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cellular basis of immunity > revision lec > Flashcards

revision lec Flashcards

1
Q

lineage of DCs

A

myeloid (they are innate cells)

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2
Q

role of DCs

A

relay information (antigen-presentation) to the adaptive immune system (CD4+ T cells) with complementary ligand

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3
Q

complement proteins are the

A

soluble components of serum (30 of them)

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4
Q

benefits of lectin pathway

A

direct recognition –> no antibodies involved

important in recognition of fungal residues

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5
Q

benefits of pentraxins

A

can activate classical complement pathway independent of antibodies

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6
Q

how does antibody binding initiate classical pathway

A

activates C1q/r/s complex

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7
Q

importance of b fragment of subunits

A

forms the next protease

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8
Q

amplification loop of alternative pathway

A

C3b required to bind directly to microbial surface to activate alternative pathway
more C3b made when C3 is broken down by C3 convertase into C3a + C3b

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9
Q

final component of complement

A

C3

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10
Q

C3a importance

A

inflammatory mediator

attracts phagocytes

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11
Q

C3b importance

A

binds to complement receptors on phagocytes (alternative pathway)
causes opsonisation of phagocytes
removal of immune complexes

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12
Q

formation of MAC complex

A

after late events of complement
C3b combines with C3 convertase to make C5 convertase
cascade of proteolytic events –> MAC

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13
Q

importance of phage display

A

alternative method to make really highly specific monoclonal antibodies
allows you to make a combinatorial library

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14
Q

method of phage display to make combinatorial library

A

take B cell source e.g. spleen from animal
extract mRNA from the B cells
convert mRNA to cDNA
PCR all the heavy and light chains
all the heavy and light chains from different B cells are mixed up and form many different combinations

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15
Q

controlling localisation of antibody expression on phage

A

express them at specific proteins e.g. pVIII or pIII

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16
Q

why use biopanning?

A

identify specific antibodies expressed on phages

17
Q

method of biopanning

A

introduce phage population to microtitre plate containing wells of antigen
wash away any unbound antibody
UV source kills any other unbound antibodies
(bound antibodies identified by marker)
elute just antibodies of interest (bound)
introduce specific antibody to E.Coli and allow phage s to infect and multiply

18
Q

what is immunofluorescence good for

A

good at looking at/identifying localisation of surface antigens (extracellular)

good at picking up antigens from live cells (not dormant cells) –> important diagnostic marker

19
Q

identifying antibodies at a higher resolution

A

use immunogold electron microscopy

20
Q

role of SDS

A

applies negative charge to molecules

gives molcules the same charge to mass ratio

21
Q

SDS-PAGE in 1D

A

separation on basis of molecular weight

highest molecular weight at top of gel

22
Q

why do we denature the polypeptides in western blotting

A

break protein up into small components to see better localisation of where the antigens are and where the antibody will bind

23
Q

SDS-PAGE in 2D

A

separate peptides on basis of isoelectric point

then apply SDS and separate on basis of molecular charge

24
Q

reason for 2D

A

allows further separation of peptides e.g. even dimers can be separated

25
Q

how do you get the proteins out the gel in western blotting

A

sandwich the gel
expose to an electric current which fixes the proteins permanently to a membrane
can then probe proteins using enzyme-linked antibodies

26
Q

what is DAS-ELISA good for

A

double antibody sandwich

good for measuring components within human serum

27
Q

method of DAS-ELISA

A

capture antibody bound to microtitre plate by its Fc domain
bathe plate in solution containing antigen of interest
antigen binds to Fab domain of capture antibody
add reporter antibody
binds to epitope of antigen

28
Q

features of reporter antibody in DAS-ELISA

A

must bind to different epitope on the antigen than the capture antibody
otherwise capture antibody will bind competitively to antigen and there will be no binding sites left for the reporter antibody

29
Q

why are glycoconjugate vaccines developed

A

there are many limitations of polysaccharide only vaccines

30
Q

limitations of polysaccharide vaccines

A

do not induce memory
induce T-cell independent immune response
immune response is immature (only IgM)
vaccines do not work well in very young or elderly (populations that most require the vaccine)

31
Q

example of protein conjugate used in chemical coupling

A

CRM197 diptheria genetic toxoid protein

32
Q

benefits of glyconjugate vaccines

A

immune response against protein/polysaccharides fully mature
immune memory induced
long-lasting

33
Q

example glycoconjugate vaccine and against what disease

A

prenvar 7

against streptococcus pneumoniae

cellular basis of immunity (21 decks)

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