1.1b Immunology MICROBIOLOGY

Dr. Fontanilla |June 18, 2013 1st 2013-2014

OUTLINE   C.  T-­‐cell  Proliferation  and  Differentiation  

I.  Cell-­‐Mediated  Immunity   III.  Complement  System  
A.  Development  of  T-­‐Cells   IV.  Cytokines   • MHC  Class  II  presentation:  2  signals  needed  to  activate  naïve  CD4  T-­‐
B.  T-­‐Cell  Receptor  For  Antigen   V.  Hypersensitivity   cells  
C.  T-­‐Cell  Proliferation  and   VI.  Deficiencies  of  the  Immune   1. T-­‐cell  receptor  interacting  with  the  MHC-­‐peptide  complex  
Differentiation   Response   presented  on  the  APC  
II.  T-­‐Effector  Cell  Function  
o CD4  glycoprotein  on  the  naïve  T-­‐cell  acts  as  co-­‐receptor  
A.  CD4  Effector  Cells  
B.  CD8  Effector  Cells   à  ensures  stability  between  T-­‐cell  and  APC  
  2. Interaction  of  B7  costimulatory  molecules  on  the  APC  with  
OBJECTIVES   CD28  on  the  T-­‐cell  
  o T-­‐cell  secretes  cytokines  IL-­‐2  and  IFN-­‐γ  
I. Describe  the  basic  principles  of  immunology,  particularly  as  they  relate  to  
o T-­‐cells  are  able  to  proliferate  and  differentiate  into  
response  to  infection.  
II. Define  the  following  terms:   effector  cells  
A. Immune  response   • MHC  Class  I  presentation:  CD8  activation  
B. Innate  immunity  
C. Adaptive  immunity   T-­‐cell  receptor  interacts  with  MHC  class  I  –  peptide  complex  on  the  
D. Antigen  
infected  cell  
E. Antibody  
F. Major  histocompatibility  complex   ⇓  
G. Complement   CD8  glycoprotein  acts  as  a  co-­‐receptor  (binding  to  MHC  class  I  
H. Cytokines   molecule  on  the  APC)  
I. Hypersensitivity   ⇓  
J. Immunodeficiency  
Once  activated  the  cytotoxic  T-­‐cell  produces  IL-­‐2  and  IFN-­‐γ,  growth  
III. Differentiate  humoral  and  cellular  immunity  
IV. Describe  the  functions  of  antibodies   and  differentiation  factors  for  T-­‐cells  (independent  of  co-­‐stimulation  
V. Enumerate  and  describe  the  different  hypersensitivity  reactions   unlike  CD4)  
VI. Describe  the  more  common  deficiencies  of  the  immune  response   ⇓  
  Virus-­‐infected  cell  is  destroyed  through  cytotoxic  granules  released  
Reference:  Jawetz,  Melnick  &  Adelberg’s  MEDICAL  MICROBIOLOGY,  26th  Edition,  
from  the  CD8  T-­‐cells  
2013,  Chapter  8,  pp  123-­‐148.  
   
 

