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[(expression)-243.4(in)-244.2(the)-240.7(lungs)-241.6(up)-238.3(to)-238.5(4)-243(weeks)-244.7(after)-244(rAAV)-243.1(delivery)-241.7(to)]TJ 0 -1.1174 TD [(fetal)-324.8(airways)-327.6(\(Carlon)]TJ /F9 1 Tf 8.9391 0 TD [(et)-326(al.)]TJ /F5 1 Tf 2.085 0 TD [(,)-326(2010\).)-325.7(In)-324.7(the)-321.4(current)-326.9(study,)-325.7(we)]TJ -11.0241 -1.1116 TD [(validated)-658.9(the)-655.4(efciency)-659.6(of)-653(rAAV2/5-mediated)-658.9(perinatal)]TJ T* [(\(fetal)-268.3(and)-263.6(neonatal\))-267.1(gene)-265.9(delivery)-264.8(to)-267.3(the)-263.8(murine)-268.3(airways)-264.2(and)]TJ 0 -1.1174 TD [(monitored)-266(long-term)-265.9(reporter)-264.6(gene)-260.2(expression.)-269.9(Transduction)]TJ 0 -1.1116 TD [(of)-405.3(actively)-404.3(dividing)-400.9(tissue,)-408.5(such)-402.8(as)-400.6(the)-402(airway)-407.3(epithelium,)]TJ T* [(with)-293.5(a)-296(nonintegrating)-300.5(rAAV)-295(vector)-297(requires)-299(repeated)-301.5(vector)]TJ 0 -1.1174 TD [(administration\(s\))-339.2(to)-336.4(achieve)-340.8(long-term)-335(gene)-335(correction.)-340.7(We)]TJ 0 -1.1116 TD [(hypothesized)-276(that,)-274.2(because)-277.1(of)-272.9(the)-269.5(immaturity)-280.1(of)-267.1(the)-275.3(immune)]TJ T* [(system)-348.6(during)-346.5(perinatal)-353.6(gene)-346.6(transfer,)-348.5(an)-348.9(immune)-350.2(response)]TJ T* [(against)-545.8(the)-540.2(vector)-544.7(particle)-548.3(could)-542.5(be)-539(prevented,)-546.8(allowing)]TJ 0 -1.1174 TD [(repeated)-629.8(administration\(s\))-627.2(in)-624.4(adult)-623.2(life.)-625.5(Our)-626.2(results)-625(in-)]TJ 0 -1.1116 TD [(deed)-335(demonstrate)-333.7(long-term)-335(gene)-335(expression)-335.6(after)-330.4(perinatal)]TJ T* [(gene)-381.1(transfer)-379.6(and)-378.8(subsequent)-381.7(readministration)-11.2(.)-377.8(Analysis)-379.3(of)]TJ 0 -1.1174 TD [(capsid-specic)-499(neutralizing)-489.5(antibody)-494.1(\(nAb\))-490.1(indicated)-491.8(low)]TJ 0 -1.1116 TD [(immunoreactivity)-456.2(against)-453.6(the)-453.8(rAAV)-456.2(vector)-452.5(after)-451.4(perinatal)]TJ T* [(gene)-254.4(transfer.)-256.3(This)-253.2(allowed)-253.1(effective)-256.7(vector)-250.9(readministration)]TJ 0 -1.1174 TD [(in)-244.2(adult)-243.1(life,)-245.4(resulting)-247.4(in)-244.2(efcient)-242.8(reporter)-247.3(gene)-242.9(expression)-249.2(in)]TJ 0 -1.1116 TD [(the)-615.1(murine)-619.6(airways)-621.3(up)-612.7(to)-618.6(7)-611.6(months.)-621.7(Because)-618(multiple)]TJ T* [(readministrations)-438.3(were)-433.8(not)-429.7(feasible,)-436.2(we)-436.2(conclude)-432.9(that)-432(im-)]TJ 0 -1.1174 TD [(munological)-373.2(ignorance)-370.6(rather)-371.9(than)-368.6(tolerance)-371.9(underlies)-369.4(suc-)]TJ 0 -1.1116 TD [(cessful)-338(readministration)-339.5(of)-330.5(an)-337.4(autologous)-334.8(vector.)]TJ /F4 1 Tf 8.9663 0 0 8.9663 313.0582 725.7825 Tm [(Materials)-339(and)-331.9(Methods)]TJ /F11 1 Tf 0 -1.8336 TD [(rAAV2/5)-337(production)]TJ /F5 1 Tf 9.843 0 0 9.843 323.0361 692.9006 Tm [(rAAV2/5-CBA-eGFP-P2A-fLUC)-395.7(\(CBA,)-385.1(chicken)]TJ /F6 1 Tf 20.4354 0 TD (b)Tj /F5 1 Tf .5529 0 TD (-actin)Tj -22.0021 -1.1116 TD [(promoter;)-289(eGFP,)-280.3(enhanced)-284.2(green)-284.3(uorescent)-285(protein;)-281(fLUC,)]TJ 0 -1.1174 TD [(rey)-720.5(luciferase\))-725.2(and)-718.7(rAAV2/5-CBA-)]TJ /F6 1 Tf 16.7204 0 TD (b)Tj /F5 1 Tf .5529 0 TD [(-Gal)-720.7(\()]TJ /F6 1 Tf 2.8338 0 TD (b)Tj /F5 1 Tf .5529 0 TD (-Gal,)Tj /F6 1 Tf 2.7474 0 TD (b)Tj /F5 1 Tf .5529 0 TD (-)Tj -23.9604 -1.1116 TD [(galactosida)-11.2(se\))-551.4(were)-549(produced)-552.5(and)-551.6(puried)-550.2(as)-556.1(previously)]TJ T* [(descri)-8.8(bed)-361.6(\(Van)-363.5(der)-367.3(Perren)]TJ /F9 1 Tf 11.1969 0 TD [(et)-366.4(al.)]TJ /F5 1 Tf 2.1253 0 TD [(,)-360.5(2011\).)-366(Vector)-367(titers)-365(were)]TJ -13.3222 -1.1174 TD [(determined)-269.4(by)-261.4(real-time)-268.4(PCR,)-262.1(using)-264.6(the)-263.8(primer)-268.3(probe)-260.1(set)-263.6(for)]TJ 0 -1.1116 TD [(the)-402(poly\(A\))-394.6(sequence)-399.2(\(Limberis)]TJ /F9 1 Tf 13.6159 0 TD [(et)-400.9(al.)]TJ /F5 1 Tf 2.1599 0 TD [(,)-395.1(2009\).)-394.8(rAAV)-398.6(titers)]TJ -15.7758 -1.1116 TD [(are)-420.3(presented)-422.2(as)-417.9(DNase-resistant)-425.8(genome)-419.1(copies)-417.8(\(GC\))-415.2(per)]TJ T* [(milli)-7.3(liter)-329.6(\(Gao)]TJ /F9 1 Tf 6.2032 0 TD [(et)-331.8(al.)]TJ /F5 1 Tf 2.0908 0 TD [(,)-331.7(2002\).)]TJ /F11 1 Tf 8.9663 0 0 8.9663 313.0582 581.329 Tm [(Vector)-334.4(delivery)-337.2(to)-336.7(perinatal)-331.5(or)-335.3(adult)-336.2(murine)-330.1(airways)]TJ /F5 1 Tf 9.843 0 0 9.843 323.0361 564.8881 Tm [(For)-296.9(all)-304.2(experiments,)-302.5(FVB/N)-296.2(mice)-301.8(were)-301.3(used)-299.1(\()-127.8(Janvier,)-301.4(Le)]TJ -1.0137 -1.1116 TD [(Genest-Saint-Isle,)-476.8(France\).)