08 August 2017

Lecture #9 : The White Blood Cells


The majority of leukocytes are formed from primitive mesenchymal cells in the hematopoietic tissues. As the cells mature from their fixed or relatively fixed “blast” and “pro” stages to mature and morphologically identifiable types, they lose their intercellular connections, and acquire amoeboid activity. These cells after reaching the more mature stage, escape from the tissue by means of their own mobility into the circulating blood.

Leukocytes are lighter than red cells and tend to accumulate at the periphery of the flowing blood adjacent to the lining of the blood vessel where they tumble along at a slower rate than the axial period of the blood column. Some of the leukocytes are continually squeezing between the endothelial cells and the walls of the capillaries into the tissue spaces. From the tissue spaces, they may pass into the lymphatic channels and thus get back into the blood stream. At any given time, there are numerous leukocytes that have escaped from the hematopoietic organs and are not in circulation. These cells are readily available on demand are called back into the active circulation when there are chemotactic substances in the lumen of the vessels which attract the leukocytes in the perivascular areas.

Leukocytes after delivery into the blood stream and after surviving for a number of days in the body, eventually become senile and die. These effete cells are phagocytized by other cells in the blood stream by the phagocytic cells of the tissue spaces, by macrophages of the splenic pulp or by the littoral cells of the liver sinuses.

Leukocytes in addition to being destroyed in the body, are continually escaping from the body by passing into secretions of various glands or by passing between the cells lining the respiratory, gastrointestinal and genitourinary tracts into the lumen of these organs. Thus, we find leukocytes in all body fluids including sweat, synovia, pancreatic juices, feces, urine and secretions from the genital tracts. In inflammatory, ulcerative and hemorrhagic lesions involving any of the excretory organs there is an increased rate of loss of leukocytes from the body.


MATERIALS NEEDED FOR FORMATION OF WHITE BLOOD CELLS


In general, the leukocytes need essentially the same vitamin and amino acids as most of the other cells of the body for their formation. Especially does lack of folic acid, a compound of the Vitamin B complex, block the formation of white blood cells as well as prevent maturation of red cells.


CHEMISTRY OF LEUKOCYTES


1. Granulocytes

Granulocytes process glucose by aerobic glycolysis to yield lactic acid. Alkaline phosphatase is present in increased concentration in granulocytes during infections and decreased in granulocytic leukemia. Acid phosphatase is increased in chronic granulocytic leukemia and normal in infestations. Phospholipids, lysozyme and phagocytin are bactericidal and may also be found in granulocytes. In addition, peroxidase, lipase, sulfhydryl groups, heparin, histaminase, glucogen, dipeptatases, tripeptases, maltose, beta glucuronidase and glucuronic acid are also present in granulocytes.

2. Lymphocytes

Glycogen and acid phosphatase are found in lymphocytes. They may also contain alkaline phosphatase, dipeptidase, oxidase and peroxidase.

3. Monocytes

Glycogen, sudanophilic substances, lysozyme, acid phosphatase, phospholipids and lipids are present in monocytes.

4. Plasma cells

Plasma cells contain large amount of RNA and DNA and are responsible for the manufacture of antibodies.


LIFE SPAN OF LEUKOCYTES


The main reason leukocytes are present in the blood is simply to be transported from the bone marrow or lymphoid tissue to the areas of the body where they are needed. Therefore, it is to be expected that the life of leukocytes in the blood would be short.

            Granulocytes              –          2 – 3 hours during serious tissue infection
                                                –          14 hours (average) usually
                                                –          several days, when not needed in the tissues.

            Monocytes                  –          still a mystery, for the wander back and forth between
                                                            the tissues and the blood

            Lymphocytes              –          in the blood, only a few hours
                                                –          using radioactive lymphocytes – 100 to 200 days
                                                            depending on the tissues need for the cells.


PROPERTIES OF LEUKOCYTES


1. Diapedesis – the leukocytes can squeeze through the pores of the blood vessels by the process of diapedesis. That is, even though a pore is much smaller than the size of the cells, a small portion of the cell slides through the pore at a time, the portion sliding through being momentarily constricted to the size of the pore.



