SERIE BLANCA NORMAL Y PATOLÓGICA Bqco. Gonzalo Ojeda Hematología Clínica Fa.C.E.N.A – U.N.N.E 2013

LEUCOPOYESIS

NUEVOS MODELOS

Leucocitos granulares

Neutrófilos( 50-70%)

Basófilos(0-2%)

Eosinófilos (0-5%)

Leucocitos agranulares

Monocitos (1-9%)

Linfocitos(20-40%)

RELATIVA %

CAYADOS

ABSOLUTA (cel/mm3)

0 a 3

0 a 300

50 a 70

3000 a 6000

EOSINOFILOS

1 a 4

40 a 400

BASOFILOS

0 a 1

0 a 100

LINFOCITOS

20 a 40

1500 a 4000

MONOCITOS

2 a 8

80 a 800

SEGMENTADOS

GRANULOCITOS

GRANULOPOYESIS NEUTRÓFILO

GRANULOPOYESIS EOSINÓFILO y BASÓFILOS

Factores quimiotácticos y activadores

Mediadores

IL-8, TNF-α, IL-1

Radicales libres de oxígeno, NO, prostaglandinas, leucotrienos, PAF, citoquinas

Eosinófilos

IL-5,C5a, MIP-1

Proteínas catiónicas, radicales libres de oxígeno, fosfolípidos, citoquinas

Monocitos

MCP-1, MIP 1.

Mediadores lipídicos, citoquinas.

Mastocitos, basófilos.

IL-8, MCP-1.

Histamina, mediadores lipídicos, citoquinas.

Célula

Neutrófilo

NEUTRÓFILO

NEUTRÓFILO

GRANULOCITO NEUTRÓFILO SP:3000 A 6000 cel/mm3 Tamaño: 12-20 μm GRANULOS 1°- AZUROFILOS INESPECIFICOS Lisosomas 1°,10 a 20% del contenido granular Mieloperoxidasa (MPO) Fosfatasa acida (FAC) Esterasas Beta glucuronidasa y beta galactosidasa Lisozima Otras proteinas básicas catiónicas

*GRANULOS SECUNDARIOS O ESPECIFICOS Constituyen el 80-90% de los gránulos, son anfóteros se tiñen lila o rojizo *Fosfatasa acida (FAC) *Lactoferrina *Fagocitina *Proteínas catiónicas leucocitaria (pirógenos) * Lisozima *NADPH oxidasa *Proteína de unión de la B12

*VESICULAS SECRETORIAS: Contienen FAL *GRANULOS GELATINOSOS Gelatinasa, lisozima *Contienen alto contenido de glucógeno citoplasmático

FUNCIONES DEL GN 1-Defensa antimicrobiana: fagocitosis, bactericida 2-Síntesis de la Proteína transportadora de B12 3- Síntesis de pirógeno leucocitario 4-Biosíntesis de nucleótidos

MECANISMOS IMPLICADOS EN FAGOCITOSIS      



ADHERENCIA AL ENDOTELIO QUIMIOTAXIS OPSONIZACION Y RECONOCIMIENTO ENDOCITOSIS O INGESTION DEGRANULACION ACTIVACION DEL METABOLISMO OXIDATIVO SISTEMAS BACTERICIDAS

MORFOLOGÍA

MIELOBLASTOS Size of the cell: 15 - 25 m Shape of the cell: oval, sometimes round Colour of cytoplasm: blue, without distinct perinuclear halo or with extended perinuclear halo Granularity: nongranular cytoplasm or a few thick azurophilic granules Nucleus' shape: usually oval, sometimes irregular, rarely round Type of chromatin: fine, with reticular appearance Nuclear/cytoplasmic ratio: high

Nucleoli: visible, medium or large size 1 to 4; brighter than chromatin

Occurrence: blood: not present marrow: < 5%

Comment: One myeloblast and two more mature neutrophilic cells (myelocyte and band neutrophil leucocyte) are seen. Platelets with small number of granules.

