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Adresa za dopisivanje: Doc.dr.sc.Srana uli, Klini ka bolni ki centar Split, Klinika za dje je bolesti, Klini ki odjel za hematologiju, onkologiju, imunologiju i medicinsku genetiku 21000 Split, Spin ieva 1. Tel. 385 21 556 256; Fax: 385 21 556 256; E-mail:  HYPERLINK "mailto:srdjana.culic@st.htnet.hr" srdjana.culic@st.htnet.hr Sa~etak: PRIMJENA CITOKINA U PEDIJATRIJSKOJ ONKOLOGIJI S uli Citokini su peptidi koje lu e limfoidne stanice i va~ni su u procesu proliferacije i diferencijacije stanica. Ovi topivi medijatori sudjeluju u razli itim procesima koji nastaju nakon stimulacije antigenom. Povezani su u citokinsku mre~u, a regulirani su genima. Lije enje citokinima se primjenjuje se kao dio specifi ne potporne terapije kod malignih i drugih proliferativnih bolesti. Najviae iskustva u lije enju danas imamo s imbenicima rasta hematopoetskih stanica, kao ato su imbenik rasta granulocita i makrofaga, eritropoietin, neki interleukini, interferon-Б i -Г. Rekombinantni humani eritropoetin se primjenjuje u lije enju anemije inducirane kemoterapijom, a reducira potrebe za transfuzijama. imbenik stumulacije granulocita primjenjujemo kod neutropenije nastale nakon kemoterapije, a posebno kod epizoda febrilne neutropenije i sepse. Ovaj stimulator hematopoeze danas se takoer primjenjuje za mobilizaciju krvotvornih mati nih stanica kod transplantacije krvotvornih mati nih stanica. Interferon-Б danas uspjeano primjenjujemo u djece s histiocitozom Langerhansovih stanica, masivnim hemangiomom i melanomom. Sve mogunosti primjene citokina u pedijatrijskoj onkologiji danas joa nisu iscrpljene. Imunoterapijski pristup lije enju malignih bolesti danas uklju uje imunomodulatorne citokine kao ato su imbenik tumorske nekroze alfa, interferoni, interleukini -2, -12, -15, -18 koji poti u antitumorsku aktivnost. Primjena novih citokina IL-21, IL-23, IL-27 s njihovim imunomodulatornim i antitumorskim u inkom nai e mjesta u buduim imunoterapijskim protokolima. Klju ne rije i: maligne bolesti, primjena citokina, dijete Summary CYTOKINE ADMINISTARTION IN PEDIATRIC ONCOLOGY PATIENTS S uli Cytokines are peptides produced by lymphoid cells and play important roles in cellular proliferation and differentiation. This soluble mediators are participating in different responses to antigenic stimulation. They are connected in cytokine network and their secretion is regulated at the molecular genetic level. Cytokine treatment has been used as a part of specific or supportive therapy in neoplastic and other proliferative diseases. Therapy with colony stimulating factors, growth factors, lymphokines and interferons today is well established. Most scientific experience is with colony stimulating factors, erythropoietin, some interleukins and interferon-alpha. Recombinant human erythropoietin is used for treatment of chemotherapy-induced anemia in children, reducing transfusion requirements. Granulocyte stimulating factor is applied for threatening neutropenia, especially episodes with febrile neutropenia and sepsis after chemotherapy. This haemathopoietic stimulator is also used for stem cell mobilisation in bone marrow transplantation. Interferon-Б to day in pediatric oncology patients is well establishet for threatening Langerhans cell hystiocytosis, massive haemanigima and melanoma. In future there will be a new possibilities in cytokine therapy using immunotherapeutic