-463.7(Fetal)-468.7(vector)-464.1(delivery)-466.4(by)-463(intra-)]TJ 0 -1.1174 TD [(amniotic)-531.1(injection)-531.1(\(60)]TJ /F6 1 Tf 9.5957 0 TD (l)Tj /F5 1 Tf .5529 0 TD [(l)-531.1(o)0(f)-526.3(vector)-527.4(suspension\))-531.2(was)-529.3(per-)]TJ -10.1486 -1.1116 TD [(forme)-7.8(d)-317.9(a)0(s)-325.7(previously)-323.2(described)-330.1(\(Carlon)]TJ /F9 1 Tf 16.4843 0 TD [(et)-320.3(al.)]TJ /F5 1 Tf 2.0793 0 TD [(,)-326(2010,)-318.9(2012\).)]TJ -18.5635 -1.1116 TD [(Ne)13(on)20.8(at)19.5(al)-268.4(ve)19.7(ct)19.5(or)-267.1(ad)19.7(mi)19.5(ni)20.7(st)15.1(ra)19.7(ti)20.6(on)-267.1(to)-267.3(th)20.7(e)-278.7(a)0(i)19.5(r)0(w)20(a)0(y)13.9(s)-271.6(wa)13(s)-277.4(p)0(e)19.7(r)0(f)20.9(o)0(r)20.9(m)0(e)18.5(d)]TJ 0 -1.1174 TD [(by)-342(sn)21(if)15(n)21(g)-346.7(o)0(n)-342(d)0(a)19.7(y)-352.4(3)-346.7(p)0(o)20.8(s)0(t)15.1(p)0(a)19.7(r)0(t)20.7(u)0(m)19.6(.)-349(Pu)21(ps)-341.8(we)18.8(re)-343.2(im)19.5(mo)19.6(bi)20.7(li)20.6(ze)18.5(d)-352.4(i)0(n)]TJ 0 -1.1116 TD [(an)-297.1(up)20.8(ri)20.7(gh)20.8(t)-300.7(p)0(o)20.8(s)0(i)15.1(t)0(i)20.6(o)0(n)-290.2(a)0(n)19.7(d)-300.6(ve)19.7(ct)19.5(or)-295.9(\(6)20.9(0-)]TJ /F6 1 Tf 13.9558 0 TD (l)Tj /F5 1 Tf .5414 0 TD -.0204 Tc [(l)-321.1(v)-20.4(ec)-20.4(t)-6.6(o)-20.4(r)-310.5(s)-20.4(us)-20.4(pe)-20.4(ns)-20.4(i)-5.3(o)-20.4(n\))-321(w)-20.4(a)-1.6(s)]TJ -14.4972 -1.1116 TD 0 Tc [(ad)13.9(mi)19.5(ni)20.7(st)20.9(er)19.7(ed)-262.5(dr)20.9(op)-255.6(by)-261.4(dr)20.9(op)-255.6(in)15(to)-255.8(on)20.8(e)-267.2(n)0(o)15.1(s)0(t)20.9(r)0(i)20.7(l)-266.2(ov)15.1(er)-256.8(a)-267.2(t)0(i)14.8(m)15(e)-273(p)0(e)19.7(r)0(i)20.7(o)0(d)]TJ 0 -1.1174 TD [(of)-319(2)-329.4(h)0(r)15.1(;)-323.8(pu)15.1(ps)-318.8(we)18.8(re)-320.1(ke)19.7(pt)-319.1(wa)18.8(rm)-320.2(by)-319(pl)20.7(ac)18.5(in)20.7(g)-329.4(t)0(h)20.7(e)0(m)-321.4(o)0(n)-319(a)-330.6(he)19.7(at)19.5(in)20.7(g)]TJ 0 -1.1116 TD [(pa)13.9(d)-675(i)0(n)-664.7(b)0(e)13.9(t)0(w)19.8(e)0(e)18.5(n)-675(in)20.7(tr)15(an)19.7(as)19.9(al)-665.9(ve)13.9(ct)19.5(or)-664.5(ad)19.7(mi)19.5(ni)14.9(st)20.9(ra)19.7(ti)20.6(on)20.8(s.)-672.5(In)20.9(tr)20.7(a-)]TJ T* [(tr)15(ac)18.5(he)19.7(al)-274.2(ve)19.7(ct)19.5(or)-272.9(in)20.7(st)20.9(il)20.6(la)13.8(ti)20.6(on)-267.1(wa)13(s)-277.4(p)0(e)19.7(r)0(f)15.1(o)0(r)20.9(m)0(e)18.4(d)-283.3(in)-267.3(ad)19.7(ul)14.9(t)-277.7(m)0(i)19.5(c)0(e)-275.2(8)0()20.8(1)0(2)]TJ T* [(we)13(ek)19.7(s)-237(o)0(f)-226.8(a)0(g)19.7(e)0(.)-230.3(A)0(f)20(t)0(e)19.5(r)-237.2(ex)13.9(po)20.8(su)21(re)-228(of)-226.8(th)15(e)-238.4(t)0(r)20.7(a)0(c)18.5(h)0(e)19.7(a)-238.4(th)20.7(ro)15.1(ug)20.8(h)-237.2(a)-238.4(mi)19.5(dl)20.7(in)20.7(e)]TJ 0 -1.1174 TD [(in)15(ci)19.5(si)20.9(on)20.8(,)-256.9(1)0(6)20.8(0)]TJ /F6 1 Tf 5.2413 0 TD (l)Tj /F5 1 Tf .5414 0 TD [(l)-254.7(o)0(f)-244.1(v)0(e)19.7(c)0(t)19.5(o)0(r)-249.8(d)0(i)20.7(l)0(u)20.7(t)0(e)19.6(d)-254.5(in)-244.2(ph)20.8(os)15.3(ph)20.8(at)19.5(e-)19.7(bu)20.8(ff)15.1(er)19.7(ed)-245.3(sa)19.9(li)20.6(ne)]TJ -5.7827 -1.1116 TD [(\(P)15.3(BS)20(\))-410(w)0(a)13(s)-409.8(in)20.7(st)15.1(il)20.6(le)19.6(d)-410(i)0(n)15(t)0(o)-399.7(t)0(h)20.7(e)-417(tr)20.7(ac)18.5(he)19.7(a)-417(w)0(i)19.8(t)0(h)-399.7(a)-417(26)20.8(-g)20.9(au)19.7(ge)-406.5(p)16.2(o)21.9(l)16.1(y)21.9(-)]TJ T* -.0185 Tc [(te)-3.5(trauoroethylene)-233.9(\(PTFE\))-232.7(c)-3.5(a)]TJ 11.4561 0 TD -.0159 Tc [(th)6(et)5.9(er)-230.1(\(B)5(D)-231(V)-.6(a)4.9(s).5(c)4.9(u).3(l)5.9(o).3(n)-230.1(P)6.2(lus)6.2(;)-230.2(VWR,)]TJ -11.4561 -1.1174 TD 0 Tc [(Vienna,)-537.5(Austria\),)-532.7(using)-535.3(a)-526.4(stereo)-8.8(scopic)-533(zoom)-530.9(microscope)]TJ 0 -1.1116 TD [(\(magnicat)-10(ion,)]TJ /F10 1 Tf 6.5257 0 TD ()Tj /F5 1 Tf .72 0 TD [(3\).)-259(All)-252.1(animal)-258.1(procedures)-256.2(were)-261(carried)-258.9(out)]TJ -7.2457 -1.1116 TD [(under)-386.8(Biosafety)-387.7(Level)-389.3(2)-381.2(conditions)-386.8(and)-384.6(were)-387.7(approved)-385.5(by)]TJ 0 -1.1174 TD [(the)-436.6(KU)-435.9(Leuven)-436.4(Biosafety)-439.5(Committee)-436.6(\(laboratory)-435.1(accredi-)]TJ 0 -1.1116 TD [(tation)-745.4(number)-746.2(LA1210579;)-744.7(ethics)-746.2(committee)-747.7(approval)]TJ T* [(number)-337.3(P065/2009\).)]TJ /F11 1 Tf 8.9663 0 0 8.9663 313.0582 321.7889 Tm [(Bioluminescence)-336.1(imaging)]TJ /F5 1 Tf 9.843 0 0 9.843 323.0361 305.348 Tm [(Bioluminescence)-445.3(imaging)-436.6(\(BLI\))-439.6(was)-437.1(performed)-433.8(as)-440.9(pre-)]TJ -1.0137 -1.1116 TD [(viously)-399.4(described)-399.2(\(Carlon)]TJ /F9 1 Tf 11.0471 0 TD [(et)-395.2(al.)]TJ /F5 1 Tf 2.1541 0 TD [(,)-395.1(2010\).)