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types of leukocytes (5)
types of granulocytes
- leukocytes containing granules in cytoplasm
How to assess WBC population (4)
- microhematocrit tubes (buffy coat size)
- Total leukocyte count (manual or automated)
- Blood smear or film (differential cell count, morphologic changes, absolute count)
- CBC (hemogram)
leukocytes and microhematocrit
Buffy coat (white between RCB and serum. Width generally correlates to WBC count.
What can you see in a blood film
- immature cells
- toxic changes
- plasma cells
- blood parasites
Absolute neutrophil (or any leukocyte) value formula
absolute neutrophil value = total WBC count x neutrophil percentage
increased. Lymphocytosis = increased lymphocyte count
decreased. Neutropenia = decreased neutrophils.
What must you always do with bird blood?
manual count. Automated machines view nucleated RBCs as WBCs.
blood cell formation and differentiation
red blood cell production
white blood cell production.
first stage where you can differentiate granulocytes.
First stage where you can differentiate granulocytes
multipotential hematopoietic stem cell (hematocytoblast)
Beginning of all WBC in the bone marrow. Splits into common lymphoid progenitor and common myeloid progenitor.
comes from common myeloid progenitor, splits into basophil, neutrophil, eosinophil, and monocyte, which becomes macrophage.
becomes thrombocyte (platelets). Comes from common myeloid progenitor
common myeloid progenitor becomes (4)
megakaryocyte (thrombocytes), erythrocytes, mast cell, myeloblast (all other WBC but lymph)
lymphocytes and monocytes. Have one, round nucleus and no granules
- all leukocytes in one animal.
- Incldues leukocyte precursers in bone marrow and leukocytes in blood and lymph (transit), and in tissues (where they live/mature)
Why are leukocytes in blood
they are in transit from bone marrow to tissue, where they live. This includes mast cells
Where do monocytes mature?
in the tissues, they become macrophages
Tissue leukocytes include
- granulocytes (neutrophils, eosinophils and basophils)
- Monocytes (become histocytes or macrophages)
- Mast cell
which cell doesn't die immediatly?
Lymphocytes. blastogenesis, immature cells proliferate, become sensitized and stay until needed. Do not die immediatly.
Which cells can divide?
Monocytes are capable of mitosis.
What do mast cells do
allergic immune cell response. Histamine granules.
Neutrophils in canine, feline, equine, bovine, and band cell
- canine is segmented, usu no more than 5
- Feline less constricted, more of a snake
- Equine has lots of segments, almost spiky
- Bovine has very separated segments, reddish cytoplasm
- Band neutrophil is immature, no separations. Looks like a monocyte, but smaller.
the neutrophil of a reptile or bird. VERY granulated with a segmented nucleus.
- phagocytosis of organisms or other foreign material by enzymatic digestion
- Recognize inflammatory signals, leave circulation, migrate into infected tissue site, neutralize bacteria.
- chemoattraction to inflammatory sites
What do neutrophils do in: bone marrow, blood, tissue
- maturation in bone marrow (2 pools, 3 days in each)
- transit in blood (5-10 hours)
- perform function in tissue (24-48 hours)
circulating neutrophil pool (the ones in the blood). Move as fast as RBCs. Count approximated by differential and WBC counts.
marginal neutrophil pool. Attached temporarily to endothelium of capillaries and venules, lungs, bone marrow, spleen and liver. Not counted in WBC count.
In what species is CNP = MNP? In what species is it different?
- MNP = CNP in dog, horse, calf
- MNP is 3x CNP in cat
Order of neutrophilic maturation in the bone marrow
- mitotic pool: myeloblast (first discernable), promyelocyte, myelocyte (first differentiation)
- post-mitotoic pool: metamyelocyte, band cell, neutrophil
- Gets smaller, cytoplasm gets more blue, nucleus more segmented, granules increase.
Two "pools" of neutrophil maturation in the bone marrow
- mitotic pool: proliferation and maturation, 3 days (myeloblasts, promyelocytes, myelocytes)
- post-mitotic pool: maturation and storage, 2-3 days (metamyelocytes, bands and segmented neutrophils)
- After these pools they enter circulation
- increased numbers of immature neutrophils in blood. Can be degenerative or regenerative.
- Caused by infectious agents (penias, viral, bacterial or fungi), noninfectious disorders (necrosis, immune-mediated disease, neoplasia), glucocorticoid hormones (stress, maybe a LITTLE), endotoxins
degenerative left shift
- INFLAMMATION without neutrophilia
- when there are more immature neutrophils (band cells) than mature neutrophils. The bone marrow can't keep up.
regenerative left shift
- when there are more mature neutrophils than band cells. Bone marrow is releasing early but keeping up. Usually band cells. Appropriate response.