approaches to enhance host immune responses to developing tumors. Immunomodulatory cytokines such as tumor necrosis factor alpha, interferons, interleukins such as IL-2, IL-12, IL-15 and IL-18 are the most potent inducers of anti-tumor activity. New cytokines IL-21, IL-23, IL-27 and their immunomodulatory and antitumor effects suggest a promise for future immunotherapy protocols. Key words: malignant disease, cytokine administration, childhood Citokini Citokini su polipeptidi ili glikopeptidi molekularne mase od 6-70 kDa, a djeluju preko specifi nih receptora u samoj stanici i na stani noj membrani. Mogu biti pozitivni i negativni regulatori imunog odgovora. Oni su glasnici koji, pored hormona i neurotransmitera, spadaju u vrlo va~ne imbenike u komunikaciji izmeu ljudskih stanica. Citokini predaju informaciju ciljnoj stanici, koja ispoljava odgovarajui receptor. Interakcija citokina s receptorima za citokine rezultira aktiviranjem kaskade signala u stanici. Aktiviraju se faktori transkripcije, koji ulaze u jezgru gdje poti u aktivaciju razli itih gena. Nastaje aktivacija gena s posljedi nim fenotipskim ili funkcionalnim promjenama ciljne stanice (1). Sintezu i otpuatanje citokina mogu zaustaviti inhibitori modulirajui bioloaku aktivnost citokina ili inhibirajui sposobnost odgovora ciljne stanice. Rije  citokini je izvedena iz dvije rije i gr kog podrijetla -  cytos , ato zna i stanica, i  kine , iz rije i  kinein , ato zna i micati se. Ovaj naziv je uveden da bi se mogla razlikovati grupa imunomodulatornih molekula od hematopoetskih imbenika stani nog rasta. U citokine spada grupa: interleukina imbenika rasta interferona Izraz interleukin je prihvaen za grupu medijatora odgovornih za meusobno komuniciranje leukocita. Danas poznajemo 29 vrsta interleukina, koji se nazivaju od IL-1 do IL-29. Efektorske funkcije ovih proteina su aktivacija i diferencijacija stanice, kemotaksija i proliferacija airokog spektra stanica. Mnogi citokini su imbenici rasta stanica, neki su hematopoetski faktori rasta, dok neki imaju antiviralnu aktivnost i nazivamo ih interferonima. Reguliraju rast i diferencijaciju stanica imunosnog sustava, a meusobno su povezani u citokinsku regulacijsku mre~u. Oni modificiraju bioloaki odgovor, a stimuliraju proliferaciju i diferencijaciju limfocita T i B, NK stanica, makrofaga i dendriti nih stanica. Njihovo djelovanje regulirano je na molekularnom nivou genima. U pedijatrijskoj onkologiji primjenjujemo danas razli ite limfokine i interferone, a naro ito eritropoetin (EPO), imbenike stimulacije kolonija (engl. colony stimulating factor  CSF), imbenik rasta granulocita i makrofaga (engl. granulocyte macrophage colony stimulating factor, G-CSF, GM-CSF) (2). Konvencionalna terapija maligne bolesti u djece uklju uje kiruraki pristup, kemoterapijski i radioterapijski, koji esto ne daju zadovoljavajui rezultat. Imunoterapijski pristup lije enju malignih bolesti danas uklju uje imunomodulatorne citokine kao ato su imbenik tumorske nekroze alfa (engl. tumor necrosis factor  TNF), interferoni tipa I i II (IFN), zatim interleukini (IL) IL-2, IL-12, IL-15, IL-18 za koje se danas smatra da najja e poti u antitumoraku aktivnost, a ispituju se u razli itim predklini kim studijama. Weiss navodi da najnovija istra~ivanja uklju uju nove citokine kao ato su IL-21, IL-23, IL-27 ukazujui da bioloaka imunomodulacija kombinacijom razli itih citokina mo~e donijeti uspjeh u lije enju maligne bolesti. IL-12 