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CHAPTER  12  –  Leukocyte  Development,   GRANULOCYTES  
Kinetics,  and  Function    
   Characterized  by  the  presence  of  
Leukocytes:   differently  staining  granules  in  their  
1. Also  known  as  white  blood  cells,   cytoplasm  when  viewed  under  light  
because  they  are  relatively  colorless   microscopy.  
compared  to  red  blood  cells.    Three  types  of  granulocytes:    
2. Can  be  identified  either  by  a   a) Neutrophils:  with  granules  that  can  
Romanowsky  stain  (5-­‐6  types)  or  by   react  to  both  acidic  and  basic  
flow  cytometry  (at  lest  10  types).   stains,  which  gives  them  a  pink  to  
  lavender  color.  
Leukocyte  development:   b) Basophils:  with  granules  containing  
1. Develop  from  primitive  stem  cells  in   acidic  proteins  stains  with  basic  
the  bone  marrow,  where  most  undergo   stains  such  as  methylene  blue.  
differentiation  and  maturation  before   c) Eosinophils:  with  granules  
they  are  released  into  circulation.   containing  basic  proteins  stains  
2. The  number  of  circulating  leukocytes   with  acidic  stains  such  as  eosin.  
varies  with:    
a) Sex,  age,  activity,  time  of  day  and   Neutrophils  
ethnicity    Also  called  polymorphonuclear  cells  
b) Reaction  of  leukocytes  to  stress   (PMNs)  because  of  their  prominent  
c) Sufficient  production  of  leukocytes   nuclear  segmentation  
in  the  bone  marrow    Most  numerous  type  of  lymphocyte  in  
3. Normal  range  of  leukocytes  in:   the  peripheral  blood  
a) Infants  –  3.8x109  cells  per  liter  to    Present  in  the  peripheral  blood  in  two  
25  or  30x109  cells  per  liter   forms:  
b) Adults  –  11.5x109  cells  per  liter   a) Segmented  (PMN)  –  old  neutrophil  
  b) Band  shape  –  young  neutrophil,  
Overall  function:   3%-­‐15%  of  leukocytes  depending  
1. Protection  from  foreign  organisms,   on  identification  criteria  
cells  or  material  by  phagocytosis.    
2. Immunity    
a) Innate  (nonspecific)  –  phagocytosis    
by  neutrophils  or  monocytes    
b) Adaptive  (specific)  –  production  of    
specific  antibodies  by  the    
lymphocytes   Amount  in  the  peripheral  blood  
3. Kinetics   –   refers   to   the   movement   of   -­‐ 50%  to  75%  of  leukocytes  in  relative  
cells   through   developmental   stages,   numbers    
into   the   circulation,   and   from   the   -­‐ 2.3  to  8.7x109cells  per  liter  in  absolute  
circulation   to   the   tissues   and   includes   terms  
the   time   spent   in   each   phase   of   the   -­‐ 18%  to  20%  in  infants    
cell’s  life    
  Diameter:  10-­‐12μm  
  Main  targets:  bacteria  fungi  
  Nucleus:  multi  lobed  
  Granules:  fine,  faintly  pink    
  Lifespan:  6hours  to  few  days  
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Neutrophil  Development:    Subdivided  into  type  I  and  type  II  
 Occurs  in  the  bone  marrow   a) Type  I  Myeloblast:  no  visible  
 Share  common  progenitor  cell  with   granules  
monocytes  known  as  the  granulocyte-­‐ b) Type  II  Myeloblast:  20  visible  
monocyte  progenitor  (GMP).   azurophilic  granules  (primary  
 Major  cytokine  responsible  for  the   granules)  
stimulation  of  neutrophil  production  is   *Primary   granules   or   nonspecific  
granulocyte  colony  stimulating  factor   granules  are  first  granules  that  appear  
(G-­‐CSF).   and   they   are   not   unique   to   any   one   of  
 Two  developmental  pools  of  the   the  cell  type.  
neutrophils:    
a) Mitotic  pool  (proliferating)     Promyelocyte  
-­‐  Consist  of  cells  that  are  dividing    1%  to  6%  of  the  nucleated  
by  mitosis   cells  in  the  bone  marrow  
b) Storage  pool  (maturation)    16  to  25μm  in  size    
-­‐  Consist  of  cells  that  are  no  longer    Nucleus  is  round  to  oval  and  often  
dividing  but  are  undergoing   eccentric  
nuclear  maturation    Nucleus  is  homogenous  with  delicate  
  euchromatin  and  two  to  four  easily  
Stages  of  neutrophilic  development:   seen  nucleoli.  
   Chromatin  clamping  
Mitotic  pool   (heterochromatin)  may  be  visible,  
1) Common  myeloid  progenitor  (CMP)   especially  around  the  edges  of  the  