Which species have the most/least neutrophils in reserve?
- so left shift in a dog is a serious condition, left shift in a cow is pretty common, even degenerative
- Dogs regenerate fast, have a lot in marrow and margins
- cows regenerate slow and don't have much in marrow or margins
Causes of neutrophilia
- Excitement response (epinephrine)
- Stress response (steroid response)
- Most common leukocyte responses. Marrow is pumping out neutrophils faster than tissue is eating them.
- Inflammation causes vasodilation, marginated neutrophils go to inflammatory site, marrow pumps out post-mitotic pool.
- When too many are pumped too fast, toxic changes occur (hyperplasia)
- Caused by infections, IMHA, necrosis (hemolysis, hemorrhage, burns, neoplasia), sterile foreign body.
- caused by release of epinephrine, very normal. Pseudoneutrophilia (no increased production, just a shift of marginated)
- Excitement, fear, exercise, convulsions, parturition, CPR
- Least common in dogs
Usually excitement-based, when marginated neutrophils mobilize and it looks like philia, but no production is happening.
- Release or administration of corticosteroids, comes from physiologic stress response (pituitary to ACTH to adrenal gland to cortisol) leads to release from bone marrow. More in blood, no tissue to go to.
- Major systemic illness, metabolic disturbance (like renal failure or diabetic ketoacidosis) or pain. Hyperadrenocorticism, steriod therapy, ACTH administration
- Common in cats
- neutrophilia, lymphopenia, monocytosis (maybe), eosinopenia (maybe)
Left shift response for: epinephrine
acute purulent inflammation
established purulent inflammation
chronic closed cavity purulent inflammation
- epinephrine (excitement): no left shift
- Corticosteroids (stress): little/no left shift
- acute purulent inflammation: left shift
- established purulent inflammation: left shift
- chronic closed cavity purulent inflammation: little or no left shift
response of Dr. to neutroPHILIA
- left shift, inflammation
- no left shift, look at lymph
- lymph normal, slightly up, excitement, pseudo
- lymphopenia = stress or both stress and inflammation
causes of neutroPENIA
- margination (not real, just more sticking, endotoxins)
- inflammation (can cause penia or philia, acute and severe causes penia)
- peripheral destruction (usually immune-mediated)
- Stem cell injury (reversible or irreversible)
- Look at other blood cells, if also penia, look at bone marrow. If not, look for left shift. Left shift = acute inflammation, none = acute viral or marrow injury, stress
- Decreased/ineffective production, check out eosin (same precurser), lymph and platelets (dif precursor) to narrow
endotoxic neutropenia, caused when a gram negative organism releases endotoxins, makes neutrophils "sticky"
severe inflammation causes overwhelming tissue demand (overwhelming bacterial infection, viral like parvo or equine influenza, or something simpler in cattle, they are often in penia)
In what animal is neutropenia not serious?
Cows. No reserve, so you see it all the time.
Peripheral destruction neutropenia
usually immune-mediated, where neturophils are phagocytized by macrophages. Very uncommon
reversible stem cell injury neutropenia
- acute, transient injury, caused by viruses that attack dividing cells. Days or longer.
- decreased production, damage, ineffective or just out of whack.
- All cell lines disrupted (neutropenia, thrombocytopenia, nonregenerative anemia)
- often no thrombocytopenia or nonregen anemia, too short
- Infectious causes include parvo, feline panleukopenia
- Phenylbutazone in horses
irreversible stem cell injury neutropenia
- Bone marrow diseases or chronic disorders.
- decreased production, damage, ineffective or just out of whack. All cell lines disrupted (neutropenia, thrombocytopenia, nonregenerative anemia)
- Infectious causes include idiopathic hyperproliferative disorders, FeLV
- toxic like chemo drugs or estrogen
- marrow necrosis
- ineffective production, precursers interrupted
- cytotoxic or cell-mediated immunity.
- activation of macrophages
- natural killer cells
- antigen-specific cytotoxic T-lymphocytes destroy invaders
- T-helper cells release cytokines in response to an antigen.
humoral immunity. Produce antibodies
- Natural killer cells (NK), large lymphocytes.
- destroy tumor and virally infected cells.
- Antibody-dependant, coated with antibodes, kill cells coated with antigen.