ima sna~nu antitumorsku aktivnost regulirajui uroeni i ste eni imunosni odgovor (3). ERITROPOETIN Vrijednosti hemoglobina odreuju kvalitetu ~ivota, koja opada svakim gramom sni~enja hemoglobina. Primjena rekombinantnog humanog eritropoetina (rHEpo) zna ajno popravlja kvalitetu ~ivota u bolesnika s malignom bolesti (4). Nacionalni institut za rak (engl. National Cancer Institute - NCI) je prihvatio klasifikaciju anemija prema vrijednostima hemoglobina i dijeli ih na blage, srednje teake, teake i po ~ivot opasne (tablica 1). Tablica 1. Klasifikacija anemija Nacionalnog Instituta za rak Te~ina anemijeVrijednosti Hemoglobina (g/L)Blaga100 - 120 Srednje teaka80 - 100Teaka65 - 80 Opasna po ~ivot< 65  Ve viae godina rHuEpo se uspjeano primjenjuje u djece koja zbog hematoloake maligne bolesti ili solidnog malignog tumora primaju kemoterapiju ili su zra ena (5). Takoer se primjenjuje kod transplantacije krvotvornih mati nih stanica (6). Anemija je o ekivana posljedica maligne bolesti i lije enja kemoterapijom, no esto se zanemari i ne lije i. esto se svrstava u anemije kroni ne bolesti, jer etiologija ove anemije nije do kraja razjaanjena. Kod bolesnika koji boluje od maligne bolesti stalno je prisutan umor uz sni~ene vrijednosti hemoglobina. Uzroci anemije u djece s malignom bolesti mogu biti supresija koatane sr~i, oateenja bubrega kemoterapijom, gubitak krvi, infiltracija koatane sr~i ili hemoliza. Aplasti na anemija nastaje zbog supresije koatane sr~i, a naj eaa je anemija vezana uz malignu bolest i primjenu kemoterapije i zra enja. Razina EPO u djece sa solidnim malignim tumorom je znatno ni~a, pa djeca imaju anemiju i prije kemoterapije. Kvaliteta ~ivota raste sa svakim gramom koncentracije Hb, a idealna vrijednost Hb koja bi se trebala postii je 120 g/L. Primjena rHuEpo smanjuje potrebe za transfuzijama i zna ajno poboljaava uspjeh lije enja kod djece s malignom bolesti (7,8,9). Yilmaz je dokazao da se primjenom rHEpo zana ajno smanjila potreba za transfuzijama kod djece s malignom bolesti (10). Primjenom zra enja u pedijatrijskoj onkologiji smanjujemo tumor prevenirajui proliferaciju tumorskih stanica, a u inak zraenja ovisi o dobroj oksigenaciji tumorskih stanica. Veina djece s malignom bolesti razvija anemiju, pa je efikasnost radioterapije kod njih smanjena. Primjena rHuEpo mo~e zna ajno poboljaati kvalitetu ~ivota ovih bolesnika, a isto tako i djelovanje radioterapije (11). RhuEpo se primjenjuje u transplantaciji autolognih i alogeni nih krvotvornih mati nih stanica. Stvaranje endogenog EPO kod primaoca alogeni nog transplantanta je neadekvatno, pa primjena rHuEpo mo~e ubrzati rekonstituciju eritroidnih kolonija nakon alogeni ne transplantacije krvotvornih mati nih stanica (12). Veliki klini ki problem predstavljai anemija kod mijelodisplasti nih sindroma (MDS), klonalnih bolesti krvotvorne mati ne stanice koje karakterizira periferna citopenija nastala zbog abnormalne diferencijacije i sazrijevanja hematopoetskih stanica. Oko 90% bolesnika s MDS imaju anemiju ve od po etka bolesti i esto zahtijevaju transfuzije. RHuEpo kao jaki stimulator eritropoeze mo~e poboljaati anemiju kod mijelodisplazije, naro ito ako se primjenjuje zajedno s citokinima koji poti u ranije faze diferencijacije mati nih stanica (13). IMBENIK RASTA GRANULOCITA I MAKROFAGA Primjenom periodi ke intenzivne kemoterapije koja se primjenjuje u lije enju malignih tumora u djece mo~emo inducirati nastanak teake