Professional Documents
Culture Documents
1: Intro
The innate immune system includes anatomical barriers against infection both physical
and chemical as well as cellular responses.
o Anatomical Barriers:
The main physical barriers are the epithelial layers of the skin and the
epithelial layers of the mucosal and glandular tissue surfaces. Epithelial
barriers prevent infection by blocking pathogens from entering the body.
The main chemical barriers at these surfaces are specialized soluble
substances that possess antimicrobial activity AND acid pH.
o Cellular Responses:
The cellular innate immune response occurs after an infectious agent
overcomes the initially epithelial barriers.
This response is rapid.
It is triggered by cell surface or intracellular receptors that recognize
conserved molecular components of pathogens.
What does it constitute? Macrophages and neutrophils (white blood cells) are
activated to engulf and destroy microbes via phagocytosis. Other receptors
induce production of proteins and other substances that have various effects
such are direct antimicrobial activity or recruitment of fluid, cells and
molecules.
The influx of fluid, cells, and molecules causes swelling inflammation.
REFER TO FIGURE 5-1
Skin has two layers: thin outer layer (epidermis) and thick inner layer (dermis)
Epidermis:
o Contains tiers of epithelial cells
o Consists of mostly dead cells filled with a waterproofing protein called keratin.
Dermis:
o Composed of connective tissues and contains blood vessels, hair follicles, sebaceous
glands, sweat glands, and scattered myeloid leukocytes such as dendritic cells,
macrophages, and mast cells.
Instead of skin, the respiratory, gastrointestinal, and urogenital tracts, as well as the ducts of
the salivary, lacrimal, and mammary glands are lined by strong barrier layers of epithelial
cells stitched together by tight junctions that prevent pathogens from squeezing between
them to enter the body.
Other Barriers
Secretions of secretory tissue (mucus, urine, saliva, tears, and milk) wash away potential
invaders and also contain antibacterial and antiviral substances.
Mucus from mucosal epithelial layers entraps foreign microorganisms.
Cilia in the respiratory tract are hairlike protrusions of the cell membrane which cover
epithelial cells. The movement of cilia propers mucus-entrapped microorganisms from the
respiratory tract.
The flow of urine sweeps bacteria from the urinary tract.
Saliva has antimicrobial compounds that attack microbes the moment we ingest food.
The vagina has vaginal secretions with an acidic pH.
Some organisms evolved to evade these barriers. For instance, influenza virus has a surface
molecule that enables it to attach firmly to cells in mucous membranes of the respiratory
tract, preventing the virus from being swept out by the ciliated epithelial cells.
Epithelial cells secrete a broad spectrum of proteins and peptides that provide protection
against pathogens.
Antimicrobial Proteins
Among the antimicrobial proteins produced by the skin and other epithelia in humans (Table
5-2), several are enzymes and binding proteins that kill or inhibit the growth of bacterial and
fungal cells.
o Lysozyme is an enzyme found in saliva, tears, and fluids of the respiratory tract that
cleaves the peptidoglycan components of bacterial cells walls.
o Lactoferrin and calprotectin are two proteins that bind and sequester metal ions
needed by bacteria and fungi, limiting their growth.
o Psoriasin is an antimicrobial protein that kills E.coli on the skin. However it cannot
kill S.auerus. This highlights the fact that antimicrobial proteins show some
specificity toward particular pathogens.
Antimicrobial Peptides
These are also antimicrobial components secreted by skin and other epithelial layers.
Antimicrobial peptides generally are cysteine-rich, cationic, and amphipathic (containing
both hydrophilic and hydrophobic regions).
Because of their positive charge and amphipathic nature, they interact with acidic
phospholipids in lipid bilayers, disrupting the membranes of bacteria, fungi, parasites, and
viruses.
They then can enter the microbes where they have other toxic effects, such as inhibiting the
synthesis of DNA, RNA, or proteins, and activating antimicrobial enzymes, resulting in
death.
Surfacants
The epithelium of the respiratory tract secretes a variety of lubricating lipids and proteins
called surfacants.
There are two: SP-A and SP-D. They are members of a class of microbe-binding proteins
called collectins.
SP-A and SP-D bind differentially to sets of carbohydrate, lipid, and protein components
of microbial surfaces and help to prevent infection by blocking and modifying surface
components and promoting pathogen clearance.
Now that weve covered the physical and chemical anatomical barriers, lets discuss the cellular
response that results when anatomic barriers fail.
1.3: Phagocytosis
A. Overview
Once pathogens penetrate through the epithelial barrier layers into the tissue spaces of the
body, an array of cellular membrane receptors and soluble proteins that recognize
microbial components play the essential roles of detecting the pathogen and triggering
effective defenses against it.
