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Haematology > Lab Investigations of White Cell Disorders > Flashcards

Lab Investigations of White Cell Disorders Flashcards

1
Q

What is a Haemocytometer Neubauer Chamber used for?

A

Haemocytometer Neubauer Chamber was used to count cells in the past

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2
Q

How is a Haemocytometer Neubauer Chamber used to count no. of cells?

A

Put blood film on cover glass mounting support and count

Would count no. of squares and multiply it out to get cell no.

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3
Q

What is the modern day technique for counting WBCs?

A

Nowadays more automation techniques are used e.g. ADVIA

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4
Q

How do modern methods distinguish between cells?

A

Results obtained are complicated dot plots distinguishing samples based on size and granules

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5
Q

What blood cells are we able to identify nowadays?

A

We identify:

  • Haemoglobin
  • White cell
  • Platelets
  • Neutrophils count
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6
Q

Describe the normal Hb levels expected of a FBC

A

Haemoglobin (Hb)
Normal Male: 130 - 180 g/L
Normal Female 120 – 160 g/L (lower due to menstruation)

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7
Q

What is the purpose of a full blood count?

A

need to know normal values to compare to

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8
Q

Once we’ve obtained abnormal Hb results from FBC how can we identify the problem?

A

Once we’ve identified Hb value we differentiate using MCV (micro/macrocytic)

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9
Q

What is a normal WBC count?

A

Normal Adult 4.0 - 11.0 x 109 / L

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10
Q

Outline a normal platelet count of an adult

A

Platelet Count Normal Adult 150 - 400 x 109 / L (Plt)

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11
Q

How many types of WBCs are there?

A

There are the 5 major groups of WBCs - their subtotals should add up to total WBC count

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12
Q

Outline the normal ranges of the 5 major WBCs

A

> Normal Range x 109 / L Adult

Neutrophils	3.5 x 109 / L		2.0 - 7.5  (most abundant)
Lymphocytes	1.7 x 109 / L 		1.5 - 4.0
Monocytes	0.3 x 109 / L 		0.2 - 0.8
Eosinophils	0.2 x 109 / L 		0.04 - 0.4
Basophils	0.0 x 109 / L 		<0.01 - 0.1
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13
Q

What is the significance of neutrophils?

A

Neutrophils are very important in bacterial infections → belong to myeloid pathway

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14
Q

What is neutrophilia?

A

Neutrophilia - high neutrophil count

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15
Q

What is neutropenia?

A

Neutropenia - low neutrophil count

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16
Q

What is a high neutrophilia indicative of?

A

A higher neutrophilia is most likely a haematological cancer not bacterial infection

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17
Q

What is the significance of lymphocytes?

A

Lymphocytes are next most common WBC and they belong to lymphoid pathway (mostly raised in viral infection)

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18
Q

What is lymphocytosis?

A

Lymphocytosis - high lymphocyte count

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19
Q

What is lymphocytopenia?

A

Lymphocytopenia - low lymphocyte count

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20
Q

Which WBCs would we not expect a high frequency?

A

Monocytes, eosinophils and basophils are minority populations in the blood.

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21
Q

What does a high basophil count tell us?

A

Basophils are rarely seen in the blood, so high no.’s indicate a problem (most commonly haematological malignancy) - nothing to do with infections

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22
Q

Describe the significance of eosinophils

A

Eosinophils important in parasitic infections - test stool sample to confirm

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23
Q

When is the eosinophil count elevated?

A

Also important in allergic reactions (ie. skin rash, asthma etc.) increase eosinophil count

Also elevated in some autoimmune diseases

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24
Q

What is the significance of monocytes?

A

Monocytes involved in TB and raised in haematological malignancies

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25
Q

How is a blood film observed in the lab?

A

blood film seen using light microscope

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26
Q

What can we see from a WBC blood film?

A

Looking at the tail end we can identify different cell populations

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27
Q

Describe the normal arrangement of neutrophils on a blood film

A

Have 3-5 lobes and granules
Larger than RBCs
Cytoplasm appears pinkier toned

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28
Q

How can we find out if a RBC is regularly sized just by looking at a blood film?