neutropenije. Prolongirana teaka neutropenija poveava rizik od smrti uzrokovane teakim septi kim infekcijama ato se dogaa kod 5% malih bolesnika. Primjena imbenika rasta granulocita (engl. granulocyte colony stimulating factor, G-CSF) i makrofaga (engl. granulocyte macrophage colony stimulating factor, GM-CSF) zna ajno ubrzava hematoloaki oporavak i izlije enje (14). Primjena G-CSF i GM-CSF skrauje trajanje neutropenije. Njegova primjena smanjuje u estalost nastanka febrilne neutropenije u djece s malignom bolesti koja se lije e kemoterapijom ili zra enjem. Izuzetak je kroni na mijeloi ka leukemija i mijelodisplasti ni sindromi, kod kojih nije dokazana u inkovitost. Primjenom ovog lijeka skrauje se trajanje neutropenije i njenih klini kih posljedica za dijete. G-CSF se takoer primjenjuje za mobilizaciju progenitorski stanica autologne periferne krvi ili za poticanje oporavka granulocitne loze nakon mijeloablativne kemoterapije. Primjena ovog lijeka kod bolesnika s malignom bolesti ne utjee na svukupni mortalitet, ali skrauje vrijeme hospitalizacije (15). Faktori koji stimuliraju granulocitopoezu su G-CSF i GM-CSF ako se daju preventivno u lije enju malignih limfoma smanjuje rizik neutropenije, febrilne neutropenije i infekcije (16). Zbog nastanka teakih aplazija limitirano je doziranje kemoterapije. Febrilna neutropenija naj eae nastaje u 25% do 40% bolesnika, a njena te~ina ovisi o primjenjenoj dozi kemoterapije pa su terapeuti esto prisiljeni odgaati kemoterapiju ili reducirati dozu iste (17). G-CSF i GM-CSF ubrzavaju nastanak neutrofila odnosno makrofaga i skrauju vrijeme febrilnih epizoda ili epizoda neutropenije. U kombinaciji s rHuEpo se koriste za mobilizaciju perifernih krvotvornih mati nih stanica nakon transplantacije koatane sr~i da bi ubrzali hematopoezu (18). INTERFERONI Interferoni nastaju kao odgovor na virus, dvostruku RNA, antigene i mitogene. U ovjeka postoje tri izoforme interferona, a to su: alfa, beta i gama. Ovi proteini brane organizam od virusa, poveavaju otpornost stanica na virusnu infekciju i djeluju kao citokini. Imaju airoki spektar djelovanja: va~an antimikrobni, imunomodulatorni, antiproliferativni uz poticanje diferencijacije stanica, kao i poticanje apoptoze (19). Inhibiraju replikaciju virusa unutar stanica i proliferaciju hematopoetskih stanica. Raefsky i suradnici su ispitivali u inak rekombinantnog IFN-Б i IFN-Г na stvaranje hematopoetskih kolonija koatane sr~i. Dokazali su da su oba rekombinantna interferona sna~ni supresori mijeloidnih kolonija nastalih iz CFU-C i da djeluju sinergisti ki. IFN-Г inhibira eritroidne progenitore i ukupne kolonije CFU-E u koatanoj sr~i (20). Stani na infekcija virusom je poticaj da se sintetiziraju neki od interferona (IFN-Б i IFN-В) zaati ujui stanicu od infekcije novim virusom. Drugu grupu interferona (IFN-В i IFN-Г) stvaraju limfociti kao odgovor na stimulaciju antigenima. Ovi interferoni imaju produ~eno antiviralno djelovanje i sudjeluju u eliminaciji virusa iz limfocita. Razlikujemo interferone tipa I i II. U tip I interferon spadaju IFN-Б, IFN-В, IFN-Г, IFN-o, IFN-Ф. Ve~u se na stani ne receptore za interferone. Razlikujema dva tipa stani nih interferonskih receptora: Receptori tip I koji prepoznaju IFN-Б i IFN-В, ali ne prepoznaju IFN-Г Receptori tip II koji prepoznaju IFN-Г, ali ne i IFN-Б i IFN-В. Interferonski sustav ima svoju vlastitu funkcionalnu organizaciju. Za stvaranje interferona potreban je induktor