2) Granulocyte-­‐monocyte  progenitor   nucleus.  
(GMP)    
3) Myeloblast   Neutrophil  Myelocyte    
4) Promyelocyte    1%  to  2%  of  the  nucleated  
5) Myelocyte     cells  in  the  bone  marrow  
   Final  stage  in  which  cell  division  
Storage  pool   occurs.  
6) Metamyelocyte    Production  of  primary  granules  ceases,  
7) Band  neutrophil   and  cell  begins  to  produce  secondary  
8) Segmented  neutrophil  (PMN)   or  specific  neutrophil  granules  
   Subdivided  into  early  Myelocyte  and  
Myeloblast   late  Myelocyte.    
 1%  to  2%  of  the   1) Early  Myelocyte    
nucleated  cells  in  the   -­‐ Also  called  dawn  of  neutrophilia    
bone  marrow     -­‐ Looks  similar  to  Promyelocytes  
 14  to  20μm  in  size   in  size  and  nuclear  
 Nucleus  is  round  to  oval  and  usually   characteristics.  
centrally  located   -­‐ Secondary  granules  slowly  
 Nucleus  is  homogenous  with  delicate   spread  through  the  cell  until  its  
euchromatin  and  two  to  four  visible   cytoplasm  is  more  lavender-­‐pink  
nucleoli.   than  blue.  
 Cytoplasm  is  slightly  basophilic   -­‐ Primary  granules  are  decreased    
 Nucleus,  cytoplasm  ratio  is  1:1  or  1:0.5   -­‐ Membrane  chemistry  is  changed  
  so  that  they  are  less  visible.  
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2) Late  Myelocyte      Secretory  granules  (secretory  vesicles)  
-­‐ 15-­‐18  μm  in  size  (somewhat   continue  to  form.  
smaller  than  the  Promyelocyte).      Nucleus  is  highly  clamped  
-­‐ The  nucleus  has  more    Presence  of  three  to  four  nuclear  
heterochromatin     segments  connected  by  threadlike  
  filaments.  
Neutrophil  Metamyelocyte      
 1%  to  2%  of  the   Neutrophil  granules:  
nucleated  marrow  cells    
 No  longer  capable  of  cell   Primary  granules  
division   -­‐ Formed  during  the  Promyelocyte  stage    
 Major  morphologic  change  in  the   -­‐ Last  to  be  released  (exocytosis)  
shape  of  the  nucleus.    
 Synthesis  of  tertiary  granules   1) Myeloperoxidase  
(gelatinase  granules)  begin   2) Acid  β-­‐glycerophosphatase  
 14-­‐16  μm  in  size  (a  little  smaller  than   3) Cathepsins  
the  Myelocyte).     4) Defensins  
 The  cytoplasm  contains  very  little   5) Elastase  
residual  RNA  and  therefore  little  or  no   6) Proteinase-­‐3  
basophilia      
 The  nucleus  is  indented  (kidney  bean   Secondary  (specific)  granules  
shape  or  peanut  shape)   -­‐ Formed  during  Myelocyte  and  
 Chromatin  is  increasingly  clumped   Metamyelocyte  stages  
 Nucleoli  are  absent   -­‐ Third  to  be  released  
 Nuclear  indentation  starts    
  1) Β2-­‐Microglobulin  
Neutrophil  band  form     2) Collagenase  
 9%  to  32%  of  the  nucleated   3) Gelatinase  
marrow  cells   4) Lactoferrin  
5) Neutrophil  gelatinase-­‐associated  
 All  evidence  of  RNA  
(cytoplasmic  basophilia)  should  be   lipocalin  
absent    
Tertiary  granules  
 Tertiary  granules  continue  to  form  
-­‐ Formed  during  Metamyelocyte  and  
 Secretory  granules  (secretory  vesicles)  
band  stages  
may  start  to  form.  
-­‐ Second  to  be  released    
 Nucleus  is  highly  clamped    
 Nuclear  indentation  now  exceeds  one   1) Gelatinase  
half  of  the  width  of  the  nucleus,  actual   2) Collagenase  
segmentation  has  not  yet  occurred.   3) Lysozyme  
  4) Acetyltransferase  
Segmented  neutrophils     5) Β2-­‐Microglobulin  
 Also  called  PMN    
neutrophils   Secretory  granules  (secretory  vesicles)  
 7%  to  30%  of  the   -­‐ Formed  during  band  and  segmented  
nucleated  marrow  cells   stages  
 All  evidence  of  RNA  (cytoplasmic   -­‐ First  to  be  released  (fused  to  plasma  
basophilia)  should  be  absent   membrane  for  adhesion)  
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1) CD11b  /  CD18    There  is  a  free  equilibrium  between  
2) Alkaline  phosphatase   the  circulating  and  marginal  pool.  
3) Vesicle-­‐associated  membrane-­‐2    Upon  antigenic  stimulation,  the  
4) CD10,  CD13,  CD14,  CD16   marginal  neutrophils  will  move  into  
5) Cytochrome  b558   the  tissues  (diapedesis).  
6) Complement  1q  receptor    The  average  neutrophil  circulates~  10  
7) Complement  receptor-­‐1   hours  before  diapedesis.  
   
