Lecture

 13  –  Review  Questions  

 
Slide  2:  
1. Describe  the  route  and  cells  in  order  of  antigen  entering  lymph  nodes  all  the  
way  through  T-­‐independent  B  cell  activation.  
a. Antigen  brought  in  by  Macrophages  
b. Antigen  handed  off  to  ______________  
c. Antigen  then  handed  off  to  _____________  
d. Antigen  then  handed  off  to  FDC  
e. B  cells  then  recognize  antigen  displayed  by  FDC  in  the  Cortex  (region  
of  lymph  node)  
f. If  no  CD40  signal,  B  cells  become  Short  lived  plasmoblasts  and  form  a    
Primary  Foci  producing  low  affinity,  pentameric  IgM  antibodies  
 
Slide  3:  
1. Describe  the  route  and  cells  in  order  of  antigen  entering  lymph  nodes  all  the  

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way  through  T-­‐dependent  B  cell  activation.  

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a. Antigen  brought  in  by_____________  

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b. Antigen  handed  off  to  ______________  
c. Antigen  then  handed  off  to  _____________  

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d. Antigen  then  handed  off  to  _____________  
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e. B  cells  then  recognize  antigen  displayed  by  FDC  in  the  __________  
(region  of  lymph  node)  
f. If  _____________  signal  received,  B  cells  form  _____________  and  undergo  
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_____________,  _______________  and  ____________  

g. B  cells  become  either  long-­‐lived  _____________  or  _____________  
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Slide  4:  
1. What  are  the  three  zones  of  a  germinal  center?  
Dark  zone,  light  zones,  mantle  zone  
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2. What  occurs  within  the  dark  zones,  which  type  of  B  cells  can  be  found  there  
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and  are  these  B  cells  dividing  or  not?  

Rapidly  dividing,  centroblasts,  somatic  hypermutation  
3. What  occurs  within  the  light  zones,  which  type  of  B  cells  can  be  found  there  
and  are  these  B  cells  dividing  or  not?  
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Non  –  dividing  ,  centrocytes,  affinity  maturation  

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4. Which  zone(s)  also  contain  FDC  and  Tfh  cells  and  what  do  these  cells  do  
there?  
Light  zones  
 
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Slide  5:  
1. Which  type  of  B  cells  express  AID  and  what  process  are  these  cells  
undergoing?  
Centroblasts,  somatic  hypermutation  
2. Where  (which  zone)  is  AID  expressed?  
Dark  zones  

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 3. Which  process  do  centrocytes  undergo  and  what  is  its  function?  
Affinity  maturation  
4. What  are  the  3  steps  of  affinity  maturation?  
• B  cell  interaction  with  Ag  on  FDC    
• B  cell  presentation  of  peptides  derived  from  Ag  to  T  cells  
• Antigen  capture  from  SCS  macrophages  
5. Somatic  hypermutation  and  affinity  maturation  will  keep  occurring  until…?  
Germinal  center  FDC  and  SCS  macrophages  run  out  of  Ag  or  B  cells  are  
sufficiently  high  affinity  
6. What  are  the  two  possible  outcomes  for  B  cells  that  have  undergone  somatic  
hypermutation  and  affinity  maturation?  
Long  lived  plasma  cells  or  memory  B  cells  
 
Slide  6:  
1. What  is  the  key  enzyme  involved  with  somatic  hypermutation?  

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AID  (  Activation  induced  deaminase)  

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2. What  is  the  enzymatic  activity  of  this  ^  enzyme?  

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Converts  cytidine  to  uridine  

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3. Which  regions  of  the  BCR  DNA  are  targeted  by  AID?  

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HVR  
 
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Slide  7:  
1. What  are  the  3  possible  ways  AID  mutations  can  be  repaired?  
Proliferation  of  mismatched  bases  
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Rev  replaces  with  random  base  

Proliferation  results  in  additional  diversification  
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2. What  is  the  role  of  UNG  in  SMH?  

Creates  abasic  site  
3. What  is  the  role  of  MSH  in  SMH?  
Fix  UàC  but  mutate  AT  pairs  
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4. Which  of  the  3  repair  mechanisms  is  predictable  and  which  are  
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unpredictable?  
 
