SERIE BLANCA NORMAL Y PATOLÓGICA Bqco. Gonzalo Ojeda Hematología Clínica Fa.C.E.N.A – U.N.N.E 2011 LEUCOPOYESIS Leucocitos granulares Neutrófilos( 50-70%) Basófilos(0-2%) Eosinófilos (0-5%) Leucocitos agranulares Monocitos (1-9%) Linfocitos(20-40%) RELATIVA % CAYADOS ABSOLUTA (mm3) 0a 3 0 a 300 50 a 70 3000 a 6000 EOSINOFILOS 1a 4 40 a 400 BASOFILOS 0a 1 0 a 100 LINFOCITOS 20 a 40 1500 a 4000 MONOCITOS 2a 8 80 a 800 SEGMENTADOS FUNCIONES DESTACADAS GRANULOCITOS : inmunidad innata MONOCITOS: inmunidad innata/adaptativa LINFOCITOS: inmunidad adaptativa 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 *VESICULAS SECRETORIAS: Contienen FAL *GRANULOS GELATINOSOS Gelatinasa, lisozima *Contienen alto contenido de glucógeno citoplasmático *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 FUNCIONES DEL GN 1-Defensa antimicrobiana: fagocitosis, bactericidia 2-Síntesis de la Proteína transportadora de B12 3- Síntesis de pirógeno leucocitario 4-Biosíntesis de nucleótidos MECANISMOS DE LA FUNCION FAGOCITARIA ADHERENCIA AL ENDOTELIO QUIMIOTAXIS OPSONIZACION Y RECONOCIMIENTO ENDOCITOSIS O INGESTION DEGRANULACION ACTIVACION DEL METABOLISMO OXIDATIVO SISTEMAS BACTERICIDAS ALTERACIONES FUNCIONALES ADHERENCIA QUIMIOTAXIS GRÁNULOS ACTIVIDAD OXIDATIVA CHEDIAK H DEFICIT DE MPO EGC DEFICT DE G6PDH CD11/CD18 CITOESQUELETO Rc DE MEMB 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 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 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 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 CHEDIAK HIGASHI ALBINISMO PARCIAL MAY HEGGLIN ALDER REILLY NEUTROFILO AGRANULAR Occurrence in blood: normally not present Comment: Granulesless segmented neutrophil. Staining: MGG Magnification: x 1000 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