Neutrophil  Kinetics:   Tissue  pool  
 Neutrophil  production  is  about  0.9  and    Life  span  is  variable  depending  on  
1.0x109cells  per  kg  per  day   whether  or  not  they  are  responding  to  
 Movement  of  the  neutrophil  through   infectious  or  inflammatory  agents.  
the  five  pools  known  as:    Some  products  of  inflammation  tend  to  
1) Mitotic  pool  in  the  bone  marrow   prolong  the  neutrophils  life  span  
2) Storage  pool  in  the  bone  marrow   through  antiapoptotic  signals.  
3) Circulating  pool  in  the  peripheral    MAC-­‐1  triggers  the  death  and  
blood   phagocytosis  of  neutrophils  that  
4) Marginated  pool  in  the  peripheral   reached  their  lifespan.  
blood    
5) Tissue  pool   Neutrophilic  Function:  
   Neutrophil  is  part  of  the  innate  
Mitotic  pool   immune  system  that  has  the  following  
 Contains  approximately  2.11x109  cells   characteristics:  
per  kg.   1) Destruction  of  foreign  organisms  is  
 Myeloblast  up  to  Myelocyte  –  takes  3-­‐6   not  antigen  specific.  
days  to  mature  to  this  stage   2) It  provides  no  protection  against  
  reexposure  to  the  same  pathogen.  
Storage  pool   3) It  relies  on  the  barriers  provided  
 Contains  approximately  5.6x109  cells   by  the  skin  and  mucous  
per  kg  or  a  5-­‐day  supply.   membranes  as  well  as  phagocytes  
 Transit  through  the  storage  pool  is   such  as  neutrophils  and  
estimately  6  to  7  days.   monocytes.  
 Granulocyte  release  from  the  bone   4) It  includes  a  humoral  component  
marrow  is  stimulated  by  the   known  as  the  complement  system.  
Granulocyte  colony  stimulating  factor    
(G-­‐CSF)    Neutrophil  has  three  function:  
 Metamyelocyte  up  the  mature   1) Phagocytosis  –  main  function  
segmented  neutrophils  –  takes  5-­‐7   2) Generation  of  neutrophil  
days  to  mature  to  this  stage   extracellular  traps  or  NETs.  
  3) Secretion  function  
Circulating  neutrophilic  pool  (CNP)      
Marginal  neutrophilic  pool  (MNP)   Phagocytosis  
 Only  about  half  of  the  peripheral    Main  function  of  the  neutrophils.  
neutrophils  are  freely  circulating  in  the    Destruction  of  foreign  material  and  
blood,  while  the  other  half  are  attached   microorganism  
to  the  vessel  walls  (called  the  marginal    
pool).      
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 Process  involves  seeking  (chemotaxis,    Pseudopodia   are   extended   around   the  
motility,  and  diapedesis)  and   foreign  particle  and  enclose  it  within  a  
destruction  (phagocytosis  and   phagosome.  
digestion)    
  C.  Killing  and  digestion  
   Formation   of   the   phagosome   allows  
Phagocytosis  process   reduced   nicotinamide   adenine  
A.  Recognition  and  attachment   dinucleotide   (NADH)   oxidize   complex  
 To   participate   in   an   inflammatory   within  the  phagosome  membrane.  
reaction,   which   is   the   body’s   response    Oxygen  dependent:  
to   infection,   neutrophils   enter   the   -­‐ Respiratory   burst   through   the  
tissues   where   they   are   attracted   by   activation   of   nicotinamide   adenine  
chemotactic   stimuli   released   after   dinucleotide   phosphate   (NADPH)  
tissue   injury   and   during   an   oxidase.   H2O2   and   hypochlorite   is  
inflammatory  response.   produced.  
 First   neutrophilic   response   is   to   roll    Oxygen  independent:  
along   endothelial   cells   using   stronger   -­‐ The   pH   within   the   phagosome  
adhesive  molecules.   becomes   alkaline   and   then   neutral,  
 Rolling   consist   of   transient   adhesive   the   pH   where   digestive   enzymes  
contacts   between   neutrophil   selectins   work.  
and  adhesive  molecules.   -­‐ Primary   and   secondary   lysosomes  
 CD11b  /  CD18  contributes  to  the  tight   (granules)   fuse   to   the   phagosome  
stationary  binding  between  neutrophil   and   empty   hydrolytic   enzymes   and  
and  endothelial  cells     other  bactericidal  molecules  into  the  
 Diapedesis   or   transmigration   of   phagosome.  
neutrophils  either  between  or  through    In  this  process  most  neutrophils  die  
endothelial   cells.   It   is   mediated   by   and  are  themselves  phagocytosed  by  
integrins   and   integrin   associated   macrophages.  
proteins.      
 Transmigrating  neutrophils  release  the   Generation   of   neutrophil   extracellular   traps  
tertiary   granules   containing   gelatinase   or  NETs  
and  collagenase.    Extracellular   threadlike   structures  
 Gelatinase   degrades   denatured   believed   to   represent   chains   of  
collagen   as   well   as   type   IV   and   V   nucleosomes   from   unfolded   nuclear  
collagen   and   activates   chemokines   chromatin  material  (DNA).  
such  as  interleukin-­‐8  (IL-­‐8).      Have   enzymes   from   neutrophilic  
 Neutrophil   then   migrate   in   a   granules   that   are   able   to   trap   Gram  
directional   manner   to   the   infected   positive   and   negative   bacteria   as   well  
area.   as  fungi  
   Generated   at   the   time   of   neutrophilic  
B.  Ingestion   death  
 Neutrophils   use   their   enormous    
inventory  of  surface  receptors  to  either   Secretory  function  
directly   recognize   the   pathogen   and    Neutrophils   are   a   source   of  
