Leukocyte Structure and Function Flashcards
What does leukocyte activation require?
A change in morphology and behavior from extracellular signal
cell surface markers expressed on surface of all blood cells (leukocytes)
<p>CD antigens (cluster of differentiation)</p>
Primary cells of innate immunity
Myeloid cells
There are about 5,000-10,000 total leukocytes (cells/ul) circulating. Which type make up the most? Which make up the least?
Neutrophils- 55-70%
Basophiles-0.5-1%
This type of leukocyte has a really round nucleus, almost same size as cytoplasm
Lymphocyte
This cell has a segmented nucleus, but it stains dark in H/S stain you probably can’t see it
basophil
What is the activated function of neutrophil?
Phagocytosis and activation of bactericidal mechanisms
Are neutrophils short lived or long lived?
Short lived; circulate for a few hours, undergo apoptosis/necrosis
How many rounds of phagocytosis do neutrophils undergo before cell death?
ONE. A single one.
What clears out dead neutrophils?
Tissue macrophages
What is the chief cell of acute inflammation?
Neutrophils
These molecules are associated with groups of pathogens, recognized by cells of innate immune system
PAMPs “pathogen associated molecular patterns”
DAMPs “danger associated”
All neutrophils express this integral membrane protein on their surface to recognize pathogens
Mannose receptor
This term describes the rapid release of reactive oxygen species and NADPH-oxidase subunites to destroy internalized bacteria
Respiratory or oxidative burst
This is a small vesicle that contains the bacterium inside
phagosome
What is a lysosome fuzed with phagosome called?
Phagolysosome
What do tertiary neutrophil granules contain?
Gelitinase (MMP-9)
What are the characteristics and role of primary / azurophilic neutrophil granules?
Dark staining, antimicrobial proteins, fuse with endosomes and empty contents. Kill ingested bacteria
What are the characteristics and role of secondary/specific/definitive neutrophil granules?
Pale, fuse with endosome, trigger respiratory burst. Release contents extracellulary
What are 3 important consequences of the Respiratory Burst?
- Increase oxygen consumption
- production of superoxide O2-
- Production of hydrogen peroxide (H2O2) and other bactericidal reactive oxygen species (ROS)
What is the primary azurophilic neutrophil granule?
What is another important one?
Myeloperoxide (MO) Sulfated Proteoglycan (Sulfated PG)
NADPH oxidase, sometimes called “phagocytic oxidase,” is in the walls of which neutrophil granule?
Secondary granules
How many subunits does NADPH oxidase have? How amny are in cytoplasm? Which ones?
5 subunits, 3 in cytoplasm: p40phox, p45, p67
What is the membrane bound component of the NADPH oxidase?
flavocytochrome b (consists of 2 subunits: gp22phox, gp91 phox)
Once activated, NADPH oxidase converts oxygen molecules to what?
Superoxide, O2-
What enzyme converts superoxide to hydrogen peroxide?
Superoxide dismutase
What enzyme breaks down hydrogen peroxide to hypochloric acid? (HOCL-)
Myeloperoxidase (MPO)-
This enzyme breaks down hydrogen peroxide to benign components such as water and oxygen
Catalase
Mutations in NADPH oxidase underlie what disease?
Chronic Granulomatous Disease (can be autosomal recessive or X-linked if mutation of gp91)
How do neutrophils aid in infection control and wound healing?
Release mediators to destroy damaged tissue; disassemble matrix proteins to allow neutrophil-microbe interaction; prevent microbe escape
How do matrix metalloproteinases (MMPs) contribute to tissue destruction from chronic inflammation?
Inflammation causes cytokines to attract more leukocytes, MMPS digest ECM»_space; tissue destruction
Both of these cells release histamine and have surface receptors for antibody
Mast cells and basophils
Basophil or Mast cell? Recruited to tissue
Basophils
Basophil or Mast cell? Source of tissue histamine
Mast cell
Basophil or Mast cell? Complete maturation in tissues
Mast cell
Basophil or Mast cell? Resident in tissues
Mast cell
Source of blood histamine
Basophil
Basophil or Mast cell? Circulate as mature cells
Basophil
Monocyte Inflammatory phase (3)
inflammatory trafficking, phagocytosis, proteolysis
Monocyte Reparative phase (2)
Patrol for signs of damage, wound clean up and repair
CD14
inflammatory phase monocyte
CD16
reparative phase monocyte
Once in damaged tissue, monocytes polarize two these two primary types
Macrophages (M1 and M2)
4 major functions of Macrophages
Scavengers, Secretory cells, Regulate, Antigen-presenting cells
(R ASS)
Inflammatory monocytes become what type of macrophage?
M1
Reparative monocytes become what type of macrophage?