-394.8(Measurements)-405.6(are)]TJ -13.2012 -1.1174 TD [(report)-7.8(ed)-291.3(as)-296.9(the)-298.3(total)-295(photon)-294.7(ux)-296.9(from)-293.5(a)-296(4.3-cm)]TJ 7.3823 0 0 6.5614 501.1085 287.8299 Tm (2)Tj 9.843 0 0 9.843 507.6849 283.4078 Tm (rectangular)Tj -19.7731 -1.1116 TD [(region)-324.6(of)-324.7(interest)-322.3(\(ROI\))-324.1(for)-325.8(the)-321.4(lung)-321.3(and)-321.2(a)-324.8(2.0-cm)]TJ 7.3823 0 0 6.5614 515.0549 276.8314 Tm (2)Tj 9.843 0 0 9.843 521.9148 272.466 Tm (circular)Tj -21.2188 -1.1116 TD [(ROI)-339.2(for)-331.5(the)-332.9(nose.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 313.0582 237.5999 Tm (5-Bromo-4-chloro-3-indolyl-)Tj /F6 1 Tf 12.1147 0 TD (b)Tj /F11 1 Tf .5501 0 TD (-)Tj /F8 1 Tf .3351 0 TD (d)Tj /F11 1 Tf .5501 0 TD (-galactopyranoside)Tj -13.55 -1.2203 TD [(staining)-337.2(for)]TJ /F6 1 Tf 5.2607 0 TD (b)Tj /F11 1 Tf .5564 0 TD [(-Gal)-329(expression)-335.9(in)-336.5(tissue)-330.8(sections)]TJ /F5 1 Tf 9.843 0 0 9.843 323.0361 210.2173 Tm -.0216 Tc [(P)-5.3(r).4(oc)-6.6(ess)-5.2(i).2(n)-5.4(g)-362.5(o)-5.4(f)-362.5(lu)-5.4(ng)-368.3(sam)-6.6(p).3(le)-6.6(s)-362.3(a)-6.6(nd)-362.5(q)-5.4(u).3(a)-6.6(n).3(ti)-5.5(ca)-6.6(tio)-5.4(n)-362.5(o)-5.4(f)-362.5(t)-5.5(ran)-5.4(s).5(du)-5.4(c-)]TJ -1.0137 -1.1116 TD -.0198 Tc [(tion)-406.8(efcie)-4.8(ncy)-406.8(i)-3.8(n)-406.8(t)2(he)-408(conduc)-4.8(ting)-406.8(a)1(irways)-406.6(wa)-4.8(s)-406.6(d)2.1(eterm)-4.8(i)2(ne)-4.8(d)-406.8(a)1(s)]TJ 0 -1.1174 TD -.017 Tc [(pr)5(ev)4.9(i)4.8(o)-.8(u)4.9(s)5.1(ly)-237(d)4.9(e)-2(s)5.1(c)3.8(ri)4.8(bed)-237(\()-.8(Carl)4.8(o)4.9(n)]TJ /F9 1 Tf 11.3466 0 TD -.0199 Tc [(et)-240(al.)]TJ /F5 1 Tf 1.9007 0 TD -.0218 Tc [(,)-244.1(201)-5.6(0\).)-244.1(B)-6.6(rie)-6.8().3(y,)-244.1(t)-5.7(r).2(an)-5.6(sdu)-5.6(c)-1(ti)-5.7(on)]TJ -13.2473 -1.1116 TD .3985 Tc [(e)413.5(f)420.5()414.8(c)419.3(i)420.3(e)413.5(n)420.4(c)419.3(yo)414.7(ft)420.3(h)414.7(e)-1.2(c)419.3(o)420.4(n)414.7(d)420.4(u)420.4(c)413.5(t)420.3(i)420.3(n)414.7(ga)419.3(i)414.5(r)420.5(w)419.6(a)413.5(y)420.4(s).2(w)419.6(a)413.5(sd)420.4(e)413.5(t)420.3(e)419.3(r)414.7(m)419.2(i)420.3(n)414.7(e)419.3(dw)419.6(i)414.6(t)420.3(ha)]TJ T* -.0216 Tc 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[(du)15.1(ct)19.5(in)20.7(g)-392.8(a)0(i)19.5(r)0(w)20(a)0(y)-389.3(r)0(e)19.7(s)0(p)21(i)0(r)20.7(a)0(t)13.8(o)0(r)20.9(y)-392.8(ep)19.7(it)20.6(he)13.9(li)20.6(um)-383.6(\(G)20(ru)20.9(bb)-388.1(an)19.7(d)-392.8(B)0(o)19.8(u)0(c)19.7(h)0(e)19.7(r)0(,)]TJ T* -.0162 Tc [(19)5.7(99\))5.8(.)]TJ /F6 1 Tf 2.8107 0 TD 0 Tc (b)Tj /F5 1 Tf .5299 0 TD -.0169 Tc [(-Gal)4.9(-p)5(osi)4.9(t)-.8(i)4.9(v)-.7(e)-335.9(c)3.9(el)4.9(ls)-334.6(wer)5.1(e)-335.9(vi)4.9(su)5(al)4.9(ized)-334.8(wit)4.9(h)-334.8(a)-335.9(L)-1.9(eica)-335.9(l)4.9(i)-.8(gh)5(t)]TJ -3.3406 -1.1174 TD -.017 Tc [(mi)4.8(cr)5(oscop)4.9(e)-284.2(equi)4.8(pp)4.9(ed)-283(wit)4.8(h)-283(a)-284.2(B)-1.8(i)4.8(o)-.8(P)5.1(o)-.8(i)4.8(n)-.8(t)-283.2(2)-283(mot)4.8(o)-.8(r)5(i)-.9(zed)-283(s)-.6(t)4.8(a)-2(g)4.9(e)-284.2(\(Lud)4.9(l)]TJ 0 -1.1116 TD -.0167 Tc [(El)5.1(ectr)5.3(oni)5.1(c)-295.4(P)-.4(r)5.3(o)-.5(d)5.2(u)-.5(cts,)-290.8(Hawt)5.1(ho)5.2(rne,)-296.6(N)4.4(Y\))5.3(.)-296.6(B)-1.5(r)5.3(i)-.6(g)5.2(h)-.5(t)5.1(n)-.5(ess)-294.1(a)-1.7(n)5.2(d)-294.3(co)5.2(nt)5.1(rast)]TJ T* [(were)-318.5(op)5.2(ti)5.1(mi)5.1(zed)-317.3(w)-1.4(i)5.1(t)-.6(h)-317.3(A)-1.4(d)5.2(o)-.5(b)5.2(e)-318.5(Ph)5.2(ot)5.1(osh)5.2(o)-.5(p)-317.3(C)4.2(S3)-317.3(\(Ado)5.2(be)-318.5(S)5.4(y)-.5(s)5.4(t)-.6(ems,)]TJ 0 -1.1174 TD -.0169 Tc [(Mo)5(un)5(tain)-329(Vi)4.9(ew,)-331.4(C)-1.7(A\).)]TJ /F11 1 Tf 8.9663 0 0 8.9663 59.7543 507.9684 Tm 0 Tc [(Immunohistochemistry)-337.7(and)-335.7(hematoxylineosin)]TJ 0 -1.2203 TD [(staining)-337.1(on)-331.3(lung)-339(sections)]TJ /F5 1 Tf 9.843 0 0 9.843 69.7323 480.5857 Tm [(Fr)21(oz)13.9(en)-216.5(se)19.9(ct)13.8(io)20.7(ns)-220.9(\(6)]TJ /F6 1 Tf 7.3321 0 TD (l)Tj /F5 1 Tf .5414 0 TD [(m\))-216.5(we)13(re)-216.4(x)21(ed)-222.2(in)-221.2(co)19.7(ld)-221.2(\()]TJ /F10 1 Tf 9.1003 0 TD (-)Tj /F5 1 Tf .7084 0 TD (20)Tj /F10 1 Tf .9791 0 TD ()Tj /F5 1 Tf .3859 0 TD [(C\))-222.1(ac)18.5(et)19.5(on)20.8(e)]TJ -20.0611 -1.1174 TD -.0157 Tc [(fo)6.2(r)-339.3(1)6.2(5)-166.5(m)5(i)6.1(n).5(.)-330.2(S).6(ec)5.1(ti)6.1(on)6.2(s)-333.4(w)-.4(e)5.1(r).5(e)-334.7(s)6.4(ta)5.1(in)6.2(ed)-333.6(w)5.4(i).4(t)6.1(h)-333.6(a)-334.7(r).5(a)5.1(t)-333.7(mo)6.2(no)6.2(cl)6.1(on)6.2(al)-333.7(a)5.1(n).5(t)6.1(i).4(-)]TJ 0 -1.1116 TD -.0167 Tc [(bo)5.2(dy)-386.4(\()5.3(m)-1.7(Ab\))-380.6(s)-.3(peci)5.1(c)-381.8(for)-380.6(CD4)-386.4(\()-.5(GK1)5.2(.)2.9(5;)-386.5(i)5.1(n)-.5(-)5.3(h)-.5(o)5.2(u)-.5(s)5.4(e)-387.6(su)5.2(pern)5.2(at)5.1(an)5.2(t\))]TJ T* 0 Tc [(and)-401.9(a)-405.4(rat)-402(mAb)-402.8(specic)-402.6(for)-400.7(CD8)-402.6(\(clone)-406.5(2.43;)-399.6(in-house)-401.5(su-)]TJ 0 -1.1174 TD [(pernatant\).)