2. Amoeboid movement – one the cells have entered the tissue spaces, the polymorphonuclear neutrophils, the lymphocytes and monocytes to a lesser degree, move through the tissues by amoeboid motion.

3. Chemotaxis – a number of different chemical substances in the tissues cause the leukocytes to move either toward or away from the source of the chemical. This phenomenon is known as chemotaxis. Degenerative products of inflamed tissues, bacterial toxins can cause chemotaxis of leukocytes.

Positive chemotaxis – when cells are attracted toward the source of the chemical substance

Negative chemotaxis – when the leukocytes are repel from the source of the chemicals.

4. Phagocytosis – ingestion of particulate matter by the cells. Whether or not phagocytosis will occur, depends upon three selective procedures:

a. If the surface of the particle is rough, like hood of phagocytosis is increased, whereas a smooth particle is resistant to phagocytosis.

b. Most natural substances of the body have electronegative surface charge, therefore are repelled from the phagocytes, which also carry electronegative surface charges. Dead tissues and foreign particles are frequently electropositive and are therefore subject to phagocytosis.

c. The body has a means for promoting phagocytosis of foreign materials by selectively combining foreign particles with globulin molecules called opsonins.


TOTAL LEUKOCYTE COUNT


Adults             5,000 – 10,000 / cu.mm         5 – 10 x 109/L
Infants            6,000 – 18,000/ cu.mm          6 – 19 x 109/L
Children          5,000 – 15,000/ cu.mm          5 – 15 x 109/L






FUNCTIONS OF NEUTROPHILS

1. Prevent or retard the intrusion of infectious agents and other foreign materials into the host environment. This is accomplished by phagocytosis and digestion of the material.

2. Secretory function – neutrophils release an enzyme known as lysozyme which acts as a hydrolyzing agent and is important in the destruction of certain bacteria.

3. They also liberate immune bodies and other enzymes that interferes bacterial growth.

4. Neutrophils exhibit amoeboid motion and play a role in inflammatory process. They release endogenous pyrogen that produces fever by acting on the hypothalamus to set the body’s thermostat at a higher level.

FUNCTIONS OF EOSINOPHILS

1. Interact with foreign protein (they detoxify protein) under the control of adrenal cortical hormone.

2. They exhibit chemotaxis. They are attracted to fibrin or proteolytic enzyme. Among the chemotactic factors that attract eosinophils is present in basophils and mast cells.

3. Eosinophils phagocytose foreign particles and antigen – antibody complexes.

4. Eosinophils contain substances that inactivate factors released by mast cells and basophils, such as histamine, slow reacting substances of anaphylaxis, and platelet – activating factor.

5. They produce antihistamine and are associated with allergy.

6. They provide some defense against helminthic parasites.

7. Eosinophils are sources of plasminogen

FUNCTIONS OF BASOPHILS

1. Basophils respond to adrenal cortex hormones in similar fashion to eosinophils.

2. They liberate heparin, histamine, hyaluronic acid and serotonin.

3. Basophils synthesize and store histamine and eosinophil chemotactic factor anaphylaxis.

4. They appear to be involved in immediate hypersensitivity reactions, such as allergic asthma.

5. Speculations suggest that basophils are associated with fibrinolysis.

6. Lipid metabolism and anaphylactoid reactions

7. Secretory functions – basophils release their granule contents outside the cells, after exposure to stimuli.

Note:   Acid mucopolysaccharide is responsible for the metachromatic staining property
            of Basophil granules

FUNCTIONS OF MONOCYTES

1. Monocytes are formed in the marrow, transported by the blood, and migrate into the tissues where they transform into histiocytes or macrophages, to spend the majority of their life span. The blood monocytes and tissue macrophages make up a mononuclear phagocyte system (reticuloendothelial system). This system has an important role in defense against microorganisms, including bacteria, fungi, viruses and Protista.

