Lecture 5 - B cell Development Flashcards

1
Q

True or false - the antigen binding site of an antibody occurs at the C-terminus.

A

False - occurs at the N-terminus. The C-terminus is the Fc region.

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2
Q

How many different light chain constant domains are there, and what are they?

A

Two - lambda and kappa

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3
Q

How many different heavy chain constant domains are there, and what are they called?

A

Five (major ones) - alpha, mu, gamma, delta and epsilon.

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4
Q

Name the process that allows such vast diversity of B cell paratopes.

A

Somatic gene rearrangement

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5
Q

How many different possible kappa segments are there? What level does Variable kappa and joining segment recombination occur at?

A

About 85

The level of the DNA

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6
Q

What level does the combination of the VJ segment with the Constant kappa segment occur at?

A

RNA (during splicing)

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7
Q

How many Variable, Diversity and Joining segments are there for the heavy chain of antibodies? At what level are these recombined?

A
V = 134
D = 13
J = 4

Level of the DNA

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8
Q

At what level is the newly re-arranged heavy chain VDJ segment combined with a mu or delta constant domain?

A

The RNA - during splicing

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9
Q

What mechanism prevents B cells producing two different antibodies?

A

Allelic exclusion - only one chromosome is subject to re-arrangment

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10
Q

Which B cell receptor chain - heavy or light - is rearranged first? What is the B cell called after that?

A

Heavy

Pre B cell

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11
Q

Once both chains of the B cell receptor have been re-arranged, what is the B cell known as? What process still has to occur before the cell is ‘mature’?

A

Immature B cell

Differential RNA splicing > IgM or IgD producing

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12
Q

Regarding B cells;

Blast cells may differentiate into what?

A

Plasma cell or memory B cell

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13
Q

How do B cells make both soluble and membrane-bound antibody?

A

Use of a different polyadenylation site to make a different mRNA for that antibody, which has 20 hydrophilic aa’s at the C-terminus, rather than the ~40 hydrophobic aa’s.

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14
Q

How is a B cell able to produce both IgM and IgD at once?

A

Splice the RNA for the antibody molecule differently, as IgD constant region is just downstream of the IgM

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15
Q

What level (DNA or RNA) does class switching from IgD to any other class occur at?

A

DNA

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16
Q

After IgD, what is the order of the subsequent constant regions along the DNA strand?

A

Gamma, epsilon, alpha

17
Q

What does ‘polyclonal antibodies’ mean?

A

Mix of antibodies collected from an animal’s sera following immunisation against something. There will be a mix of isotypes and paratopes.

18
Q

True or false - polyclonal antibodies will be slightly different for different animal donors immunised against the same pathogen.

A

True

19
Q

Briefly, how are monoclonal antibodies produced?

A

Immortalise a B cell by fusing with a myeloma cell, work out its antibody paratope and isotype, collect and purify it.

20
Q

What is an indirect ELISA looking for ? (Antibody or antigen)

A

Antibody

21
Q

What is a sandwich ELISA looking for? (Antibody or antigen) Name one veterinary application.

A

Antigen

E.g detection of IFN-gamma produced by T cells stimulated by bovine tb antigen to determine if cow has seen this bacteria before.