A

To identify a regular RBC size, compare to lymphocytes (neutrophils are larger than lymphocytes)

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29
Q

Outline normal morphology of lymphocytes on a blood film

A

Similar size to RBC, and contain hardly any cytoplasm (bluer) mostly occupied by large, dense nucleus - no granules

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30
Q

Describe the difference between T and B lymphocytes on a blood film

A

Can’t distinguish between T or B lymphocytes from a blood film

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31
Q

What is the main role of lymphocytes?

A

Major role is in Adaptive Immune Response

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32
Q

How do lymphocytes respond to viral attack?

A

Differentiation between self and non-self e.g. Viral attack
- B Lymphocytes produce antibodies to destroy that
specific virus
- Memory cells

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33
Q

What are the 3 types of lymphocytes?

A

T-cells
B-cells
Natural Killer (NK) cells

34
Q

What is the role of T cells?

A

T-cells are involved in Cell-Mediated Immunity (eat up cells)

35
Q

What are the common types of T cells?

A

CD4+ T-Helper cells

CD8+ Cytotoxic T-cells

36
Q

What is the role of B cells?

A

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

37
Q

What is the role of NK cells?

A

Natural Killer cells are part of the Innate Immune System attacking virally infected cells and tumour cells without developing specific cell mediated response

38
Q

Describe the morphology of a large granular lymphocyte

A

Granular but unlike neutrophils has only one lobe

Cytoplasm is paler

39
Q

What is the morphology of monocytes on a blood film?

A

Biggest cells in a blood film (almost 3x size of RBCs)
Dumbbell shaped nucleus
Bilobed vacuoles also present

40
Q

How do we differentiate between eosinophils and basophils on a blood film?

A

Eosinophils are very similar to neutrophils

But eosinophil granules are a lot denser and orange in colour - looks as though its ready to degranulate (allergic response)

41
Q

Describe basophil morphology on a blood film

A

Thicker, darker granules that hide the nucleus

Very distinctive

42
Q

Where are blood cells formed?

A

Bone marrow produces all blood cells

43
Q

Describe the haematopoietic pathway WBCs belong to

A

Most WBCs belong to the myeloid progenitor pathway

44
Q

Which cells belong to the lymphoid pathway?

A

Lymphoid pathway produces T lymphs , B lymphs and NK cells

45
Q

What is leukocytosis?

A

Leukocytosis: Increase in white cells

46
Q

What is leukocytopenia?

A

Leucopenia: Decrease in white cells

47
Q

How do toxic granulation cells appear on a blood film?

A

Granules are no longer hazy, they are darker and denser (similar to basophil) - basophils are still much thicker

48
Q

Why are toxic granulation cells less developed?

A

Nucleus is less developed (less lobed) as hasn’t had the time to form correct lobed structure as body responding to stress quickly

49
Q

When does myeloid maturation occur?

A

Shift to the LEFT in neutrophil development

Starts with blast cells in bone marrow - should not be seen on blood film - indicative of leukaemia

50
Q

What causes a hypersegmented neutrophil to be present on blood film?

A

shift to the RIGHT

9-12 lobes commonly due to B12 / folate deficiency

51
Q

What are dohle bodies?

A

Toxic granulations form a prominent single rod shaped dohle body - not v. common

52
Q

When is neutrophilia expected to occur?

A

Normal Physiological Reaction
Post-operative
Pregnancy

53
Q

What are the pathological causes o

A
  • Bacterial Infection
  • Inflammation e.g. vasculitis
  • myocardial infarction
  • Carcinoma
  • Steroid treatment - v. common
    neutrophils have steroid receptors
  • Myeloproliferative disorders
  • Treatment with myeloid growth factors
54
Q

How do we investigate neutrophilia?

A
  1. FBC and differential white cell count
  2. Blood film examination
  3. Bacterial culture screen for infection
  4. Bone marrow examination + chromosome analysis
55
Q

How can we investigate neutrophilia using FBC?

A
  • if all cells affected = bone marrow issue
  • RBC = anaemia /iron
  • White cells = leukaemia / infections
56
Q

How is a bacterial culture screen taken in neutrophilia patients?