2. The cells are motile and respond to chemotactic factors.

3. They engage in phagocytosis, a process that is enhanced if the particle is coated by IgG or complement for which the macrophages have membrane receptors. These mononuclear phagocytes are an integral part of both humoral and cell – mediated immunity.

4. Play a role in the synthesis and secretion of transferrin, interferon, endogenous pyrogen, lysozyme.

5. Macrophages remove and process senescent cells and debris through phagocytosis and digestion.

6. They act as feeder cells (trephocytes) supplying iron to red cells and protein to antigenic sites to plasmacytes and lymphocytes.

7.     They play an important role in the regulation of hematopoietic activity.

FUNCTIONS OF LYMPHOCYTES

1. The lymphocyte has a primary function in cell–mediated immunity, which includes delayed hypersensitivity; graft rejection, graft versus host reactions, defense against intracellular organisms such as tubercle bacilli and Brucella and probably defense against neoplasms.

2. B cells and their pyrogeny perform in humoral immunity, or in the production of antibodies, either as a lymphocyte or after transformation into plasmacyte.

3. They act as feeder cells (trephocytes) – play a role in the synthesis of protein, enzymes, minerals and transport them to the sites of cellular growth.

4. Play a role in chronic inflammatory process.

T cells – lymphocytes influenced by the thymus (thymus dependent)
B cells – lymphocytes influenced by the bursal equivalent organ (bursa dependent)

FUNCTIONS OF PLASMACYTES

1. Plasmacytes play a role in antibody formation. They produce immunoglobulin for secretion outside of the cell.

The so called antigen–antibody quartet:

a.     Lymphocytes
b.     Monocytes
c.      Eosinophils
d.     Plasma cells


MORPHOLOGIC ABNORMALITIES OF LEUKOCYTES


Hereditary conditions affecting leukocytes:

1. Pelger–Huet anomaly – also called “hereditary hyposegmentation” hereditary autosomal dominant condition characterized by hypolobulation of the granulocyte or failure of normal segmentation of granulocytic nuclei. Most nuclei are band–shaped, rod –like dumbells or peanut–shaped, “spectacle–like” or “pince-nez” nuclei with smooth, round or oval individual lobes and pyknotic nuclear chromatin.


2. Alder’s anomaly or Alder–Reilly anomaly – recessive trait characterized by the presence of larger than normal azurophilic and basophilic granules confused with toxic granulation but is unrelated to infection and is not transient. Cells usually affected are PMN, lymphocyte and monocyte.



3. May–Hegglin anomaly – rare autosomal dominant condition characterized by the presence of pale blue inclusions resembling Dohle bodies in neutrophils, giant platelets and in some persons, thrombocytopenia. The inclusions are larger and more prominent than the Dohle bodies found in infections. Cells usually infected are eosinophils, basophils, monocytes and neutrophils.


4. Chediak–Higashi anomaly – also known as Chediak–Steinbrick Anomaly
– congenital gigantism of peroxidase granules.
– autosomal recessive disorder characterized by partial albinism,
            photophobia, Increased susceptibility to infection and presence
            of very large granules which appear to be abnormal lysosomes.
– cells affected are granulocytes, monocytes and lymphocytes



5.     Jordan’s anomaly – characterized by vacuolization of leukocytes
– vacuoles are present in the cytoplasm of granulocytes, monocytes and
occasionally lymphocytes and plasma cells of patients with progressive muscular dystrophy.



6. Hereditary neutrophilic hypersegmentation – hyperlobulation of the nuclei of the granulocytes from 4 – 6 lobes or more with no evidence of Vitamin B12 and / or folic acid deficiency

7. Hereditary giant neutrophilic leukocytosis – rare hereditary disorder where there is a tendency to produce polyploid cells. Characterized by presence of neutrophils and hyperlobulation of nuclei.



ABNORMAL INCLUSION BODIES FOUND IN LEUKOCYTES


1. Auer bodies or Auer rods – with Romanowsky stains, Auer rods are linear or spindle–shaped red–purple inclusions in myeloblast and monoblast. Auer rods are derivatives of azurophilic granules and stain positively for Sudan Black B, myeloperoxidase, chloroacetate, esterase and acid phosphatase. They are caused by unusual development of lysozymes.



