趨化因子與共刺激分子聯(lián)合基因治療肝癌的實驗研究.pdf

1、第二軍醫(yī)大學(xué)博士學(xué)位論文趨化因子與共刺激分子聯(lián)合基因治療肝癌的實驗研究姓名：覃林花申請學(xué)位級別：博士專業(yè)：生物化學(xué)和分子生物學(xué)指導(dǎo)教師：郭亞軍時玉舫2001.5.1AbstractAbstractAnimportantaspectofimmunityistherecruitmentandaccumulationoflymphocytesintotargettissuesAllchemokinefamilymembersarechemot

2、acticforselectedpopulationsofieukocytesAnotherimportantaspectofimmunityistoinduceproliferationandcytokineproductioninTcells，andcostimulatorymoleculesarethoughttoplayanimportantroleinprimingTcellsGeneticmodificationofmany

3、typesoftumorcellstoexpresseithercostimulatorymolecules，suchasB71，4一lBBL，orchemokines，suchasMIP—let，increasestheirimmunogenicityToaugmentantitumoreffectinvivo，thecombinationtreatmentofhepatocellularcarcinomacellswithcosti

4、mulatorymoleculegenesandchemokinegenesmediatedbyretroviruswasinvestigatedInthisworkwetransfectedhepal6celllinewithmMIPltx，m4—1BBLormB7—1generespectively，moreoverwealsotransfectthetumorcellswithbothmMIPletandm41BBLgenesTh

5、eexpressionofmMIPletwasconfirmedbyRT—PCRandimmunohistochemistry；whiletheexpressionofm4一lBBLormB7—1wasconfirmedbyflowcytometryWhenm41BBLormB71transfectedcellswereusedasstimulatorsofallogeneicspleenlymphocytesinmixedlympho

6、cyte—tumorculture(MLTC)，T—cellproliferationweresignificantlyincreasedChemotaxisassayshowedthatmMIP一1dmodifiedcellscallsecretedsubstantiallevelsofchemokineactivityforactivatedmousespleenlymphocytesNoneofthetransfectants，i

7、ncludingthemulti—genetranfectantshowedalteredgrowrateinvitroTheinjectionofmMIP1stransfectedtumorcellssubcutaneously(sc)retardedtumourgrowth，whiletheinjectionofm41BBLormB71transfectantsprolongedtheanimaltumorbearinglifesp

8、anAlltumorfreeanimalsdevelopedlong—termimmunityagainstwild—typeHepal6butnotagainstEL4MiceinoculatedwiththemixtureofwtHepaI6andthechemokine／costimulatorymoleculeproducinghepal一6didn’tdeveloptumorInflammatorycellsinfiltrat

9、ionwerefoundintheinoculationsite_sofhepal一6transfectants，whilebothCD8andCD4cellsweredetectedTheseresultssuggestthatthecombinationofcostimulatorygeneandchemokinegenetherapycanaugmentantitumoreffectinvivoKeywords：chemokine