Slide  8-­‐9:  
1. Be  able  to  list/draw  the  steps  of  the  cyclic  re-­‐entry  method  of  somatic  
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hypermutation/affinity  maturation:  
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a. B  cells  receive  CD40  signal  from  CD4  T  cells  

b. B  cells  then  formGerminal  centers  in  the  Dark  zones  
c. B  cells  in  the  dark  zones  become  centroblasts  
d. Centroblasts  in  the  dark  zones  undergo  Somatic  hypermutation  of  
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their  BCR  genes  

e. After  the  cells  divide,  they  move  into  the  _Light    zone  
f. Cells  that  have  reached  the  light  zone  are  then  called  _Centrocytes  
g. Centrocytes  are  non  dividing_  (dividing/nondividing)  cells  
h. Centrocytes  interact  with  Ag:iC3b  complexes  on  FDC  

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 i. They  then  present  Ag  peptides  to  T  FH  
j. If  they  receive  a  good  signal  and  CD4  T  cell  help  they  will  Survival  /  
activation  signal  
k. If  they  receive  a  bad  signal  and/or  no  CD4  T  cell  help  they  will  
_Apoptosis  
l. If  the  cells  received  a  good  signal  +  CD4  T  cell  help  they  will  then  
capture  antigen  from  _SCS  macrophages  and  go  back  to  the  _Dark  zone  
and  repeat  the  entire  SMH+AM  process  
m. When  _Antigen  runs  out_  or  _B  cells  recievd  stron  BCR  signal__  the  B  
cells  will  exit  to  the  periphery  and  differentiate  into  either  long  lived  
_antibody  secreting  plasma  cells_  or  _memory___  B  cells  
 
Slide  10:  
1. What  does  the  process  of  isotype  switching  result  in?  
B  cells  change  the  function  of  the  antibody  produced  
2. Which  isotypes  are  ALWAYS  initially  produced?  

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IgM  (with  minimal  IgD)  

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3. What’s  the  main  function  of  the  following  isotypes  and  where  are  they  found:  

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a. IgM  (IgD):    
Complement  activation  

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b. IgG:  
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Serum:  neutralization,  opsonization  for  degradation,  NK  cell  
activation,  complement  activation  
c. IgA:  
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Mucosal:  microbial  neutralization  in  mucosa  

d. IgE:  
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Epithelial  surface:  bind  to  Fc  receptors  on  mast  cells  

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Slide  11:  
1. Which  region  (viable/constant,  heavy/light)  of  an  antibody  determines  the  
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isotype?  
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Viable,  constant  
Slide  12-­‐13:  
1. What  is  the  role  of  switch  regions  in  isotype  switching  and  where  are  they  
found  (i.e.  where  are  they  found  within  the  DNA  seq)?  
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Occur  upstream  of  the  first  exon  for  each  C  gene  segment  
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2. What  do  S-­‐transcripts  form,  where  do  they  form  it  and  how  does  this  allow  
AID  to  begin  isotype  switching?  
Recombination  loops  
3. What  are  the  three  (key)  enzymes  involved  with  isotype  switching  and  what  
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is  their  function?  
AID  demaminates  many  CàU  
UNG  cleaves  Uà  abasic  sites  
4. Which  of  the  three  enzymes  involved  in  isotype  switching  is  NOT  involved  in  
SMH?  
APE  1  

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Slide  14:  
1. After  Ape-­‐1  creates  nicks  in  DNA,  which  two  enzymes  are  involved  with  
repairing  the  nicks  during  isotype  switching?  
Ku  70/80  and  DNA  ligase  
2. What  other  process  (that  we  have  discussed)  are  these  two  enzymes  
involved  in?  
Somatic  Recombination  
3. Why  is  APE-­‐1  double-­‐stranded  nicking  favored  over  nucleotide  repair  (i.e.  
from  MSH/REV)  during  isotype  switching?  
Stable  R  loops  favor  double  stranded  nicks  by  APE-­‐1  
Slide  15:  
1. In  which  type  of  B  cells  does  somatic  recombination  occur  and  what  is  this  
process  controlled  by?  
Developing  B  cells:    
Controlled  by  recombina8on  signal  sequences  RSS  (12/23  rule)  

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2. In  which  type  of  B  cells  does  somatic  hypermutation  occur  and  what  is  this  

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process  controlled  by?  

