IL-18DNA免疫對(duì)HIV-1核酸疫苗誘導(dǎo)的免疫應(yīng)答的影響.

1、* 19 a 2 mqi M 4如年“V1ROLOGICA StNICAIL-18 DNA免疫對(duì)H1V.I核酸疫苗誘導(dǎo)的免疫應(yīng)答的影響王福祥，孫水灣孫永年.徐 哲，王臨旭，劉 娟康文球Modulation of Immune Responses to a HIV-1 Nucleic Acid Vaccine byIntcrlcukindS DNA ImmunizationWANG Ri-xiamg. SUN Yotig W. SUN Yong-ntan. XU Zhc. WANG Lin xu UU Jiun KANG Wcn /henfCrwr q /Mcwcmi PruMmi仙仙Mrwwr

2、wiof PtA. 3灑fhtpiraf .TV FownA M金金MrafUSnyvcirx Xi *R7I(K佩AbMract: To incuifiie ihr effect of interleukin*18 (IL-1S DNA immunixatian on immune rcponc induced byHuman immurnnkfioeniry virus liHIV-l) nucleic wid vaccine (DNA vKcinc). the rcvo( (xte expression vector pVAXl. BAIM: mkcwere immunizai with

3、 pCI-neoGAG alone or CO-admtmMcrvd with the DNA enoudinf for IL-18. Thru cra werecollected for analyzingAMI-HIV antibody and IHN-y by F.LISA and tplecn ceils were isolated for dctcirtmgantigcn-pevifk Iy*m()()hoprviifertirespung and specific CTL reipomc by MTT aseuy and I.DH aviay. ropoclivdy.Robinet

4、MXIenzyma dicuioa aaal) )*Ms jnd DNA tequenemg rcwlts repealed ttuu the rofntxruni cipro%KMi vectorpVAXML-18 hm been comiructcd wccevifully. ThcMiiHIV antibody tiieni of mice coimmuni/rd with pCI-ncoGAGund the DNA encoding for II.-18 uerr lower chim that of mux immunized with pCI*neoGAG akme (P. (n

5、contrmt.lhe IFN-y level of mice co-immunized with pCl-rwoGAG and the DNA encoding for IL IR higher than that of miceimmunized with pC?l-nenGAG alone Furthermore, compared with mice mjccttd with pCI- nccGAG alone (bespecific CTL cjtoioxrtyMMI antifen-specific lymphopmlifentirwpong 4 mice immunirai wi

6、th pCI-oeoGAG jrxi theDNA encoding for (L-IR woe Mgmlkanily enhanced P. The DNA cncrxling foe IL- IR together with IIIV DNAveccinc may enhance pcrafic Tb-I rcpony and cellular immune rc%ponc elicited in mice However, the DNAencoding for II.-I8 may dnwnrcguljic the humonl ropoogKty wonk: Human jmmunn

7、ddkMTKy vinu l(HIV-l); DNA Axinaom. imakuiun lMIl.-18; Imrnuni/jtfim.為/坍充fl窮介仁血口兇軸人Q咬敵HIV hW4f64#免吸的形將人他#VWWXMftH*l|.pVAXIfpVAXl IL-IB：m pci eoGAG ft pVAX I (L. 18 |序ra*尿割r&Q金徇S耳pCI-fieoGAG (Cl neoGAG Uft pVAXI IL IK霓*朋金MlfclRE颯V lp4秋*ri rtftj rCl neoGAG鼻lifl比斂pH teoGAG Uft pVAXI fVlS fclftffl+R

8、tlrt的fFN-V ftft Pcooi 4VAXI IL II巴饗*儀金ISO uMttWBm.一一 二二oauai. awes, Finan nni ion iH Illh 豐.敏 1 斤不 R I K.H- tl-穴.ttl. 0ME-wAvmghMrHve二二 i付i %l. KCWnUMrE trad冷( (2):V7-IOO AralXKMT pCI-mcKJAGIM)011. IL I YNft：性峻中分鬟號(hào)RJ739jtMtiiRW：A文量號(hào)，1001-51 $2iVMi02M)Q7OI個(gè)CjikM人免疫y臥訓(xùn)4| I Human immuno還0、2 HIV吸洶必復(fù)紇紗請(qǐng)9

9、和堆卄鯛則ty ttCTL biff. DNA疫苗可以調(diào)勸機(jī)仏的- ?1HIV I*心11的林疫（6共免蛙小歐 許價(jià)人IL W Efi/cafii仇対HJV核展儀伯講詔免 夠?yàn)榻袼庡麡?shù)幾效的治 件Hiv HIW冷伯披供實(shí)附和丹論舉Mi 1材料與方法1.1實(shí)色材Hnn jMio9i5wiibi及樓act俾PVAXI為本卒?！安挪既薎L IM移囚的廣斡PgEM- I II 18為安交通人學(xué)直flU晴穴宇I(lǐng) fltt授便常色有中厲人HIV-I Aftlrlt Gag農(nóng)達(dá)MTox96RNofbRadMMKtive CyKMoiiciiy Axsay試刊魚(yú)為X：U1Protnc?!公對(duì)八U1.2 pVA

