Immunology & serology

INNATE IMMUNITY
Professor: Ruby Meim

Immune system Natural Immunity

 Includes all parts of the body that help in the recognition  Also called innate immunity
and destruction of foreign materials.  Non specific/non-adaptive
 White blood cells, phagocytes and lymphocytes, bone  Ability to resist infection by means of normally present
marrow, lymph nodes, tonsils, thymus, and your spleen are body functions
all part of the immune system.  Same response for all pathogens
o response does not change upon repeated exposure
 No prior exposure is required
 Influenced by factors like nutrients, age, fatigue, stress and
genetic determinants
o Genetic determinants: HLA / MHC inherited
from parents; increase the probability of having
disease
LYMPHOCYTES  Ex: HLA-DR3 – increased risk for
Diabetes Mellitus
 External defense mechanism - keep microorganisms from
entering the body
 Internal defense mechanism - both cellular and humoral
factors

Innate immune system

Immunity  physical barriers eg. skin and mucous membranes
 Immunity is the body's ability to fight off PATHOGENS  chemical barriers eg. stomach acidity, secreted
that invade it. antimicrobial peptides
o Fungi, protozoans, bacteria, and viruses are all  cellular barriers eg. macrophages, neutrophils
 Innate immune response activation occurs within minutes
potential pathogens.
of pathogen recognition
 The immune system produces antibodies or cells that can o Rapid bur non specific response
1st line of
deactivate pathogens.
defense o Same magnitude of response regardless of
 Two types:
2nd line of pathogen
o Innate / Natural / Non specific
o Adaptive / Specific / Acquired defense
External defense system
 Structural barriers that prevent microorganisms from
entering the body
o Intact skin
o Mucous membranes
o Secretions
 Examples of secretions
o lactic acid (sweat, vagina)
o fatty acids (sebaceous glands)
o lysozyme (saliva, tears): attacks gram + bacteria
 Active – body synthesizes antibody o acid pH (stomach)
 Active (immunization) – o anti-fungal peptides called alpha-defensins
attenuated/weakened organism (intestinal tract)
 Passive – antibody is transferred o anti-microbial peptides called beta-defensins
 Passive (maternal) – breast feeding (respiratory, urogenital tract)
o surfactant-A and -D proteins opsonize pathogens
 Passive (antibody transfer) – serum gamma
immunoglobulin for enhanced phagocytosis (lung)

Active immunity epithelial defense mechanisms

 Occurs when one makes his/her own antibodies.  Ciliated epithelium
 This type of immunity is long term. (due to memory B  Flushing action of urine (also acid pH)
cell)  Motion of the cilia
 Infectious disease stimulates the production of MEMORY  Presence of normal flora (competitive exclusion)
cells which are then stored to prevent the infection in the
future First line defenses
 The body's first line of defense against pathogens uses Serum amyloid a
mostly physical and chemical barriers  Apolipoprotein
 Normal circulating levels = 30 μg/mL
second line defenses  Associated w/ HDL-cholesterol
 If a pathogen is able to get past the body's first line of  Functions:
defense, and an infection starts, the body can rely on its o Cleaning up cholesterol from macrophages at the
second line of defense. site of tissue injuries
o WBCs except lymphocytes o Recycles cell membrane cholesterol

