I&I Week 8 Leukocytes and inflammation

What are the three methods of counting leukocytes?

Impedance, Optical, and Manual.

What is the principle of leukocyte counting for the impedance method?

Each nucleus of the leukocyte impedes electron flow and is counted.

What is the principle of leukocyte counting for the optical method?

Each whole cell intersects a laser beam, and light scatter defines their size, granularity, and nuclear shape.

What is the principle of leukocyte counting for the manual method?

100-leukocyte subsets stained on a blood film are counted using a microscope and cell counter.

Which leukocyte counting method provides the most accurate blood leukocyte subset concentrations?

Manual.

Name a positive acute phase protein that increases in concentration in the blood less than 2 days after inflammation.

Fibrinogen.

Name another positive acute phase protein that increases in concentration in the blood less than 2 days after inflammation.

Haptoglobin.

Name a negative acute phase protein that decreases in concentration in the blood less than 2 days after inflammation.

Transferrin.

Name another negative acute phase protein that decreases in concentration in the blood less than 2 days after inflammation.

Albumin.

What type of proteins increases in concentration after 1-3 weeks of inflammation?

Immunoglobulins.

What analytical method detects acute phase proteins?

Copper dye.

What analytical method detects delayed phase proteins?

Copper dye and Serum protein electrophoresis (SPE).

What analytical method is used for Serum ALB (Albumin)?

BCG dye.

How are Serum GLB (Globulins) measured?

Subtraction (TP – Alb).

What analytical method is used for Albumin and globulin fractions (α1, α2, β1, β2, γ)?

Serum protein electrophoresis (SPE).

What analytical method is used for Plasma TP?

Refractometry.

What terminology is used when TP is above the reference interval?

Hyperproteinemia.

What terminology is used when ALB is below the reference interval?

Hypoalbuminemia.

What terminology is used when GLB is above the reference interval?

Hyperglobulinemia.

What terminology is used when Plasma TP is above the reference interval?

Hyperproteinemia.

What terminology is used when Fibrinogen is above the reference interval?

Hyperfibrinogenemia.

What does the Widow Maker's serum protein electrophoresis (SPE) represent?

Inflammation (polyclonal gammopathy).

What causes the increase in positive acute phase proteins?

Production by the liver increases by cytokines.

What causes the decrease in negative acute phase proteins?

Production by the liver decreases by cytokines.

What causes the increase in delayed phase proteins?

Production in lymphocytes in lymph nodes increased by cytokines.

What is seen on SPE pattern for B-lymphocyte neoplasia?

A narrow-based peak in the gamma fraction indicating B-cell neoplasia and production of immunoglobulins by B-cell clones (lymphoma).

What are the learning outcomes for leukocyte patterns in inflammation?

Identify leukocytosis, differentiate acute and chronic inflammatory leukograms from cortisol (stress) and catecholamine leukogram patterns, and identify a leukopenia pattern induced by inflammation.

What abnormalities should you recognize in the leukogram?

Leukocytosis patterns.

What is a left shift?

Increase in Band neutrophils (non-segmented neutrophils).

Why is it called a left shift?

Because band or stab neutrophils are left of segmented neutrophils on a manual counter.

What does an increase in WBC indicate?

Leukocytosis.

What does an increase in Segs indicate?

Neutrophilia.

How long do neutrophils typically stay in the blood in healthy animals?

Approximately 8 hours.

How long do neutrophils stay in the blood during inflammation?

Less than 8 hours.

How long does it take to make a new supply of neutrophils during inflammation?

Five days or sooner.

What are the classifications of a left shift?

Regenerative and Degenerative.

What are the characteristics of a regenerative left shift?

Leukocytosis due to neutrophilia with an increase in bands, and [segs] > [bands].

What are the characteristics of a degenerative left shift?

[bands] > [segs], usually decreased [WBC] or leukopenia, but WBC may be within the reference interval.

What is the clinical significance of a degenerative left shift in cats?

Cats with a degenerative left shift are 1.5 times more at risk to be euthanized in the hospital.