10、XMI. IHMIX*達(dá)tH的mjn內(nèi)切酬AurnH k EmR I燈*切質(zhì)外pGEMT-1L-I8. KJt中的/L/& WIUWHH切儀帰的曲檢衣達(dá)戦體pXI P 翔埶介&IL U兇的典林衣仏H體pVAXI IH8） .UlUftft步課見(jiàn)分了加 實(shí)於指南）4 tJ ficyiHL *l &含疲片的6幾培養(yǎng)第中祝種傳代的p8l5 豈 X 細(xì)倉(cāng)卞長(zhǎng)年90%T5%電令時(shí)fKtttt用A.mm和扶牛蟻的DMhM汛2劇毎孔1U入2mL DMEM 12配A：Hi.5min內(nèi)轄A和B思勻屯泓作用20min.（3） W 522pL （Tjtf |0 lipofccuminc起合輸

11、加入軻 孔含細(xì)曲的DMEM中減勻.U） ttMir 379. SCO?WnMtt 6h.10%FCS的IMO 4 2mL h站并以G4I83C0nu/mL 4 4 WJ同H設(shè)空朋槍対開(kāi)#L 1.4質(zhì)符DNA的大l和代化Mktrff試列盒W.*J-f5tfNA用TW1*W巾噥浚章WEL 201*帀憐用.1.5小I*的DNA ft6 WM01） BaINc小MHUnmMTIK人7實(shí)阱動(dòng)H4 ar-j if 2ogigse.Ms只.A ffl：t合豹妙m UllMit劎取淪pCI-ccoGAG lOOpL/R和pVAXML-18 lOOpU只i B41 i PCIFCOGAG肌pCIcoGAGIOO

12、MU只:C虬對(duì)恿細(xì)注創(chuàng)空爪忖pCI o lOOplJ R. IvHbM4：g次住片削24h劉Sl“ 03的 比*兇lOOpLNftft.件2. 4 WHNITMM加令M. 2姐尚樸齊次免稷HU俱展取tl 6 Ml時(shí)18球 取血及分測(cè)免疫功!t 1.6E1JSA小特異性杭H1Vp24|ASF1.ISA山.Kttttlft人騰樸曲灰達(dá)的廈爼HIV-lp24蛍白.3%BSA刃WJfrifcl入適勺轉(zhuǎn)軒的小 乂血溝祈件洗板GUI入化期標(biāo)記的BE優(yōu)板所抽&第H?色tfHtt450nmM9?$/lODffi.只休慢竹步StWJt紋叫 免疫辦Mdn和p IFNFI4SA方法.W1KUM食晁供的比明L

13、* 卞II DNA扯*畸1丫I慮倩4#的純蟲(chóng)幷彌電17免侵小RRiM巴細(xì)般增旳實(shí)轉(zhuǎn)口?:檢解別免疫爾fcllW 100 IIWMWfl 收 #? 以Y渣JUtBi潯滾(NH4CI 155mmo4/L. KHCOi lOmmold. EDTA 0 1 mmol/L去球紅細(xì)泓.UXAlg FCS W RPMI-IMO林統(tǒng)副MP沫復(fù)個(gè)2XIG/ml.,収此WmfiM液IGOpL. tUX96yttt. Cttiflill入HIV lp24iTI ma為i皿.njRflt不XHIV.IIA*卜 林齦人遊胛增眼的/l18小Mcn,&QJIJ力仏鴻;赫實(shí)驗(yàn).x HIV.|p24 ififi. n*

14、lj為免瞪介fcOW淋巴i 37V S*Or堆弄54麻件為毀&煥 以pa-M0GAGnis第貝廉応4U1G作為BURI效 副9削以50 2S:I和115:13個(gè)盤(pán)IR比淤介竦掙5h G.以1.DH cn.明竹堆”H時(shí)設(shè)效孔.杞聖電ri發(fā)科放孔和代mififtxwttfl. cn.朋性 i丈 -效應(yīng)魅Ift白發(fā)發(fā)fftfU八比紙覚 發(fā)*族)xian.19嫌計(jì)學(xué)方法SPSSrt.計(jì)用林眈.2.1 pVAXML達(dá)啟體耐殮及蛙定pVAXl IL IB MB定麗itXJa I pVAXI-II.-I8用SH I：UpR I刃曲切H 町切岀SOObp1( (niinn.豪劇殘?bào)w構(gòu)議近( (w.R

15、WitM ttw UWITj n aenBink宇。介 金H.氏I AmlyiHMpWAXl M川切enzyme*血唄I DI3MQ X RVAXI IL- IS dfCMfd *! * I2 2免疫小Ml*中椀H【V p24敏體測(cè)違m ELISA沈分KHftAHta 5 p24 t*體淘度熱云計(jì)WfK/tfl MiRtum 2 網(wǎng)網(wǎng)rn p24加你淌般簡(jiǎn)IH間堆Efutlr%免復(fù)而堆島. 對(duì)曲粗均尢轉(zhuǎn)牙件悅停&出.免幣JG 2別時(shí).pCI ncoCAG pVAXI-IL-IS IXC他疫細(xì)4號(hào)出小 紋的加p24加休屮為冷腫為l/368 pCl ncoGAG免 繽姐為卿爼相比浪仆乂井如