The next line of defense after the anatomic barriers are the phagocytic cells. Phagocytosis is
the cellular uptake (eating) of particulate materials such as bacteria. Phagocytes are nonspecific.
o In the tissues, the phagocytic cells are macrophages, neutrophils, and dendritic cells.
o In the blood, the phagocytic cells are the monocytes.
Phagocytes have a variety of surface receptors. We can organize them into two categories (1)
Pattern recognition receptors, OR (2) Opsonin receptors.
REFER TO TABLE 5-3
REFER TO FIGURE 5-6
Opsonin Receptors
o Instead of directly activating phagocytosis (the receptor on the phagocyte (the PRR)
directly binds to the PAMPs on the microbe and results in phagocytosis), we can
activate phagocytosis indirectly.
o There are soluble proteins that can bind to microbial surface and can be easily
recognized by phagocytes. These soluble phagocytosis-enhancing proteins (called
opsonins) will bind to carbohydrate structures, lipopolysaccharides, and viral proteins
on the microbe.
o Once bound to microbe surfaces, opsonins are recognized by membrane opsonin
receptors on phagocytes, thereby activating phagocytosis.
Opsonins
o A variety of soluble proteins function as opsonins; many play other roles as well in
innate immunity. Produced elsewhere and found floating around.
o Lets look at an example: SP-A and SP-D.
We said earlier that SP-A and SP-D are anatomic barriers, specifically,
surfacants that lubricate.
They are ALSO found in the blood in addition to the mucosal secretions
throughout the body.
In the blood, they can function as opsonins.
After binding to microbes, they are recognized by the CD91 opsonin receptor
on alveolar and other phagocyte (in this case macrophages) populations.
Binding activates phagocytosis.
This function of SP-A and SP-D contributes to the clearance of the fungal
respiratory pathogen Pneumocystis carinii, a major cause of pneumonia.
o Lets look at another example: Mannose-binding lectin (MBL)
This is a third collectin with opsonizing activity.
It is found in the blood and respiratory fluids.
o Lets look at another example: L-fociolin
A member of the ficolin family that is related to MBL and other collectins.
It is found in the blood where it binds to acetylated sugars on microbes,
including some streptococcal bacteria.
o Lets look at another example: C1q
o MBL (and other collectins), ficolins, and C1q share structural features, including
similar polymeric structures with collagen-like shafts, but have recognition regions
with different binding specificities
Because of their structural similarities, they are all received by the same
receptor on a phagocyte (the CD91 opsonin receptor):
INSERT FIGURE 5-7
o Another opsonin, C-reactive protein (CRP), regnozies phosphorylcholine and
carbohydrates on bacteria, fungi, and parasites, and is then bound by Fc receptors
(FcRs) for IgG found on most phagocytes.
FcRs are also important for the opsonizing activity of IgA antibodies and
some IgG antibody subclasses.
After binding specifically to antigens on microbe surfaces, the Fc regions of
these antibodies can be recognized by specific FcRs, triggering phagocytosis.
o Collectively the ROS and RNS are highly toxic to phagocytosed microbes due to the
alteration of microbial molecules through oxidation, hydroxylation, chlorination,
nitration, and S-nitrosylation, along with formation of sulfonic acids and destruction
of iron-sulfur clusters in proteins.
o For example, oxidation by ROS of cysteine sulfhydryls that are present in the active
site of many enzymes in pathogens results in the inactivation of those enzymes.
Collectively the components of dead/dying cells and damaged tissues that are recognized by
PRRs leading to their clearance are referred to as damage-associated molecular patterns
(DAMPs).
Phagocytosis is the major mode of clearance of cells that have undergone apoptosis as part of
developmental remodeling of tissues, normal cell turnover, or killing of pathogen-infected or
tumor cells by innate or adaptive immune responses.
Apoptic cells attract phagocytes by releasing the lipid mediator lysophosphatidic acid, which
functions as a chemoattractant.
o These dying cells facilitate their own phagocytosis by expressing on their surfaces an
array of molecules not expressed on healthy cells, including phospholipids (such as
phosphatidyl serine and lysophosphatidyl choline), proteins (annexin I), and altered
carbohydrates.
An important additional activity of macrophages in the spleen and those in the liver (Kupffer
cells) is to recognize, phagocytose, and degrade aging and damaged red blood cells.
o As the RBCs age, specific molecules accumulate on their membranes.
o For instance, phosphatidyl serine flips from the inner to the outer leaflet on the
bilayer and is recognized by phosphatidyl serine receptors on the phagocytes.
There are several families of pattern recognition receptors (PRRs) that bind to PAMPs as
well as to some endogenous (self) DAMPs and trigger signal-transduction pathways that turn
on the expression of genes with important functions in innate immunity.