- Larger, more cytoplasm that is granular. Not lymphoblasts because they have no granules.
Maturation of B-lymphocytes
mature in bone marrow, proliferate in lymphoid tissue such as lymph nodes, spleen, tonsils BALT (bronchial-associate lymph tissue), Peyer's patches in ILEUM, Bursa of Fabricius in birds.
Maturation of T-lymphocytes
produced in marrow, mature in thymus, early in life. Accumulate in lymph nodes, spleen and jujunal peyer's patches.
Where lymphocytes hang out
- 2-5% circulate in blood (of that, 50-75% is T, 10-40% is B and 5-10% is NK)
- Rest of lymphocytes live in lymphoid tissue like lymph nodes, thymus, spleen, bone marrow, GALT(gastrointestinal-associate) and BALT (bronchial-associated)
Recirculation of leukocytes
only leukocytes do it to sensitize themselves, learn antigens, primed to fight that particular antigen.
Causes of lymphocytosis
- excitement: shift from MLP to CLP, with and like neutrophils. Seen a lot in cats.
- Lymphoproliferative neoplasia: cancer (bovine leukemia virus and FeLV, lymphoma, thymoma, lumphoid leukemia)
- chronic inflammation: ehrlichia can cause true lymphoid hyperplasia (NK)
- hypoadrenocorticism (Addison's): stressed with neutropenia
- young animals: physiologic after vaccs
- Infections: bacteria, rickettsia, viral, protozoal, deep mycosis
- Extreme lymphocytosis with immature or abnormal morphology. IF you see a LOT of lymph or abnormal, think of this. (not abnormal or lower = ehrlichia or excitement)
- chronic lymphocytic leukemia causes small cell expansion
- acute lymphoblastic leukemia has huge nucleus
- leukemic manifestation of lymphoma is tissue cancer in lymph that can get into marrow.
new test that can tell by chemicals what kind of cell is proliferating
causes of lymphopenia
- STEROID/STRESS response most common, caused by endogenous and exogenous glucocorticoids or hyperadrenocorticism (Cushings)
- acute inflammation (bacterial,viral infections or endotoxins)
- uncommon like depletion, hypoplasia or aplasia or lymphoma
Monocytes are related to what other WBC and how?
share a common bipotential stem cell (myeloblast) with neutrophils
Monocytes become what, where?
marrow->blood->tissue, develop into macrophages and dendritic cells
macrophages live how long? And come from where?
- 3 months in tissues after developing from monocytes.
- Mononuclear phagocytes, eat viral, fungal, protozoal and helminth infections
- Stimulate T helper cells, and recycle RBCs.
Dendritic cells live how long and come from where?
monocytes develop into dendritic cells, which live 10-14 days in lymphoid organs
- Become macrophages, which eat infections (viral, fungal, protozoal, and helminth)
- stimulate T helpers
- recycle erythrocytes.
- too many monocytes
- Cause by inflammatory disease (acute and chronic), steroid response (stress if no left shift) or neoplastic (cancer).
- Clinically not that big a deal.
decreased monocyte concentration. Not diagnostic. Hard to document, as healthy animals have few/no monocytes in blood.
pink granules, rod shaped in cat, obscure nucleus in cow, raspberry in horse, white vacuoles in greyhounds.
- in marrow, granules at myelocyte stage, 2-6 days to produce, body stores a lot but they're in blood 30 minutes (VERY short).
- Live in subepithelial tissues in skin, lung, GI, endometrium, minimal recirculation.
- Allergies, IBS, asthma, skin allergies, parasites
- ALLERGIES AND PARASITES
- phagocytic and bacteriocidal (like neutrophils)
- kill helminths (worms)
- suppress hypersensitivity (inactivate mast cells)
- promote inflammation in allergies and asthma (IgE, release granules, contribute to tissue damage)
eosinophilia = mast cell tumor, t-cell lymphoma, Fibrosarcoma, carcinoma.
- increased eosinophils in blood.
- ALLERGY AND PARASITE
- Parasites, allergies (skin, lungs, GI), neoplasia (lymphoma or mast cell tumor as mast cells attract eosinophils).
- Eosinophilic leukemia/hypereosinophilia Syndrome
can be stress or inflammation, not diagnostic, not a big deal
- swirly nucleus, less segmented. Dark granules, purple. Equine has largest granules, bovine granules obscure nucleus, feline nucleus and granules are dark, canine lighter.
- Usually accompany eosinophils.
