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2.1 Molecular Immunology > Greg - Clinical Immunology (Allergy) > Flashcards

Greg - Clinical Immunology (Allergy) Flashcards

1
Q

What are the clinical areas in the immunology lab?
(3)

A

Autoimmunity

Primary/secondary immunodeficiency

Allergy

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2
Q

What is done in an autoimmunity lab?

A

Screening for autoantibodies which are indicators for disease

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3
Q

Give an example of some antibodies measured in an autoimmunity lab
(5)

A

Antinuclear antibodies

Anti-neutrophil cytoplasmic antibodies

tTG antibodies

Rheumatoid factor

Anti-cyclic citrullinated peptide antibodies

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4
Q

List some methods of detecting antibodies in the autoimmunity lab
(3)

A

ELISA/immunoassay

Indirect immunofluorescence

Direct immunofluorescence

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5
Q

What is done in a primary/secondary immunodeficiency laboratory

A

Detecting dysfunction in the immune system caused by inherited (primary) or non-inherited (secondary) immunodeficiency

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6
Q

What methods are used in an immunodeficiency laboratory?

A

Flow cytometry
Cell function assays
Complement assays
Serum protein electrophoresis

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7
Q

What is carried out in an allergy laboratory?

A

Detecting allergies against environmental antigens

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8
Q

What methods are used in an allergy laboratory?
(3)

A

Specific IgE

Mast cell tryptase

Basophil activation

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9
Q

Why would we need to measure specific antibody responses?

A

To test for vaccination

To test for allergens

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10
Q

Why would we need to test for antinuclear antibodies

A

To test for lupus

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11
Q

What antibodies do we test for in coeliac disease?

A

Tissue transglutaminate antibodies (tTG antibodies)

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12
Q

What antibodies do we test for in lupus?

A

Antinuclear antibodies

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13
Q

What antibodies do we test for in rheumatoid arthritis?

A

Anti-cyclic citrullinated peptide antibodies

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14
Q

What is a primary immunodeficiency and give an example?

A

An inherited condition you are born with

e.g. X linked agammaglobulinaemia

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15
Q

What is a secondary immunodeficiency and give an example?

A

Immunodeficiency developed in life

e.g. HIV

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16
Q

What can we determine in an allergy lab?

A

Diagnosis and prognosis of an allergy

How severe your allergy will be and what causes the allergy

17
Q

What is an allergy?

A

An inappropriate immune response to a harmless environmental antigen

18
Q

List the components that are mainly involved in allergy
(5)

A

Mast cells

T cells

B cells

IgE

Histamine

19
Q

What are the differences between an allergy and an intollerance?

A

Allergies are rapid onset, reproductible reactions

Intolerances are usually slower in onset and are not life-threatening

20
Q

List the main symptoms of allergies
(6)

A

Asthma

Eczema

Rhinoconjunctivitis

Urticaria

Vomiting/diarrhoea

Systemic anaphylaxis/Death

21
Q

What test is usually carried out within 6 hours after an allergic reaction?

A

Test for mast cell tryptase

Which detects products of mast cell degranulation

22
Q

What are the requirements of a type 1 hypersensitivity?
(4)

A

Requires sensitisation

Multiple genetic factors contribute

Environmental factors contribute

Risk is conferred by many genes including HLA and cytokines

23
Q

What usually happens to IgE after a while?

A

It degrades

24
Q

What happens to IgE when it binds to mast cells?

A

IgE doesn’t break down and can remain stable

25
Q

What happens when IgE bound to mast cells become cross linked?

A

It results in histamine being released (mast cell degranulation) and thus an allergy

26
Q

What are the two mechanisms of diagnosing an allergy?

A

Measurement of IgE

Measurement of mast cell products such as tryptase

27
Q

How is total IgE measured?

A

By immunoassay via chemiluminescence or fluorescence based means

28
Q

Why is high IgE regarded as non specific?
(3)

A

It doesnt just indicate allergy

Indicates parasitic diseases

Or conditions such as hyper-IgE syndrome

29
Q

What is a specific IgE test?

A

Measurement of the level of IgE directed against the suspected allergen

30
Q

What is ELISA?

A

Enzyme-linked immunosorbent assay

A method which utilises the specificity of antibodies to quantify the level of a soluble analyte of interest

31
Q

How is ELISA usually carried out?
(2)

A

Over multi-well plates or tubes (immunocap)

A labelled commercially available antibody is used to detect the molecule of interest

32
Q

What are the three different types of reporter molecules?

A

Colorimetric

Chemiluminescence

Fluorescence

33
Q

Whats the difference between an indirect and a direct ELISA?

A

Indirect -> antigen coats well, serum sample added, if antibodies present they bind to antigen, labelled antibody is added, amount of fluorescence is labelled

Direct -> labelled antibody binds to antigen

34
Q

How do we use ELISA?

A

A series of standards with known concentration of antibody come with the bought in labelled antibody are run alongside the unknowns

Form standard curve

Measure sample against curve

35
Q

How do you detect allergen specific IgE using an immunocap?
(7)

A

Antigen is pre-coated on a ‘cap’

Diluted patient serum is added and if the specific IgE is present will bind

Unbound serum proteins are washed away

A fluorescent-labelled secondary anti-IgE is added

Unbound secondary antibody is washed away

Fluorescence signal is measured

Fluorescence is proportional to anti-peanut antibody concentration

2.1 Molecular Immunology (19 decks)

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