apoptotic   cell   or   to   recognize   opsonic   transcobalamin   I   or   R   binder   protein  
molecules  attached  to  foreign  particles   which   is   necessary   for   absorption   of  
such  as  antibodies.    
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Vitamin  B12.  Also  a  source  of  variety  of    Hematologic  Disorders  
cytokines   a) Pernicious  anemia  
  b) Aplastic  anemia  
Neutrophilia     c) Hypochromic  anemia  
 Describes   high   level   of   neutrophilic   d) Agranulocytosis    
granulocyte  in  the  blood    
  Eosinophils  
Causes  of  neutrophilia    Make   up   between   1%   to   5%   of   the  
 Neutrophils   are   the   primary  white   peripheral   blood   leukocytes,   with   an  
blood   cells  that   respond   to   a   bacterial   absolute  number  between  40  t  550  per  
infection,  so  the  most  common  cause  of   microliter  of  blood  
neutrophilia   is   a   bacterial   infection,    
especially  pyogenic  infections.    
 Neutrophils   are   also   increased   in   any    
acute  inflammation,   so   will   be   raised    
after   a  heart   attack,   other   infarct   or    
burns.    
 Some   drugs   have   effects   that   cause   Diameter:  10-­‐12μm  
marginated   neutrophils   to   enter   the   Main  targets:  Larger  parasites  and  modulate  
blood  stream.     allergic  inflammatory  response  
 Nervousness  will  very  slightly  raise  the   Nucleus:  bi  lobed  
neutrophil  count  because  of  this  effect.   Granules:  full  of  pink-­‐orange    
 Post   surgery   can   also   cause   high   Lifespan:  8  to  12  days  (circulate  for  4-­‐5  
neutrophil  in  the  blood   hours)  
 Other  factors  are:    
a) Environmental  factors   Eosinophil  development  
b) Emotional  factors    Eosinophil   development   is   similar   to  
c) Acute  hemorrhage   neutrophil.  
d) Acute  hemolysis    Evidence   of   common   precursor   of  
e) Intoxication   eosinophil  and  basophil  includes:  
f) Myeloproliferative  Disorders   1) The   granules   of   both   eosinophil  
g) Leukemoid  Reaction   and   basophil   contain   major   basic  
h) Chronic  Myelogenous  Leukemia   protein  and  lysophospholipase.  
i) Malignant  Neoplasm   2) Both   have   receptors   for   IL-­‐3,   IL-­‐5  
  and   granulocyte-­‐macrophage  
Neutropenia   colony  stimulating  factor  (GM-­‐CSF)  
 Describes   high   level   of   neutrophilic   3) Individuals   with   increased  
granulocyte  in  the  blood   basophils   often   have   increased  
  eosinophil  
Causes  of  neutropenia   4) There   is   a   rare   syndrome  
 Congenital   decrease   in   production   in   characterized  solely  by  the  absence  
bone  marrow   of  both  cell  types  
 Increased  loss  of  WBC   5) Cells   with   both   eosinophil   and  
 Chronic   Bacterial   and   Rickettsial   basophil   granules   are   sometimes  
Infections   seen   in   patients   with  
 Chronic   Chlamydial   and   Viral   myeloproliferative  syndrome  
Infections   6) Hybrid   eosinophil-­‐basophil   cells  
have  been  grown  in  culture  
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 Eosinophil   lineage   is   established   Secondary  granules  
through   the   interaction   between   the   -­‐ Formed  throughout  the  maturation    
cytokines   IL-­‐3,   IL-­‐5   and   GM-­‐CSF   and    
the  three  transcription  factors  (GATA-­‐ 1) Major  basic  protein  (core)  
1,  PU.1,  c/EBP)   2) Eosinophil  cationic  protein  (matrix)  
 Eosinophilic   Promyelocytes   can   be   3) Eosinophil  derived  neurotoxin  
identified   by   the   presence   of   the   (matrix)  
Charcot-­‐Leyden   crystal   protein   in   4) Eosinophil  peroxidase  (matrix)  
their  primary  granules.   5) Lysosome  (matrix)  
  6) Catalase  (core  and  matrix)  
Stages  of  Eosinophil  development   7) β-­‐Glucuronidase  (core  and  matrix)  
1) Eosinophil  Myelocyte     8) Cathepsin  D  (core  and  matrix)  
2) Eosinophil   Metamyelocyte   and   9) Interleukins  2,  4  and  5  (core)  
band  form   10)Interleukins  6  (matrix)  
3) Mature  Eosinophil   11)Granulocyte-­‐macrophage  colony  
  stimulating  factor  (GM-­‐CSF)  (core)  
Eosinophil  Myelocyte    
 Presence  of  large,  pale,  reddish,  orange   Small  lysosomal  granules    
secondary   granules   along   with   azure    
granules  in  blue  cytoplasm.   1) Acid  phosphatase  
 Secondary  Eosinophil  granules  contain   2) Arylsulfatase  B  
an  electron-­‐dense  crystalline  core   3) Catalase  
 Nucleus   is   similar   to   neutrophil   4) Cytochrome  b558  
counterpart.   5) Elastase    
  6) Eosinophil  cationic  protein  
Eosinophil  Metamyelocyte  and  band  form    
 Secondary   granules   increase   in   Lipid  bodies  
number   and   also   becomes   more    
distinct  and  refractory  while  Secretory   1) Cyclooxygenase  
granules  start  to  generate   2) 5-­‐Lipoxygenase  
 Electron  microscopy  reveals  two  other   3) 15-­‐Lipoxygenase  
organelles   that   are   lipid   bodies   and   4) Leukotriene  C4  synthase  
small  granules.   5) Eosinophil  peroxidase  
  Nucleus   is   similar   to   neutrophil   6) Esterase  
counterpart.    
  Storage  vesicles  
Mature  eosinophil     -­‐ Carry  proteins  from  secondary  
 Nuclei  has  two  segments   granules  to  be  released  into  the  
extracellular  medium.  
 Cytoplasm   is   full   of   the   characteristic  
 