ADAPTIVE  IMMUNITY  
I.  CELL-­‐MEDIATED  IMMUNITY  
A.  Development  of  T-­‐Cells  
• T-­‐cell  progenitor  cells  undergo  differentiation  in  the  thymus  
• Under  the  influence  of  thymic  hormones,  T-­‐cells  differentiate  into  
committed  cells  expressing  a  specific  T-­‐cell  receptor  (TCR)  
• Most  T-­‐cells  with  TCR  become  positive  for  both  CD4  and  CD8  (co-­‐
receptor  molecules)  
• T-­‐cells  undergo  selection  process  that  result  in  the  retention  only  of  
those  cells  with  antigen  receptors  for  nonself  or  foreign  antigens  –  
99%  die  in  the  thymus  
• Those  that  survive  undergo  final  maturation  process  which  express  
either  CD4  or  CD8    
• Exit  into  the  periphery  to  mature  into  effector  cells   Figure  1.  Activation  of  Naïve  T-­‐cell.  The  red  triangle  is  the  antigen,  together  
with  MHC  II  on  the  surface  of  APC.  On  the  surface  of  Naïve  T  helper  cell  is  the  T-­‐
B.  T-­‐cell  Receptor  for  Antigen   cell  receptor  (TCR)  and  a  co-­‐stimulatiry  receptor,  CD28.  
• T-­‐cell  receptor  proteins  have  variable  and  constant  regions  similar  to    
antibodies   II.  T-­‐EFFECTOR  CELL  FUNCTION  
• Variable  regions  are  located  at  the  amino  terminals  of  the   A.  CD4  Effector  Cells  
polypeptide  chain  farthest  away  from  the  cell  membrane    
• Both  chains  contribute  to  the  variable  domain  that  interact  with   Table  1.  CD4  Effector  Cells  
antigen  presented  by  MHC  complex   Th1  Cells   • Differentiate  in  the  presence  of  IFN-­‐γ  
• CD4  and  CD8  function  as  co-­‐receptors   • Activate  macrophages  
• During  recognition  of  antigen,  the  CD4  and  CD8  molecules  interact   • Cause  B  cells  to  switch  to  produce  different  subclasses  
with  T-­‐cell  receptor  complex  and  MHC  molecules  on  the  APC   of  IgG  
• CD4  binds  to  MHC  class  II  and  CD8  binds  to  MHC  class  I   • Promote  clearance  of  bacteria  by  direct  destruction  in  
  the  IFN-­‐γ  activated  macrophage  or  by  destruction  

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after  phagocytosis  of    opsonized  particles  
• Produce  IL-­‐2  and  IFN-­‐γ  
Th2  Cells   • Predominate  in  the  presence  of  IL-­‐4  
• Activate  mast  cells  and  eosinophils  and  cause  B  cells  
to  synthesize  IgE  
• Respond  to  helminthic  infections  
• Secrete  IL-­‐4,  IL-­‐5,  IL-­‐9  and  IL-­‐1  
Th17  Cells   • Produced  when  TGF-­‐β  and  IL-­‐6  come  together  
• Produce  IL-­‐17  and  IL-­‐22  
• IL-­‐17  –  cytokine  that  induces  stromal  and  epithelial  
cells  to  produce  IL-­‐8  
• IL-­‐8  –  a  potent  chemokine  that  recruits  neutrophils  
and  macrophages  to  infected  tissues  
Regulatory   • CD4  T  cells  become  Treg  when  exposed  to  TGF-­‐β  
T     • Function  by  suppressing  T-­‐cell  responses  
(Treg)   • Produce  TGF-­‐β  and  IL-­‐10  which  suppress  immune  
Cells   responses   Figure  3.  HIV  infects  CD4+  T  helper  cells  which  acts  on  the  dentritic  cell  which  in  
turn  stimulates  the  development  of  CD8+  T  cells;  end  result  of  HIV  is  
  immunosuppression  because  of  impaired  release  of  cytokines,  chemokines,  and  
delayed  lysis  of  infected  cells.  

III.  COMPLEMENT  SYSTEM  

A.  Major  Pathways  that  Activate  Complement  
   
Table  2.  Major  Pathways  that  Activate  Complement  
Classical  Pathway  
Alternative  Pathway  
Mannan-­‐binding  Lectin  (MBL)  Pathway  
 

 
Figure  2.  T-­‐cell  Proliferation  and  Differentiation  to  different  CD4  Effector  cells  
and  its  secretions.  

B.  CD8  Effector  Cells  

• CD8  cells  become  effector  cytotoxic  cells  through  engagement  of  
TCR(T-­‐cell  receptor)  and  recognition  of  class  I  MHC-­‐peptide  complex  
on  the  surface  of  an  infected  cell  
• CD8  T-­‐cells  kill  infected  cells  
o Cytotoxic  granules  containing  perforin,  granzymes  and  
granulysin    
o Releases  perforin  that  helps  granzyme  and  granulysin  to  enter  
the  infected  cell;  perforin  creates  pores  on  the  cell  membranes  
of  infected  cells  
o Granzyme  initiates  apoptosis  (programmed  cell  death)  
o CD8  T  cells  can  also  be  activated  via  the  help  of  CD4  T  cells  that  
are  interacting  with  the  same  antigen  
 