-484.7(For)-481.2(immunouorescence,)-485.2(the)-476.9(following)-483.4(detec-)]TJ 0 -1.1116 TD [(tion)-304.2(antibodies)-307.3(were)-307.1(used:)-306.1(Alexa)-308.3(Fluor)-299.2(488donkey)-305.9(anti-rat)]TJ T* [(\(A21208;)-683.7(Molecular)-688.4(Probes,)-684.7(Gent,)-683.9(Belgium\))-686.4(and)-684.1(4)]TJ /F10 1 Tf 22.9582 0 TD ()Tj /F5 1 Tf .2534 0 TD (,6-)Tj -23.2116 -1.1174 TD -.0217 Tc [(d)-5.5(i).1(am)-6.7(idi)-5.6(no-)-5.5(2-)-5.5(phe)-6.7(nyl)-5.6(ind)-5.5(o).2(le)-283.1(\()-5.5(DAP)-5.4(I\))-282(\()-5.5(D)-.6(13)-5.5(06;)-282.1(Mo)-5.5(le)-6.7(cul)-5.6(a)-.9(r)-282(P).4(r)-5.5(obe)-6.7(s\).)]TJ 0 -1.1116 TD -.0199 Tc [(Images)-406.7(w)-4.6(e).9(re)-413.8(acquired)-406.9(w)-4.6(ith)-412.7(an)-406.9(LSM)-407.8(5)2(10)-406.9(Me)-4.9(ta)-408.1(confoca)-4.9(l)-407(m)6.6(i)1.9(-)]TJ T* -.022 Tc [(cro)5.7(s).1(co)5.7(pe)-329.5(\(Zeiss)5.9(\).)-330.7(Ad)5.7(jacent)-328.5(lu)5.7(ng)-328.4(sectio)5.7(ns)-328.2(of)-328.3(6)]TJ /F6 1 Tf 17.7917 0 TD 0 Tc (l)Tj /F5 1 Tf .5299 0 TD -.022 Tc [(m)-329.6(w)4.8(ere)-329.5(s).1(tain)5.7(ed)]TJ -18.3216 -1.1116 TD -.0243 Tc [(w)-3.2(i)-2.5(th)-399.8(he)-3.5(ma)-3.5(toxylin)-399.8(and)-399.8(e)2.2(osin)-399.8(\(H)-3.2(&E)-3.6(\))-399.7(t)3.3(o)-399.8(v)-2.4(isua)-3.5(lize)-400.9(im)-3.6(mune)-400.9(c)-3.5(e)2.2(ll)]TJ 0 -1.1174 TD -.0215 Tc [(in)6.4(ltr)6.2(ati)6.1(on,)-330.2(u)6.2(s).6(ing)-327.9(a)-329(Lei)6.1(ca)-329(Bio)6.2(P).6(oi)6.1(nt)-328(2)-333.6(l).3(i)6.1(ght)-328(mi)6.1(cro)6.2(s).6(co)6.2(pe.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 59.7543 340.1007 Tm 0 Tc [(rAAV)-337.3(transduction)-335.2(inhibition)-336(assay)]TJ /F5 1 Tf 9.843 0 0 9.843 69.7323 323.6598 Tm [(In)20.9(d)16.2(i)0(v)20.7(i)0(d)26.5(u)0(a)19.7(l)-439(bl)26.5(oo)20.8(d)-438.8(s)0(a)25.6(m)0(p)19.6(l)16.1(es)-429.4(we)24.6(re)-429.5(co)19.7(l)16.1(l)0(e)19.6(c)0(t)25.3(e)0(d)-429.6(a)0(t)-429.7(v)16.2(ar)19.7(io)26.5(us)-428.2(ti)20.6(m)15(e)]TJ -1.0137 -1.1174 TD [(po)20.8(in)26.5(ts)-727.9(fr)26.7(om)-729.1(a)15(n)0(i)20.7(m)15(al)19.5(s)-732.4(b)0(y)-727.9(r)0(e)25.5(t)0(r)20.7(o)0(-)26.6(o)0(r)20.9(b)0(i)26.5(t)0(a)19.6(l)-738.5(b)16.2(l)0(e)19.6(e)15(di)20.7(ng)26.6(.)-740.7(B)15.2(ro)20.9(nc)25.4(h-)]TJ 0 -1.1116 TD [(oa)19.7(lv)26.5(eo)19.7(la)25.3(r)-323.6(l)0(a)19.6(v)0(a)25.4(g)0(e)-314.4()0(u)21(i)16.1(d)-323.6(\()0(B)19.8(A)0(L)24.5(F)0(\))-313(s)0(a)19.9(m)15(pl)20.7(e)15(s)-329.2(w)15.3(e)0(r)19.7(e)-324.8(co)25.4(ll)20.6(ec)24.3(te)19.6(d)-323.6(a)0(t)-314.5(t)16.1(he)]TJ T* [(ti)20.6(m)15(e)-549.4(po)20.8(in)26.5(t)-548.4(o)0(f)-537.8(h)0(a)25.4(r)0(v)20.9(e)0(s)19.9(t)-542.6(\(1)-537.8(mo)25.4(nt)20.7(h)-548.3(a)0(f)25.5(t)0(e)19.6(r)-548.2(th)26.5(e)-549.4()0(r)21(s)0(t)-537.8(o)16.2(r)-548.2(s)0(e)19.9(c)0(o)25.4(n)0(d)]TJ T* [(re)19.7(ad)25.4(mi)19.5(n)16.2(i)0(s)20.9(t)0(r)20.7(a)15(ti)20.6(on)26.6(\))-231.5(b)0(y)-215.3()0(u)21(s)0(h)26.8(i)0(n)20.7(g)-231.5(2)-150.8(m)0(l)-222.4(o)16.2(f)-231.4(P)0(B)25.7(S)-231.3(th)20.7(r)16.2(o)0(u)20.8(g)0(h)-215.3(t)0(h)20.7(e)-226.9(tr)20.7(ac)24.3(he)19.7(a)15(.)]TJ 0 -1.1174 TD [(Se)19.8(ru)26.6(m)-244.2(a)0(n)25.4(d)-243(BA)24.7(LF)-227.9(sa)19.9(mp)25.4(le)19.6(s)-237(w)0(e)18.8(r)0(e)-228(h)0(e)19.7(a)15(t-)20.7(in)26.5(ac)18.5(ti)26.3(va)19.7(te)25.3(d)-243(f)0(o)26.6(r)-243(45)-140.4(mi)19.5(n)-237.2(a)0(t)]TJ 0 -1.1116 TD (56)Tj /F10 1 Tf .9792 0 TD ()Tj /F5 1 Tf .3859 0 TD [(C)15.2(.)-297.2(H)15.3(E)0(K)18.8(2)0(9)26.6(3)0(T)-285.6(c)0(e)24.3(l)0(l)20.6(s)-294.6(we)24.6(re)-285.6(se)25.6(ed)19.7(ed)-285.6(a)15(t)-295(5.)24.3(0)]TJ /F10 1 Tf 15.2402 0 TD ()Tj /F5 1 Tf .7084 0 TD -.0162 Tc (10)Tj 7.3823 0 0 6.5614 239.7543 262.2614 Tm 0 Tc (4)Tj 9.843 0 0 9.843 246.3873 257.896 Tm [(c)15(e)0(l)19.5(l)16.1(s)-294.6(p)0(e)19.7(r)-294.8(we)24.6(ll)]TJ -18.961 -1.1116 TD [(in)20.7(to)-244.2(96)26.6(-w)20(el)25.3(l)-260.4(p)0(l)26.5(a)0(t)19.5(e)15(s)-260.1(o)0(n)-244.1(d)0(a)25.4(y)-260.3(1)16.2(.)-256.9(Th)19.6(e)-255.7(n)0(e)25.4(x)0(t)-250(d)16.2(ay)19.7(,)-256.9(4)16.2(-f)20.9(ol)20.7(d)-254.5(d)0(i)26.5(l)0(u)20.7(t)0(i)26.3(o)0(n)20.8(s)-254.3(of)]TJ 0 -1.1174 TD [(se)19.9(ru)26.6(m)-538(o)0(r)-520.5(B)0(A)18.9(L)15(F)-536.6(s)0(a)19.9(m)15(pl)20.7(e)15(s)-536.6(in)-520.7(Du)20(lb)26.5(ec)18.5(c)15(o)0()20.9(s)-536.6(m)15(o)0(d)20.8(i)0()26.6(e)0(d)-527.5(E)15(ag)19.7(l)16.1(e)0()19.7(s)]TJ 0 -1.1116 TD [(me)18.4(d)16.2(i)0(u)20.7(m)-359.4(\()16.2(D)0(M)19.1(E)15(M\))-348.7(wi)25.6(th)20.7(ou)26.6(t)-358.3(f)0(e)19.7(t)0(a)25.3(l)-358.3(ca)24.3(lf)-347.9(se)19.9(r)16.2(u)0(m)-349(\()0(F)26.8(C)0(S)20(\))-358.2(w)15.3(e)0(r)19.7(e)-359.4(pr)26.6(e-)]TJ T* [(pa)19.7(re)25.5(d.)