M2
M1 or M2? Remove dead cells
M1
M1 or M2? Suppress inflammation
M2
M1 or M2? Pro-inflammatory cytokines
M1
M1 or M2? Stimulate inflammation
M1
M1 or M2? Anti-inflammatory cytokines
M2
M1 or M2? Promote angiogenesis
M2
Monocyte/Macrophage or Neutrophil?
Long lived-weeks/months
M/M
Monocyte/Macrophage or Neutrophil?
Short lived-hours
Neutrophil
Monocyte/Macrophage or Neutrophil?
Self-replicating and adaptable
M/M
Monocyte/Macrophage or Neutrophil?
End stage cells with a single function
Neutrophil
Monocyte/Macrophage or Neutrophil?
Accumulate slowly, persist longer
M/M
Monocyte/Macrophage or Neutrophil?
Accumulate quickly, short lived
Neutrophil
Monocyte/Macrophage or Neutrophil?
Modest metabolic burst
M/M
Monocyte/Macrophage or Neutrophil?
Stronger metabolic burst
Neutrohil
Reticulo-endothelial system
Resident macrophages
Site of fluid exchange and inflammatory trafficking (leukocyte migration)
Capillary beds of veins
What causes leukocytes to stick and roll along vessel walls?
Selectins expressed by endothelial cells
What are the four adhesion molecule CAM families? Which bind with which?
SelectinsMucin-like CAMs
Integrins Ig-superfamily CAMs
Soluble cytokines
Chemokines
Inflammation is controlled by what interaction?
Cell surface adhesion molecules (CAMs) and chemokines and their receptors on endothelial cells and leukocytes regulate migration in and out of tissue
Proteins are classified as chemokines due to shared small shape. What is important in forming their 3-D shape?
Presence of 4 cysteine residues
Inflammation exacerbates disease. List 8 (A CAN PACT)
Alzheimers, Cancer, Arthritis, Neurological diseases, Pulmonary diseases, Autoimmune diseases, cardiovascular diseases, Type II diabetes
These G-protein coupled receptor’s main function is to attract leukocytes and activate adhesion
Chemokine
a tetrasaccharide carbohydrate that is attached to O-glycans on the surface of endothelial cells or ECM molecules, involved in capture/tethering and rolling part of trafficking
Sialyl Lewis X (s-Lex)
This protein functions as a CAM on the surfaces of activated endothelial cells (or ECM), and expressed very early in rolling stage
P selectin (along with E-selectin and s-Lex)
True or false: in the multi step model for leukocyte trafficking, the steps are overlapping and combinatorial
TRUE.
True or false: in the mutli step model for leukocyte trafficking, the components play only one role in a single step
FALSE. They can play more than one role in multiple steps
What accounts for the specificity and diversity in migration in the multi-step model?
It is due to the differential expression and modification of the components
What are the 7 steps of mult-step model of Leukocyte trafficking?
Capture, Rolling, Activation, Adhesion, Diapedesis, Traversing Basal Lamina, Migration through ECM
What are the interactions in rolling? What is the strength and why
Selectins (E and P) interact with leukocyte sialyl-lewis-x, it is a weak interaction to allow rolling
What adhesion molecules (ligands) of the endothelium will interact with leukocyte’s integrin LFA-1 during adhesion?
ICAM-1 and ICAM-2
In capturing phase, what part of leukocyte binds with what part of endothelial cell?
L-selectin binds with s-Lex
Strong interactions are achieved by what two types of signaling?
Outside-in signaling, and inside-out
What makes up outside-in signaling?
Chemokines from endothelium signal through G-protein coupled receptors
What make up inside-out signaling?
Integrins are activated and allow for tight interaction with ICAMs on endothelium
Takes 90 seconds and requires disassembly/reassembly of both leukocytes and endothelial cytoskeleton
Diapedesis
Describe the adhesion molecule zipper
Endothelial tight junctions open up like a zipper, form new junctions between leukocyte and endothelium without any leak
During intercellular diapedesis, the “transmigratory cup” forms. What are the three important molecules involved?
Integrin, ICAM-1, VCAM-1
During transcellular diapedesis, what are the three important molecules involved?
VE-cadherin (vascular endothelial, CD144)
ICAM-1
LFA-1 (lymphocyte function-associated antigen 1)
In lymph nodes, what do T-cells require to be activated and go into the specific T-cell area?
Chemokines
When macrophages attack the pathogen/bacteria that has entered at the site of injury/inflammation, it releases this important cytokine
TNFa (tumor-necrosis factor alpha) (along with IL-6, IL-1, and IL-8)
Why is TNF-a considered a pro-inflammatory cytokine?
It activates receptors on the blood vessel endothelium