Staining: MGG Magnification: x 1000

1. myeloblast 2. neutrophil myelocyte 3. band neutrophil

MIELOBLASTOS Size of the cell: 15 - 25 m Shape of the cell: oval, sometimes round Colour of cytoplasm: blue, without distinct perinuclear halo or with extended perinuclear halo Granularity: nongranular cytoplasm or a few thick azurophilic granules Nucleus' shape: usually oval, sometimes irregular, rarely round Type of chromatin: fine, with reticular appearance Nuclear/cytoplasmic ratio: high or relatively high Nucleoli: visible, medium or large size 1 to 4; brighter than chromatin

Occurrence: blood: not present marrow: < 5%

Comment: Early myeloblast with very high cytoplasm - nucleus ratio, without granules. In the picture there are numerous other cells representing next stages of maturation of the series of granulopoiesis.

Staining: MGG Magnification: x 1000

1. myeloblast 2. promyelocyte 3. neutrophil myelocyte 4. neutrophil metamyelocyte 5. band neutrophil 6. segmented neutrophil 7. pycnotic normoblast 8. polychromatic normoblast 9. basophilic normoblast 10. proerythroblast

PROMIELOCITOS Size of the cell: 15 - 30 m Shape of the cell: oval or round Colour of cytoplasm: light-blue, with distinct halo Granularity: thick, azurophilic abundant or very abundant Nucleus' shape: oval Type of chromatin: start of condensation Nuclear/cytoplasmic ratio: moderate, low or very low Nucleoli: visible, medium or large size, brighter than chromatin, 1-2. Sometimes not visible.

Occurrence: blood: not present marrow: < 5 %

Comment: The arrow indicates one promyelocyte, which is the only promyelocyte in the field. The nearby large cell of the granulopoiesis series is not a completely differentiated promyelocyte (lack of perinuclear zone, and not abundant granules).

Staining: MGG Magnification: x 1000

1. neutrophil myelocyte 2. neutrophil metamyelocyte 3. band neutrophil 4. segmented neutrophil 5. plasmocyte 6. eosinophil 7. megakaryoblast

Size of the cell: 15 - 30 m

PROMIELOCITOS

Shape of the cell: oval or round Colour of cytoplasm: light-blue, with distinct halo Granularity: thick, azurophilic abundant or very abundant Nucleus' shape: oval Type of chromatin: start of condensation Nuclear/cytoplasmic ratio: moderate, low or very low Nucleoli: visible, medium or large size, brighter than chromatin, 1-2. Sometimes not visible. Occurrence: blood: not present marrow: < 5 %

Comment: The promyelocyte contains very abundant primary granules and a distinct zone of perinuclear halo. Degranulated platelets and discrete anisocytosis of the erythrocytes are also seen.

Staining: MGG Magnification: × 1000

PROMIELOCITOS

Size of the cell: 15 - 30 m Shape of the cell: oval or round Colour of cytoplasm: light-blue, with distinct halo Granularity: thick, azurophilic abundant or very abundant Nucleus' shape: oval Type of chromatin: start of condensation Nuclear/cytoplasmic ratio: moderate, low or very low Nucleoli: visible, medium or large size, brighter than chromatin, 1-2. Sometimes not visible.

Occurrence: blood: not present marrow: < 5 %

Comment: The promyelocyte contains abundant primary granules and a distinct zone of perinuclear halo. Also distinct anisocytosis of the erythrocytes.

Staining: MGG

Magnification: x 1000

Size of the cell: 15 - 25 m Shape of the cell: oval or round Colour of cytoplasm: light-blue or of pale pink colour undiscernible halo Granularity: abundant, thick azurophilic and neutrophilic granulation Nucleus' shape: oval or kidney shaped Type of chromatin: partially condensed Nuclear/cytoplasmic ratio: low or very low Nucleoli: not visible

MIELOCITOS NEUTRÓFILOS Occurrence: blood: not present marrow: 5 - 20 %

Comment: The arrow indicates neutrophil myelocyte with pink cytoplasm and disappearing primary granules. In the field there are also four other cells at a similar stage of maturation and numerous other maturating neutrophil cells.