-298.3(W)13.9(e)0(l)19.5(l)0(s)-295.9(w)0(i)25.6(t)0(h)-296.1(t)0(h)20.7(e)-307.5(sa)25.6(me)-298.3(vo)26.6(lu)20.7(me)-298.3(o)16.2(f)-312.1(D)15.3(M)0(E)18.7(M)-307.3(se)25.6(rv)20.9(e)15(d)-312.1(a)15(s)-311.9(n)16.2(o)0(-)]TJ 0 -1.1174 TD [(se)19.9(ru)26.6(m)-330.6(o)0(r)-318.9(n)0(o)20.8(-)16.2(BA)18.9(LF)-320(c)15(o)0(n)20.8(t)0(r)26.5(o)0(l)20.7(s)0(.)-315.4(A)0(p)20(p)0(r)26.6(o)0(x)20.8(i)0(m)25.2(a)0(t)19.5(e)15(ly)-319.1(1.)24.3(0)]TJ /F10 1 Tf 19.3814 0 TD ()Tj /F5 1 Tf .7084 0 TD -.0162 Tc (10)Tj 7.3823 0 0 6.5614 267.0802 207.496 Tm 0 Tc (9)Tj 9.843 0 0 9.843 274.0535 203.0739 Tm -.0153 Tc [(GC)-345.7(o)-15.3(f)]TJ -21.7717 -1.1116 TD 0 Tc [(rA)20(AV)24.9(2/)20.7(5-)26.6(CB)18.8(A-)25.8(eG)18.8(FP)26.9(-P)21(2A)25.7(-f)20.9(LU)24.5(C)-370.7(i)0(n)-353.7(D)0(M)19.1(E)15(M)-370.6(w)15.3(as)-360.3(a)15(d)0(d)20.8(e)0(d)25.4(,)-372.1(p)16.2(e)0(r)]TJ T* [(we)18.8(ll)26.3(,)-268.4(t)16.1(o)-266(s)0(e)19.9(r)16.2(um)-256.8(o)16.2(r)-266(BA)18.9(L)15(F)-265.9(sa)25.6(mp)19.6(l)16.1(e)-267.2(or)-255.6(c)15(o)0(n)20.8(t)0(r)26.5(o)0(l)-255.8(w)0(e)24.6(l)0(l)20.6(s)0(,)-252(i)0(n)26.5(c)0(u)19.7(b)0(a)25.4(t)0(e)19.6(d)-260.3(at)]TJ T* (37)Tj /F10 1 Tf .9792 0 TD ()Tj /F5 1 Tf .3859 0 TD [(C)-244(f)0(o)20.9(r)-248.7(6)16.2(0)-156.6(mi)25.2(n,)-240.7(a)15(n)0(d)-238.3(a)15(dd)20.8(ed)-233.7(to)-238.5(th)26.5(e)-249.9(c)0(e)18.5(l)16.1(ls)-238.3(\(1)20.9(0)16.2(0)0(-)]TJ /F6 1 Tf 16.3576 0 TD (l)Tj /F5 1 Tf .5414 0 TD .2431 Tc [(l)243.1(n)264.1(a)243.1(l)9.2(v)243.1(o)263.9(l)243.1(u)269.6(m)243.1(e)261.5(\))259.3(.)]TJ -18.264 -1.1174 TD 0 Tc [(On)20(e)-428.5(h)0(o)26.6(u)0(r)-416.9(l)0(a)19.6(t)16.1(er)19.7(,)-429.6(1)16.2(00)]TJ /F6 1 Tf 8.3401 0 TD (l)Tj /F5 1 Tf .5357 0 TD [(l)-433.2(o)16.2(f)-427.3(D)0(M)19.1(E)15(M)19.9(20)26.6(%)-428.4(F)0(C)20(S)-427.2(wa)24.6(s)-427.1(a)0(d)19.7(d)0(e)25.4(d)-427.3(to)]TJ -8.8757 -1.1116 TD [(ea)18.5(c)15(h)-225.7(we)24.6(ll)20.6(.)-222.3(T)0(w)24.5(e)0(n)19.7(t)0(y)26.5(-)0(f)20.9(o)0(u)26.6(r)-225.7(ho)26.6(ur)20.9(s)-225.5(l)16.1(at)19.5(er)25.5(,)-222.3(t)0(h)20.7(e)-221.1(tr)20.7(an)25.4(sd)21(uc)25.4(ti)20.6(on)-209.5(ef)19.7()16.3(c)0(i)19.5(e)0(n)25.4(c)0(y)]TJ T* [(wa)18.8(s)-358(a)0(n)19.7(a)15(ly)20.7(ze)24.3(d)-364(b)16.2(y)-364(q)16.2(ua)19.7(nt)26.5(if)20.7(yi)26.5(ng)-353.5(f)16.2(L)0(U)18.8(C)-359.2(ac)24.3(ti)20.6(vi)26.5(ty)20.7(.)-360.5(T)15(he)-354.7(p)16.2(e)0(r)19.7(c)15(en)19.7(ta)25.3(ge)]TJ 0 -1.1174 TD [(of)-232.6(i)16.1(n)0(h)20.8(i)0(b)26.5(i)0(t)20.6(i)0(o)26.5(n)-243(w)15.3(a)0(s)-233.6(c)15(al)19.5(cu)25.4(la)19.6(t)16.1(e)0(d)-233.7(r)16.2(el)19.5(at)25.3(iv)20.7(e)-238.4(t)0(o)-227(p)0(o)20.8(s)0(i)26.7(t)0(i)20.6(v)0(e)-228(\()0(i)20.7(.)13.9(e.)23.1(,)-239.6(n)0(o)26.6(-)0(s)21.1(e)0(r)19.7(u)16.2(m)]TJ 0 -1.1116 TD [(or)-313.2(n)16.2(o)0(-)20.9(B)0(A)24.7(L)0(F)19.8(\))-317.8(co)19.7(nt)26.5(ro)20.9(ls)20.9(.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 59.7543 95.0173 Tm [(Luciferase)-333.6(assay)]TJ /F5 1 Tf 9.843 0 0 9.843 69.7323 78.5763 Tm [(Ce)18.6(ll)20.6(s)-335(w)0(e)18.8(r)0(e)-320.1(l)0(y)15(s)0(e)19.9(d)-329.4(in)-324.9(70)]TJ /F6 1 Tf 9.2616 0 TD (l)Tj /F5 1 Tf .5414 0 TD [(l)-329.5(o)0(f)-324.7(l)0(y)20.7(s)0(i)20.9(s)-335(bu)20.8(ff)20.9(er)-325.9(\(5)20.9(0)-156.6(m)]TJ /F9 1 Tf 8.8642 0 TD (M)Tj /F5 1 Tf 1.1635 0 TD [(Tr)19.7(is)-324.7([p)20.9(H)]TJ -20.8444 -1.1174 TD [(7.)18.5(5])20.9(,)-660(2)0(0)20.8(0)-156.6(m)]TJ /F9 1 Tf 4.8612 0 TD (M)Tj /F5 1 Tf 1.486 0 TD [(Na)18.8(Cl)19.7(,)-660(0)0(.)24.3(2)0(%)-648.3(N)0(o)20(n)0(i)20.7(d)0(e)19.7(t)-657.8(P-)21(40)-647.3([N)20(P-)21(40)15.1(],)-643.8(10)20.8(%)]TJ -6.3472 -1.1116 TD [(gl)14.9(yc)19.7(er)19.7(ol)20.7(\))-369.7(a)0(n)19.7(d)-375.5(th)20.7(e)-370.9(l)0(y)20.7(s)0(a)19.9(t)0(e)-360.6(w)0(a)13(s)-369.5(as)19.9(sa)19.9(ye)19.7(d)-375.5(a)0(c)18.5(c)0(o)19.7(r)0(d)20.9(i)0(n)20.7(g)-369.7(to)-365.2(th)20.7(e)-370.9(m)0(a)18.4(n)0(-)]TJ 25.5097 67.9184 TD [(uf)20.9(ac)18.5(tu)20.7(re)19.7(r)15.1(s)-329.2(p)0(r)20.9(o)0(t)14.9(o)0(c)19.7(o)0(l)-319.1(\()0(O)20(N)0(E)18.8(-)0(G)14.2(l)0(o)-319.1(l)0(u)20.7(c)0(i)19.5(f)0(e)14(r)0(a)19.7(s)0(e)-319.9(a)0(s)14.1(s)0(a)19.9(y)-329.4(sy)21(st)15.1(em)18.5(;)-329.5(P)0(r)21(o)0(-)]TJ 0 -1.1174 TD [(me)18.4(ga)19.7(,)-366.3(M)0(a)18.8(d)0(i)20.7(s)0(o)21(n)0(,)-355.9(W)0(I)18.6(\))0(.)