- originate in marrow, like neutrophils. Granules at myelocyte stage. Production takes 2.5 days, minimal storage.
- Circulate half-life in blood, 6 hours.
- Survive up to 2 weeks in tissues.
- Very few. Like mast cells, granules release substances that contribute to inflammation response, enhance allergic reactions and attract eosinophils.
- Initiate inflammatory response (histamine, serotonin, bradykinin, lysosomal enzymes)
increased concentration of hypersegmented neutrophils in blood (over 5 for dogs, over 7 for horses). Older, or longer production time.
- hematopoietic neoplasia (affecting production, cause proliferation of any WBC).
- Can be in circulation or in bone marrow.
- Chronic lymphocytic leukemia (CLL) (small lymphocytes, lOTS)
- acute lymphoblastic leukemia (ALL) HUGE cells, LOTS
- Leukemic Lumphoma (one enormous cell)
- Toxic when marrow is trying to rapidly produce.
- Can be inflammatory conditions.
- Can see cytoplasmic basophilia (granules), dohle bodies (blue clumps in cytoplasm), cytoplasmic vacuolation, larger size
- blue clumps in cytoplasm of toxic neutrophil.
- Clumps of RNA, indicating rapid production
aging or slow production.
Immune reaction. Large, HUGE nucleus, very little, dark blue cytoplasm. Can be confused with lymphoblast.
Natural Killer cells
granular lymphocytes, large lymphocytes.
lymphocyte with lots of cytoplasm and white dots in cytoplasm. Variety of disorders, some inherited. Lysosomal storage disease.
- Genetic disorder, big blue granules in leukocytes, melanocytes and platelets.
- Can include partial albinism in oculocutaneous membranes (lips, vulva), increased susceptibility to infections and hemmorhagic tendencies.
- Silver-blue coat. Persian cats, cattle, beige mice and rats, mink, foxes and killer whales.
- Inherited condition characterized by neutrophil hyposegmentation. Shows up as neutropenia but does not impair function.
- Occurs in dogs, cats, horses and humans. Very common in Aussies, in Coonhounds, Foxhounds.
- Acquired and transient, not inherited.
- Associated with chronic infections (cattle) and certain drugs.
- Neutrophil hyposegmentation
Lysosomal Storage Disease
- Inherited enzyme deficiency resulting in accumulation of undigestable substances within lysosomes.
- Can't digest, so make vacuoles.
Tick-borne, Morula in monocyte. Big spot in cytoplasm of monocyte, actually a pack of organisms. Bacteria
Morula in neutrophil (big spot in neutrophil cytoplasm). Likes joints, can cause arthritis. Bacteria
Morula in neutrophil. Bacteria
Yeast stage in neutrophil. Two dots in cell, big and dark. Fungal infection.
Viral inclusions in neutrophil. Dark spot in cytoplasm.
Protozoa in neutrophil. Big white bubble with a nucleus inside cell.
extremely immature neutrophils, before band cells. Extreme left shift.
Neutrophil consumption = production
- mild to moderate neutrophilia with a variable left shift. Some regenerative, appropriate response.
- Inflammation of soft tissue, like cellulitis associate with a bite wound.
neutrophil consuption > production
- neutropenia with prominent left shift. May be degenerative.
- Acute peritonitis, infection.
neutrophil production > consumption
- neutrophilia. Body makes neutrophils to deal with damage, but damage cannot be accessed.
- chronic walled-off inflammatory lesion.
- Marked neutrophilia with severe left shift (metamyelocytes) indicative of serious inflammatory disease.
- resembles granulocytic leukemia (pyometra, pleuritis, pyothorax, pneumonia, large absesses)
Ill animal without stress response
hypoadrenocorticism (Addison's). Or chronic.
- increased basophils in blood (200-300/uL).
- Usually accompanies eosinophilia
- Allergic, parasitic (heartworm esp) or neoplastic.
- Blue granules, not round nucleus. Machines think it's a neutrophil.
hard to tell, 0 can be normal. not clinically significant.
fake heartworm, has a blunt head and a hook. Used to be called dipetalonema
fake heartworm, blunt head, shepherd's crook. Now called acanthocheilonema
- degenerative left shift
- persistent lumphopenia
- persistent eosinopenia
- toxic neutrophils (dohle, vacuoles, basophilic cytoplasm)
- decreased leukocytosis with return of lumphocytes and eosinophils
- decreased immature neutrophils
- decrease in toxic neutrophils
Order of looking at a hemogram
- retic index, absolute retic, corrected index
- MCV, MCHC