Internal defense system complement

 Cells and soluble factors (cell secretions) play essential  Series of naturally occurring serum proteins that are
parts normally present
 Recognize molecules  Major functions include:
o CHO mannose o Opsonization – Facilitated phagocytosis
 WBCs, phagocytosis, inflammation o Chemotaxis – attract cells to the site iof infection
 Acute phase reactants: enhances phagocytosis o Cytolysis – Complement causes holes and
bursting of target cell
Acute phase reactants
 Normal serum constituents Mannose-binding protein
 Increase rapidly due to infection, trauma, injury  Aka mannose-binding lectin
 Produced by hepatocytes  Trimer
 12-24 hours in response to cytokines  Acts as an opsonin
o Cytokines are signaling peptides produced by  Ca-dependent
macrophages at the site of inflammation  Recognize foreign CHO (mannose) found on bacteria,
 Examples yeasts, viruses, parasites
o C-RP  Activates complement, helps promote phagocytosis
o serum amyloid A  Normal concentration = 10 μg/mL
o complement components
o alpha 1-antitrypsin Alpha1 - antitrypsin
o MBP  Major component of the alpha band in serum protein
o Fibrinogen electrophoresis (SPE)
o haptoglobin  Plasma inhibitor of proteases released from WBC (eg.
o ceruloplasmin elastase)
o Elastase : degrade elastin and collagen
C-reactive protein  Acts a “mop-up” or counteracts the effect of neutrophil
 Non specific inflammatory marker invasion during inflammation
 Discovered by Tillet and Francis  Regulates expression of pro-inflammatory cytokines
 Used to monitor INFLAMMATION  Can also react w/ serine proteases triggering C’ cascade or
 Trace constituent of serum fibrinolysis
 Thought to be an antibody to the c-polysaccharide of
pneumococci haptoglobin
 Increases w/in 4-6 hours following infection, surgery,
trauma
 Peaks w/in 48 hours  Acute phase reactant that DROPS in level
 Declines rapidly w/ cessation of stimulus  Alpha2- globulin
 Half life= 19 hours  Binds free hemoglobin (released by intravascular
 Belongs to the family of pentraxins – 5 subunits hemolysis) irreversibly
 Acts like an antibody (primitive ab)  Complex is cleared by the liver
 Capable of opsonization, agglutination, precipitation,  Levels may drop because of intravascular hemolysis
Complement (C’) activation o Since they complex with free hemoglobin
 Ca dependent, non-specific  Normal plasma conc= 40 to 290 mg/dL
 Substrate is phosphocholine (microbial membrane  Plays a role in protecting kidneys from damage
constituent) o Instead of hemoglobin getting deposited in the
 USES: kidneys, it goes to the liver for further catabolism
o Indicator of acute inflammation  Prevents loss of iron
o Non-specific indicator of disease or trauma  Prevents oxidative damage of free hemoglobin
o Following course of malignancy and organ  Binds unfolded CHONs (proteins)
o transplantation
o Significant risk factor for MI (due to low grade Fibrinogen / Factor 1
infections in arteries)  Most abundant coagulation factor
o hs-CRP (Highly Sensitive CRP)  Forms fibrin clot
 can measure minute amounts of CRP  Dimer
 predict persons at risk for MI  Normal levels= 100 to 400 mg/dL
 Increases the strength of the wound
 Stimulates endothelial cell adhesion and proliferation
 Forms a clot; a barrier to prevent spread of microbes
 Promotes aggregation of RBCs
ceruloplasmin
 Copper containing protein
 Consists of single polypeptide chain
 Binds 90-95% copper found in the plasma
 Acts as ferroxidase
 Wilson’s disease: autosomal recessive genetic disorder
o Ceruloplasmin deficiency
o Copper accumulation in organs
Cellular defense mechanism
 Myeloid cells ; participate in phagocytosis
o Granulocytes and monocytes
 Lymphocytes : acquired immune response
o 3rd line of defense
neutrophils
 Approx.50-70% of the total peripheral WBCs
 10-15 μ in diameter
 2-5 lobes (nucleus)
 w/ primary, secondary, and tertiary granules
 Azurophilic granules: antibacterial
 Frustrated phagocytosis
o Cannot fully engulf bacteria so it will degranulate
 granules are toxic to bacteria
o Will die afterwards, forming pus cells
 Half is marginating (endothelial lining), the other 50% is
circulating for 6 to 10 hours
 Capable of diapedesis: movement through blood vessel
walls to tissue.
 Selectins: help make neutrophils sticky and enhance
adherence to endothelial cells
 Form pseudopods, which squeeze through junctions
 Attracted to chemotactic factors
 Life span of neutrophil in the tissue: 5 days
eosinophils
 Phagocytize Ag-Ab complex
 Approximately 12-15 um in diameter
 1-3 % of the circulating WBCs in non allergic person
 Increases in allergic or parasitic infections
 Nucleus – usually bilobed, eccentrical
 Acidophile
 w/ primary and specific granules
 Capable of phagocytosis (Ag-Ab complex) but not as
efficient as neutrophil
 Neutralizing basophils and mast cells
 Killing parasites
basophils
 Less than 1 %
 10-15 um (smallest granulocyte)
 w/ histamine (vasoactive amine that contracts smooth
muscle
 w/ heparin (anticoagulant)
 w/ eosinophil chemotactic factor A
 IgE binds to surface of basophil
o Causes degranulation and release of vasoactive
amines, triggering infection
 Exist only for a few hours in the blood
 MAST CELLS – believed to be basophils that migrated to
the tissues
monocytes
 Largest in the peripheral blood
 12-22 um in diameter
 w/ peroxidase, acid phos, arylsulfatase (similar lysosomes
of neutrophil)
 Other type of granules (beta-glucuronidase, lysozyme,
lipase)
 Stay in peripheral blood up to 70 hours
 Long lived but slow in motility
 Function:
o microbial killing
o tumoricidal
o intracellular parasites eradication
o phagocytosis
o secretion of cytokines
o Antigen presentation
Tissue macrophages
 w/ progressive cellular enlargement to b/w 25-80 um
 Increase in ER, lysosomes, and mitochondria
 Lungs: alveolar macrophages
 Liver: Kupffer cells
 Brain: microglial cells
 Connective tissue: histiocytes
 Bone: Osteoclast
Dendritic cells
 Covered by long membranous extensions
 ANTIGEN PRESENTING CELL: Phagocytose antigen
and present it to T helper cells
 Langerhans cells: skin/mucous membrane
 Interstitial cell: organs like heart
 Interdigitating cell: T lymphocyte areas in secondary
lymphoid organs
 Innate immune receptors  Fusion = phagolysosome
 Digestion and release of debris
 Innate immune receptors are not clonally distributed
 Binding of receptors results in rapid response
 Innate immune receptors mediate three functions:
o phagocytic receptors to stimulate pathogen
uptake
o chemotactic receptors that guide phagocytes to
site of infection
o stimulate production of effector molecules and
cytokines that induce innate responses and also
influence downstream adaptive immune
responses

