Mashayekhi, Sahar

Simorgh Research Repository

,(P*rne'0"'r7,r8,.,_a"' t, -aau1{r,a}'o0r;,rS.ii orS*.illo.,*rLi.i,,j.o-il dir.iJ *i,tl ,sJ,,lLi.iJtS Cbs.o a,tti,,ll*k:rrlJils,PSA 0j L 4iirJli o.r.i u.ir. . 'i JgjU Lrt o.i! J<11 t5.rl*! .+lfBALB/c oigo l,rt luJiLF*t -L J*.,:t o.ial; ll:''.f,S*.ill.l .r.r.os;5.r:p3'r Laial.; llir"f,i*?f< 6;rt .;jsr\flt-\flv:o{ n3 Jt*rKerman University of Medical SciencesFaculty of MedicineIn Partial Fulfillment of the Requirementsfor the Degree MSc ImmunologyTitle:Evaluation of stabilify of the attenautcd line ofLeishmania major transfected withPSA gene in BALB/c mouseBy:Sahar MashayekhiSupervisors:l-Dr. Hamid Daneshvar2-Dr. Mehdi GolchinYear: 2017Dr.('tr 9 Sf U$ &-lPl L .c,*,1 jL* eLl J-tt i,.+9r ol;-'9y 'V--:r-ilr-al9 4"Ji"U: "rr;1U+ cr'Ll) ,ilt \jlJ , :l+f S5 ,is;;L ;r 9 crpl+ ol;l t oV*'9y uV*, ,rilJ,51*or5fl;-,1 ;l .r9-i .,- d"L.,,,,I ok*,91, ,tV* alr. 6 i .tL,f cG ,jy L- ,J ;L oljlol;-5y\ lo.jr crLr Oj dl r*-.* Jf*s Alp--l.sLb,-it.r 1lorti:*,I{..j irl.1r o.r52l9ir^l,slr. 4 iy 6{) ,#\ 6ls-; u,al;. !-$lt rr;lr* ,91L $1" ;rl o$ , io-,:: ,f51;l .c.-,1 PSA ;r;LLeishmania )-<.;l J f, .llL ar-l.rr.r.JLt GAA) Tumor-associated antigen I Lr O#1r," Ol*r.-,1*r .*-,1&Jlt" 6--l .i.:-r .c -,1 o, t).f o:lil*.,1 PSA ,-1j ,l+l qa (f{-line) o.L.i, .4*:: maior.rilr.r BALB/c &r i Dr; &(lil,; JS.;l.s.,lr,-t" d)t e JS:;lrxlp PSA gjip-l .rj o:ti;-*l pLEXSY-PSA-neo2 4 ii +r*,t )l 4 iy JSt qly c-a; :t-l u,i91PSA-Neo 2 .-Jn; J-6b o+ 219 kb J* *..51 aJ! JL I Swa I.r^,-.ijl ,-Fr\ 4ii +-)!"o.r-.i.:(i*il1;.st-.-!S:;l .r--:f :119 18 srRNAa-qU i #-swflljl.r-," cs-ti J.!lJ.s,[r+orli;*rl )rqr d;l+ J-|r..:+a LEXSY-Neo .fll- eL L*r. (3il )-2glJc;l 9 LSF..y )l ,-r-"rl;-s ,-*l; 9 *-, .4-:19-" oV-1 .)P1,PCR .gL-o .(-r(; t *'f PSA aj aL* t )p .si)g )lP;l l- ,jf p 9 v;, F-ty BALB/c csLr ,j:,- a1 orj c,5.i*,1, "fS:;l .5rll-! ,-*:y.,Sly..4fa+lf ,jf ,rrb- o;L;lo.l.l,:-,4.5Lrjt!9y't U J-(;I,J*-+.n, $r-j L+)y,.-i;:L-x,-lc^qe.ri a.-t-" lajGgy't u$t9Jl.t-rt 9 t)Lieshmania major o$ , i*-.'i Jftl .p PSA r,j .jl-r , ,,f1' o),tr ;,i-l 9 PCR Al; :tl arib-lAjl!9)tt ! J51;l oralr e BALB/c ,-is *. e;.12li 55! , io-.ii ,-|6Jl ,y.:i, J-t' 6_.-l;; ;;3^a .r';tr cr)y. ,-Fs".,: ;'i ,1111 cSrl-*l f.rs r JS:;l U-l .S,,h'-l', or:'o) gLij.r.al .r-s6.-i- Ol* q J,,,1? 6. abr-)lio.r.i cryii Jftl ot.:1.: ;l,i-; a,Jlt a,-l A-f; :.5.;f a$.ri.SLJumor-associated antigen (TAA) lla *sy r:V. rsly.'&Lieshmania major,PSA ;,,; at*r ,.alfr,-l,oV-9y uVy ,Ss#t.5ta 6Jl9 :ojt9 {5WAbstractBackround and Airns: Therc are several known mcthods fbr cancer treatrncnts which arc onernethod of treatment is itnmunotherapy. Live attenuated recorlbinant vector vaccine is one ofthe methods of immunotherapy. Tltis vectors can bc used for a delivery system for tumor-associated atltigen in tumor itnruunotherapy. Prostate-specific antigcn (PSA) incrcase inpatients rvith prostate cancer and could be a target fbr treatment of proslate cancer. In thisartiole, we used the attcnuated line o{'leishnrania major (H-line) that previously attenuateclwith gentamicin by l-lamid Daneshvar. He