MOLECULAR MECHNISMS OF MICROBIAL PATHOGENESIS
Processes of Infection and Disease
1. Microbial encounter and entry
2. Microbial growth after entry
3. Avoid innate host defenses
4. Tissue invasion and tropism
5. Tissue damage
6. Transmission to new host
VIRULENCE – measure of the capacity of the organism to
cause a disease (pathogenic factors)
COLONIZATION – presence of the pathogenic microbe in a
host
INFECTION – attachment and growth of pathogens and
avoidance of host defenses
DISEASE – often, BUT NOT ALWAYS, the result of the action
of elaborated toxins and toxic metabolites
MICROBIAL ENTRY AND ADHERENCE
Entry Sites
 Microorganisms can gain access to a host virtually by
any means. But the most common site of entry are
MUCOSAL SURFACES and SKIN.
 Typical ROUTES OF ENTRY include ingestion, inhalation
and sexual contact.
 A few pathogens (Schistosoma) can penetrate unbroken
skin.
 Routes of entry are also dependent on host factors:
INGESTION: Requires the organism to survive low gastric
pH and high intestinal bile content.
INHALATION: Survive in small moist droplets
SEXUAL CONTACT: Survive best in warm moist environment
of the urogenital mucosa and have restricted host ranges
(N. gonorrhoeae, T. pallidum, HIV)
 Some organisms can survive the salivary or alimentary
tract of arthropods before entering the definite host.
(Plasmodia, Trypanosomes, Leishmania)
 Most microbes that land directly on skin are DESTINED
TO DIE, because of:
• Fatty acid content
• Low pH (Acid)
• Antimicrobial factors on the skin (IgA)
 Damaged skin (particularly necrotic) can invite
pathogenic organisms to settle and elaborate toxic
products (Tetanus in burn victims)
 Rabies: Animal bite (saliva)
Microbial Adherence
 Microbes must anchor themselves to tissue of tissue
factor when they need to survive.
 Specific ligands are elaborated by the microbe that
bind to host receptors. Multiple adhesins may be made to
attach to multiple host receptors.
 Some microbes can absorb host proteins on their
surface and use this to attach.
A. Viral Adhesins
 All viruses must adhere to host cell, enter them and
replicate.
 Viral coat proteins serve as the ligands for viral entry.
B. Bacterial Adhesins
 Pili or fimbriae is mainly used by GRAM-NEGATIVE
bacteria. They are hairlike structures that may be polar
(attached to one end) or scattered evenly over the
surface.
 E. coli has a mannose binding type 1 pili which is
inhibited by D-mannose found in the host.
 Other strains have a P pilus adhesin that bind to human
P blood group.
 Flagella are long appendages attached to one or both
ends of the bacteria. They may also be distributed along
the surface of the bacteria (peritrichous). They are
mainly used for locomotion (Spirochetes) but can also be
needed for attachment (P. aeruginosa)
 Staphylococcal or streptococcal proteins that bind to
extracellular matrix protein like fibronectin, laminin or
collagen.
 Surface polysaccharide poly-N-acetyl glucosamine
appears to be responsible for binding of pathogens on
prosthetic devices.
C. Fungal Adhesins
 Binds to extracellular matrix proteins on the host
(fibronectin, laminin or collagen).
 C. albicans INT1 gene is responsible for binding to host,
but its elimination will NOT prevent binding to epithelial
surfaces. Thus there appears to be another adhesin that
mediates binding to epithelial cells.
 Blastomyces dermatitidis appears to elaborate WI-1
120Kd surface protein that binds to CD 11b/CD18 integrins
on the host.
D. Eukaryotic (Parasitic) Adhesins
 Complicated surface glycoproteins some of which are
lectins
Host Receptors
 These are found on target cells and within the mucus
layer that invests the target cell.
 Selective loss in host receptors may confide a natural
form of resistance to infection:
Ex. 70% of West Africans have no Fy antigen on the blood
conferring resistance to P. vivax.
Ex. Patients with cystic fibrosis have mutated CFTR which
is needed by S. typhi for adhesion.
 Overlap of certain virus with regards to their receptors.
One receptor may not be specific for one antigen.
MICROBIAL GROWTH AFTER ENTRY
 Prior to manifestation of a disease, the pathogen has to
replicate.
 For viruses, they disrupt nucleic acid synthesis
 For bacteria they must acquire specific nutrients or
synthesize them from precursors found in the host cell.
 Certain pathogens have predilection to certain sites
because the nutrients they need may be found at these
locations:
Influenza – respiratory tract
Gonorrhea – urogenital tract
Shigellosis – GIT
 Temperature plays also a role in the growth of some
pathogens:
Rhinovirus – 33 0C/cooler areas of the nasal tissues.