A

collect samples to test for bacteria:

  • Urine sample
  • Blood sample
  • Sputum sample
  • Skin swab
57
Q

What is the purpose of a bone marrow exam and chromosomal analysis?

A

Testing for (CML) chronic myeloid leukaemia

Philadelphia chromosome: translocation between chromosomes 9 and 22

Molecular analysis for BCR-ABL oncogene

58
Q

What form of white cell disorder is acute myeloid leukaemia?

A

Acute myeloid leukaemia is increased blast cells not mature neutrophilia

59
Q

What causes neutropenia?

A

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

60
Q

What is a non-pathological cause of neutropenia?

A

Racial: ‘benign ethnic neutropenia’ - in afro caribbean and asian populations lower neutrophil count - exclusion diagnosis (last resort)

61
Q

What causes eosinophilia?

A
  • Allergic diseases e.g. asthma, hay fever
  • Parasitic infections
  • Drug sensitivity
  • Myeloproliferative diseases e.g CML
  • Hodgkin’s Lymphoma
62
Q

How can we investigate eosinophilia?

A
  1. FBC and differential white cell count
  2. Blood film examination
  3. Stool examination for ova and parasites
63
Q

What are the causes of monocytosis?

A

Tuberculosis (TB)
Acute and chronic monocytic and myelomonocytic leukaemia
Malaria

64
Q

Outline the investigation of monocytosis

A
  1. FBC and differential white cell count
  2. Blood film examination:
    • for abnormal white blood cells
    • for malarial parasites
  3. Bone marrow examination - leukaemia
  4. TB cultures
65
Q

When is lymphocytosis a normaloccurence

A

Normal Physiological Status in childhood (1-6 years)

Normal: 5.5 – 8.5 x 109/L

66
Q

How does lymphocytosis appear on a blood film?

A

Lymphocyte no. almost equates RBC no. = lymphocytosis

67
Q

What are the pathological causes of lymphocytosis?

A 
Bacterial Infection
Viral Infections e.g.      		     
- Hepatitis
- Mumps 
- Rubella		
- Pertussis
- Glandular Fever    
- (Infectious Mononucleosis)
68
Q

What malignancies is lymphocytosis indicative of?

A

Leukaemia

Lymphoma

69
Q

Which cells become tumorogenic in lymphocytosis?

A

acute lymphoid leukaemia = blast cells

chronic lymphoid leukaemia = normal lymphocytes

70
Q

Where do lymphocytes originate?

A

Lymphomas originate from lymph glands

71
Q

How do we investigate lymphocytosis?

A
  1. FBC and differential white cell count
  2. Blood film examination
    - Atypical mononuclear cells
    - ?Glandular Fever caused by EBV
    (Infectious Mononucleosis; Kissing Disease)
  3. Monospot, Paul Bunnell Test, Clearview
  4. Infectious Mononucleosis Test
72
Q

Describe morphology of atypical mononuclear cells

A

Big = blasts or monocytes
V. pale cytoplasm
Scalloping - seen to just border RBCs

73
Q

What type of cells are atypical mononuclear cells?

A

Resemble monocytes to some extent but actually are lymphocytes that have been transformed due to infection

74
Q

What pathological condition shows atypical mononuclear cells?

A

V. common in infectious mononucleosis or glandular fever

75
Q

What are heterophile antibodies?

A

Heterophile antibodies are antibodies which react against an antigen which is completely unrelated to the antigen which originally stimulated it

76
Q

Give an example of heterophile antibodies?

A

Human antibodies reacting against sheep or horse or bovine cells

77
Q

What is chronic lymphocytic leukaemia?

A

cancer of bone marrow lymphoid cells

78
Q

Which malignancies are mature lymphocytes present in

A

Chronic lymphocytic leukaemia

Lymphoma

79
Q

When is immunophenotyping used to investigate lymphocytosis?

A

Immunophenotyping to determine if lymphocytes are:
B-cells:
- Demonstrate clonality by light chain restriction

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

80
Q

What cytokines are secreted by T cells?

A

T cells always express CD3, 4 & 8

81
Q

Which cytokines do B cells release?

A

B cells express CD 19 & 20

Haematology (8 decks)

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