-355.8(L)0(u)19.6(c)0(i)19.5(f)0(e)19.7(r)0(a)19.7(s)0(e)-360.3(a)0(c)18.5(t)0(i)20.6(v)0(i)20.7(t)0(y)-353.7(w)0(a)18.8(s)-369.5(no)20.8(rm)19.6(al)19.5(iz)19.5(ed)-354.7(to)]TJ 0 -1.1116 TD [(to)20.7(ta)19.5(l)-364.1(p)0(r)20.9(o)0(t)20.7(e)0(i)19.5(n)-364(de)19.7(te)19.5(rm)19.6(in)20.7(ed)-354.7(by)-353.5(bi)20.7(ci)19.5(nc)19.7(ho)20.8(ni)14.9(ni)20.7(c)-359.4(a)0(c)18.5(i)0(d)-353.7(\()0(B)19.8(C)0(A)18.9(\))-363.9(as)19.9(sa)19.9(y)]TJ T* [(\(P)21(ie)19.5(rc)19.7(e)-324.8(B)0(i)19.7(o)0(t)14.9(e)0(c)18.5(h)0(n)20.8(o)0(l)20.7(o)0(g)20.8(y)0(,)-315.6(R)0(o)19.8(c)0(k)19.7(f)0(o)20.9(r)0(d)15.1(,)-320.2(IL)19.7(\).)]TJ /F11 1 Tf 8.9663 0 0 8.9663 310.8472 670.3369 Tm [(Fluorescence-activated)-337.2(cell-sorting)-331.6(analysis)]TJ 0 -1.2203 TD [(of)-330.4(BALF)-334.7(samples)]TJ /F5 1 Tf 9.843 0 0 9.843 320.8251 642.9542 Tm -.017 Tc [(Aft)4.8(e)-2(r)-271.5(B)-1.8(ALF)-271.4(c)-2(o)4.9(l)-.9(l)4.8(e)-2(cti)4.8(on,)-273.9(s)5.1(a)-2(mpl)4.8(e)-2(s)-271.3(w)-1.7(ere)-272.7(c)-2(entr)5(if)5(ug)4.9(ed)-271.5(at)-271.7(400)]TJ /F10 1 Tf 22.077 0 TD 0 Tc ()Tj /F9 1 Tf .6969 0 TD (g)Tj /F5 1 Tf -23.7876 -1.1116 TD [(for)-348.8(5)-168.1(min)-344.5(to)-353.7(collect)-351.4(cell)-352.6(pellets.)-347.7(Lymphocytes)-356.6(were)-347.4(surface)]TJ 0 -1.1174 TD [(stained)-522.7(for)-515.9(30)-169.2(min)-517.3(at)-521.9(4)]TJ /F10 1 Tf 10.0449 0 TD ()Tj /F5 1 Tf .3917 0 TD [(C)-520.5(with)-518.1(phycoerythr)-8.8(in-conjugated)]TJ -10.4366 -1.1116 TD [(an)19.7(ti)20.6(-C)19.8(D4)20(5R)-268.2(\(B)19.8(22)20.8(0;)-261.5(RA)18.9(3-)15.1(6B)19.8(2;)-261.5(eB)18.7(io)20.7(sc)19.9(ie)13.8(nc)19.7(e,)-264.9(Vi)19.8(en)19.7(na)19.7(,)-274.1(A)0(u)20(s)0(t)20.9(r)0(i)20.7(a)0(\))]TJ T* [(an)19.7(d)-317.9(A)0(P)20.1(C)0(-)19.8(H)0(7)20(-)0(c)19.7(o)0(n)15.1(j)0(u)20.7(g)0(a)19.7(t)0(e)19.5(d)-317.9(an)19.7(ti)20.6(-C)19.8(D4)-308.3(\(G)20(K1)14.2(.)13.8(5)0(;)-307.6(B)0(D)-309.4(B)0(i)19.7(o)0(s)21(c)0(i)19.5(e)0(n)13.9(c)0(e)18.5(s)0(,)]TJ 0 -1.1174 TD [(Erembodegem,)-489.2(Belgium\),)-494(before)-486.9(xation)-487(and)-488.3(permeabili-)]TJ 0 -1.1116 TD [(zation)-320.2(with)-316.6(an)-320.1(eBioscience)-324.3(Foxp3)-322.1(xation/permeabilization)]TJ T* -.0171 Tc [(k)4.8(i)-1.1(t)-283.3(\()4.9(cat.)-279.7(no)4.8(.)-285.5(0)4.8(0-)4.9(55)4.8(23)4.8(-00)4.8(;)-283.3(e)3.7(Bi)4.7(oscience\))4.9(.)-285.5(Cel)4.7(l)-1.1(s)-282.9(w)4(er)4.9(e)-284.3(t)4.7(hen)-283.1(s)-.7(t)4.7(a)-2.1(i)4.7(n)-.9(ed)]TJ T* 0 Tc [(for)-492.8(30)-163.5(min)-494.2(at)-487.3(4)]TJ /F10 1 Tf 6.5949 0 TD ()Tj /F5 1 Tf .3974 0 TD [(C)-491.7(with)-489.3(anti-Foxp3-APC)-496.4(\(FJK-16s;)-494.1(eBio-)]TJ -6.9923 -1.1174 TD [(science\).)-299(Analysis)-298.7(was)-293.1(performed)-295.6(with)-299.3(a)-296(B)0(D)-292.1(FACSCanto)-301.8(II)]TJ 0 -1.1116 TD [(o)21(w)-272.7(c)0(y)19.7(t)0(o)20.7(m)0(e)18.4(t)0(e)13.8(r)-271.8(\(B)19.8(D)-272.7(B)0(i)19.7(o)0(s)21(c)0(i)19.5(e)0(n)13.9(c)0(e)18.5(s)0(\))-261.2(w)0(i)19.8(t)0(h)-261.5(F)0(l)15.1(o)0(w)20(J)0(o)-261.2(v)0(e)19.7(r)0(s)15.3(i)0(o)20.7(n)-271.8(10)20.8(.0)18.5(.)13.8(6)]TJ T* [(an)19.7(al)19.5(ys)21(is)-313.2(so)15.3(ft)20.7(wa)18.8(re)-314.4(\(T)19.7(re)19.7(e)-324.8(S)0(t)15.1(a)0(r)19.7(,)-320.2(As)20.2(hl)14.9(an)19.7(d,)-309.8(OR)18.9(\).)]TJ /F11 1 Tf 8.9663 0 0 8.9663 310.8472 500.4849 Tm [(Statistical)-336.1(analysis)]TJ /F5 1 Tf 9.843 0 0 9.843 320.8251 484.0439 Tm [(Ge)18.8(ne)19.7(ra)19.7(l)-260.4(c)0(h)19.7(a)0(n)19.7(g)0(e)13.9(s)-260.1(in)-250(BL)18.6(I)-266(s)0(i)20.9(g)0(n)20.8(a)0(l)-251.2(o)0(v)15.1(e)0(r)-251(t)0(i)20.6(m)0(e)-252.3(w)0(e)18.8(r)0(e)-256.8(a)0(n)19.7(a)0(l)19.5(y)0(z)19.7(e)0(d)-251(b)0(y)]TJ -1.0137 -1.1116 TD [(re)19.7(pe)19.7(at)19.5(ed)19.7(-m)19.6(ea)12.8(su)21(re)19.7(s)-346.5(a)0(n)19.7(a)0(l)19.6(y)0(s)21(i)0(s)-336.2(o)0(f)-330.5(v)0(a)13.9(r)0(i)20.7(a)0(n)19.7(c)0(e)-332.8(\()0(A)14.2(N)0(O)19.1(V)0(A)19.1(\))0(.)-332.8(C)0(o)19.8(m)0(p)19.6(a)0(r)19.7(-)]TJ 0 -1.1174 TD [(is)20.9(on)20.8(s)-306.2(b)0(e)19.7(t)0(w)19.8(e)0(e)18.5(n)-306.4(gr)20.9(ou)20.8(ps)-295.7(at)-291.5(sp)15.3(ec)18.5(i)20.9(c)-301.8(t)0(i)14.8(m)0(e)-292.6(p)0(o)20.8(i)0(n)20.7(t)0(s)-295.9(w)0(e)18.8(r)0(e)-297.1(p)0(e)19.7(r)0(f)20.9(o)0(r)15.1(m)15(ed)]TJ 0 -1.1116 TD [(by)-491.8(on)15.1(e-)19.7(wa)18.8(y)-507.9(A)0(N)19.1(O)0(V)19.1(A)-508.8(fo)20.9(ll)20.6(ow)20(ed)-498.7(by)-497.5(a)-503.4(T)0(u)19.6(k)0(e)19.7(y)-507.9(HS)20.1(D)-508.8(\()0(h)20.9(o)0(n)20.8(e)0(s)19.9(t)]TJ T* [(si)20.9(gn)20.8(i)20.9(ca)12.8(nt)-290.3(di)20.7(ff)20.9(er)14(en)19.7(ce)18.5(\))-300.6(p)0(o)20.8(s)0(t)15.1(-)0(h)20.9(o)0(c)-291.4(t)0(e)19.6(s)0(t)20.9(.)