension
- thrombosis - multiple organ failure.
Endotoxins

Figure 15.6
Difference between exotoxin and endotoxin

Endotoxin
• Protein polypeptide.
Exotoxin • Lipopolysaccharides complexes.
• Secreted by living cells into • Present in cell wall and released
medium from cytoplasm. only on disruption of cell.
• Heat-labile. • Heat-stable
• Not pyrogenic • Progenic
• Can be converted into toxoid • Cannot be converted into toxoid
by heat or chemical
• Poorly antigenic.
• Highly antigenic
• Produced mainly by Gram - org.
• Produced Gram + and – org.
• Non-specific in action
• Specific for particular tissue
eg. Tetanus toxin for N.S • Located on the chromosomal
gene
• Located on etrachromosme
Plasmid-mediated phenotypic
expression
• Plasmids are extra-chromosomal genetic material
• DNA fragments carrying genes for extra resistance
known as R-factors.
• Easily transferrable by conjugation
• Codes for pathogenic mechanism
-Colonization factors eg biofilm
- Enterotoxin production
- Siderophore synthesis.
- Antimicrobial resistance
HOST DEFENSES
• Resistance
- Ability to ward off disease
- Varies among organisms and individuals within the
same species
• Immunity
- Mechanisms used by the body as protection against
microbes and other foreign agents
Nonspecific immunity (innate, natural, inborn)
• Defenses against any pathogen
Specific immunity ( adaptive)
• Resistance to a specific pathogen
INNATE IMMUNITY
(NON SPECIFIC)
 Introduction
• Most of our encounters with pathogens and toxins are
taken care of by the innate immune system
• Refers to the fact that the responses do not require
time to develop
• Act within minutes of the host being infected
• Attack pathogens without specificity
 Factors influencing effectiveness of innate immunity

• Species
- mammals may contract anthrax but, birds cannot
• Racial/genetic basis
- Black are more to susceptible to TB
• Hormone-related resistance
• Nutrition
• Sex
• Age
• Stress
 FIRST LINE OF DEFENSE
Physical chemical barrier
Skin
• Tightly associated epithelia cells covered by highly cross-
linked keratin layer
• Antimicrobial properties- broad spectrum defensive chemicals
- Proteases
- Beta defensin
- Cathelicidins
• Desiccating effect
• Acidity- pH 5-6
• Presence of normal flora
• Desquamation of skin
SKIN
 Mucous membrane

• Line body cavities that open to the outside (GIT,

genitourinary and RES tracts)
- Mucus is produced by the mucosal cells
- Antimicrobial substance such as lysozymes
- Sequester iron- Lactoferrin
• - Mucosal cells are rapidly dividing and
desquamation of cells along with attached bacteria
• Immunoglobulin- IgG and secretory IgA
 Respiratory tract

• Hair- traps the organisms

• Muco-cilliary action transport invading organisms
away from the lungs
• Cough reflexes- aid in expulsion of invading
organisms
• Mucus secretions - phagocytes and antibacterial
enzymes
• Surfactant- Binds and opsonize the organism
• Alveolar macrophages and histiocytes
 GIT

• Saliva - lysozyme
• Stomach acids
• Mucus secretion
• Peristalsis
• Bile (alkaline) in small intestine
• Normal bowel microbiota
• Defecation
 Genitourinary tract

• Urination
• Length of the male urethra
• Vaginal and seminal fluid secretion
• Low pH of vagina
 OTHERS……

• Eyes
- Blinking of eyelids
- Tears containing lysozymes and the also dilutes the
invading agents
• Outer ear canal
• Wax contains antibacterial components
 SECOND LINE OF DEFENSE
• Inflammation – a series of events that removes or
contain the offending agent and repair the damage

• Chemotaxis – movement of cells toward a chemical

influence (chemokines or chemotatic agents)

• Phagocytosis – process in which cell ingest foreign

particulate matter like microbes
- WBC
 Second line of defense…….
• Acute phase proteins
• set of plasma proteins whose level increases during
infection to enhance host defense mechanisms
• Complement proteins, coagulating factors, C-reactive
proteins

• Cytokines
• small secreted proteins produced by cells
• Communication between different defense systems
• interleukins, interferon
 FEVER

• Pyrogens are substances that stimulate fever

• Exogenous- bacterial endotoxin
• Endogenous - interleukins (IL-1)

• Functions
- Stimulate WBC to deploy & destroy microbes
- increase in immunological response (e.g.
proliferation and activation of lymphocytes)
-Slow down growth of or kill pathogens
ACQUIRED IMMUNITY
 Introduction
• Ability of the body to distinguish between self and
non-self
• Specific and inducible (response after exposure to a
foreign substance)
• Humoral (antibody-mediated) response – antibody
attack free microbes in the body
• Cell-mediated response – specialized cells attack
infected or abnormal (cancer) cells

• Lymphocytes and macrophages are important to the

development of acquired immunity.

• If over-stimulated, can cause harm to host

(hypersensitivity)