dcrnonstratcd that attenuated strain of leishrnanianrajor is safe.'fhc airn of this studyrvas to cloning PSA genc into attcnuated lcishnrarrianrajor(II-linc) and evaluate o1'the safty of rcconrbinant lcishmania nrajor in BALB/c mousc.N{aterial and Methods: In the prcsent study, rvc cloncd human PSA genc into attenuatedleisltrnania lnajor and lve detectcd exprcssion of I'}SA gcuc in recourbinanl Leishrnania ma.iorby wcstern blotting. To exatrine the saltty of the transgenic parasite , groups of 5 BALLI/cntice were injectcd subcutanconsly rvith stationary-phase L. rnajor lvild , IJ-line, recombiuantpromastigotes. 'fhc [esiott volutne \,vas lllcasllred rveekly rvith a capitlary micrometer.'['o investigate the ability of promastigotes to inl'cct macrophages and thcir survival rvithinmacrophages, peritoneal lnacropharrcs were cxposcd to stationarl,-phasc promastigotes. Thecultureslidesthenrvereincubatcdin5'/oCO2,95o airforB.48,96hat37'C..Attheenclofthe incubation period. the macrophages rvere fixcd and stained with Giernsa stain ,ancl theinfection status of the rnacrophages was detcnnined by rnicroscopy.Results: The PSA gene cloned irtto leishrnania maior Il-line successlully for first time and wecould detected expression of'PSA genc in leshrnania major I-l-line. Saf-ety evaluated by 2criteria: first .that promastigotes o1' the recombinant leshrnania rnajor coulcl enter but notsurvive in ntacrophages, and second, that uricc inoculatcd subcutaneously r.vith attenuatedparasites did not develop skin lesions.Conclusions: f'hese finding represent that lcishrrania rnajor I-l-line could be used as a llewvector vaccinc {br cxpression of 'l'AAs and othcr proteins that can not express to otherdelivery systems. The use of live attenuatcd Lcishmania m{or preparations as candidatevaccines is very prornising because thcy most close ly lnintic the natural course of infectionand may, therefore, elicit similar irnmune rcsponscs.keywords: L.ntajor I I-line. vector vaccine, prostate cancer, PSA..'.-..'.'.'.'." " C1;",.....,,.'.'.,...0;L.5,..,.,.,.,...,.,.;du",Jl,j as*ra.U gt-Q;l 913.1 *-Ja.i;1ot$t y'crr, tJ&olii,lrattills*$so:!a;*(;;..r(sfr ,rf-iy, o*tJ o*u- I t-\ r-41 $-tJ,}f.93*.;b .reo-ts yar flifj s!-.,J qf ..rL-! \o6: ql+{-d jrr 1c.rtd ,,p "PSAgj L +-3,l,-"/j o.r-i ue'-"d'rj1r $1c*! ;l.St .5.1Lu-lr, ,s'--4 " }trr :*; J\,f ;32 f*Jr $* ;1.,r: c>rp Sp- 6*>t .11.> U lV i /lf i-.r15$b *iaa;rf* 661utq..-*lf , c.rs .ifa, eET urt>l3ra., jlirJfsr+ll.:{f:" (_ .b-l), Jl t t,) uU *4 .tL-rllit.rr{ db &,;.2$r iStrl.fUa (sr;l-),i1.1-.1: *|ra $2" :Itj oo*1i \.y. f o LSIiT iilt" (*;-.1-) J!11: lta^ ).;e: t$$j-F OnY J;!.i rrli:15-:,/r f'Se 6d\n rLiol 9;9 Vg\ait#Iodsii9/'t>*t \ . .r.rlclt> pti r pu,\a \:..ndr----K-/t ,\({l) L41;:L-!lgJit-v-- (;::)'..:e1; :L-l :.;>*-:\"/l-:<: - '+\1-=-:\$

بررسی پایداری انگل لیشمانیا ماژور تضعیف شده تراریخته با ژن PSA در موش BALB/c

http://eprints.kmu.ac.ir/26980/1/6431.pdf

بررسی پایداری انگل لیشمانیا ماژور تضعیف شده تراریخته با ژن PSA در موش BALB/c

Abstract

Similar works

Full text

Available Versions

Simorgh Research Repository