koji je naj eae virus, a proizvoa  je humana stanica. Interferoni moduliraju imunosni sustav indukcijom MHC i HLA sustava. Ovisno o tipu stanice koja ga proizvodi interferone dijelimo na IFN-Б kojeg stvaraju leukociti, IFN-В kojeg stvaraju fibroblasti, IFN-Г kojeg stvaraju imunokompetentne stanice. IFN-Б i IFN-В (Interferoni tipa I) su dosta srodni, a stvaraju ih leukociti i fibroblasti kao odgovor na virus ili dvostruku RNA. Suprotno tome IFN-Г (interferon tipa II) je vrlo malo sli an ostalim interferonima, a stvaraju ga najprije limfociti T kao odgovor na mitogene ili izlaganje antigenu. Utje u na regulaciju stani ne funkcije kao ato je fagocitoza, ali i poveavaju citotoksi nost limfocita T (IFN-Г), kao i aktivnost makrofaga i NK-stanica (21). Interferon-( IFN-( je jedan od prvih citokina koji nastaje kao odgovor na virusnu infekciju (slika 1). Ima va~na antiviralna i imunoregulatorna svojstva i uspjeano se upotrebljava u lije enju razli itih oboljenja. Aktivira citotoksi nost NK stanica i pospjeauje lizu stanica inficiranih virusom. IFN-( poti e ekspresiju MHC I. IFN-( mo~e se primijeniti kod djece s Hodgkin limfomom nakon autologne transplantacije krvotvornih mati nih stanica. Petropoulos sa suradnicima opisuje uspjeanu primjenu IFN-( u trajanju od 4 do 24 mjeseca kod djece nakon autologne transplantacije krvotvornih mati nih stanica i to 10 dana nakon hematoloakog oporavka (22). IFN-Б se takoer primjenjuje kod djece s histiocitozom Langerhansovih stanica (LCH) ako imaju multisistemsku bolest. Brojni izvjeataji govore o primjeni IFN-Б kao terapije spaaavanja u slu aju kada djeca nisu odgovorila na konvencionalnu kemoterapiju ili su imala rekurentnu bolest. Bolesnici s LCH koji su odgovorili na lije enje primali su IFN-Б odreeni vremenski period (23,24). Slika 1. Nastanak interferona u stanicama ubojicama nakon infekcije virusom  INCLUDEPICTURE "http://www.mfi.ku.dk/ppaulev/chapter32/images/32-4.jpg" \* MERGEFORMATINET  Preuzeto iz: http://www.mfi.ku.dk/ppaulev/chapter32/images/32-4.jpg Kod nekih bolesnika je primijenjen kao terapija odr~avanja u parcijalnoj remisiji bolesti (LCH). uli i suradnici su prikazali tri bolesnika s LCH kod kojih je uspjeano primijenjen IFN-( kao terapija odr~avanja, kod bolesnika s multisistamskom bolesti kod kojih je rizik od relapsa bio vrlo visok. Od tri bolesnika kod jednog je na konvencionalnu kemoterapiju postignuta djelomi na remisija bolesti. Svo troje djece je postiglo remisiju bolesti. Njegova primjena kod visoko rizi nih bolesnika mo~e prevenirati relaps bolesti (25). Timke sa suradnicima opisuje slu aj 11 godianjeg dje aka s diseminiranom lymphangiomatozom, koji je kroz dvije godine uspjeano lije en s IFN-( (26). Terapijska aktivnost IFN-( u onkologiji mo~e biti potencirana kemoterapijom. Uglavnom je primjenjivan kod neoplazmi koje ne odgovaraju na konvencionalnu kemoterapiju. U 14 od 24 bolesnika kod kojih je primijenjen IFN-( postignut je odgovor na lije enje. Od 14 bolesnika 8 ih je pre~ivjelo. Potpuni ili djelomi ni odgovor na lije enje primjeen je kod bolesnika s astrocitomom, neuroblastomom i ganglioneuroblastomom (27). Terapija IFN-( indicirana je kod djece s mijeloproliferativnim sindromom, koja nemaju kompatibilnog donatora hematopoetskih mati nih stanica. Djeca s metastatskm neuroblastomom, primitivnim neuroektodermalnim tumorom, inoperabilnim gliomom, relapsom leukemije ili limfoma kao i s diseminiranom LCH su kandidati za primjenu