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• These  three  pathways  lead  to  release  of  C5  convertase  which  breaks   Table  5.  Clinical  Application  
down  C5  into  C5a  and  C5b   Biomarkers  of   • Provide  clues  for  mechanisms  of  disease  
• C5a  is  an  anaphylatoxin  as  well  as  a  chemotactic  factor   disease   • Pro-­‐inflammatory  cytokines  TNF-­‐α,  Il-­‐1  and  IL-­‐6  can  
• C5b  binds  to  C6  and  C7  to  form  a  complex  that  inserts  into  the   be  detected  in  the  sera  of  patients  with  septic  shock  
membrane  bilayer     Measurement   • Useful  monitor  of  immune  status  
• C8  then  binds  to  the  C5b-­‐C6-­‐C7  complex  followed  by  polymerization   of  cytokine   • T-­‐cell  function  can  be  monitored  by  the  ability  of  T-­‐
of  up  to  16  C9  molecules  to  produce  MAC   production  in   cells  to  produce    
• The  MAC  generates  a  channel  pore  in  the  membrane  cytolysis  by   vitro   IFN-­‐γ  (eg.  Identify  TB  reactivity)  
allowing  free  passage  of  water  across  the  cell  membrane   Recombinant   • Key  therapeutic  agents  
cytokines   • INF-­‐α  –  hepatitis  C  infection  
B.  Regulation  of  The  Complement  System   • INF-­‐β  –  multiple  sclerosis  
• C1  inhibitor  binds  to  and  inactivates  the  serine  protease  activity  of   Targets  of   • Cytokine  receptor  antagonists  and  anti-­‐cytokine  
C1r  and  C1s   therapeutics   monoclonal  antibodies    
• Factor  I  cleaves  C3b  and  c4b  –  reducing  amount  of  C5  convertase   • Downgrade  pathogenic  responses  to  exaggerated  
available   cytokine  production  
• Factor  H  enhances  the  effect  of  factor  I  on  C3b   • Eg.  TNF-­‐α  inhibitors  –  used  to  manage  rheumatoid  
• Factor  P  (properdin)  protects  C3b  and  stabilizes  the  C3  convertase  of   arthritis  
the  alternative  pathway    IL-­‐2  and  IL-­‐15  inhibitors  –  for  transplantation  and  
• Decay-­‐accelerating  factor  –  a  membrane  ound  protein  found  on   cancer  
most  blood  cell  surfaces  –  accelerate  the  dissociation  of  C3    
convertase  on  all  3  pathways   V.  HYPERSENSITIVITY  
  • A  condition  in  which  an  immune  response  results  in  exaggerated  or  
Table  3.  Complement  Deficiencies   inappropriate  reactions  that  are  harmful  to  the  host  
C2  Deficiency   Serious  pyogenic  infections   • The  first  contact  with  an  antigen  induces  a  sensitization  to  that  
MAC  components  deficiency   Enhances  susceptibility  to  Neisserial   allergen  
infection   • The  second  contact,  a  hypersensitivity  reaction  can  occur  
Properdin  deficiency   Susceptibility  to  meningococcal  
disease   A.  Type  I:  Immediate  Hypersensitivity  (Allergy)  
C1  inhibitor  protein  deficiency   Hereditary  angioedema  
• Occurs  within  seconds  after  antigen  combines  with  the  specific  IgE  
 
o Intravenous  administration  –  systemic  
C.  Pathogen  Evasion   o Atopic  allergy  –  local  reaction  
• Some  microbes  have  developed  surfaces  that  interfere  with  
opsonization  by  C3b  or  interfere  with  insertion  of  the  MAC  
• Complement  activation  inhibited  by  presence  of  microbial  generated  
proteins    
o Protein  A  and  Protein  C  both  bind  IgE  Fc  
• Microbes  can  generate  enzymes  that  degrade  complement  
component  
 