Inflammatory cytokines
 These cytokines are critical for host defense.
 Examples
o TNFa activates macrophage and PMN
phagocytosis and killing
o IFNab activates anti-viral mechanisms
o IL-1 stimulates inflammation and fever
 Cytokines
o induce response by binding to specific receptors
o can function in autocrine or paracrine manner
o cytokines (and their receptors) are clustered
according to structural similarities
o critical cytokines secreted by macrophages
following activation include TNFa, IL-1, IL-6,
IL-12 to stimulate inflammation
phagocytosis/killing
 Chemokines
o diverse family of chemotactic cytokines, induce
directed chemotaxis of cells
o all related in amino acid structure
o certain chemokines induce cell activation in
addition to cell recruitment
o promiscuous in receptor usage, each can bind
more than one receptor
 likewise, receptors are promiscuous
o Infection induces the release of various
chemokines
o These substances bind specific and sometimes
shared receptors to recruit various types of
immune cells to the site of infection
phagocytosis
 Physical contact between WBC and foreign particle
 Formation of phagosomes
inflammation
 Reaction to an injury
 Both cellular and humoral mechanisms
 Increased blood supply in the area
 Increased capillary permeability
 Migration of WBCs
 Migration of macrophages
and  Cardinal signs
o Rubor
o Tumor
o Calor
o Dolor
o Functio laesa
 Symptoms of Inflammation
o Increased vascular diameter, increased blood
flow (heat, redness)
o Activation of vascular endothelium to express
adhesion molecules, increases leukocyte binding
o PMNs are first cell type recruited to site,
followed later by monocytes
o Increased vascular permeability results in local
swelling and pain
Summary of innate immunity
 Rapid
 Does not generate immunologic memory
 Dependent upon germline encoded receptors recognizing
structures common to many pathogens