The proteins that are encoded by these genes are:
o Antimicrobial peptides and interferons
o Chemokines and cytokines that recruit and activate other cells, enzymes such as
iNOS that generate antimicrobial molecules
o Proinflammatory mediators (i.e., components that promote inflammation.
A. Cellular Pattern Recognition Receptors Activated Responses to Microbes and Cell Damage
PRRs are expressed either on the surface of the plasma membrane OR inside the cell
itself (in the endosomes/lysosomes or in the cytosol).
By having so many different PRRs in two locaations it ensure that the cell can recognize
PAMPs on both extracellular and intracellular pathogens.
Also, its not just PAMPs that are recognized. You can have DAMPs that are released by
damaged cells or tissue which can be recognized.
There are multiple cells that express these intracellular and extracellular PRRs:
o The myeloid white blood cells:
Monocytes
Macrophages
Neutrophils
Eosinophils
Mast Cells
Basophils
Dendritic Cells
o And Subsets of the tree types of lymphocytes:
B cells
T cells
NK cells
o As well as other cell types, especially those exposed to infectious agents:
Skin cells
Mucosal cells
Glandular epithelial cells
Vascular endothelial cells that line blood vessels
Fibroblasts and stromal support cells in various tissues.
We will now look at one of four main families of mammalian PRRs. We wont look at the
signaling pathways that these PRRs activate and ultimate cause a protective response.
Mice homozygous for a mutant form of a gene called lps were resistant to the
harmful responses induced by lipopolysaccharide (aka endotoxin), a major
component of the cell walls of Gram-negative bacteria.
In humans, a buildup of endotoxin from severe bacterial infection can induce
too strong of an innate immune response, causing septic shock, a lifethreatening condition in which vital organs such as the brain, heart, kidney,
and liver may fail.
Beutler found that the defective mouse lps gene encodaed a mutant form of
one TLR, TLR4, which differed from the normal formb y a single amino acid
so that it was no longer activated by LPS.
Thus, he showed that TLR4 is the cellular innate pattern recognition receptor
that recognizes LPS.
Antimicrobial peptides
Type I Interferons
Cytokines
Chemokines
Enzymes: iNOS and COX2
When an innate immune response occurs it can induce a complex cascade of events known as
the inflammatory response.
Inflammation can be acute
o This means there are short-term effects contributing to combating infection, followed
by healing.
Inflammation can also be chronic
o This means that it is long term and not resolved.
o It can contribute to conditions such as arthritis, inflammatory bowel disease,
cardiovascular disease, type 2 diabetes.
What does the inflammatory response consists of?
o For one, there is an increase in vascular diameter (vasodilation), which results in a
rise of blood volume in the area of injury/infection.
o Higher blood volume heats the tissue and causes it to redden.
o Vascular permeability also increase, leading to leakage of fluid from the blood
vessels, resulting in accumulation of fluid (edema) that swells the tissue.
o Within a few hours, leukocytes also enter the tissue from the local blood vessels.
These leukocytes area activated to phagocytose bacteria and debris and to amplify the
response by producing additional mediators.
How does the inflammatory response end?
o Resolution of acute inflammatory response includes the clearance of invading
pathogens, dead cells, and damaged tissue; the activation of the systemic acute phase
response and additional physiological responses, including the initiation of wound
healing; and the induction of adaptive immune responses.
o If the infection or tissue damage is not resolved, it can lead to a chronic inflammatory
state that can cause more local tissue damage and potentially have systemic
consequences for the affected individual.
When there is local infection, tissue damage, or exposure to some harmful substances,
sentinel cells residing in the epithelial layer (basically, macrophages, mast cells, and dendritic
cells) are activated by PAMPs, DAMPs, crystals, and so on to starts phagocytosing the
offending invaders.
o The cells are also activated to release innate immunity mediators that trigger a series
of processes that collectively constitute the inflammatory response.
The recruitment of various leukocyte populations to the site of infection or damage is a
critical early component of inflammatory responses.
o PRR signaling activates resident macrophages, dendritic cells, and mast cells to
release the initial components of cellular innate immune responses.
o These components act on the vascular endothelial cells of local blood vessels,
increasing the vascular permeability and the expression of cell adhesion molecules
(CAMs) and chemokines. This epithelium is said to be inflamed or activated.
o Cells flowing through local capillaries are induced by chemoattractants and adhesion
molecule interaction to adhere to vascular endothelial cells in the inflamed region and
pass through walls of capillaries and into the tissue spaces, a process called
extravasation.
o Neutrophils are the first to be recruited to a site of infection where they enhance local
innate responses, followed by monocytes that differentiate into macrophages that
participate in pathogen clearance and help initiate wound healing.