● Second Line of Defense (Internal)

○ designed to recognize molecules that are unique to the infectious agent

(mannose) - direct
■ Mannose - carbohydrate of the antigen recognized by the immune
system like phagocytes
■ Without recognition, the infectious agent cannot be removed
○ initiates the adaptive immune system
○ The Process of Phagocytosis
■ Importance:
■ destroys foreign materials that enter the body
■ dispose damaged/dying cells
■ remove tissue debris from repairing wounds or growing
tissues
■ remove/supress arising tumors
○ Phagocytes
■ Neutrophil - granulated, nucleus has lobes
■ able to phagocytose but does not remove dying cells
■ Eosinophil - not effective phagocytes
■ has affinity for the acidic dye, eosin
■ the granular contents of eosin are toxic to parasites
■ granular contents are 1) major basic proteins and 2)
eosinophil cationic protein
■ Basophils - has a phagocytic ability but that is not its main function
■ Monocyte - called macrophage once it reaches the tissue
■
■ Mast Cells - important in the initial part of phagocytosis though it is
NOT a phagocyte
■ Dendritic cells
■ function and name is based on location ‒rem reference
"location" to see notes‒
■ function: present antigens to the lymphocytes, phagocytose
○ Process of Phagocytosis
■ Initiation/Diapedesis - when cells (especially phagocytes) in the
bloodstream travels and enters tissue without any break in the
blood vessels
■ Interleukin 1 and Tumor Necrosis Factor targets the
endothelial cells of the blood vessels ⇒ Endothelial cells will
expose adhesion molecules, selectins ⇒blood cells will be able to
grab on to and slow down in the blood stream
■ Integrin - WBC adhesion molecule
■ Chemotaxis - movement towards area where there is
more chemicals
■ Attachment of Phagocytes
■ Pattern-recognition receptors - toll-like receptors, considered
a direct recognition
 ■ Pathogen-associated molecular patterns - found on the
surface of microorganisms
■ Phagocytosis
■ Indirect Recognition - antibodies coats the bacterial cell so
that the phagocytes can recognize it
■ this process also neutralizes whatever is on the
surface of the microorganism
■ Opsonin - prepares the target for the phagocyte to
engulf
■ Opsonization - when bacterial cell is coated by
opsonin, forms pseudopods
■ Respiratory Burst - oxidative process wherein the hexose-
monophosphate shunt will help in the release of energy and
NADP that will help destroy microorganism
■ the process needed to generate enough energy and
oxygen 1) for the cell to undergo phagocytosis and 2)
for it to kill microorganisms
■ Engulfment
■ Release of Lysosomal Contents
■ Granular contents:
■ Myeloperoxidase from primary granules
■ Lysozyme
■ Lactoferrin
■ Elastase
■ Lipase (monocyte)
■ Enxymes from lysosomes
■ NADHP oxidase - converts NADP to NADPH
■ helps in the release of oxygen generated in the
respiratory burst
■ Inducible nitric oxide synthase (iNOS) - converts
arginine to nitric oxide
■ Lysosomal proteases - breaks down protein
■
■ whole process usually takes 10-30 minutes at pH 4.
Neutrophils die after phagocytosis. Macrophages and
dendritic cells process the fragments after the process
■ Neutrophil Extracellular Traps (NETs)
■ Released when neutrophils are destroyed
■ Neutrophil nuclear contents - chromatin components,
histones, and antimicrobial proteins are released and
it helps in making the microorganisms stay in place,
trapping it.
■ microbes are trapped in NETs
■ Assessing Phagocyte Dysfunction
■ used in research and specialized labs
 ■ Ingestion of bacteria - fluorescent dye is used in this test.
Mononuclear cells are isolated from the blood and exposed
to the bacteria, placed in a petri dish. Phagocytosis may be
measured by measuring the fluorescence of bacteria before
and after the addition of mononuclear cells.
■ Does not test if bacteria is killed
■ Bacterial killing - MTT dye is used. This dye precipitates in
the presence of bacteria and can be measured
spectrophotometrically.
■ if the phagocyte is able to kill the bacteria, there
would be less precipitate
■ Phagocyte migration
■ a. Boyden micropore chamber - medium is in liquid
form
■ b. Agarose - medium is agar form
■ c. Rebuck skin window - in vivo test. The skin is
abraded and a slide is placed on the skin for 24
hours. After that, the slide is removed and viewed
under the microscope for phagocytes.
