Název:
Vlastnosti myších buněk, které byly modifikovány geny pro imunomodulační faktory a endostatin
Překlad názvu:
Characteristics of HPV16 transformed mice cells, transduced with genes for immunomodulating factors and endostatin
Autoři:
Lakatošová, Monika ; Vonka, Vladimír (vedoucí práce) ; Reiniš, Milan (oponent) ; Forstová, Jitka (oponent) Typ dokumentu: Disertační práce
Rok:
2011
Jazyk:
cze
Abstrakt: [cze][eng] Souhrn Vt6toprAcijsemzkoumalavlastnostimy3lchbun6ktransformovanfchviremHPV16'kfer6 byly genetickymodifikovfny vnesenimgent pro cytokiny' chemokin ajeden angiostaticki gen' Byly testovAnyhlavnlcharakteristikytransdukovanfchbun€kvporovnAnisrodidovkjminddorovfmi buikami in vitro a in viYo VprvnldristijsempouZilabunddnouliniidefrcientnlnabunddnoutymidinkin6zu'ozratenou jakol23IA,kter6bylaodvozenazmy5ich(c57BLl6)ledvinnjchbun€ktransformovarrjchonkogeny E6lETHPV16aaktivovanfmonkogenemH.rasomaEenlchjakoMK16.Builkyl23IAjsem transfekovala bicistronickfmi plazmidy nesoucimi geny pro herpetickou tymidin kin6zu (HSv-Tn, kostimulaini molekulu B7-,1(cD 80) nebochemokin - protein 1 chemoatrahujicimonocyty (MCP-L)' ho kontrolni idely jsem pouZilaplazmid obsahujicljen genpro I/SY-IK' Do testfijsem zahrnulatak€ diive izolovan6 bufiky Bg, kterd byly transdukov6ny geny pro faktor stimulujicl tvorbu kolonii granulocytiamonocytft(GM-cSnaHSV-TK'VSechnyizolovan6liniebylycitliv6kegancikloviru' prokazujice tak piitomnost HSV-TK. po inokulaci syngennimmystm, byly bunky exprimujici GM- CSFa B7-l neonkogenni.Po odkov6nibunEkprodukujicichMCP-I vfvofily t6m6l v5echnymy3i fl(dory. Zvl?rltaimtmizovani bunkami produkujici GM-CSF a B7-l byla chr6ndnapled n6slednou...Genesfor two cytokines,one chemokineandgenecoding for oneangiostaticfactor were used in the presentwork for tansfection ofmouse IIPV-16- transformedtumor cells. Main characteristics oftransduced cells werc t€stedrnraTo and,in vivo and,cnrnparedwith theparentaltumor cells. In the first part I useda thymidine-kinase deficient (cTK) cell line designated123IA, which had beenderived from IIPVI6 transformedmouse(C57BL/6) cells MK16. To obain genetically modified cells, 1231Acells were transfectedwith bicistronic plasmids carrying the herpessimplex type 1thymidine kinase(HSV-TK) gen,eandeitherthe genefor the mouseB7-l (CD80) co-stimulatory molecule or the gene for the monocyte-chemoattractantprotsin 1 (MCP-|). For control putposes,a plasmid vector carrying only the HSV-TK genewas used.For comparativepurposesI also used89 cells, previously isolated in our laboratory, which expressthe mousegranulocyte-macrophagecolony stimulation factor (GM-CSF) andHSV-TK gene.All the cell lines testedwere found to be sensitiveto minute amountsof ganciclovir, revealing the production of functional HSV-TK. When inoculatedinto syngeneic mice, cells expressing either GM-CSF or B7-1 were non-oncogenic. Nearly all mice inoculatedwith MCP-l-producingcellsdevelopedtumors.Animalsinjectedwith GM-CSF or B7-l- producing cells were...