Eosinophilic  secondary  granules.  
Eosinophil  kinetics  
 
Primary  granules    Approximately   3%   of   the   nucleated  
-­‐ Formed  during  the  Promyelocyte  stage     bone  marrow  cells  are  eosinophils  
   One   third   of   the   total   eosinophil   is  
1) Charcot-­‐Leyden  crystal  proteins   mature,   a   quarter   is   Metamyelocyte  
  and   the   remainders   are   Promyelocyte  
  or  Myelocyte.  
 NQS  
8  
 There   is   a   large   storage   pool   of   to   their   ability   to   rapidly   secrete  
eosinophils   in   the   marrow   consisting   performed   cytokines   in   a   stimulus  
of  9  to  14x108  cells  per  kg   specific  manner  
 Half   life   of   circulating   eosinophil   is    Regulate   mast   cell   function   through  
roughly  18  hours   the  release  of  basic  protein  that  causes  
 Tissue   destination   of   eosinophils   mast   cell   degranulation   as   well   as  
appears   to   be   underlying   columnar   cytokine  production.  
epithelial   surfaces   in   the   respiratory,    Produce   nerve   growth   factor   that  
gastrointestinal,   and   genitourinary   promotes   mast   cell   survival   and  
tracts.   activation  
   
Eosinophil  function   Regulation  of  parasitic  infection  
 Eosinophil   granules   are   full   of    Capable   of   destroying   tissue-­‐invading  
previously   synthesized   proteins   helminthes   through   the   secretion   of  
including   cytokines,   chemokines,   major   basic   protein   and   eosinophil  
growth  factor,  and  cationic  proteins.   cationic  protein.  
 Basophil  has  four  functions    
1) Degranulation  of  eosinophil   Hallmark  of  allergic  disorder  
2) Immune  regulation    The  number  of  eosinophil  in  the  blood  
3) Regulation  of  parasitic  infection   and   sputum   correlates   with   disease  
4) Hallmark  of  allergic  disorder   severity.  
   Implicate   din   airway   remodeling  
Degranulation  of  eosinophil   through   eosinophil-­‐derived   fibrogenic  
 Classical  exocytosis   –  granules  move  to   growth  factors.    
the   plasma   membrane,   fuses   with   the    Accumulation   of   eosinophil   in   the  
plasma   membrane   and   empty   their   gastrointestinal   tract   occurs   in   food  
contents  into  the  extracellular  space.   allergy,   allergic   coitus,   and  
 Compound   exocytosis   -­‐   granules   fuse   inflammatory  bowel  disease.  
together  within  the  eosinophil  prior  to    
fusing  with  the  plasma  membrane   Eosinophilia  
 Piecemeal   degranulation   –   secretory    Describes   high   level   of   eosinophil  
vesicles   remove   specific   proteins   from   granulocyte  in  the  blood  
the  secondary  granules.  These  vesicles    
he   migrates   to   the   plasma   membrane   Causes  of  eosinophilia  
and  fuse  to  empty  the  specific  proteins    Eosinophilic  leukemoid  reaction  
into  the  extracellular  space    GI  disorder  
   Loeffler’s   syndrome   or   pulmonary  
Immune  regulation   infiltrate  
 Transmigration   to   the   thymus   of   new    
born   and   is   involve   in   the   deletion   of   Eosinopenia  
double  positive  thymocytes    Describes   low   level   of   eosinophil  
 Capable  of  acting  as  antigen  presenting   granulocyte  in  the  blood  
cells      
 Capable  of  promoting  the  proliferation   Causes  of  eosinophilia  
of  effector  T  cells    Acute  stressful  condition  
 Implicated   in   the   initiation   of   either    Hyper  secretion  of  ACTH  
type  1  or  type  2  immune  response  due    Acute  bacterial  infection  
 NQS  
9  
Basophils   Mature  basophil  
 Basophils   are   true   leukocytes    Contains  lobulated  nuclei  
compared  to  mast  cells.      Chromatin  pattern  is  clumped    
 Basophils   mature   in   the   bone   marrow    Nuclear   segmentation   with   visible  
and   circulate   in   the   blood   as   mature   filaments  occurs  rarely  
cells  with  granules.    Cytoplasm   is   colorless   and   contains  
 Basophils   are   the   least   numerous   of   large   number   of   the   characteristic  
the   circulating   WBCs   making   up   metachromatic  granules  
between   0.5%   and   1.5%   of   circulating    
leukocytes.   Basophilic  kinetics  
   Time  for  bone  marrow  development  is  
  4.3  days  +  11  hours    
   Mean   transit   time   in   the   peripheral  
  blood  is  3.7  days  +  21  hours  
   Life   span   is   significantly   longer   than  
  other   granulocyte   because   of  
Diameter:  12  -­‐  15μm   activation   by   the   cytokine   IL-­‐3   and  
Main  targets:  Release  histamine  for   antiapoptotic  pathways.  
inflammatory  response      
Nucleus:  bi  lobed  or  tri  lobed   Secondary  granules  
Granules:  Large  blue      
Lifespan:  a  few  hours  to  a  few  days   1) Histamine  
  2) Platelet  activating  factor  
Basophilic  Development   3) Leukotriene  C4  
 Basophilic   development   is   similar   to   4) Interleukin-­‐4  
eosinophil   development   in   that   it   5) Interleukin-­‐13  
requires   the   cytokines   IL-­‐3,   IL-­‐5   and   6) Vascular  endothelial  growth  factor  A  
GM-­‐CSF.   7) Vascular  endothelial  growth  factor  B  
 Transforming   growth   factor   β   8) Chondroitin  sulfates  (e.g.  heparin)  
suppresses   eosinophil   differentiation    
and  enhances  basophil  differentiation.   Basophilic  function  
 Due   to   small   number   basophil    Capable   of   releasing   large   quantity   of  
development   is   subdivided   into   subtype   2   helper   T   cells   (Th2)  
immature  and  mature  basophils.   cytokines   such   as   IL-­‐4   and   IL-­‐3   that  
  regulates  the  Th2  immune  response  
Immature  basophil    Functions   as   initiators   of   the   allergic  
 Round   to   somewhat   lobulated   nuclei   inflammation   through   the   release   of  
with  slightly  condensed  chromatin   preformed  cytokines  
 Cytoplasm   is   blue   and   contains   large    Basophils  are  not  restricted  to  antigen-­‐
blue-­‐black  secondary  granules  that  are   specific   IgE   cross-­‐linking,   but   can   also  
metachromatic   (stain   purple   with   be   prompted   in   nonsensitized  
toluidine  blue)   individuals  by  growing  list  of  parasitic  
 Primary   granules   may   or   may   not   be   antigens,   lectins,   and   viral   super  
seen   yet.   Basophilic   granules   are   antigens,   binding   to   nonspecific   IgE  
water-­‐soluble.   antibodies  
 
 
 NQS  
10  
 Mature  basophils  are  evidently  capable    Mast   cell   progenitor   originate   in   the  
of  synthesizing  granule  proteins  based   bone  marrow  
on  activation  signals.    The   major   cytokine   responsible   for  
 Basophils   are   also   involved   in   control   mast   cell   maturation   and  
of  helminth  infections.   differentiation   is   stem   cell   factor   also  
  known  as  kit  ligand    
Basophilia    Two  types  of  mast  cell  according  to  the  
 Describes   high   level   of   basophil   presence  of  chymase  and  tryptase.  
granulocyte  in  the  blood   1) MCtc  –  mast  cell  with  both  chymase  
  and  tryptase  
Causes  of  eosinophilia   2) MCt  –  mast  cell  with  only  tryptase  
 Chronic  myeloproliferative  disorder    
 Inflammatory  bowel  disease   Mast  cell  function  
 Hypersensitivity    Mast   cell   function   as   effector   cells   in  
 Hypothyroidism   allergic   reactions   through   the   release  
 Estrogen  therapy   of   a   wide   variety   of   lipid   mediators,  
 Radiation  exposure   proteases,   proteoglycans   and  
  cytokines.  
Basopenia    Mast   cell   can   be   activated  
 Describes   low   level   of   basophil   independently   by   IgE,   which   leads   to  
granulocyte  in  the  blood   inflammatory  response  
   Mast   cell   can   function   as   antigen-­‐
Causes  of  eosinophilia   presenting   cells   to   induce   the  
 Stress   differentiation  of  Th2  cells  
 Hyperthyroidism    Mast   cell   can   act   in   both   innate   and  
 After  anaphylaxis   adaptive  immunity  
 Glucocorticoid  therapy    Mast   cells   can   have   anti-­‐inflammatory  
 Urticaria   and   immunosuppressant   functions,  
 Acute  infections   and   thus   they   can   both   enhance   and  
  suppress   features   of   the   immune  
Mast  cells     response  
 