Leprosy – cooler body sites
Dermatophytes – cooler, exterior and keratinized areas
 BIOFILMS – growth of the microorganisms in
multicellular masses. If the microbe appears individually,
they are termed PLANKTONIC CELLS.
 Growth in biofilms is advantageous for the microbe:
• Altered metabolism
• Produce extracellular virulence factors
• Decreased susceptibility to biocides, antibiotics,
host defenses and cells.
MOLECULAR MECHNISMS OF MICROBIAL PATHOGENESIS
AVOIDANCE OF INNATE HOST DEFENSES
 The skin is acidic and is bathed in fatty acids toxic to
many microbes.
 Thick mucus secretions can trap microorganisms and
aid in their elimination.
 Mucocilliary apparatus, coughing and micturition are
mechanical clearance mechanisms.
 Lysozymes, antimicrobial peptides and interferons may
be found in saliva and tears.
 Gastric acidity kills ingested pathogens
 Certain areas contain resident commensals that
compete with the pathogen for growth:
• Nasophraynx
• Vaginal tract
• GIT
 If the pathogen is able to survive these areas, they still
have to face the host endocytic, phagocytic and
inflammatory responses.
 Host genetic factors dictate the degree to which a
pathogen can survive and grow.
Encounters with Epithelial Cells
 Bacteria seem to enter the epithelial cells by
specialized surface receptors.
 This mechanism is needed as a means of dissemination
of the pathogen to nearby tissues or deeper tissues.
 It may also serve as a means to escape phagocyte
encounter.
 Less virulent strains are more capable of entering
epithelial cells compared to more virulent strains.
Variants lacking capsule are more capable of penetrating
epithelial cells compared to wild-type strains.
 It may also be viewed as a form of host defense,
because after epithelial invasion, follows epithelial
shedding and initiation of subclinical inflammatory
response.
Encounters with Phagocytes
 Phagocytosis seem to be the major innate host defense
that limits growth and spread of pathogens.
 The cells appear rapidly in sites of infection in
conjunction to initiation of inflammation.
 Collectins, soluble defense collagens or pattern
recognition molecules found in the blood, lungs and in
other tissues bind to CHO on microbial surface to promote
phagocytic clearance.
 PMN’s are the principal phagocytes, while eosinophils
appear to ingest protozoans and multicellular parasites.
 A SUCCESSFUL PATHOGEN is termed as any pathogen
that is able to escape pathogen ingestion.
 Antiphagocytic mechanisms can be employed by some
pathogens:
• C. neoformans elaborate antiphagocytic capsule
made of low-molecular weight CHON antigen.
 Best studied system of microbial pathogenesis is the
interaction of LPS and GPI-anchored membrane protein
CD14 found on the surface of professional phagocytes.
 Peptidoglycan and lipoteichoic acid from Gram positive
bacteria and cell surface products found on
Mycobacterium can also bind to CD14.
 Toll-like receptors are responsible for initiation of
cellular activation that leads to nuclear transolocation of
transcription factor NF-κB.
 NF-κB is the master switch for production of important
inflammatory cytokines (TNF-α, IL-1)
 Bacterial flagella can also induce inflammation by
binding with TLR5 receptor.
 MyD88 is a generalized adaptor protein for the binding
of all cytoplasmic domains of all known TRL’s.
 Additional means by which pathogens can avoid
phagocytosis is the production of factors toxic to the
phagocytes:
• Hemolysin
Ex. Streptolysin O (Streptococci) binds to cholesterol
in the phagocyte membrane to initiate degranulation
reaction.
• Leukocidins
 E. hystolitica can likewise cause phagocyte destruction
by elaboration of protozoal phospolipase A and pore-
forming peptides.
 M. tuberculosis, S. typhi, and T. gondii can survive
INSIDE the phagocyte by preventing fusion of the
phagocytic vacuoles or phagosome formation. The
phagosome contains the pathogen with lysosomal granules
containing lytic material.
 L. monocytogenes can escape out of the phagocyte
cytoplasm and infect nearby cells.
TISSUE INVASION
 Viruses can invade tissues through the nerves (as in
Rabies), plasma (as in picornaviruses) or by migratory
blood cells (polio, EBV, etc.)
 Specific viral genes determine the mode of spread.
 Bacteria invade the depper mucosal layers by
• intracellular uptake by epithelial cells
• Traversal of epithelial cell junctions
• Penetration through denuded epithelium
 Among virulent Shigella and E.coli, outer membrane
proteins are needed for epithelial cell uptake and
multiplication.
 Staphylococci and Streptococci elaborate certain
substances (hyaluronidase, lipases, nucleases and
hemolysins) which are helpful in breaking matrix and
cellular structures to facilitate their invasion.