2. Toxic granules – toxic granules are dark blue to purple cytoplasmic granules in the metamyelocyte, band or neutrophilic stage. These are characteristics of bacterial infections and are frequently seen in aplastic anemia and also in myelosclerosis.



3. Dohle bodies – these are small round or oval bodies up to 2 – 3 micrometers in size, usually in the periphery of the cytoplasm of neutrophils, which stain blue–gray with Romanowsky dye. These are mostly seen in bacterial infections, severe burns, exposure to cytotoxic agents and uncomplicated pregnancy. They are remnants of free ribosomes or rough surfaced endoplasmic reticulum persistent from an earlier stage of development.


4. Snapper–Schneid bodies – inclusion bodies found in the cytoplasm of multiple myeloma and plasma cells after therapy with amidine drugs.

5. Russell or Fuch’s bodies – gamma globulin bodies in the cytoplasm of plasma cells and inflamed tissue. The bodies give the cells a grape or berry or morula cell appearance.



OTHER ABNORMALITIES IN THE NUCLEUS AND CYTOPLASM OF LEUKOCYTES


1. Hypersegmented neutrophil – also called P.A. polycell of macropolycyte; larger than normal neutrophil and has 5 – 10 segments; seen in pernicious anemia

2. Polycyte – has a normal size but with 4 – 6 lobes in the nucleus; found in stage of recovery from infection.

3. Pyknotic cell – cell whose nucleus becomes smaller and denser, nuclear segments disappear, leaving several balls of dense chromatin.

4. Virocyte or atypical lymphocyte or Downey type cell or Turk irritation cell – cell has a chromatin arrangement which gives the cell a “moth–eaten” or “tunneled” appearance or “Swiss–cheese” form; vacuolated which gives the cell a “foamy” or “bubbly” appearance; cell has prominent axurophilic granules

Downey type I – cell with deeply indented nuclei

Downey type II – cell with smooth cytoplasm with patchy peripheral and radial basophilia has been called “stress” lymphocyte; seen in infectious mononucleosis, viral pneumonia, herpes zoster, herpes simplex and other viral infections

5. Rieder cell – myeloblast that is characterized by having a nucleus with deep indentations often suggesting lobulations; seen in acute myeloid leukemia.


6. Vacuolated cell – cell with holes or vacuoles in the cytoplasm; vacuoles are signs of degeneration in severe infection, chemical poisoning, leukemia; if seen in normal blood, indicative of smear prepared from over two – hour old oxalated blood.



7. Cells exhibiting phagocytosis – cells that are endowed with the ability to engulf particles are called phagocytes. In general, these include the polymorphonuclear granulocytes (neutrophils and eosinophils), the monocytes in the formation of Lupus erythematosus (L.E.) cells and Tart cells.

8. Basket cell – net like nucleus from a ruptured white cell; in normal blood it is believed to be the older form of smudge cell.



IRREGULARITIES IN THE BLOOD SMEAR PREPARATION AFFECTING LEUKOCYTES


1. Squashed or distorted lymphocytes – caused by excessive pressure on the cell during the process of preparation.

2. Accumulated white cell – a bunch of white cells seen on the edge of a blood smear and caused by improper spreading technique during smear preparation.

3. Smudge cell – a bare nucleus of a ruptured white cell caused by excessive pressure on the cells during smear preparation. This is indicative of increased fragility of cells or abnormal destruction of cell.

4. Disintegrated or ruptured cell – this is caused by excessive pressure on the cells during smear preparation and is found on the smear prepared from old blood. It is also found in toxic conditions.

5. Poorly stained leukocytes – this maybe caused by:

a.     Incorrect pH of buffer
b.     Improper mixing of stain and buffer
c.      Too short staining period.

6. Precipitated stain – this is caused by the failure to flush properly the excess stain on the side.

                                     





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