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16 Cards in this Set
- Front
- Back
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Too much ultraviolet light can do what |
Cause damage to tissues |
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Whats the two types of immunisation mechanism of protection |
Passive immunisation Adoptive immunisationhow |
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How to get passive immunisation |
Inject killed pathogen (test vaccine( in mice) Challange after 10 days- animal remains healthy, active immunisation Transfer serum to other mice and challenge with lethal dose of pathogen Animal remains healthy so passive immunisation |
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How to get adoptive immunisation |
Inject killed pathogen and challange after 10 days, animal remains healthy so active immunisation, Transfer spleen cells to another mouse and challange with lethal dose of pathogen, animal remains healthy so adoptive immunization |
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Targeting specific molecules in vivo allows us to identify specific molecules to inhibit in patients using targeted therapeutics, give an example |
In arthritis we have confirmed a important role for TNF-a so now we usr a specific antibody to inhibit in patients |
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How can we develop therapeutics in vivo experiments |
Identify specific molecules important in a given disease so we can target them |
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What is ficoll histopaque |
Used in cell isolation you have diluted blood on top half of test tube and ficoll hypaque specificity on bottom Certrifugation of this leads to peripheral blood mononuclear cells on top and red blood cells and granulates on bottom |
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Using antibodies and magnets we can be more specific and isolate a certain type or subset of cells for further analysis, how does this work |
Heterogenous population of lymphocytes is mixed with antibodies coupled to paramagnetic particles or beads and poured over and iron wool mesh When a magnetic field is applied the coupled cells stick to iron wool, unlabelled cells are washed out, then magnetic field is removed releasing the Coupled cells |
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How could we identify how many cells we have isolated are B cells and how many are T cells |
flow cytometry |
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For each mitogen whats the responding cells PHA, Concanavalin (conA), PWM and lipopolysaccharide |
Pha- T cell. ConA-T cell. PWM- T and B cell. LPS- B cells(mouse) |
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What are examples of mitogens |
Phytogenagglutinin PHA, Concanavalin (conA), pokeweed mitogen PWM and lipopolysaccharide LPS. |
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Using an enzymatic approch for cell proliferation whats a MTT assay |
Relies on enzymes in the mitochondria converting salt into coloured product Only living cells have active mitochondria, amount of colour is proportional to number of cells which is proportional to amount of proliferation. |
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Whats the radioactive approach of cell proliferation |
3H-thymidine incorportation Source of radioactive thymidine is added to cell culture As cells proliferate they incorporate this into the DNA of daughter cells resulting in radioactive cells Measure the amount of crll associated radioactivity, proportional to the amount of cell division |
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What is the fluorescence approche for measuring cell proliferation |
Label proteins of cells with a fluorescent die If cell divides, each daughter cell is half as bright Allows accurate quantification of the number of cells divisions within the population |
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Give an example of the vital dyes that enter the cell and stain them |
Trypan blue, can see down conventional microscope Propidium iodide- becomes fluorescent when binds DNA |
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Measuring cell functions-give an example of how to measure cell killing using radioactivity or fluorescece |
Label target cells with Na2 51CrO4 Add cytotoxic T cells to labelled target cells Killed cells release this radioactive chromium |