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Activated  centroblast  B  cells  
Controlled  by  open  DA  structures  during  active  transcription  of  Ig  

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3. In  which  type  of  B  cells  does  isotype  switching  occur  and  what  is  this  process  
ou urc
controlled  by?  
Activated  centroblast  B  cells  
Controlled  by  stable  open  DNA  structures  (R-­‐loops)  formed  from  low  level  
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transcripts  of  C-­‐region  RNA  pairing  with  DNA  and  Ku70/80  

 
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Slide  16:  
vi y re

1. For  the  following  cytokines  describe  what  they  are  produced  in  response  to,  
what  they  signal  through,  what  they  induce  and  what  they  result  in:  
Cytokine   Produced  in   Signals   Induces:   Results  in:  
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response  to:   through:  

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IFNγ   Systemic   STAT1  dimers   Ig  gamma-­‐S-­‐ Systemic  

infections   transcripts   antibodies  
TGFβ   T  cells  in   Smad2/3   Ig  alpha  S-­‐   Mucosal  
mucosal   transcripts   antibodies  
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lymphoid  
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tissues  
IL-­‐4   Extracellular   STAT3/6   Ig  epsilon  S-­‐   Immunity  at  
pathogens  at   transcripts   epithelia  
epithelial   surfaces  
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surfaces  
 
Slide  17:  
1. What  is  the  main  function  of  plasma  cells?  
Produce  large  amounts  of  antibody  

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 2. Where  do  plasma  cells  exist?  
Bone  marrow  
3. Describe  the  difference  (i.e.  isotypes)  of  plasma  cell  Abs  and  initial  short  lived  
Ab-­‐producing  cells  during  an  infection?  
Antibodies  are  proteins  and  have  short  half  lives  so  continually  needs  to  be  
replenished  
Plasma  cells  have  long  half  lives  but  still  needs  to  be  replenished  over  time  by  
memory  B  cells  
4. About  how  much  antibody  can  be  produced  by  a  single  plasma  cell  per  day?  
1  mg    
5. Why  does  so  much  antibody  need  to  be  produced  daily?  
Short  half  lives  
6. Are  plasma  cells  short-­‐  or  long-­‐lived,  and  how  are  they  eventually  
replenished?  
Ling  lived  by  memory  B  cells  
 

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Slide  18:  

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1. What  are  the  two  main  functions  of  memory  B  cells?  

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Replace  plasma  cells  
Provide  rapid  recall  B  cell  response  

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2. Where  do  memory  B  cells  exist?  
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Circulate  In  the  blood  
3. Describe  two  ways  that  memory  B  cells  are  different  from  naïve  B  cells.  
Memory  B  cells  have  higher  affinity  BCR  and  can  acquire  antigen  directly  for  
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internalization,  processing  and  presentation  to  T  cells  

 
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Slide  19:  
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1. Which  provides  a  faster  immune  response  to  a  given  antigen,  naïve  B  cells  or  
memory  B  cells  and  why?  
Memory  B  cells  
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Don’t  require  innate  signals  for  activation  

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Can  directly  bind  and  internalize  antigens  for  presentation  to  T  cells  to  
initiate  formation  of  germinal  centers  
 
Slide  20:  
is

1. What  cell  type  (centroblasts/centrocytes)  forms  primary  foci  and  what  do  
Th

these  cells  produce?  

 
2. Which  type(s)  of  cells  produce  IgG/IgA/IgE  antibodies?  
3. Be  able  to  draw  a  graph  displaying  the  difference  in  IgM  and  IgG  production  
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during  both  a  primary  infection  and  a  secondary  infection  (hint:  each  isotype  
should  have  two  peaks,  be  able  to  explain  what  the  peaks  are  in  response  to  
and  how  they  are  different  for  IgM  and  IgG).  
 
 
 

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