IV.  CYTOKINES  
• Potent  low-­‐molecular-­‐weight  protein  cell  regulators  
• Produced  transiently  and  locally  by  numerous  cell  types  
• Multifunctional  proteins  
o Hematopoiesis,  immunity,  infectious  disease  tumorigenesis,  
homeostasis,  tissue  repair  and  cellular  development  and  growth  
• Act  as  signaling  molecules  by  binding  to  their  own  glycoprotein  
receptors  on  cell  membranes  
Figure  4.  Pathogenesis  of  Type  I  Hypersensitivity  Allergen  on  the  antigen-­‐
• Followed  by  a  relay  of  the  signal  to  the  cell  nucleus  
presenting  cell  àinteraction  triggers  memory  cells  to  produce  plasma  cells,  
  which  produce  immunoglobulins  à  stimulate/sensitize  mast  cells  by  second  
Table  4.  Classification  and  Functions   exposure  to  allergen  à  release  of  vasoactive  amines  which  affect  smooth  
CLASSIFICATION   FUNCTION   muscle,  blood  vessel,  mucous  glands,  platelets,  sensory  nerve  endings,  
Immunoregulatory:  IFN-­‐γ   Role  in  antigen  presentation   eosinophils.  
Proinflammatory:  IL-­‐11,  IL-­‐6,   Seen  in  infectious  diseases  
TNF-­‐α  and  IFNs     Table  6.  Mediators  of  Type  I:  Immediate  Hypersensitivity  
Anti-­‐inflammatory:  TGF-­‐β,  IL-­‐10,   Down  regulate  an  overactive   HISTAMINE   PROSTRAGLANDINS  &  
IL-­‐11  and  IFN-­‐β   inflammatory  response   LEUKOTRIENES  
Growth  and  Differentiation     Colony  stimulation  factor  (CSFS)   • Primary  mediator   • Secondary  mediators  
and  stem  cell  factor   • Causes  vasodilation,   • Derived  from  arachidonic  acid  via  
  increased  capillary   cyclooxygenase  pathway  
  permeability,  and  smooth   • PG  à  bronchoconstriction  
  muscle  contraction  causing   • Leukotrienes  à  increased  

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bronchospasm   permeability  of  capillaries   Systemic   Ex:  Acute  Poststreptococcal  glomerulonephritis  
  Immune  
Complex  
Treatment   Prevention   disease  
• Reverse  action  of  mediators  by   • Prevention  by  identification    
maintaining  airway,  artificial   of  allergen  
ventilation,  supporting  cardiac   • Avoidance    
function    
• Give  epinephrine,  antihistamines,  
and  corticosteroids  
   
• Atopy  –  strong  familial  predisposition;  elevated  IgE  levels  
o Symptoms  induced  by  exposure  to  specific  allergens:  pollen,  
house  dust,  food  (shellfish)  
o Manifestations:  Hay  fever,  asthma,  eczema,  urticarial  

B.  Type  II:  Hypersensitivity    

Figure  6.  Pathogenesis  of  Type  III  Hypersensitivity  

D.  Type  IV:  Cell-­‐Mediated  (Delayed)  Hypersensitivity    

• A  function  of  specifically  sensitized  T-­‐lymphocytes  that  activate  
macrophages  to  cause  an  inflammatory  response  
• Response  is  delayed  –  starts  2-­‐3  days  after  contact  with  antigen  and  
  lasts  for  days  
Figure  5.  Type  2  Hypersensitivity  Reaction  
Table  8.  Two  Types  of  Type  IV  Hypersensitivity  
• Involves  binding  of  IgG  antibodies  to  cell  surface  antigens  or  ECM  
CONTACT  HYPERSENSITIVITY   TYBERCULIN-­‐TYPE  
molecules  
HYPERSENSITIVITY  
• Antibodies  directed  at  cell  surface  antigens  can  activate  complement  
• Sensitization  with  simple   • Little  immediate  reaction  after  
to  damage  cells  (ie.  hemolytic  anemias,  ABO  transfusion  reactions  
chemical  (eg.  Formaldehyde)   small  amount  of  tuberculin  is  
and  Rh  hemolytic  disease  
• Plant  materials  (poison  ivy)   injected  into  the  epidermis  of  a  
• Autoimmune  antibodies  can  combine  with  cell  surface  with  
topically  applied  drugs   patient    previously  exposed  to  
resulting  hemolysis  
(sulfonamides)  some  cosmetic   mycobacterium  tuberculosis  
o Examples  
    • Induration  and  redness  develop  
§ Penicillin  on  surface  of  RBC  
  and  reach  a  peak  in  24-­‐72  hrs  
§ Goodpasture  Syndrome  (antibody  forms  against  basement  
membrane  of  kidneys  and  lungs)   • That  person  has  been  infected  
§ Graves  disease  (without  cell  injury,  autoantibody  binds  to   with  the  agent  but  does  not  
imply  the  presence  of  current  
TSH  receptor  and  stimulates  the  thyroid  causing  
disease  
hyperthyroidism)  
 