IFN-(. Kombinacija IFN-( s kemoterapijom poboljaava prognozu bolesti, a primjena IFN-( u fazi odr~avanja poma~e boljoj kontroli minimalne ostatne bolesti (28). Intratumoralnu primjenu IFN-( kod troje djece s kraniofaringeomom prikazao je Pancucci. Ova primjena bila je korisna u cilju odgaanja operacijskog zahvata do sazrijevanja hipotlami ko-hipofizarne osovine (29). Takoer je uspjeano primjenjen intralezionalno u lije enju velikih hemangioma naro ito onih koji ugro~avaju ~ivot djeteta (30). Greinwald je prikazao kod 24 pedijatrijska bolesnika s masivnim hemangiomom koji je ugro~avao ~ivot djece pozitivan ishod lije enja s interferon alfa-2a u dozi veoj od 3 miliona U/m2 dnevno minimalno 4 mjeseca. Potpuni odgovor na lije enje postignut je kod (42%), zna ajan kod 16%, srednji kod 26%, a bez odgovora bilo je 16% bolesnika (31). French je prikazao iskustva u lije enju pedijatrijskog melanoma kod kojeg je primjenjeno adjuvantno lije enje kod 2 bolesnika s visokim rizikom povrata bolesti. Autori preporu aju da se kod visoko rizi nog melanoma nakon kirurakog odstranjenja nastavi adjuvantna terapija s interferonom alpha-2b. Prikazani bolesnici nisu imali povrat bolesti (32). Interferon-( IFN-Г je citokin kojeg stvaraju aktivirani limfociti T i limfociti T4, NK stanice, te monociti i makrofazi. Igra vrlo va~nu ulogu u obrani domaina ispoljavajui svoju aktivnost na airokom dijapazonu stanica, regulirajui aktivnost imunih stanica. Ima vrlo va~nu ulogu u kontroli bakterijskih, virusnih i parazitnih infekcija. Stani ni odgovor na IFN-Г odreuje receptor (IFN-ГR) koji aktivira signal prema stani noj jezgri i kona no regulira ekspresiju gena (33). To je najva~niji proinflamatorni citokin odgovoran za aktivaciju i reguliranje fagocitne funkcije mononuklearnih stanica. Regulira stvaranje nekoliko imunomodulacijskih i proinflamatornih citokina kao ato su IL-12 i TNF-(. Sni~ene vrijednosti nalazimo kod RA, SLE, limfoma, kongenitalne rubeole ili citomegalije, malarije, herpes simpleks infekcije, AIDS-a, leishmanije i TBC. Neposredno potiskuje eritropoezu, a visoke koncentracije EPO ne mogu poniatiti njegov inhibitorni u inak (34). Funkcionalna interakcija IFN-Г s molekulama cjelokupne interleukinske i citokinske mre~e danas je interes brojnih znanstvenih istra~ivanja. Koliko je meusobno povezana interleukinska mre~a pokazao je Hsu sa suradnicima, kada je dokazao da antiinflamatorni citokini IL-4 i IL-10 inhibiraju IL-2 induciranu sintezu IFN-Г (35). Recombinantni interferon gamma (USA Actimunne) je rekombinantni preparat koji je kemijski identi an ili sli an endogenom IFN-Г i ima antineoplasti ne, imunoregulatorne i antiviralne sposobnosti. Ve~e se za stani ni povrainski receptor za IFN-Г stimulirajui protutijelima specifi nu citotoksi nost i poja ava prijanjanje NK stanica za tumorske stanice. Aktivira kaspaze i na taj na in inducira apoptozu malignih stanica. He i koautori su pokazali da IFN-Г stimulira sazrijevanje dendriti nih stanica i na taj na in mo~e potaknuti antigen specifi ni imunosni odgovor i antitumorsku imunost (36). Stanice Ewingova sarkoma jako su podlo~ne apoptozi putem inducirajueg liganda imbenika tumorske nekroze (engl. tumor necrosis factor apoptosis-inducing ligand - TRAIL). Kombinacija s IFN-Г poja ava ovaj u inak (37). Apoptoza povezana s imbenikom tumorske nekroze javlja se selektivno kod razli itih tumora uklju ujui veinu stani nih linija kod Ewingovog sarkoma, agresivnog tumora