Staining: MGG Magnification: x 1000

1. neutrophil myelocyte 2. neutrophil metamyelocyte 3. band neutrophil 4. segmented neutrophil 5. lymphocyte 6. plasmocyte 7. proerythroblast 8. polychromatic normoblast 9. pycnotic normoblast

Size of the cell: 15 - 25 m

MIELOCITOS NEUTRÓFILOS

Shape of the cell: oval or round Colour of cytoplasm: light-blue or of pale pink colour undiscernible halo Granularity: abundant, thick azurophilic and neutrophilic granulation Nucleus' shape: oval or kidney shaped Type of chromatin: partially condensed Nuclear/cytoplasmic ratio: low or very low

Nucleoli: not visible

Occurrence: blood: not present marrow: 5 - 20 %

Comment: Early neutrophil myelocyte in the blood. Also two matured neutrophilic leucocytes, a lymphocyte and platelets.

Staining: MGG Magnification: x 1000

METAMIELOCITOS NEUTRÓFILOS Size of the cell: 14 - 20 m Shape of the cell: oval or round Colour of cytoplasm: pink Granularity: a few azurophilic and neutrophilic, different in number Nucleus' shape: elongated, semicircular Type of chromatin: condensed Nuclear/cytoplasmic ratio: low or very low

Nucleoli: not visible

Occurrence: blood: not present marrow: 10 - 25 %

Comment: The arrow indicates a neutrophil metamyelocyte, one of seven present in the picture. Also other forms of maturating granulopoiesis are seen.

Staining: MGG Magnification: x 1000

1. neutrophil metamyelocyte 2. neutrophil myelocyte 3. promonocyte 4. promyelocyte 5. plasmocyte 6. basophilic normoblast 7. polychromatic normoblast 8. pycnotic normoblast

Size of the cell: 14 - 20 m

METAMIELOCITOS NEUTRÓFILOS Shape of the cell: oval or round Colour of cytoplasm: pink Granularity: a few azurophilic and neutrophilic, different in number Nucleus' shape: elongated, semicircular Type of chromatin: condensed Nuclear/cytoplasmic ratio: low or very low

Nucleoli: not visible

Occurrence: blood: not present marrow: 10 - 25 %

Comment: Neutrophil metamyelocyte indicated by the arrow is present in blood. Besides, neutrophil segmented and band-forms leucocytes are seen. Platelets not rich in granules.

Staining: MGG Magnification: x 1000

Size of the cell: 14 - 20 m

Shape of the cell: oval or round

CAYADOS

Colour of cytoplasm: pink Granularity: a few azurophilic and neutrophilic, different in number Nucleus' shape: semicircular Type of chromatin: condensed Nuclear/cytoplasmic ratio: low or very low

Nucleoli: not visible

Occurrence: blood: < 5% marrow: 5 - 20 %

Comment: Two band forms and one segmented neutrophil leucocytes in the blood. Also crenated blood cells and

Staining: MGG

platelets without granules are seen. Magnification: x 1000

CRITERIOS PARA CLASIFICAR UN NEUTRÓFILO COMO “EN CAYADO” -CUANDO NO SE OBSERVA LOBULACIÓN EVIDENTE -CUANDO EL NÚCLEO TIENE DIÁMETRO UNIFORME SIN ESTRANGULACIÓN EVIDENTE -SI EL NÚCLEO PRESENTA UNA PARTE MÁS DELGADA, ESTA NO DEBE SER MENOR A 1/3 DE LA PARTE DE MAYOR GROSOR

Size of the cell: 14 - 20 m

Shape of the cell: oval or round Colour of cytoplasm: pink

NEUTRÓFILO SEGMENTADO

Granularity: a few azurophilic and neutrophilic, different in number granulation Nucleus' shape: lobulated (normally less than 5 lobes) Type of chromatin: condensed Nuclear/cytoplasmic ratio: low or very low

Nucleoli: not visible

Occurrence: blood: 40 - 75 % marrow: 5 - 20 %

Comment: Three-lobulated segmented neutrophil leucocyte with fine neutrophil granularity.

