Granulocytes and Mononuclear Cells

 Granulocytic Cells
 Neutrophils
-a.k.a Polymorphonuclear (PMN)
-effective host defense against bacterial and fungal infections
-principal leukocyte associated with phagocytosis and a localized inflammatory response
-exudate (pus) develops rapidly in an inflammatory response and is composed primarily of neutrophils and
monocytes
-prolong inflammation by the release of cytokines and chemokines
-influence the adaptive immune response
-two pools of mature neutrophils:
a. Marginating pool - adhere to the vascular endothelium
b. Circulating pool - transit to their potential sites of action
-diapedesis - movement of granulocytes from the circulating pool to the peripheral tissues
-neutrophils contain various antibacterial substances
 Eosinophils
-homeostatic regulator of inflammation
-ability to kill the larval stages of some helminth parasites through oxidative mechanisms
 Basophils
-high concentrations of heparin and histamine in their granules
-degranulation results in:
1. increased vascular permeability
2. smooth muscle spasm
3. vasodilation
-leukotrienes mediates the inflammatory functions of leukocytes

 Process of Phagocytosis
1. Chemotaxis
-movement towards the site of infection or injury
-neutrophils arrive at the site of injury and can be found in the initial exudate in less than 1 hour
-segmented neutrophils are able to gather quickly at the site of injury because they are actively motile
-monocytes are slower in moving to the inflammatory site
-macrophages resident in the tissues of the body are already in place to deal with an intruding agent
-Mediators produced by microorganisms and by cells in the inflammatory process include:
a. interleukin-1 (IL-1)
-released by macrophages in response to infection or tissue injury
b. histamine
-released by circulating basophils, tissue mast cells, and blood platelets

 Chemoattractant/Chemotaxins
-substance that guide cells to the site of injury through chemotaxis
-induce a positive movement toward and a negative movement away from a chemotactic response

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 -attachment devices that influences the speed of phagocytosis:
i. Fc receptor—binds the Fc portion of antibody molecules, chiefly immunoglobulin G (IgG). The IgG
attaches to the organism through its Fab site.
ii. Complement receptor—the third component of complement, C3, also binds to organisms and
then attaches to the complement receptor.
 Opsonization
-coating of the organisms by molecules that speed up phagocytosis
-Fc portions of antibody and C3 are called opsonins
-steps in opsonization:
a. Antibody attached to the surface of a bacterium minimally binds the Fc phagocyte receptor.
b. Complement C3b is attached to the surface of the bacterium and binds loosely to the phagocyte
C3b receptor.
c. Both antibody and C3b are attached to the surface of the bacterium and bound tightly to the
phagocyte, allowing greater opportunity for the phagocyte to engulf the bacterium.
 Necrotic cells
-release an independent chemoattractant of necrotaxis signal
-promote localization of neutrophils directly into existing areas of injury to focus the innate immune
response on damaged areas and away from healthy tissue, which provides an additional safeguard against
collateral damage during sterile inflammatory responses.
2. Adherence
-five steps for effective leukocyte recruitment to the site of injury:
a. Capture
b. Rolling
c. slow rolling
d. firm adhesion
e. transmigration
 Capture (tethering)
-represents the first contact of a leukocyte with the activated endothelium
-Cell Adhesion Molecules (CAMs):
 P-selectin
-found on endothelial cells and the primary adhesion molecule for capture and the initiation of rolling
 E-selectin
-include CD44
 L-selectin
-also has an important role in capture
 PECAM-1
 ICAM-1
 VE-cadherin
 LFA-1 [CD11a/CD18]
 IAP [CD47]
 VLA-4 [4β1–integrin]
3. Engulfment
-on reaching the site of infection, phagocytes engulf and destroy the foreign matter
-bacteria can be engulfed through active membrane invagination

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  Hydrophobicity
-principal factor in determining whether phagocytosis can occur is the physical nature of the surface of
the bacteria and phagocytic cell
4. Digestion
-granules in the phagocyte cytosol then migrate to and fuse with the phagosome to form the phagolysosome
-granules contain degradatory enzymes of the following three types:
a. Primary, or azurophilic, granules containing enzymes (e.g., lysozyme, myeloperoxidase)
b. Secondary, or specific, granules containing substances such as lactoferrin
c. Tertiary granules containing substances such as caspases
 Myeloperoxidase granules
-responsible for the action of the oxygen-dependent, myeloperoxidase-mediated system
-Hydrogen peroxide (H2O2) and an oxidizable cofactor serve as major factors in the actual killing of
bacteria
 Monocytes
-are particularly effective as phagocytic cells because of the large amounts of lipase in their cytoplasm
 Lipase
-able to attack bacteria with a lipid capsule, such as Mycobacterium tuberculosis
 Release of lytic enzymes results in the destruction of neutrophils
 Macrophage digestion proceeds without risk to the cell
 Respiratory burst
-cells demonstrate increased metabolic activity
-production of large quantities of reactive oxygen species (ROS)
 Nicotinamide-Adenine DinucleotidePhosphate (NADPH) oxidase
-together with the granules elicit microbicidal results
-forms the centerpiece of the phagocyte-killing mechanism and is activated in about 2 seconds
-generates ROS by generating the superoxide radical (O2−)
5. Exocytosis
-excretion of the digested/killed bacteria
-some peptides instead of being eliminated, they attach to a host molecule called major histocompatibility
complex (MHC) class II and are expressed on the surface of the cell