koji se javlja kod djece i mladih ljudi. Primjena IFN-Г ili primjena IFN-Г u kombinaciji s agonistom TRAIL receptor zna ajno smanjuje u estalost metastatske bolesti. Ovom kombinacijom posti~e se impresivan efekt na primarni tumor kao i na tumorske metastaze Merchant sa suradnicima smatra da se ovom kombinacijom lije enja mo~e kontrolirati primarna i metastatska bolest kod Ewingovog sarkoma (38). INTERLEUKINI Interleukin 3 IL-3 je citokin koji primarno nastaje iz aktiviranih limfocita T. Stvaraju ga i limfociti T8. Gen za IL-3 nalazi se na humanom kromosomu 5 samo 9 kilobaza daleko od GM-CSF gena. Ovaj pleiotropni imbenik mo~e stimulirati proliferaciju i diferencijaciju pluripotentnih krvotvornih mati nih stanica, kao i razli itih hematopoetskih progenitora. Djelujui sinergisti ki s ostalim ranim hematopoetskim imbenicima rasta (IL-1, IL-6, SCF, G-CSF), poti e proliferaciju multipotentnih progenitora samo u ranom stadiju njihovog razvoja (39). GM-CSF, IL-5, M-CSF i EPO su potrebni za kona nu diferencijaciju ovih s IL-3 induciranih progenitora (40,41). IL-3 svoju bioloaku aktivnost provodi putem receptora na povraini stanice. U inci humanog i miajeg rekombinantnog IL-3 na hematopoezu in vivo prou eni su kod raznih ~ivotinjskih modela, koji su pokazali da IL-3 mo~e poja ati hematopoezu in vivo. IL-3 ima autokrini u inak na megakariocitopoezu djelujui zna ajno na megakariocite da otpuataju imbenik rasta deriviranog iz trombocita (engl. platelet-derived growth factor - PDGF) i transformirajueg imbenika rasta В1 (engl. transforming growth factor - TGF). Mobilizacija perifernih krvotvornih mati nih stanica mo~e se uspjeano napraviti primjenom kombinacije IL-3 i GM-CSF-a ato su opisali Brugger i suradnici (42). Brugger sa suradnicima ispitivao je utjecaj primjene rhGM-CSF-a na oporavak neutrofila i primjene rekombinantnoh humanog IL-3 (rhIL-3) na oporavak trombocita koji su primjenjeni nakon standardne doze kemoterapije. Brzi oporavak neutrofila primjeen je kod svih bolesnika, a oporavak trombocita samo kod grupe bolesnika koji su primali intenzivni kemoterapijski tretman (43). Lemoli je prikazao kako se uspjeano mo~e primijeniti kombinacija rhIL-3 i rhG-CSF-a da bi ubrzali hematopoetski oporavak nakon autologne transplantacije krvotvornih mati nih stanica kod bolesnika s malignim limfomom, koji su predhodno lije eni visokim dozama kemoterapije. Takvi bolesnici su imali manje potrebe za transfuzijama, a navedenu kombinaciju su primili bez ne~eljenih posljedica (44). Isti autor ponovo dokazuje da je proliferacija i koncentracija progenitora iz koatane sr~i vea primjenom dvojne terapije IL-3 s G-CSF nego kod primjene samog G-CSF-a (45). Kombinacija viae faktora rasta hematopoetskih stanica ubrzava oporavak bolesnika podvrgnutih transplantaciji hematopoetskih mati nih stanica zbog maligne bolesti (46). Interleukin-2 Autologna transplantacija krvotvornih mati nih stanica kod bolesnika s leukemijom ili limfomom esto je povezana s relapsom bolesti. Primjena rIL-2 u vrijeme transplantacije, naro ito ako je dokazana minimalna rezidualna bolest mo~e smanjiti rizik od relapsa hematoloake maligne bolesti. Fefer sa suradnicima je dokazao da primjena rIL-2 nakon autologne transplantacije krvotvornih mati nih stanica reducira u estalost posttransplantacijskog relapsa i poboljaava pre~ivljenje bolesnika s hematoloakim malignim bolestima kao ato su leukemije i limfomi (47). Primjena rIL-2 potie razli ite