Pripravki levkocitov. Dr. Dragoslav Domanovič, dr.med. Zavod RS za transfuzijsko medicino Ljubljana
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1 Pripravki levkocitov Dr. Dragoslav Domanovič, dr.med. Zavod RS za transfuzijsko medicino Ljubljana
2 Razdelitev levkocitov Granulociti polimorfonuklearni levkociti Nevtrofilci Bazofilci Eozinofilci Agranulociti mononuklearni levkociti Limfociti Monociti Makrofagi Dendritske celice Celice naravne ubijalke
3 Pripravki levkocitov Pripravki granulocitov Pripravki mononuklaernih celic Monociti Limfociti Celice naravne ubijalke Dendritske celice
4 Pripravki granulocitov Komponenta krvi, ki vsebuje predvsem v plazmi suspendirane granulocite, pridobljene z granulocitaferezo posameznega dajalca.
5 Funkcija granulocitov Antimikrobno delovanje Fagocitoza Degranulacija NET (neutrophyl extracelullar traps)
6 Kinetika granulocitov Dinamična zaloga - 9x10 9 nevtrofilcev/kg TM Prehod v krvni obtok 16,3x10 8 nevtrofilcev/kg/dan Od celotne količine granulocitov -v krvi le 5 10 % (4 x ) Cirkulirajoči pool 40 % Marginalni pool 60 % Dnevno jih nastane 1 x Razpolovni čas je 6 h
7 Zgodovina transfuzije granulocitov Strumia 1934 (i.m. aplikacija granulocitov) Freireich, Judson, Levin 1965 (granulocitafereza) Besinger 1993 (G-CSF) Freireich EJ, Judson G, Levine RM: Separation and collection of leukocytes. Cancer Res 25: ,1965.
8 Granulociti načini priprave Polna kri Trombocitno levkocitna plast ( buffy-coat ) Celični ločevalci Levkocitafereza Centrifugiranje Filtracija
9 Granulociti načini priprave Priprava granulocitov s celičnimi ločevalci Postopek citafereze s centrifugiranjem in ločevanjem na podlagi različne velikosti, specifične teže in fleksibilnosti celic. Postopek je lahko: Neprekinjen Prekinjen Mehanizem ločevanja: Consko centrifugiranje Pretočno centrifugiranje Elutriacija
10 Granulociti načini priprave Izboljšanje ločevanja med eritrociti in levkociti: Hidroksietilni škrob 6% Nizkomolekularni dekstran Modificirana tekoča želatina Povečanje izplavljanja levkocitov iz marginalnega poola in kostnega mozga Kortikosteroidi pred odvzemom rhu G-CSF
11 Granulociti lastnosti pripravka Granulociti (zlitje trombocitno levkocitnih plasti) Volumen ene trombocitno levkocitne plasti ml Število granulocitov 0,5 do 1 x 10 9 Hematokrit 0,3 do 0,5 Potrebno 10 krvodajalcev za terapevtski odmerek Granulociti (granulocitafereza) Volumen < 500 ml Število granulocitov > 1 x Hematokrit ~ 0,1 Hranjenje največ 24 h od priprave pri +20 do 24 C Potreben 1 krvodajalec za terapevtski odmerek
12 Granulociti kazalniki kakovosti Parameter Zahteva kakovosti Pogostost preverjanja AB0, RhD, K Določitev krvne skupine Vse enote Anti HIV I/II Negativen Vse enote HBs Ag Negativen Vse enote Anti HCV Negativen Vse enote Anti Lues Negativen Vse enote Anti CMV Negativen Po potrebi NAT RNA HIV Negativen Vse enote NAT DNA HBV Negativen Vse enote NAT RNA HCV Negativen Vse enote HLA Določitev HLA Po potrebi Volumen < 500 ml Vse enote Število granulocitov > 1 x Vse enote
13 Neželeni učinki - darovalec Aferezni postopek Flebotomija Citrat Izventelesni volumen HES, Cortikosteroidi G-CSF
14 Granulociti izbira pripravka ABO in Rh Zaradi prisotnosti velikega števila eritrocitov mora biti pripravek ABO in Rh skladen Potreben je navzkrižni preizkus HLA V primeru prisotnih HLA protiteles so potrebni HLA skladni pripravki enega dajalca (CMV negativen)
15 Granulociti - transfuzija Doziranje: >1 x granulocitov na dan, vsaj 4 dni (novorojenčki 1 2 x 10 9 / kg / dan) Pripravek mora biti obsevan Počasna transfuzija skozi 170 do 200 μm filter
16 Neželeni učinki - prejemnik HLA aloimunizacija Prenos povzročiteljev nalezljivih bolezni (CMV) Bolezen presadka proti gostitelju Akutna poškodba pljuč po transfuziji Nehemolitična reakcija po trasnfuziji Potransfuzijska purpura Nakopičenje hidroksietilnega škroba Alergične reakcije
17 Letno število granulocitaferez N = 214; povprečje = 10 / leto; Mediana = 6 (0 32) / leto število posegov leto število granulocitaferez
18 Zaključek Pripravke granulocitov lahko pripravimo z levkocitaferezami v zadostnih odmerkih Uporaba je redka Potrebno je določiti smernice za njihovo zagotavljanje in indikacije za njihovo smotrno uporabo.
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