■ Activation of NADPH Oxidase - test for the respiratory burst
■ Nitroblue tetrazolium (NBT) dye - yellow and becomes
blue or black when respiratory burst takes place
■ screening for chronic granulomatous disease
○ Immunologic Surveillance
■ activation of natural killer cells (mostly intracellular)
■ NK cells - has several receptors that identifies target cells called
antibody dependent cell mediated cytotoxicity
■ should be able to identify its target
■ NK cell Receptors
■ FcyR
■ Complement Receptors for C3 and C4
■ Killer activation receptor - recognizes stress molecules on
the surface of the cell
■ Killer inhibitory receptor - measures the level of MHC 1
(Major Histocompatibility Complex) molecule that the cell is
expressing
■ MHC 1 - mobilizes cytotoxic C cells
■ How NK cell works
■
■ It first identifies through the antibodies that coats the antigen,
or complement protein or if its expressing stress molecules
■ granular content - perforin (creates holes into the target cells
to expose the microorganism)
■ water enters through the holes and the cell bursts, allowing
other cells to neutralize the microorganism
■ NK cell Function Test
 ■ virally infected cells with Cr51 + patient NK cells
■ Increased radioactivity = NK cells lysed the target cell
■ No radioactivity = NK cells unable to lyse the target cells
○ Interferon
■ Type 1 - alpha and beta, main substances that helps inhibit viral
replication
■ Type 2 - gamma interferon, for T-cell function
■ released from macrophages, lymphocytes, and tissue cells already
infected by the virus unable to defend itself; the effect would be for
the uninfected cells
■ interferons are like a signal for other cells to get ready to protect the
body
○ Inflammation
■ "The battlefield where total war is being waged; Where the host
forces are throwing all of their weaponry at the invader."
■
■ Goals of Inflammation
■ to perform temporary repair and prevent additional pathogen
entry
■ slow the spread of pathogens away from the area
■ mobilize local, regional, and systemic defenses that can
overcome the pathogens and facilitate permanent repair
■ to repair/replace tissue damaged by theinjurious agent or its
byproducts
■ Causes of Inflammation
■ Microbial infections
■ Hypersensitivity reactions
■ Physical agents
■ Irritant/Corrosive chemicals
■ Tissue necrosis
■ Hypoxia - lack of oxygen
■ Process of Inflammation
■ Tissue injury ⇒ increased blood supply to help facilitate the
transport of WBC
■ C5a - complement protein that is a chemotoxinx`
■ induces the phagocytes to the site of inflammation
■ C3b - complement protein that is an opsonin
■ could attach to host cells but there is complement
control. But if it is not working, your cells can also be
destroyed
■
■
■ Pus - dead WBCs
■ Neutrophils - fast white blood cells because it reaches the
site of infection within 40-60 minutes
 ■ If you have to open an inflamed site because of pus build up,
it has to be sterile
■ Systemic inflammatory response syndrome (SIRS)
■ harder to control if found late, could cause death
■ when inflammation overwhelms the whole body
■ criteria
■ alteration of body temperature
■ increased heart rate
■ increased respiratory rate (>20 per minute)
■ total WBC count of >12.0 x 10^9/L (or >10% immature
forms)
■ stages
■ sepsis - when there is SIRS + infection
■ severe sepsis - when there is sepsis + organ
dysfunction
■ septic shock - when there is severe drop in blood
pressure
■ Fever
■ heat is one of the classic signs of inflammation; calor or heat
■ more of a systemic response
■
■ cytokines in fever: IL-1, TNF-a, IL-6
■ the liver produces cytokines and induces the release of
acute phase proteins
■
■ C-reactive protein - good marker for acute inflammation;
nonspecific, used only for monitoring
■ Women usually have higher CRP, especially post
menopausal women.
■ Serum amyloid A - protein that causes adhesion and
chemotaxis of phagocytic cell; acute phase reactant
■ Mannose-binding lectin - activates the lectin pathway,
opsonin, calcium dependent
■ Alpha 1 Antitrypsin - acute phase protein that is part of the
alpha band of proteins; control of neutrophils
■ Haptoglobin - binds the free hemoglobin, brings it to the liver
for hemostasis
■ Fibrinogen - acute phase protein
■ Ceruloplasmin - copper transport protein in the plasma,
neutralizes any superoxide ions during phagocytosis
■ Wilson's disease - accumulation of copper
■ Procalcitonin - biomarker for sepsis