 Monocytes-Macrophages
-macrophage and its precursors are widely distributed throughout the body
-rapid phagocytosis mediated by receptors for IgG and the major fragment of C3
-migrate freely into the tissues from the blood to replenish and reinforce the macrophage population
-Macrophages exist as fixed or wandering cells
-macrophage-activating cytokines:
a. interferon-gamma (IFN-γ)
b. granulocyte colony-stimulating factor (G-CSF)
c. tumor necrosis factor α (TNF-α, cachectin)

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 Location of Macrophage Name of Macrophage
Adipose Tissue Adipose Tissue Macrophage
Peripheral Blood Monocyte
Liver Kupffer cell
Lymph node Sinus histiocyte
Lungs Dust cells
Skin and Mucosa Langerhans cell
Central Nervous system Microglia
Placenta Hofbauer cell
Kidney Mesangial cell
Bone Osteoclasts

 Acute Inflammation
-primary objective of inflammation is to localize and eradicate the irritant and repair the surrounding tissue
 Celsus
-a practitioner of Greek medicine
-credited with recording the cardinal signs of inflammation:
a. rubor (redness)
b. calor (heat)
c. dolor (pain)
d. tumor (swelling)
e. Functio laesa (loss of function) Galen
-inflammatory response involves the following three major stages:
1. Dilation of capillaries to increase blood flow
2. Microvascular structural changes and escape of plasma proteins from the bloodstream
3. Leukocyte transmigration through endothelium and accumulation at the site of injury

 Sepsis
-systemic inflammatory response syndrome (SIRS) + infection
-severe sepsis is defined as sepsis + evidence of organ dysfunction
-criteria for SIRS require two or more conditions:
a. Increased heart rate
b. increased respiratory rate
c. total leukocyte count of >12.0 × 10(9)/L (or >10% immature forms)
-begins when the innate immune system responds aggressively to the presence of bacteria
-Biochemical markers associated with sepsis include:
a. tumor necrosis factor (TNF)
b. IL-1
c. IL-6
d. procalcitonin
e. chemokine
f. C-reactive protein (C-RP)

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 Disorders of Neutrophils
 Noninfectious Neutrophil-Mediated Inflammatory Disease
-the same oxidative and non-oxidative processes that destroy microorganisms can affect adjacent host tissues
-occurs when phagocytes attempt to engulf particles that are too large then releases oxygen radicals and
granule contents onto the particle, but these escape into the surrounding tissues, generating tissue damage
 Congenital Neutrophil Abnormalities
a. Chédiak-Higashi Syndrome
-qualitative disorder of neutrophils
-autosomal recessive trait
-giant granules, display impaired chemotaxis and delayed killing of ingested bacteria
b. Chronic Granulomatous Disease
-genetically heterogeneous group of disorders of oxidative metabolism affecting the cascade of
events required for H2O2 production by phagocytes
-defect of neutrophil microbicidal ROS generation resulting from gp91phox deficiency
-caused by a missense, nonsense, frameshift, splice, or deletion mutation in the genes for p22 phox,
p40 phox, p47 phox, p67 phox (autosomal CGD), or gy91phox (X-linked CGD)
-infections with catalase-positive bacteria and fungi affecting the skin, lungs, liver, and bones
-develop granulomas that impairs some physiologic processes (e.g., obstruction of the esophagus or
urinary tract)
i. X-linked CGD (X-CGD)
-mutation in CYBB encoding the transmembrane gp91phox
ii. Autosomal recessive CGD
-have a less severe clinical course than patients with the X-linked form
-Laboratory evaluation of CGD:
 Nitroblue Tetrazolium test (NBT)
 serum quantitative immunoglobulin
 complement activity enzyme immunoassay
 CBC with differential
 Myeloperoxidase stain
 neutrophil receptor profile
c. Complement Receptor 3 Deficiency
-rare autosomal recessive trait
-presents as a leukocyte adhesion deficiency
-abnormalities of adherence-related functions:
 Including decreased aggregation of neutrophils to each other after activation
 decreased adherence of neutrophils to endothelial cells
 poor adherence and phagocytosis of opsonized microorganisms
 defective spreading
 decreased diapedesis and chemotaxis
i. Leukocyte adhesion deficiency type 1 (LAD-1)
-caused by a deficiency of CD18
ii. Leukocyte adhesion deficiency type 2 (LAD-2)
- is caused by the absence of sialyl–Lewis X (CD15s) blood group antigen
d. Myeloperoxidase Deficiency
-inherited as an autosomal recessive trait on chromosome 17
-azurophilic granules are present, but myeloperoxidase is decreased or absent

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  Myeloperoxidase
-an iron-containing heme protein
-responsible for the peroxidase activity characteristic of azurophilic granules
-it accounts for the greenish color of pus

 Monocyte-Macrophage Disorders
a. Gaucher’s Disease
-disturbance in cellular lipid metabolism
-most frequently affects children
-deficiency of β-glucocerebrosidase, the enzyme that normally splits glucose from its parent
sphingolipid, glucosylceramide
 Gaucher’s cells
-are rarely found in the circulating blood
-cell is large, with one to three eccentric nuclei and a characteristically wrinkled cytoplasm
b. Niemann-Pick Disease
-abnormality of lipid metabolism
-affects infants and children, with an average life expectancy of 5 years
-rare autosomal recessive deficiency of the enzyme sphingomyelinase
-characterized by massive accumulation of sphingomyelin in the mononuclear phagocytes
 Pick’s cell
-is similar in appearance to Gaucher’s cell, although the cytoplasm of the cell is foamy

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