C.  Type  III:  Immune  Complex  Hypersensitivity    
• Formation  of  immune  complexes  
• Antibody  combines  with  its  specific  antigen  
• Deposited  in  tissues  
• In  persistent  microbial  or  viral  infections,  immune  complexes  
deposited  in  organs  (kidneys)  
• Autoimmune  disorders  –  self  antigens  elicit  antibodies  that  bind  to  
organs  and  tissues  (joints,  kidneys,  blood  vessels)  
• Environmental  antigens  can  cause  immune  complex  formation  with  
disease  

Table  7.  Two  Major  Forms  of  Type  III:  Immune  Complex  Hypersensitivity  
Arthus  reaction   • local,  typically  elicited  on  skin  when  a  low  dose  
of  antigen  is  injected  and  immune  complex  form  
locally;  occurs  within  12  hours    
Figure  7.  Type  4  Hypersensitivity  Reaction  
• IgG  is  formed;  activation  of  complement  leads  to  
activation  of  mast  cells  and  neutrophils,  
 
mediator  release  and  vascular  permeability    
 

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VI.  DEFICIENCIES  OF  THE  IMMUNE  RESPONSE   • HIV  –  uses  CD4  molecules  as  the  virus  receptor  and  chemokine  
A.  Primary  Immunodeficiency   receptor;  progressive  loss  of  CD4  T-­‐cells  
o Develop  multiple  opportunistic  infections  (TB,  oral  candidiasis,  
• Disorders  of  the  immune  system  in  which  the  defect  is  intrinsic  to  
cryptococcocis,  pneumocystis  jirovecii)  
the  cells  of  the  immune  system  
• Have  a  genetic  basis  
D.  Malignancy  
• Loss  of  number  or  function  of  b  cells,  t  cells  or  phagocytic  cells,  
complement  components,  cytokines  or  tlrs  lead  to  increased   • Leukemias:  Deficiency  in  functional  neutrophils,  loss  of  phagocytosis  
susceptibility  to  infections   (increased  production  of  non-­‐functional  leukocytes)  
• Example:  Chronic  Granulomatous  Disease  (CGD)   o Increased  infections  with  bacteria  and  fungi  
o Impaired  phagocytic  function   • Lymphomas  and  multiple  myeloma  
o Normal  levels  of  igs,  t-­‐  and  b-­‐  cells  and  phagocytic  cells   • Some  tumors  secrete  high  levels  of  tgf-­‐β  that  suppress  a  variety  of  
o Genetic  defect  in  cytochrome  b-­‐558   responses  including  th1  responses  
o Defect  in  the  ability  of  phagocytes  to  produce  peroxide  and  
superoxide,  which  in  turn  calls  on  other  macrophages  and  they   E.  Drugs  
coalesce  to  become  multi-­‐nucleated   • Cytotoxic  drugs  used  to  treat  cancer  (cisplatin)  
o Usually  seen  in  areas  with  lymph  nodes  (cervical,  axillary,   • Immunosuppressive  drugs  (cyclosporine)  –  transplantation  
inguinal  areas)   • Treatment  of  autoimmune  diseases  (ra)  
    • Patients  become  more  prone  to  infection  

Figure  8.  Chronic  Granulomatous  Disease  

• Nitro  Blue-­‐Tetraolium  Test  (NBT)  

o Semi-­‐quantitative  assay  
o Tests  the  phagocytic  function  of  polymorphonuclear  leukocytes  
(PMNL)  
o Cells  reduce  a  colorless  NBT  into  a  blue-­‐black  deposit  within  the  
cell  
o In  CGD,  the  PMNLs  will  not  turn  blue  because  they  have  no  
superoxide  inside  their  granules  

B.  Secondary  Immunodeficiency  
• Disorders  of  immune  system  in  which  the  defect  is  induced  by  
external  factors  such  as  viruses  (HIV,  Herpes),  malignancy  and  drugs  

C.  Infections  
• Viral  infections  :  Measles  (Rubeola)  and  Ebstein  Barr  Virus  (EBV)    
• You  become  more  susceptible  to  secondary  bacterial  infections  
(bacterial  pneumonia  in  children  with  measles)  

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