-302.9(Ch)19.8(an)13.9(ge)19.7(s)-300.4(i)0(n)]TJ /F6 1 Tf 19.3469 0 TD (b)Tj /F5 1 Tf .5414 0 TD [(-G)14.2(al)-291.5(si)20.9(gn)20.8(al)]TJ -19.8883 -1.1116 TD [(in)-468.9(ea)18.5(ch)-464.1(gr)15.1(ou)20.8(p)-473.4(s)0(e)14.1(p)0(a)19.7(r)0(a)19.7(t)0(e)19.6(l)0(y)-468.9(o)0(v)20.8(e)0(r)-469.9(t)0(i)20.6(m)0(e)-471.1(w)0(e)18.8(r)0(e)-464.1(a)0(n)13.9(a)0(l)19.6(y)0(z)19.7(e)0(d)-469.9(w)0(i)19.8(t)0(h)-463.1(a)]TJ 0 -1.1174 TD [(mi)19.5(xe)19.7(d-)20.9(mo)19.6(de)19.7(l)-398.6(A)0(N)19.1(O)0(V)19.1(A)0(,)-391.3(i)0(n)20.7(c)0(l)19.6(u)0(d)20.8(i)0(n)15(g)-398.5(an)19.7(im)19.5(al)-389.4(id)20.7(en)19.7(ti)20.6(c)19.8(at)19.5(io)15(n)-398.5(a)0(s)-383.3(a)]TJ 0 -1.1116 TD [(ra)19.7(nd)20.8(om)-216.5(ef)19.7(fe)19.7(ct)19.6(,)-228.1(g)0(r)20.9(o)0(u)20.8(p)-225.7(as)-216.3(a)-226.9()0(x)21(e)0(d)-216.5(e)0(f)19.7(f)0(e)19.7(c)0(t)19.5(,)-228.1(an)19.7(d)-225.7(t)0(i)20.6(m)0(e)-217.7(a)0(s)-210.5(a)-226.9(co)19.7(va)19.7(ri)15(at)19.5(e.)]TJ T* [(Fo)21(r)-398.5()0(u)21(o)0(r)20.9(e)0(s)14.1(c)0(e)18.5(n)0(c)19.7(e)0(-)19.7(a)0(c)18.5(t)0(i)20.6(v)0(a)19.7(t)0(e)13.8(d)-392.8(ce)18.5(ll)14.8(-s)21.1(or)20.9(ti)20.6(ng)-388.1(\(F)15.3(AC)18.9(S\))-387.9(an)19.7(al)19.5(ys)21(is)-388(an)19.7(d)]TJ 0 -1.1174 TD [(rA)20(AV)-217(tr)20.7(an)19.7(sd)15.3(uc)19.7(ti)20.6(on)-215.3(in)20.7(hi)20.7(bi)14.9(ti)20.6(on)-215.3(da)19.7(ta)19.5(,)-228.1(s)0(t)20.9(a)0(t)19.5(i)0(s)20.9(t)0(i)20.6(c)0(a)12.8(l)-220.1(si)15.1(gn)20.8(i)20.9(ca)18.5(nc)19.7(e)-226.9(w)0(a)18.8(s)]TJ 0 -1.1116 TD [(ev)19.7(al)19.5(ua)19.7(te)19.6(d)-260.3(b)0(y)-244.1(A)0(N)19.1(O)0(V)19.1(A)-255.4(to)-250(co)19.7(mp)19.6(ar)19.7(e)-255.7(d)0(i)14.9(f)0(f)20.9(e)0(r)19.7(e)0(n)19.7(t)-254.7(tr)15(ea)18.5(tm)19.5(en)19.7(t)-254.7(g)0(r)20.9(o)0(u)15.1(p)0(s)21(.)]TJ T* .4051 Tc [(AS)405.1(t)426(u)405.1(d)425.9(e)405.1(n)424.8(t)]TJ /F9 1 Tf 4.5271 0 TD 0 Tc (t)Tj /F5 1 Tf .6854 0 TD [(te)19.6(st)-399.6(wa)18.8(s)-404.1(s)0(u)21(b)0(s)21(e)0(q)19.7(u)0(e)13.9(n)0(t)20.7(l)0(y)-394(u)0(s)21(e)0(d)-395(f)0(o)20.9(r)-404.2(pa)13.9(ir)20.7(-w)20(is)20.9(e)-405.4(c)0(o)19.7(m)0(-)]TJ -5.2125 -1.1174 TD [(pa)19.7(ri)20.7(so)21(ns)15.3(.)-251.1(A)]TJ /F9 1 Tf 4.7287 0 TD (p)Tj /F5 1 Tf .7545 0 TD [(va)19.7(lu)15(e)-255.7(l)0(e)19.5(s)0(s)-243.7(t)0(h)20.7(a)0(n)-245.3(0)0(.)18.5(0)0(5)-244.1(w)0(a)18.8(s)-254.3(co)13.9(ns)21(id)20.7(er)19.7(ed)-245.3(st)15.1(at)19.5(is)20.9(ti)20.6(ca)18.5(ll)14.8(y)]TJ -5.4832 -1.1116 TD [(si)20.9(gn)20.8(i)20.9(ca)12.8(nt)20.7(.)-268.4(D)0(a)18.8(t)0(a)-262.7(a)0(r)19.7(e)-273(pr)20.9(es)14.1(en)19.7(te)19.5(d)-271.8(a)0(s)-262.4(m)0(e)18.4(a)0(n)19.7(s)]TJ /F10 1 Tf 16.0235 0 TD ()Tj /F5 1 Tf .7084 0 TD [(SE)19.8(M,)-264.6(un)20.8(le)19.5(ss)-261(st)20.9(at)19.5(ed)]TJ -16.7319 -1.1116 TD [(ot)20.7(he)19.7(rw)20(is)15.1(e.)-535.6(ST)19.8(AT)18.8(IS)15.3(TI)19.7(CA)-539.8(\(v)20.9(er)19.7(si)20.9(on)-537.8(8.)18.5(0\))-537.8(so)21(ft)15(wa)18.8(re)-539(\(S)21(ta)19.5(tS)15.1(of)20.9(t,)]TJ T* [(Tu)19.6(ls)20.9(a,)-316.7(OK)19.1(\))-323.6(w)0(a)18.8(s)-323.4(us)21(ed)-314.4(fo)20.9(r)-323.6(s)0(t)15.1(a)0(t)19.5(i)0(s)20.9(t)0(i)20.6(c)0(a)18.5(l)-323.8(an)19.7(al)13.8(ys)21(is)20.9(.)]TJ /F4 1 Tf 8.9663 0 0 8.9663 310.8472 297.7511 Tm (Results)Tj /F11 1 Tf 0 -1.8336 TD [(Efcient)-332.3(but)-334.8(transient)-333.2(gene)-333.8(expression)-335.9(in)-330.2(murine)]TJ 0 -1.2203 TD [(airways)-332.2(after)-334.2(perinatal)-337.9(rAAV2/5-based)-331(gene)-333.8(transfer)]TJ /F5 1 Tf 9.843 0 0 9.843 320.8251 253.9275 Tm [(T)15(o)-698(s)16.4(t)0(u)20.7(d)16.2(y)-698(s)16.4(ta)19.6(b)16.2(i)0(l)26.3(i)0(t)26.3(y)-698(of)-687.6(g)16.2(e)0(n)25.4(e)-699.2(e)15(x)0(p)20.8(r)16.2(es)25.6(si)20.9(o)16.2(n)-698(a)15(f)0(t)20.7(e)15(r)-698(p)16.2(er)19.7(i)16(n)0(a)25.4(t)0(a)25.3(l)]TJ -1.0137 -1.1174 TD [(r)16.2(A)0(A)24.9(V)0(2)20(/)16.1(5)-542.5(d)0(e)19.7(l)16.1(iv)26.5(er)25.5(y)-548.3(i)16.1(n)-548.3(t)16(he)-533.3(up)26.6(pe)25.4(r)-548.2(\()16.2(no)26.6(se)19.9(\))-542.5(a)15(nd)-532.1(lo)26.5(we)24.6(r)-548.2(\()16.2(lu)20.7(n)16.2(g)0(s)26.8(\))]TJ 0 -1.1116 TD [(m)15(u)0(r)26.6(i)0(n)20.7(e)-238.4(a)15(i)0(r)26.5(w)0(a)24.6(y)0(s)21(,)-239.6(w)15.3(e)-238.4(co)25.4(mp)25.4(ar)25.5(ed)-228(ge)25.4(ne)-228(ex)25.4(pr)26.6(es)25.6(si)20.9(o)16.2(n)-237.2(ov)20.8(e)15(r)-237.2(ti)20.6(m)15(e)-238.4(af)25.5(te)25.3(r)]TJ T* -.0249 Tc [(f)-8.7(e)-24.9(t)-5.3(a)-9.9(l)-279.6(a)-24.9(nd)-285.2(n)-8.7(e)-24.9(on)-24.9(at)-24.9(al)-285.3(r)-8.7(A)-24.9(AV)-24.9(2)-4.9(/)-8.9(5)-279.4(d)-24.9(el)-24.9(iv)-8.7(e)-24.9(r).6(y)-285.2(o)-8.7(f)-279.4(f)-24.9(L)-5.2(U)-9.6(C)-280.5(\()-24.9(1.)-24.9(5)]TJ /F10 1 Tf 19.9747 0 TD 0 Tc ()Tj /F5 1 Tf .7084 0 TD -.0162 Tc (10)Tj 7.3823 0 0 6.5614 524.0125 225.411 Tm (10)Tj 9.843 0 0 9.843 533.7637 221.0456 Tm 0 Tc [(GC)24.7(/)]TJ -22.6472 -1.1174 TD -.015 Tc [(an)-15(i)11.5(m)-15(a)9.2(l)-15(\))-345.6(a)-15(n)10.4(d)]TJ /F6 1 Tf 5.0973 0 TD 0 Tc (b)Tj /F5 1 Tf .5414 0 TD -.0162 Tc [(-G)-16.2(a)8.4(l)-357.3(\()-16.2(1)]TJ /F10 1 Tf 3.0584 0 TD 0 Tc ()Tj /F5 1 Tf .7027 0 TD (10)Tj 7.3823 0 0 6.5614 412.951 214.4692 Tm -.0162 Tc (10)Tj 9.843 0 0 9.843 423.6093 210.0472 Tm -.0253 Tc [(G)-10(C)-25.3(/a)-25.3(ni)-25.3(ma)-25.3(l\))-371.9(\()]TJ /F9 1 Tf 5.2817 0 TD 0 Tc (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7027 0 TD .3409 Tc [(3o)340.9(r)10.4(4)0(p)357.1(e)340.9(r)9.3(t)340.9(i)367.2(m)340.9(e)]TJ -18.097 -1.1116 TD 0 Tc [(p)16.2(o)0(i)26.5(n)0(t)-497.7(p)0(e)25.4(r)-513.7(g)16.2(r)0(o)20.9(u)16.2(p\))26.6(.)