immunology

Question 1

what is attached to section of antibody that doesn’t bind to antigen

Answer 1

reporter or drug

Question 2

examples of reporter

Answer 2

enzymes, fluorescent probes, magnetic beads

Question 3

examples of enzymes

Answer 3

peroxidase, alkaline phosphatase

Question 4

examples of fluorescent probes

Answer 4

dyes, beads of different sizes

Question 5

what are magnetic beads used for

Answer 5

purification of cell types

Question 6

2 examples of drugs

Answer 6

Kadcyla, anti-HER2 linked to emtansine

Question 7

what allows antibodies to target antigens for diagnostic tests

Answer 7

unique specificity

Question 8

what can antibodies bind to

Answer 8

antigens, other antibodies

Question 9

what are anti-antibodies

Answer 9

target primary antibodies of other species

Question 10

when are antibodies produced by patient

Answer 10

in autoimmune disease, for defence against infection

Question 11

3 techniques to manufacture antibodies

Answer 11

antisera from immunised animals (polyclonal), monoclonal antibodies, genetically engineered antibodies

Question 12

disadvantage of antisera from immunised animals

Answer 12

limited amount produced by animal

Question 13

how are monoclonal antibodies generated

Answer 13

immunise animal with antigen -> remove spleen cells for antibody-producing B-cells -> remove myeloma cells (continue deviding indefinitely) -> fuse B-cells and myeloma cells with detergent to produce hybridomas -> culture in HAT medium and select for positive cells -> clone by isolating each cell (limiting dilution) -> harvest monoclonal antibodies

Question 14

production of antibodies using recombinant DNA technology

Answer 14

place in bacteriophage -> displayed on surface of bacteriophage -> diverse range, and one with specific binding will bind and others will wash off MORE (SLIDE 11)

Question 15

benefit of production of antibodies using recombinant DNA technology

Answer 15

expand massively when growing in bacteria

Question 16

therapeutic uses of manufacture antibodies

Answer 16

prophylactic protection vs microbial infection, anti-cancer therapy, removal of T-cells from bone marrow grafts, block cytokine activity

Question 17

example of antibodies used for prophylactic protection vs microbial infection

Answer 17

IVIG, synagis (anti-RSV)

Question 18

example of antibodies used for anti-cancer therapy

Answer 18

anti-HER2

Question 19

example of antibodies used for removal of T-cells from bone marrow grafts

Answer 19

anti-CD3

Question 20

example of antibodies used for blocking cytokine activity

Answer 20

anti-TNFa

Question 21

issue of using antibodies as treatment

Answer 21

very expensive

Question 22

diagnostic uses of manufactured antibodies

Answer 22

blood group serology, immunoasays, immunodiagnosis

Question 23

what immunoasssays are produced using antibodies

Answer 23

hormones, antibodies, antigens

Question 24

4 things detected by antibodies in immunodiagnosis

Answer 24

infectious diseases, autoimmunity, allergy (IgE), malignancy (myeloma)

Question 25

what does ELISA stand for

Answer 25

enzyme linked immunosorbent assay

Question 26

process of ELISA

Answer 26

add anti-antigen antibody covalently linked to enzyme -> wash away unbound antibody -> enzyme makes coloured product from added colourless substrate -> measure absorbance of light by coloured product

Question 27

process of rapid testing

Answer 27

lateral flow assay architecture; at first strip if molecule present will bind to antibody; excess antibody will go to second strip to bind to anti-antibody to make mark to show test worked

Question 28

why might serum antibodies against HIV be present without the disease

Answer 28

if mother had HIV, antibodies crossed placenta

Question 29

immunological concern about vague aches and pains

Answer 29

immune complexes

Question 30

immunological concern about loss of appetite and weight loss

Answer 30

effect of poor nutrition on bone marrow cells

Question 31

immunological concern about swollen lymph nodes in neck

Answer 31

immune activation

Question 32

immunological concern about fever, rash, small red patches (some lumpy)

Answer 32

acute phase, activation of complement, immune complexes

Question 33

what might immune complexes cause

Answer 33

inflammation and complement activation; serum sickness (in circulation); glomerulonephritis (kidney); deposition at skin, joints and lungs

Question 34

what might look for if immunodeficiency concern and how

Answer 34

serum Ig levels (serum electrophoresis, nephelometry or ELISA), specific antibodies (ELISA), lymphocyte subsets (flow cytometry)

Question 35

serum electrophoresis

Answer 35

compare with healthy control and active immune response; if monoclonal expansion of B cells indicates malignancy so investigate if myeloma

Question 36

lymphocyte flow cytometry

Answer 36

mixture of cells labelled with fluorescent antibody -> stream of fluid containing antibody-labelled cells -> laser to detect which antibody present on lymphocyte to know type and number (CD4+ T cell important in HIV)

Question 37

natural history of HIV infection

Answer 37

slide 27

Question 38

HIV first and second line therapy

Answer 38

every 3 months monitor CD4+ count and viral load, then lower by using anti-HIV drug; second line therapy if first line doesn’t work