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I need to learen > Immunology Lab > Flashcards

Immunology Lab Flashcards

1
Q

Describe the diagnostic uses of antibodies

A

Determining blood groups
Immunoassays
Immunodiagnosis

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2
Q

What can immunoassays be used to measure

A

Hormones
Antibodies
Antigens

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3
Q

What can immunodiagnosis assist in diagnosing

A

Infectious diseases
Autoimmunity
Allergy (IgE)
Malignancy (myeloma)

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4
Q

What are the therapeutic uses of manufactured antibodies and give examples of each

A

Prophylactic protection against microbial infection e.g. IV-IG , synagis (anti-RSV)

Anti-cancer therapy e.g. anti-HER2

Removal of T-cells from bone marrow grafts e.g. anti-CD3

Block cytokine activity e.g. TNF-α in rheumatoid arthritis

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5
Q

What can be attached to antibodies’ constant region for diagnostic testing

A

Enzymes: e.g. peroxidase, alkaline phosphatase

Fluorescent probes: dyes, beads of different sizes

Magnetic beads: e.g. purification of cell types

Drugs: e.g. Kadcyla, anti-HER2 linked to emtansine

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6
Q

What feature of antibodies make them useful in diagnostics

A

unique specificity of antibodies for their target antigens

Can be raised against almost any antigen (often but not always proteins), including immunoglobulins from other species (anti-antibodies)

Can perform indirect labelling using anti-antibody

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7
Q

What are the sources of antibodies

A

Produced by the patient (autoimmune disease or defence against infection)

Manufactured
antisera from immunised animals (polyclonal)
Monoclonal antibodies
Genetically engineered antibodies

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8
Q

Explain how monoclonal antibodies are generated

A
  1. Immunise an animal with the antigen for the desired antibody
  2. Take the spleen cells – source of antibody-producing b cells
  3. Fusion of the B cells and myeloma cells to form hybridomas
  4. Culture in HAT medium,
  5. Select for the positive cells and grow hybridomas with enzyme
  6. Clone by limiting dilution: put single cells in wells à screen clones for those that produce antibody
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9
Q

How are antibodies produced using recombinant DNA technology

A
  1. Isolate population of genes encoding antibody variable regions
  2. Construct fusion protein of V region with a bacteriophage coat protein
  3. Cloning a random population of variable regions to give rise to a mixture of bacteriophages (phase-displat library)
  4. Select phage with desired V regions by specific binding to antigen
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10
Q

What is ELISA

A

Enzyme Linked Immunosorbent Assay

Use of anti-antibodies for rapid diagnosis

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11
Q

Describe rapid ELISA testing

A
  1. Antigen/antibody is immobilised in a well
  2. Antibodies added to bind to antigen
  3. Complex moves down and binds a second antibody on the test line
  4. Complex moves further to the control line (where only 1st antibodies bind to show sample has moved down)
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12
Q

What feature of myeloma cells make them useful for monoclonal antibody production

A

Grow indefinitely and do not produce own antibodies, no particular enzymes

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13
Q

What are the features of hybridomas used in monoclonal antibody production

A

Immortal
Have enzyme
Produce desired antibodies

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14
Q

What does the suffix -omab suggest

A

mouse monoclonal

e.g. Muronomab = anti-CD3, transplant immunosuppression

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15
Q

What does the suffix -imab suggest

A

chimeric or partly humanised

e.g. Infliximab (Remicade) = anti-TNFa, Rituximab = anti-CD20

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16
Q

What does the suffix -umab suggest

A

fully human antibody

e.g. Palivizumab, anti-RSV (Synagis)

17
Q

Which tests are carried out if immunodeficiency is suspected

A

Serum immunoglobulin levels
Tests for specific antibodies (ELISA)
Lymphocyte subsets

18
Q

When is a test for specific antibodies carried out and what could be checked for

A

After a given vaccine, antibodies should be investigated

If immunodeficient, check for protein antigens (Tetanus & Haemophilus)
Or polysaccharides antigens (Pneumococcus)

19
Q

Which markers are found on the T cells, T helper cells, cytotoxic T cells, B cells and NK cells

A 
CD3+ found on all T cells 
CD4+ = T helper cells
 CD8+ = cytotoxic T cells
CD19+ = B cells
CD56+ = NK cells
20
Q

Which markers are found on the T cells, T helper cells, cytotoxic T cells, B cells and NK cells

A 
CD3+ found on all T cells 
CD4+ = T helper cells
CD8+ = cytotoxic T cells
CD19+ = B cells
CD56+ = NK cells
21
Q

Describe the history of HIV infections

A

Initial wide dissemination of the virus, seeding of lymphoid organs
CD4 T cell decreases during clinical latency
Increase after around 7 years

22
Q

Describe ART treatment for HIV patients

A

HIV patients get CD4 count + viral load measured, before + after receiving ART
CD4 T cell count defines extent of immune damage - predicts short-term outlook in HIV-1 patients w/o ART are more susceptible to infections e.g. opportunistic infections or Kaposi’s sarcoma

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