MONONUCLEAR  CELLS  
 Mast   cells   are   not   lymphocytes   they  
are  tissue  cells.    Also  called  agranulocyte  
 They   are   grouped   with   the    Leukocytes   characterized   by   the  
lymphocytes  because:   apparent   absence   of   granules   in   their  
1) Their   precursors   circulate   in   the   cytoplasm.   Although   the   name   implies  
peripheral   blood   for   a   brief   period   a  lack  of  granules  these  cells  do  contain  
on   its   way   to   their   tissue   non-­‐specific   azurophilic   granules,  
destination   which  are  lysosomes.  
2) Mast   cells   have   special   phenotypic    Two  types  of  mononuclear  cells  
and   functional   similarities   with   a) Monocytes  
both  eosinophil  and  basophil.   b) Lymphocytes  
   
   
   
   
 NQS  
11  
Monocyte   Dendritic  cells  
 Monocytes   make   up   between   2%   to    
10%  of  circulating  leukocytes,  with  96    
to   1100   monocytes   per   microliter   of    
whole  blood.    
 Progenitor   cell   for   Macrophage,    
Osteoclast  and  Dendritic  cells.    
  Main  targets:  Can  be  myeloid  or  lymphoid  
Monocyte   derived.  Main  function  is  as  an  antigen-­‐
  presenting  cell  (APC)  that  activates  T  
  lymphocytes.  
  Granules:  None    
  Lifespan:  activated:  days  
  immune:  months  to  years  
   
  Monocyte  Development  
Diameter:  12  -­‐  20μm    Development  is  similar  to  Neutrophilic  
Main  targets:  Monocytes  migrate  from  the   development,   because   the   two   share  
bloodstream  to  other  tissues  and  differentiate   the  Granulocyte-­‐monocyte  Progenitor  
into  tissue  resident    Macrophage   colony   stimulating   factor  
Nucleus:  kidney  shaped   is   the   major   cytokine   responsible   for  
Granules:  None     the   growth   and   differentiation   of  
Lifespan:  hours  to  a  few  days   monocyte.  
   
Macrophage   Stages  of  Monocyte  development  
  1) Monoblast  
  2) Promonocytes  
  3) Monocytes  
   
  Monoblast  
   12  to  20  µm  in  diameter  
Diameter:  40  -­‐  50μm  Has  a  nuclear  to  cytoplasm  ratio  of  4:1  
Main  targets:  Phagocytosis  of  cellular  debris   to  3:1  
and  pathogens  and  stimulation  of    Has  a  round  to  oval  nucleus  with  fine  
lymphocytes  and  other  immune  cells  that   chromatin  structure  
respond  to  the  pathogen    One  to  four  nucleoli  are  usually  visible  
Nucleus:  Oval  with  at  least  one  prominent    The  nucleus  can  be  central  or  eccentric  
nucleolus   and  it  can  show  evidence  of  
Chromatin  pattern:  netlike  (reticulated)   indentation  or  folding  
Cytoplasm:  Pale,  frequently  vacuolated    The  cytoplasm  is  agranular,  stains  
Granules:  None     moderately  to  lightly  basophilic  
Lifespan:  activated:  days    
immune:  months  to  years   Promonocytes  
   12  to  18  µm  in  diameter  
   Nucleus  is  large  in  relation  to  its  size  
   Nucleus  may  be  round  or  slightly  
  indented  or  folded  
 
 NQS  
12  
 Chromatin  pattern  is  delicate    Marginal   pool   of   monocytes   is   3.5  
 Cytoplasm  is  blue  and  contains   times  the  circulating  pool  
scattered  azure  granules    Monocytes   remain   in   circulation   for  
 Azure  granules  are  heterogeneous   approximately  30  hours  
with  regards  to  their  content  of    Monocyte   derived   Kupffer   cells   have   a  
lysosomal  enzymes,  peroxidase,   life  span  of  approximately  21  days  
nonspecific  esterases  and  lysozyme    Inflammatory   macrophages   have   a   life  
  span  measured  in  hours.  
Monocyte    
 15  to  20  µm  in  diameter   Specific  inflammatory  macrophages:  
 Monocytes  are  slightly  immature  cells   1) Kupffer  cells-­‐  liver  
whose  ultimate  goal  is  to  is  to  enter  the   2) Alveolar  macrophage  –  lungs  
tissue  and  mature  into  macrophage,   3) Microglia  –  brain  
osteoclast  and  dendritic  cells   4) Langerhans  cells  –  skin  
 Nucleus  cytoplasm  ratio  is  1   5) Splenic  macrophages  –  spleen  
 Nucleus  may  be  round  or  slightly   6) Intestinal  macrophages  –  intestine  
indented  or  folded  (horse  shoe  shape)   7) Peritoneal  macrophages  –  peritoneum    
 Chromatin  pattern  is  loose  and   8) Osteoclast  –  bone  
described  as  lacelike  or  stingy   9) Synovial  macrophages  -­‐  type  A  cells  
 Cytoplasm  is  blue  grey  with  fine  azure   10)  Renal  macrophages  –  kidneys  
granules  referred  as  azure  dust  or  a   11)  Reproductive   organ   macrophages   –  
ground  glass  appearance   genitals  
 Small  cytoplasmic  pseudo  pods  or   12)  Dendritic  cells  –  Lymph  nodes  
blebs  may  be  seen    
 Cytoplasmic  and  nuclear  vacuoles  may   Monocyte  function  
also  be  present    Function   of   monocyte   can   be   divided  
 Two  subsets  of  monocytes  are  CD16+   into:  
and  CD16-­‐   a) Innate  immunity  
  b) Adaptive  immunity  
Monocyte  kinetics   c) Housekeeping  functions  
 Promonocytes   pool   approximately    
contains   6x108   cells   per   kg   and   they   Innate  immunity  
produce   7x106   monocytes   per   kg   per    Macrophages   recognize   a   wide   range  
hour.   of   bacterial   pathogens   by   means   of  
 Normally   Promonocytes   undergo   two   pattern   recognition   receptors   that  
mitotic  cycle  per  60  hours  to  produce  4   stimulate   inflammatory   cytokine  
Promonocytes   but   if   there   is   an   production  and  phagocytosis  
increased   demand   it   can   undergo   4    Can   synthesize   nitric   oxide,   which   is  
mitotic   cycle   which   would   yield   16   cytotoxic   against   viruses,   bacteria,  
Promonocytes   fungi,   protozoa,   helminth,   and   tumor  
 Monocytes   in   the   splenic   reservoir   cells  
appear   to   respond   to   tissue   injury   to    Have   Fc   receptors   and   complement  
participate  in  wound  healing   receptors   to   phagocytize   foreign  
 Monocytes  in  the  peripheral  blood  can   organisms   and   material   that   has   been  
be   found   in   the   marginal   pool   and   coated   with   antibodies   and  
circulating  pool.   complements.  
 
