Lab Investigation of White Cell Disorders

Question 1

how were WBC’s numbers measures in the past?

Answer 1

Haemocytometer

• Neubauer Chamber
• Way of counting manually
• Grid system, count cells within the grid and get an idea of the cell concentration and number of cells

still used for bone marrow aspirates

Question 2

what is used now to measure WBC numbers?

Answer 2

Advia

-Machine gives you a print out

• Based on size and granularity and thus you can characterise most cells
• Plotting density/granularity vs size

Question 3

with a Full Blood Count, what 3 things do we look at?

Answer 3

Haemoglobin (Hb)
White Cell (Blood) Count (WBC)
Platelet Count (Plt)

Question 4

out of all the WBC types, which is most frequently found in the body?

Answer 4

neutrophils (and then LYMPHOCYTES)

Question 5

what is a Romanowsky Stain

Answer 5

2 main components:

1) Azure B or Methylene Blue (Basic Dyes)
2) Eosin Y (Acidic Dye)

Buffer - pH 6.8

used to differentiate cells for microscopic examination in pathologic specimens, especially blood and bone marrow films

Question 6

what stains purple?

Answer 6

All WBC’s have a nucleus that will stain purple – multi lobed nucleus

Question 7

another word for neutrophil?

Answer 7

granulocyte, contains granzymes and perphorin

Question 8

lymphocyte nucleus?

Answer 8

round, occupies majority of the cell

Question 9

types of lymphocytes?

Answer 9

3
T and B cells
Natural Killer (NK) cells

Question 10

major role of lymphocytes?

Answer 10

in Adaptive Immune Response
Differentiation between self and non-self

e.g. Viral attack

Lymphocytes produce antibodies to destroy that virus

Memory cells

Question 11

lymphocytes vs granulocytes (in terms of their function)

Answer 11

Lymphocytes mainly involved in viruses whereas granulocytes (aka neutrophils) are more involved in bacteria

Question 12

lymphocyte functions

Answer 12

T-cells are involved in Cell-Mediated Immunity - CD4+ T-Helper cells, CD8+ Cytotoxic T-cells

B-cells in Humoral Immunity i.e. Antibody Production

NK cells are part of the Innate Immune System attacking virally infected cells and tumour cells

Question 13

how many lineages arise in the bone marrow?

Answer 13

2
Myeloid Progenitor Cell
Lymphoid Progenitor Cell

Question 14

another name for WBC?

Answer 14

leucocyte

Question 15

Leucocytosis vs Leucopenia

Answer 15

cytosis - Increase in white cells

penia - Decrease in white cells

Question 16

Neutrophilia

Answer 16

Increase in neutrophils

Question 17

Neutropenia

Answer 17

Decrease in neutrophils

Question 18

steroids can cause what?

Answer 18

an increase in neutrophils

Question 19

what is Toxic Granulation?

Answer 19

in response to infection the granules become more toxic and larger
-in terms of myeloid maturation, the pathway shifts to the left and you will see a lot more of the immature cells present in the blood (You should only see the mature neutrophils in the blood)

Question 20

indication for myeloid leukemia?

Answer 20

Lots of myeloblasts present in a blood film

Question 21

when might Neutrophilia occur?

Answer 21

as part of a normal physiological reaction (post-operative, pregnancy)

or because of inflammation or bacterial infection

or because of carcinoma, steroid treatment, myeloproliferative disorders, or treatment with myeloid growth factors

Question 22

how is neutrophilia investigated?

Answer 22

FBC and differential white cell count
Blood film examination

Bone marrow examination + chromosome analysis for chronic myeloid leukaemia (Philadelphia
chromosome: translocation between ch 9 and 22)

Molecular analysis for BCR-ABL oncogene

Question 23

when might Neutropenia occur?

Answer 23

Viral Infection
Drug Induced e.g. sulphonamides
Radiotherapy and chemotherapy
Part of a pancytopenia in bone marrow failure (aplastic anaemia) or infiltration e.g. leukaemia

Question 24

causes of Eosinophilia?

Answer 24

Allergic diseases (asthma/hayfever)
Parasitic infections
Drug sensitivity
Myeloproliferative diseases (chronic myeloid leukaemia)
Hodgkin’s Lymphoma

Question 25

Investigation of Eosinophilia?

Answer 25

FBC and differential white cell count
Blood film examination

Stool examination for ova and parasites

Question 26

causes of Monocytosis

Answer 26

Tuberculosis (TB)

Acute and chronic monocytic and myelomonocytic leukaemia

Malaria

Question 27

Investigation of Monocytosis

Answer 27

FBC and differential white cell count

Blood film examination:

 - for abnormal white blood cells
 - for malarial parasites

Bone marrow examination - leukaemia

TB cultures

Question 28

causes of Lymphocytosis

Answer 28

lymphocytosis of childhood, 1-6 years

Bacterial Infection

Viral Infections   e.g.      Hepatitis
				      Mumps
				      Rubella
				      Pertussis
				      Glandular Fever

Leukaemias and lymphomas

Question 29

Investigation of Lymphocytosis?

Answer 29

FBC and differential white cell count

Blood film examination

Question 30

Infectious Mononucleosis (Glandular Fever)

Answer 30

Heterophile antibodies are antibodies which react against an antigen which is completely unrelated to the antigen which originally stimulated them
e.g. Human antibodies reacting against sheep
or horse or bovine cells

Question 31

Immunophenotyping use?

Answer 31

to determine if lymphocytes are:

 B-cells: 	Demonstrate clonality by light 				chain restriction

or T-cells: Demonstrate clonality by T-cell Receptor Gene Rearrangement Studies