efektorske limfocite T i NK stanice i graft versus tumor efekt (48). Kontinuirana infuzija niskih doza rIL-2 poveava anti tumorsku citotoksi nost zna ajno poveavajui broj i aktivnost NK stanica nakon transplantacije hematopoetskih mati nih stanica (49). Interleukin-12 Izgleda da je ovaj interleukin odlu ujui faktor uroenog i steenog imunosnog odgovora. Sam po sebi mo~e potaknuti sna~an antitumorski efekt djelujui sinergisti ki s nekoliko drugih citokina. Na isti na in mo~e poja ati imunoregulatornu i antitumorsku aktivnost regulirajui interakciju domaina i tumora (3). Za sada znamo da su mogunosti njegovog djelovanja ograni ene samo na odreene maligne tumore izmeu ostalih melanom. IMUNOMODULACIJA CITOKINA Talidomid Talidomid i njegovi analozi lenalidomide, Revlimid, ACTIMID djeluju na pojedine citokine u citokinskoj mre~i stimulirajui humani imunosni sustav inhibiraji brojne citokine kao ato su TNF-Б, IL-1В, IL-6,-12 i GM-CSF. Ovi preparati mogu inducirati proliferaciju limfocita T, poticati stvaranje citokina IL-2 i IFN-Г te na taj nain poveavati antitumorsku aktivnost limfocita T. Takoer pokazuju odreenu aktivnost protiv nekih solidnih tumora. Iako mehanizam njihovog djelovanja nije do kraja jasan smatra se da anti tumorski efekt potsti~u pomou imunomodulatornog i neimunomodulatornog djelovanja. Smatra se da poti u antitumorsku aktivnost putem Th1 citokina IL-2 and IFN-Г. Takoer stimuliraju makrofage i NK stanice. Antiangiogeni i proapoptotski efekt primjene ovih preparata pokazao se efikasan kod dobro vaskulariziranih tumora inhibirajui vaskularni endotelni imbenik rasta (engl. vascular endothelial growth factor  VEGF) (50). Chintagumpala je proveo studiju kombinacije thalidomida s carboplatinumom u djece s refrakternim solidnim malignim tumorom (maligni gliom, meduloblastom, anaplasti ni ependimom, Ewingov sarkom, neuroblastom, angiosarkom, osteosarkom, nefroblastom). Studija je pokazala dobru podnoaljivost u djece uz produ~eno odr~avanje stabilne bolesti (51). ZAKLJU AK Imunoterapijski pristup lije enju malignih bolesti danas uklju uje imunomodulatorne citokine kao ato su imbenik tumorske nekroze alfa, interferoni, interleukini -2, -12, -15, -18 koji poti u antitumorsku aktivnost. Primjena novih citokina IL-21, IL-23, IL-27 s njihovim imunomodulatornim i antitumorskim efektom nai e mjesta u buduim imunoterapijskim protokolima (52,53). LITERATURA uli S. Serumski citokini u djece i odraslih s autoimunom trombocitopenijom. Doktorska disertacija. Medicinski fakultet Sveu iliata u Zagrebu, 2004.  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5X5Y5j5k5~557686>6?6’6“6п6р6ё6ђ6777У7Ф7Ю7Я7б7в78Ž8—8˜8š8›8Y9Z9f9g9i9j9&:':1:љюљюоюљюљюоюљюљюоюљюљюоюљюљюоюљеШеШЙШеюљюоюљШеШЙШељюљюоюљюљюоюљюљюоюљюљюоhkmLhЉbп0J>*B*phџjhkmLhЉbп0JUhkmLhЉbп0JhkmLhЉbп0J>*B*\phџjhkmLhЉbпU hkmLhЉbпI1:2:4:5:ё:ђ:ќ:§:џ:;О;П;Ы;Ь;Ю;Я;‡<ˆ<Ž<<‘<’<P=Q=[=\=™=š=т=у=ђ=ѓ=>>У>Ф>Э>Ю>а>б>?‘?ž?Ÿ?Ё?Ђ?]@^@g@h@j@k@&A'A0A1AŠA‹AŒAзAиAшAщABBBдBеBфBхBчBшBЂCєэєэєнєэєэєнєэєэєнєэєэєнєэаЧаИаЧєэєнєэєэєнєэєэєнєэєэєнєэЧаЧаИаЧэєэєнєэєэhkmLhЉbп0J>*B*phџhkmLhЉbп0JjhkmLhЉbп0JUhkmLhЉbп0J>*B*\phџ hkmLhЉbпjhkmLhЉbпUHЂCЃCЋCЌCЎCЏClDmDxDyD{D|D7E8EAEBEDEEEџEFРР РYРZР[РЙРКРйРкРэРюРЉСЊСГСДСЖСЗСpТqТxТyТ{Т|Т4У5У;У<У>У?УљУњУФФOФPФQФ”Ф•ФЁФЂФИФЙФХєфєнєнєфєнєнєфєнєнєлфєнвХвХЖХвєнєфєнєнєфєнєнєфєнєнєфєнвХвХЖХвАЅhfDхh^(РmHsH h^(Р0JhkmLhЉbп0J>*B*phџjhkmLhЉbп0JUhkmLhЉbп0JU hkmLhЉbпhkmLhЉbп0J>*B*\phџjhkmLhЉbпU?Smith RJ. 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