 NQS  
13  
Adaptive  immunity    Adaptive   immunity   has   the   following  
 Macrophage   and   dendritic   cells   both   functions:  
degrade   antigen   and   present   antigen   a) Relies   on   an   enormous   number   of  
fragments  on  their  surface   distinct   lymphocyte   each   having  
 They   interact   and   activate   both   T-­‐ surface   receptors   for   a   different  
lymphocyte   and   B-­‐lymphocyte   to   specific   molecular   structure   on   a  
initiate  adaptive  immune  response     foreign  antigen  
 Dendritic   cells   are   most   efficient   and   b) After   an   encounter   with   a  
potent  of  the  antigen  presenting  cells   particular   antigen,   memory   cells  
  are   produced   that   will   react   faster  
Housekeeping  functions   and   more   vigorous   to   the   same  
 Removal  of  debris  and  dead  cells  at  site   antigen  upon  reexposure  
of  infection  or  tissue  damage   c) Self-­‐antigens   are   ignored   under  
 Destruction   of   senescent   red   cells   and   normal   circumstances   referred   to  
maintenance   of   a   storage   pool   of   iron   as  tolerance  
for  erythropoiesis    Lymphocytes   are   different   from   other  
 Synthesis   of   a   wide   variety   of   protein,   leukocytes   in   several   ways,   including  
including   complements,   coagulation   the  following:  
factors,   prostaglandins,   leukotrienes   a) Lymphocytes  are  not  end  cell.  They  
growth   factors,   and   binding   protein   are   resting   blast   forms   capable,  
such  as  transferrin   when   stimulated,   of   transforming  
  into   active   blast   that   undergo  
Lymphocytes     mitosis   to   produce   both   memory  
 Lymphocyte  make  up  20  to  40%  of  the   and  effector  cells  
circulating   leukocytes   with   960   to   b) Lymphocytes   recirculate   from   the  
4400   lymphocytes   per   microliter   of   blood  to  the  tissue  and  back  to  the  
whole  blood   blood  
  c) B   and   T   cells   are   capable   of  
  recombining   gene   segments   to  
  produce   surface   receptors   and  
  antibodies  
  d) T   and   NK   cells   usually   develop  
  outside  the  bone  marrow  
 Lymphocytes   are   divided   into   three    
major  groups:   Gross  Morphological  Stages  
a) T   cell   or   T   lymphocytes   (Thymus):    Lymphoblast    
One   of   the   major   players   in    Prolymphocyte    
adaptive  immunity.    Lymphocyte    
b) B   cell   or   B   lymphocytes   (Bone):    
One   of   the   major   players   in   Lymphoblast    
adaptive  immunity.    10-­‐18  μm  in  size  
c) Natural   killer   (NK)   cells:   Makes   up    Coarse  chromatin  pattern  
the   small   part   of   the   lymphocytes    Round  or  oval  nucleus  
and  are  part  of  the  innate  immunity    1-­‐2  distinct  nucleoli  
   
   
 NQS  
14  
 Cytoplasm   is   scanty   with   no   granules,   Lymphocyte  development  
described   as   smooth,   moderate   to   dark    B  and  T  cells  are  subdivided  into:  
blue,   deeper   blue   in   periphery   and   a) Antigen   independent   –   occurs   in  
lighter  near  the  nucleus   the   bone   marrow   and   thymus  
  (primary  lymphatic  organs)  
Prolymphocyte     b) Antigen   independent   –   occurs   in  
 Nucleus   is   round,   oval,   or   slightly   spleen,   lymph   nodes,   tonsils,   and  
indented   with   more   clumped   non   encapsulated   aggregates   of  
chromatin  pattern.   lymphocytes   such   as   the   peyer  
 Cytoplasm   is   moderate   to   dark   blue   patches   in   the   intestinal   wall  
usually   non   granular,   but   may   contain   (Peripheral   or   secondary   lymphatic  
small  azurophilic  granules   organs)  
   