Staining: MGG Magnification: x 1000

ALTERACIONES LEUCOCITARIAS CUANTITATIVAS CUALITATIVAS LEUCOPENIAS F(X)

NUCLEARES

CITOPLASMÁTICAS

LEUCOCITOSIS

ALTERACIONES FUNCIONALES ADHERENCIA

QUIMIOTAXIS

GRÁNULOS

ACTIVIDAD OXIDATIVA

CHEDIAK H DEFICIT DE MPO

EGC DEFICT DE G6PDH

CD11/CD18 CITOESQUELETO Rc DE MEMB

NUCLEARES OTROS

HIPOSEG

CONGENITA

PELGER HÜET

ADQUIRIDA

PSEUDO PELGER O PELGUEROIDE SMD EMBARAZADAS FARMACOS SEPSIS DE MALA EV LMA-LMC

HIPERSEG

CONGÉNITA

ADQUIRIDA

UNDRITZ DÉFICIT DE B12 SMD

CITOPLASMÁTICAS

CONGÉNITAS

-A. DE CHEDIAK-HIGASHI -A. DE MAY HEGGLIN -A. DE ALDER -DEGRANULADOS

ADQUIRIDAS

-GRAN. TÓXICAS -VACUOLAS -CUERPOS DE DöHLE -DEGRANULADOS

ALTERACIONES NUCLEARES

HIPOSEGMENTACIÓN NUCLEAR 

NO CONFUNDIR PELGER CON CAYADOS

CROMATINA MEDIANAMENTE CONDENSADA CROMATINA HIPERCONDENSADA LLEGA A MADURO SIN SEGMENTAR

PELGER HÜET   

AUTOSÓMICO DOMINATE PUEDE SER HOMO O HETEROCIGOTA PUEDE AFECTAR TB A EOS Y BASOFILOS HETEROCIGOTA HOMOCIGOTA

STODSMEYSTER

PELGER

Dynamics of nuclear envelope assembly in embryos and somatic cells, hilippe Collas ISBN: 978-0-306-47439-2 January 1, 2003

HIPERSEGMENTACIÓN   

 

> A 5 LÓBULOS LIGERO AUMENTO DE TAMAÑO CELULAR CONGÉNITA (UNDRITZ) – AUTOSÓMICA DOMINANTE- MUY RARA-80 A 90 % NEUTRÓFILOS CON MÁS DE 5 LÓBULOS PUEDE AFECTAR A EOSINÓFILOS ADQUIRIDA- BASTANTE FRECUENTESMD-ANEMIAS MEGALOBLÁSTICAS ALCOHOLISMO

HIPERSEGMENTANCIÓN

HIPERSEGMENTANCIÓN

ALTERACIONES CITOPLASMÁTICAS

ALTERACIONES CITOPLASMÁTICAS 

CHEDIAK HIGASHI

ALBINISMO PARCIAL

FISIOPATOLOGÍA MOLECULAR Mutación en el gen LYST (“lysosomal trafficking” regulator gen) o CHS1 que se encuentra en el cromosoma 1q42.1-q42.2 

MAY HEGGLIN

ALDER REILLY

NEUTROFILO AGRANULAR

GRANULACIONES TOXICAS

Granularity: thick granules, more eosinophilic stained than typical neutrophilic granularity. Single granules with a tendency to aggregate.

Occurrence in blood: normally not present

Comment: Band neutrophil leucocyte with dark, very abundant toxic granulation. Also anisocytosis of erythrocytes. Numerous ovalocytes. Normal platelets.

Staining: MGG Magnification: x 1000

CUERPOS DE DÖHLE

Definition: Dotted inclusions of blue, non-granular cytoplasm in pink cytoplasm of mature neutrophil leucocyte.

Occurrence in blood: normally not present.

Comment: The arrow points Döhle’s body in the granulocyte. Erythrocytes difficult to assess.

Staining: MGG Magnification: x 1000

Definition: Dotted inclusions of blue, nongranular cytoplasm in pink cytoplasm of mature neutrophil leucocyte.

Occurrence in blood: normally not present.

Comment: Döhle’s body pointed by the arrow. Also anisocytosis of erythrocytes.

Staining: MGG Magnification: x 1000

VACUOLAS

Occurrence: normal mature neutrophil leucocytes may contain single small vacuoles in cytoplasm

Comment: Distinct vacuoles changes in the neutrophilic leucocytes. Two target cells and one ovalocyte are present.

Staining: MGG Magnification: x 1000

VACUOLAS

Occurrence: normal mature neutrophil leucocytes may contain single small vacuoles in cytoplasm

Comment: Small vacuoles in a neutrophil leucocyte with fine neutrophilic granulation. Also anizocytosis of erythrocytes. A single spherocyte and a polychromatophilic cell are seen.

Staining: MGG Magnification: x 1000

NEUTRÓFILO APOPTÓTICO/MUESTRA VIEJA