What is the clinical significance of a degenerative left shift in dogs?

Dogs with a degenerative left shift are 1.9 times more at risk to be euthanized in the hospital.

What is important to remember about Bands?

Bands are immature and less functional, and in a degenerative left shift, Bands > Segs.

What is the best interpretation of leukocytosis, neutrophilia, and regenerative left shift?

Segs > bands.

What is the hallmark of an acute inflammatory reaction?

Left shift.

What characterizes a regenerative left shift?

Adequate response to inflammatory disorder ([Segs > Bands]).

What is the role of cytokines in a regenerative left shift?

Cytokines cause the need for segs in inflamed tissues, and the marrow responds by releasing segs cells from the SNP.

Where do segs cells and bands cells release from during a regenerative left shift?

Segs cells release from SNP and bands cells release from maturation pool.

What stimulates the release of cytokines from tissue macrophages?

Bacteria.

What does inflammatory disorder mean in terms of time?

Acute = hours to days.

What are the causes of inflammatory disorder?

Infectious (bacterial, other) and noninfectious (necrosis, immune).

What stimulates the release of neutrophils from the SNP of bone marrow?

Cytokines.

What stimulates the release of bands from the MatNP of bone marrow?

Cytokines.

What are toxic neutrophils associated with?

Severe inflammatory disease.

What are common causes of toxic neutrophils?

Commonly bacterial infections (i.e., endotoxemia) and noninfectious inflammatory disorders.

What is the pathogenesis of lymphopenia in an acute inflammatory leukogram?

Cytokines from inflammatory site increase migration to inflamed tissues and 'homing' to lymph nodes, decreasing efflux of lymphocytes from LN due to antigenic stimulation in LN.

What should you expect to see concurrently with lymphopenia?

Neutrophilia with a left shift.

How does cortisol affect adhesion molecules on blood vessels?

Cortisol decreases expression of adhesion molecules on blood vessels.

Where do lymphocytes go in a cortisol stress leukogram?

From blood to bone marrow.

What are the causes of chronic inflammatory neutrophilia?

Established, persistent inflammation.

What do macrophages and lymphocytes release in chronic inflammatory neutrophilia?

Cytokines.

What do cytokines stimulate in chronic inflammatory neutrophilia?

Production of neutrophils.

What characterizes chronic inflammatory neutrophilia?

Mostly mature neutrophils.

What is the pathogenesis of chronic inflammatory lymphocytosis?

Chronic antigenic or cytokine stimulation leads to increased lymphocyte production.

What are reactive lymphocytes?

Stimulated B-cells or T-cells, usually due to infectious disorders.

What are common causes of monocytosis?

Acute or chronic inflammation, steroids (stress), or catecholamine (shifting from MP to CP).

What are common causes of eosinophilia?

Hypersensitivity/Allergic disorders, parasitism (tissue migration of larvae), and hypoadrenocorticism.

What are common causes of basophilia?

Hypersensitivity/Allergic disorders and parasitism (tissue migration).

What does a catecholamine leukogram look like?

Chronic inflammatory leukogram.

What causes a catecholamine leukogram?

Fight or Flight stimulus (excitement, fright, flight, exercise), increased catecholamines, and increased blood flow rate through small vessels.

What are the characteristics of a catecholamine leukogram?

Neutrophilia without a left shift, lymphocytosis, and monocytosis.

What is the clinical significance of a degenerative left shift?

Inadequate response to inflammatory disorder ([bands] > [segs]).

What are possible reasons for a degenerative left shift?

Marrow is not keeping up with the demand, severe bacterial infection, poorer prognosis, and cytokines leading to depletion of cells.

What does the term acute overwhelming inflammatory mean in terms of time?

Hours to days.

What stimulates the migration of neutrophils to inflamed tissue in acute overwhelming inflammatory neutropenia?

Cytokines.

What does cytokines stimulate in acute overwhelming inflammatory neutropenia?

Release of bands from Maturation NP.

What characterizes an acute overwhelming inflammatory leukogram?

Leukopenia, neutropenia with + a degenerative left shift with toxic.