Lymphocyte  (small,  medium,  large)   B  lymphocyte  
Small  lymphocyte      Approximately  10%  to  15%  of  
 8-­‐10  um  in  size   lymphocytes  
 Cytoplasm   has   thin   rim    9  μm  in  diameter  in  resting  state  
around   nucleus   which   is    Nucleus:  Cytoplasm  ratio  is  1:1  
moderate   blue   in   color   with    Chromatin  is  arranged  in  blocks  
few  azurophilic  granules    Have  three  stages  known  as  pro-­‐B,  pre-­‐
 Nucleus   is   dense   and   clumped   B  and  immature-­‐B  cells  
chromatin  pattern,  round  or  oval,  may    During   the   three   stages   gene  
be  slightly  indented  with  no  nucleoli   rearrangement   occurs   to   produce  
  unique  immunoglobulin  chains  
Medium  lymphocyte      Immature   B-­‐cells   referred   to   as   naive  
 11-­‐14  um  in  size   B-­‐cells   leave   the   bone   marrow   to  
 Cytoplasm  is  more   migrate   to   secondary   lymphatic  
abundant,  pale  to  moderate   organs,   where   they   take   residence   in  
blue     specific   zones   such   as   lymph   nodes  
 Nucleus   is   same   as   small   lymphocyte,   follicle    
chromatin  is  not  as  dense  as  the  small    B-­‐cells  also  known  as  hematogones  are  
lymphocyte   cells   with   homogenous   nuclear  
  chromatin   pattern   and   extremely  
Large  lymphocyte     scanty   (small   or   insufficient   quantity)  
 12  –  16  um  in  size   cytoplasm  
 Cytoplasm  is  abundant    Normally   seen   in   new   born   peripheral  
which   is   clear,   deep   blood  and  bone  marrow  
blue   in   color   with    B-­‐cells   in   the   secondary   lymphatic  
adequate  granulation   organs  or  in  the  peripheral  blood  come  
 Nucleus   is   same   as   small   with   coarse   in   contact   with   the   antigen   resulting   to  
chromatin,   may   be   centrally   or   cell   activation,   cell   division  
eccentrically  located       (blastogenesis)   and   the   production   of  
  either  memory  cells  or  effector  B-­‐cells  
   Effector  B-­‐cells  are  antibody  producing  
  cells   known   as   plasma   cells   and  
  plasmacytoid  lymphocytes  
   
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T  lymphocytes   referred   to   as   large   granular  
 Approximately   85%   of   the   circulating   lymphocyte  
lymphocyte    
 Develop   initially   in   the   thymus   Lymphocyte  function  
(thymic  cortex)     B-­‐cells  
 Have   three   stages   known   as   pro-­‐T,    Essential  for  antibody  production  
pre-­‐T  and  immature-­‐T  cells    Have   a   role   in   antigen   presentation   to  
 During   the   three   stages   gene   T-­‐cell  
rearrangement   occurs   to   produce   T-­‐  Necessary  for  optimal  CD4activation  
cell   receptors   that   are   unique   to   each    Produce   cytokines   that   regulate   a  
T-­‐cell   variety  of  T-­‐cell  and  antigen  presenting  
 T-­‐cells   are   subdivided   into   two   major   cell  factor  
category  depending  on  the  presence  of   T-­‐cells  
CD4  or  CD8  antigen  on  their  surface   1)  CD4+  T-­‐cells  (effector  lymphocytes)  
 Immature   T-­‐cells   proceed   to   the   a)  TH1  
thymic  medulla  for  further  removal  of   -­‐   Mediate   immune   response   against  
self  reactive  T-­‐cells   intercellular  pathogens  
 Remaining   immature   T-­‐cells   referred   b)  Th2  
to  as  naive  T-­‐cells  leave  the  thymus  to   -­‐   Mediate   host   defense   against  
migrate   to   secondary   lymphatic   extracellular   parasites   including  
organs,   where   they   take   residence   in   helminthes  
specific   zones   such   as   paracortical   -­‐  Important  in  the  induction  of  asthma  
areas     and  other  allergic  disease    
 Morphology   of   effector   T-­‐cells   varies   c)  Th17  
with   the   subtype   of   T-­‐cell   involve   they   -­‐   Involve   in   the   immune   responses  
are   often   referred   as   reactive   variant   against  extracellular  bacteria  and  fungi  
lymphocyte   d)  Treg  (CD4+  CD25+  regulatory  T-­‐cells)  
 T-­‐cells   in   the   secondary   lymphatic   -­‐   Play   a   role   in   maintaining   self-­‐
organs   or   in   the   peripheral   blood   tolerance   by   regulating   immune  
come   in   contact   with   the   antigen   responses  
resulting   to   cell   activation,   cell    
division   (blastogenesis)   and   the   2)  CD8+  T-­‐cells  (effector  lymphocytes)  
production   of   either   memory   cells   or    Also   referred   as   cytotoxic   T  
effector  T-­‐cells   lymphocyte  
   Capable   of   killing   target   cells   by  
NK  cells   secreting   granules   containing  
 Approximately   2%   of   circulating   granzyme  and  perforin  
lymphocyte    
 Heterogeneous   group   of   cells   with   NK  cells  
respect  to  their  surface  antigen    Functions   as   part   of   innate   immunity  
 Majority  are  CD56+,  CD16+,  CD3-­‐,  CD8-­‐   in   that   they   are   capable   of   killing  
 Mature   NK   cells   are   relatively   large   certain   tumor   cells   and   virus-­‐infected  
because   of   their   increased   amount   of   cells  without  prior  sensitization  
cytoplasm    
 Cytoplasm   contains   azurophilic    
granules   that   are   peroxidase   negative    
 
 NQS  
16  
Plasma  cell  
 Plasmablast  
 Proplasmacyte  
 Plasmacyte  (Plasma  Cell)  
 
Plasmablast  
 18  –  25  um  in  size  
 Cytoplasm   is   basophilic,   abundant,   non  
granular  
 Nucleus  has  more  clumped  chromatin    
 Nucleus  is  eccentric  with  round  or  oval  
in  shape  
 Perinuclear   halo   may   or   may   not   be  
present  in  the  nucleus  
 Multiple   nucleoli   may   or   may   not   be  
visible  
 
Proplasmacyte  
 15  –  25  um  in  size  
 Cytoplasm   is   intensely   basophilic,   non  
granular,   more   abundant   which   has  
perinuclear  halo  
 Nucleus   is   round   or   oval   and   is  
eccentric  which  is  coarser  and  densely  
stained,  occasional  nuclei  may  be  seen    
 
Plasmacyte  (Plasma  Cell)  
 8  –  20  um  in  size  
 Cytoplasm   is   moderately   abundant,  
deeply   basophilic,   well-­‐defined   hof  
next   to   nucleus   and   usually   non-­‐
granular  
 Nucleus   is   8   –   2   um   in   size   with  
condensed   and   coarse   chromatin,  
round   or   oval,   eccentric   with   no  
nucleoli  
 Nucleus  has  “Cartwheel”  like  pattern