Immunological Methods for Diagnostics and Therapeutics

Tutorial 11: Immunological Methods for Diagnostics and Therapeutics

Antibodies

1. Describe the properties and immunological mechanism of action of the therapeutic antibodies Trastuzumab and Rituximab

Flow Cytometry

1. Using the information shown in the dual-parameter histogram of leukocytes from peripheral blood (below). Describe the characteristics of the cells shown in relation to their forward and side scatter.

2. Using the single parameter histogram shown below of fluorescently labelled cells describe the characteristics of the cells in this population. What does the area under the peak signify?

3. You have undertaken an experiment to look the activation status of T cells in a population. The CD4 T cell marker is identified using a red dye, the activation status CD69 is identified using a green dye. A colleague gives you a breakdown of the data as follows.

CD4 +ve-CD69 –ve = 26% cells

CD4-ve  – CD69-ve = 18% cells

CD4-ve – CD69 +ve = 22% cells

CD4 +ve-CD69 +ve = 34% cells

Put the % cell number values in the histogram shown overleaf as they would appear on the FACS machine.

Green

Red

Diagnose the patient!

1. Sam, a two-year-old child has failed to thrive. He has had allergic reactions at injection sites and showed a poor response to his childhood vaccinations. Blood tests are shown in Table 1:

	Reference Range	Patient
White blood cell count	6.0 – 14.0 x 109/L	3.00 x 109/L
Neutrophils	2.0 – 6.0 x 109/L	2.50 x 109/L
Lymphocytes	2.5 – 8.0 x 109/L	0.60 x 109/L
Monocytes	0.2 – 1.0 x 109/L	0.30 x 109/L
Haemoglobin	115 -140 g/dL	90 g/dL
Platelet count	150 – 400  x 109/L	125x 109/L
C-reactive Protein	<10 mg/L	14 mg/L
Erythrocyte Sedimentation Rate	<15.0 mm/h	20mm/h
IgA	0.40 – 2.0 mg/dL	0.2 mg/dL
IgM	39 – 240 mg/dL	2.5 mg/dL
IgG	4.9 – 16.1 mg/dL	1.5 mg/dL

• A five year old child was admitted to hospital following repeated viral infections. She had recurrent respiratory infections since birth. Blood test results are shown in Table 2:

	Reference Range	Patient
White blood cell count	6.0 – 14.0 x 109/L	3.00 x 109/L
NK cells	2.0 – 6.0 x 109/L	0.50 x 109/L
Lymphocytes	2.5 – 8.0 x 109/L	0.50 x 109/L
Haemoglobin	115 -140 g/dL	105 g/dL
Platelet count	150 – 400  x 109/L	135 x 109/L
C-reactive Protein	<10 mg/L	15 mg/L
Erythrocyte Sedimentation Rate	<15.0 mm/h	20 mm/h
IgA	0.40 – 2.0 mg/dL	0.1 mg/dL
IgM	39 – 240 mg/dL	0.8 mg/dL
IgG	4.9 – 16.1 mg/dL	0.1 mg/dL

• Patient 3, a 3-year-old child, was examined by his GP following reports of repeated bacterial infections. His blood test results showed a deficiency in C3b.

Exam essay topics

In last week’s tutorial, you were provided with your exam topics. To prepare for your exam revision, write a plan for your chosen topic area (choose one):

• Primary immunodeficiencies
• CAR-T-cells
• Checkpoint inhibitors

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