 Y. enterocolitica use invasin proteins.
 Brucella is carried from the mucosal site to a distant
site by a PMN that failed to neutralize it.
 Fungal pathogens can take advantage of the immune
status of the host and is able to spread hematogenously.
TISSUE TROPISM
 Known as the propensity of certain microbes to cause
disease by infecting specific tissues.
 Principles are more understood among viruses
compared to bacteria.
 Specific receptor-ligand interactions underlie the
ability of certain pathogens to invade cells, cause a
disease. But mere presence of receptors is not enough for
tissue tropism.
 N. gonorrhea vs. N. meningiditis (with capsular
polysaccharide)
TISSUE DAMAGE AND DISEASE
 Viruses in general cause diseases thru their cytopathic
effect and inhibition of host defense.
MOLECULAR MECHNISMS OF MICROBIAL PATHOGENESIS
 Growth of bacteria, protozoa or fungus in tissues, even  Bacteria avoid phagocytes and other defenses by
without toxin elaboration, may cause tissue compromise manifesting cell surface polysaccharides (capsular or long
 Elaboration of toxins is the best studied mechanism of O-side chain antigens)
pathogenesis among bacteria and fungi.  These molecules inhibit the activation or deposition of
 Host factors, including mediators of inflammation, complement opsonins or limit the access of phagocytic
contribute to disease severity. cells with receptors for complement opsonins.
 Presentation of the capsule as an apparent self-antigen
Viral Diseases through molecular mimicry.
 Immune response can be affected by: Host Response
• Downregulating production of MHC molecules  Inflammation is necessary for interruption and
• Diminish cytotoxic T-cell recognition resolution of the infectious process, but is also responsible
• Producing virus-encoded complement receptor for the SIGNS and SYMPTOMS of the disease.
proteins (that protect virus from action of  Effects of IL-1 and TNF-α
complements) • Fever
• Proteins that interfere with interferon • Muscle proteolysis
• Produce superantigen-like proteins  An inability to kill the pathogen will result to more
 Production of peptide growth factors, that disrupt host disease and progression of inflammation.
cell function, growth, proliferation and differentiation.  Chronic inflammation in any tissue can lead to
 Interfere with functions of neurons: destruction of that particular tissue and loss of organ
• Interfere with levels of NT release w/o function.
necessarily destroying the cells.  Local inflammation = tissue damage
• Induce apoptosis  Systemic inflammation (sepsis) = septic shock
• Inhibit apoptosis so that they can prolong their  If there is intracellular parasitism, there is granuloma
formation in an attempt to wall of the parasite in the
stay and prolong the infection.
fibrotic lesion. The epithelial cells fuse forming
Bacterial Toxins
multinucleated giant cells.
 Diptheria, botulism and tetanus toxins were responsible
 Zwitterionic surface polysaccharides provoke the
for diseases associated with local infections.
formation of abscesses (particularly among anaerobes)
 E. coli, Salmonella, Shigella, V. cholera and
Staphylococcus elaborate ENTEROTOXINS that mediate
TRANSMISSION TO ANOTHER HOST
diarrhea
 Transmissibility may not necessarily be high among
 Staphylococcus, Streptococcus, P. aeruginosa and
some severely infected hosts.
Bordetella produce a variety of toxins:
 Most pathogens exit the host by similar way they have
• TSST-1
entered. (Ex. Aerosols by sneezing and coughing due to
• Erythrogenic toxin respiratory pathogens)
• Exotoxins A, S and U  Ability to produce phenotypic variants that resist
• Pertussis toxin hostile environmental destruction (ex. E. histolytica in
 Some toxins enzymatically catalyze the transfer of the cyst form is not easily destroyed by the environment.
ADP-ribosyl portion of NAD to target CHON and inactivate
them. VINCE
 Staphylococcal enterotoxin, TSST-1 and the
streptococcal pyogenic toxin behave as superantigen
(activate T-cells without the processing of protein toxin by
antigen presenting cells).
Endotoxin
 Lipid A portion of the Gram negative LPS have potent
biologic activities that cause manifestations of Gram
negative sepsis.
 LPS binding to CD14 and signal transduction via TLR’s.
 Effects of the resulting cytokines:
• Potent hypothermic effect
• Increase vascular permeability
• Induce endothelial procoagulant activities
Invasion
 Invading pathogens are found in places that are
normally sterile (ex. Blood, CSF)
 S. pneumonia + production of pneumolysin is partly
responsible for growth in the lung.
 Melanism has been shown to protect the fungal cell
against death caused by phagocyte factors such as NO,
superoxide and hypochlorite.
 Morphogenic variation + Proteases: implicated in fungal
invasion of host tissues.