16、瞪匕4 Ml時(shí).pCI-ncoGAG pVAXI-IHS聯(lián)合俺悵細(xì)*廿出小K的Mp24 JtttTiAn為I/III4.而pCbneoGAG為1/2229 (IM)01 ). 5tG 6刪時(shí)pCI-ncoGAG* pVAXI-IL-1&介免論細(xì)*甘出小MtFJU p24E體為1什20而pCI ncoGAG$ttg為1/2560 P the HIV-1 DNAvAcclne dutin diffcraM tifiw2.3覽疫小KUim*IFN.ttJI)tS檢訊ffi呆恨不pCI-ieoGAG pVAXI-IL-l IX介咬細(xì)小紋血“中的IE Y (424 4土6 0pgAiiL)A J

17、 pTI.nmCAG龜雜細(xì) 小喊血血中的IFN- Y4IM8 65AnlJ 兩也比ttflWMrtP00| h24克ffi小實(shí)匕栓測(cè)結(jié)*HIV p24 pa*MoGAG4. pVAXi n-18QtH艇初皿巴IBMtL. SI為ll.42069.llb pCI ncoGAG免疫til為4 94031.州饑比?P00|).2.5 CTI.ft他疫爾絃釘符升件CTL反燉加曲3 ft. W比為30I W. pCI ncoGAG* pVAXI-D. l Rft5L疫姐和pCI nctsGAG免疫姐的CT1殺傷*分別為M0in. 4i isiO JoaipcoGAG-l8IXimIPVAXIn.-i?r

18、it4trtffllDpC|.ncuGAO的CTI.累仿乍分刖為589%2J1. 31.62%0.3( po00l)：4ft：是比Aj I23i IIt pCbncoGAG* pVAXI-lL IB lf)t合免ttfflWpCI ncciGAG免疫組的CTL余飭*分料X/4858% I 44% 21 MO.I8%ppiNMc* ttf miiv oirMRinJ HlV-IGa。出Acte ee fur IL-183討論OS的iwiftW子展粒yti緞咬苗共免熨足捉島 的 個(gè)*!&將細(xì)粗兇子噴什打檢嘏oJUtt機(jī)體產(chǎn)牛的細(xì)柜岡保持艷定 的木屮.以出有散地促遇免amzw的櫛孔件HLH.

19、arm于金功注射刖胞兇十帝* 的不Q反應(yīng).并町丸恥堆昭兇了字賣(mài)期歸的從點(diǎn)IL-I8的1ZH功能為滴化T堀龍和NK M產(chǎn)牛IFN.r.4f IL-I2有協(xié)HManitii IL-I8可以上NKtftHL的的ft達(dá)達(dá).IL I8所 構(gòu)H的I卜衣 Y又可以刺融E種地馳賦的您達(dá)A aFM/FML的相耳作用發(fā)萍ihi紙昭的ttrr用；比外.IL I8還可以促iThl屏的分化.K町促遏Thl娥覚的m*UWIL-2的 分 海 . 押 制 活 化TMmtiu( (x4|.iL48MWttKMMMMb其 應(yīng)H!傾 城 包 知 坑 騎矗.MftZlrfinWAWfitft工反應(yīng)第.其中 ttiaa分1桿慚.蟲(chóng)尋覚

20、內(nèi)妨蛙Krancr龍Hdwm可以侃追m乜紹世免堆很W*偉獨(dú)免堆純?？梢詍于斤發(fā)KVffi免儀冷什塗略.以控制分 Bilhut Muloi A理 坍代中1?不7A ifKRirW66 1L I8鼻因笛Afi護(hù)袋夠帕Mi講#CTL反曲的反曲的時(shí)何. 抗皈分*IL-2 Hl IW r 性卅巴席0性fiin. ft (VI8兇ft炳舟低門(mén)執(zhí)體FJT* 本請(qǐng)究結(jié)果竄明5 pCI-ncoGAG A免檢小載 細(xì)比牧n.-IJ V兇敘合pCIoeoGAG免疫細(xì)小ftl所誘9的林只性枕HIVP24坑侔*廈澤低迖IL-I8酒化7W紹電功能.2細(xì)臉妙ttWX. WA.降低runt免疫尿衿此休研究*SJK IL.I8pCbneoGAG *ffl小W 1L iH P tn IFN- ( 424.4 6.0pfAnL) )高于pClncGAG單M免疫小歟級(jí)血的中的IFN- V l68865pgAnL)(P00l). iAif實(shí)f IH8在促iini霹分化中 的重餐性甘枝祗化T*H#1產(chǎn)主IFN Y本麗憲址Wi*RH IL