-510.3(A)0(n)25.7(a)0(l)25.3(y)0(s)21(i)16.1(s)-513.5(o)16.2(f)-513.7(l)16.1(un)26.6(g)-513.7(s)16.4(ec)24.3(ti)26.3(on)20.8(s)-507.8(s)0(h)26.8(o)0(w)25.7(e)0(d)-498.7(a)0(v)25.4(e)0(r)25.5(-)]TJ T* [(a)15(g)0(e)-297.1(t)0(r)26.5(a)0(n)19.7(s)16.4(du)26.6(ct)25.3(io)20.7(n)-306.4(e)0(f)25.5()0(c)25.6(i)0(e)25.3(n)0(c)25.4(i)0(e)19.5(s)-306.2(\(p)26.6(er)25.5(ce)24.3(nt)]TJ /F6 1 Tf 15.0616 0 TD (b)Tj /F5 1 Tf .5414 0 TD [(-G)25.8(al)25.3(-p)20.9(o)16.2(s)0(i)26.7(t)0(i)20.6(v)16.2(e)-307.5(c)0(e)18.5(l)16(ls)26.7(\))-312.1(o)16.2(f)]TJ -15.603 -1.1116 TD -.0151 Tc (6.6)Tj /F10 1 Tf 1.3766 0 TD 0 Tc ()Tj /F5 1 Tf .7084 0 TD [(1)16.2(.)13.9(1%)-360.4(i)16.1(n)-369.7(th)20.7(e)-370.9(f)16.2(et)25.3(al)-360.6(gr)26.6(ou)26.6(p)-369.7(a)0(n)19.7(d)-369.7(8)16.2(.)13.8(9)]TJ /F10 1 Tf 13.0573 0 TD ()Tj /F5 1 Tf .7027 0 TD [(1.)24.3(5%)-360.4(i)16(n)-369.7(th)20.7(e)-370.9(n)16.2(eo)25.4(na)25.4(ta)25.3(l)]TJ -15.8449 -1.1174 TD [(g)16.2(r)0(o)20.9(u)16.2(p)-352.4(a)15(t)-352.6(1)-352.4(m)15(o)16.2(nt)20.7(h)-346.7(p)0(o)26.6(s)0(t)20.9(i)16.1(nj)26.5(ec)24.3(ti)20.6(o)16.2(n)-352.4(\()16.2(F)0(i)20.9(g)16.2(.)-349(1)0(A)20(\))16.2(.)-349(A)-353.3(6-)26.6(fo)26.6(ld)-342.2(d)16.2(e)0(c)24.3(r)0(e)25.5(a)0(s)19.9(e)]TJ 0 -1.1116 TD [(w)15.3(a)0(s)-435.1(n)0(o)26.6(t)0(e)25.3(d)-444.6(be)25.4(tw)25.6(ee)18.5(n)-444.6(1)-444.6(a)15(nd)-434.2(3)-444.6(m)15(on)26.6(th)26.5(s)-444.4(i)0(n)-434.3(t)0(h)26.5(e)-445.8(ne)25.4(on)26.6(at)19.5(a)15(l)-444.7(gr)26.6(ou)26.6(p)]TJ T* (\()Tj /F9 1 Tf .4838 0 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7027 0 TD [(0.)24.3(00)26.6(1\))26.6(,)-446.9(i)0(n)-440.1(c)15(on)26.6(tr)26.5(as)19.9(t)-444.7(t)0(o)-434.3(t)0(h)26.5(e)-451.5(f)16.2(e)0(t)25.3(a)0(l)-435.5(g)0(r)20.9(o)16.2(up)-434.2(wh)25.7(er)25.5(e)-451.5(t)16(he)-435.3(tr)20.7(a)15(n)0(s)26.8(-)]TJ -1.8431 -1.1174 TD [(d)16.2(u)0(c)25.4(t)0(i)20.6(o)16.2(n)-387(e)0(f)25.5()0(c)19.8(i)16.1(en)25.4(cy)-377.7(d)16.2(i)0(d)-376.7(n)0(o)26.6(t)-387.1(ch)19.7(a)15(n)0(g)26.6(e)-388.2(si)20.9(g)16.2(n)0(i)26.5()0(c)25.6(a)0(n)19.7(t)16.1(ly)-376.7(\()16.2(1)0(.)24.3(7)0(-)26.6(f)0(o)20.9(l)16.1(d)-387(d)0(e)25.4(-)]TJ 0 -1.1116 TD -.0162 Tc [(cr)-16.2(e)9.3(a)-16.2(se;)]TJ /F9 1 Tf 2.9835 0 TD 0 Tc (p)Tj /F10 1 Tf .6508 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0)16.2(.)0(1)24.3(8)0(\))26.6(.)-274.1(S)16.3(i)0(m)25.2(i)0(l)26.3(a)0(r)-256.8(d)0(a)19.7(t)16.1(a)-273(w)15.3(er)25.5(e)-273(o)0(b)26.6(t)0(a)25.3(i)0(n)26.5(e)0(d)-262.5(f)16.2(or)-255.6(fL)25.4(UC)-263.3(u)16.2(n)0(t)26.5(i)0(l)-255.9(3)]TJ -4.3428 -1.1116 TD [(m)15(o)0(n)26.6(t)0(h)26.5(s)-311.9(p)16.2(o)0(s)26.8(t)0(i)20.6(n)16.2(je)25.3(ct)25.3(io)20.7(n)-306.4(\()16.2(Su)26.8(pp)20.8(l)16.1(e)0(m)24.2(e)0(n)25.4(t)0(a)25.3(r)0(y)-295.9(F)0(i)26.6(g)0(.)-292.5(S)0(1)21(;)-306.5(s)16.4(u)0(p)20.8(p)16.2(le)25.3(me)24.2(nt)26.5(ar)25.5(y)]TJ 0 -1.1174 TD [(d)16.2(a)0(t)25.3(a)-526.4(a)15(r)0(e)-510.2(a)0(v)25.4(a)0(i)25.3(l)0(a)25.3(b)0(l)20.7(e)-520.6(on)26.6(li)26.3(ne)-510.2(at)-510.3(ww)24.9(w.)23.4(l)16(i)0(e)25.3(b)0(e)19.7(r)16.2(tp)26.5(ub)20.8(.)13.8(c)15(om)25.4(/h)26.5(um)25.4(\).)-511.3(I)16.2(n)]TJ 0 -1.1116 TD [(p)16.2(a)0(r)25.5(a)0(l)19.5(l)16(el)25.3(,)-481.5(w)0(e)-470.8(q)16.2(ua)25.4(nt)26.5(i)20.9(e)15(d)-479.1(th)26.5(e)-480.3(e)0(f)19.7()16.3(ci)25.3(en)25.4(cy)-469.9(of)-468.7(n)16.2(a)0(s)25.6(a)0(l)-470(t)0(r)26.5(a)0(n)25.4(s)0(d)21(u)16.2(ct)25.3(io)26.5(n)]TJ T* [(\()16.2(F)0(i)20.9(g)16.2(.)-521.8(1)0(B)25.6(\))0(.)-511.4(S)0(t)26.6(a)0(r)25.5(t)0(i)20.6(n)16.2(g)-525.2(a)15(t)-525.4(1)16.2(1.)24.3(6)]TJ /F10 1 Tf 11.3351 0 TD ()Tj /F5 1 Tf .7027 0 TD [(1.)24.3(5)16.2(%)-526.3(a)15(n)0(d)-509.1(2)0(0)20.8(.)13.9(5)]TJ /F10 1 Tf 6.376 0 TD ()Tj /F5 1 Tf .7084 0 TD -.0162 Tc [(3.)-16.2(6)8.1(%)]TJ /F6 1 Tf 2.5631 0 TD 0 Tc (b)Tj /F5 1 Tf .5414 0 TD [(-G)25.8(al)25.3(-)]TJ -22.2267 -1.1174 TD [(p)16.2(o)0(s)21(i)16(ti)26.3(ve)-343.2(c)15(e)0(l)25.3(l)0(s)-342(1)-346.7(mo)25.4(nt)26.5(h)-352.4(a)0(f)25.5(t)0(e)25.3(r)-352.4(ge)25.4(ne)-337.4(de)25.4(li)20.6(v)16.2(e)0(r)25.5(y)-352.4(fo)26.6(r)-352.4(f)0(e)25.5(t)0(a)25.3(l)-352.6(a)15(n)0(d)-342(n)16.2(eo)25.4(-)]TJ 0 -1.1116 TD [(n)16.2(a)0(t)25.3(a)0(l)-343.3(t)16(re)19.7(a)15(t)0(m)25.2(e)0(n)25.4(t)0(,)-338.7(r)0(e)25.5(s)0(p)26.8(e)0(c)18.5(t)16.1(iv)26.5(el)25.3(y,)-338.6(th)26.5(e)-353.6(n)0(u)26.6(m)0(b)25.4(e)0(r)-343.2(o)16.2(f)-352.4(t)0(r)26.5(a)0(n)25.4(s)0(d)21(u)16.2(ce)24.3(d)-352.4(c)0(e)24.3(l)0(l)26.3(s)]TJ /F4 1 Tf 8.9663 0 0 8.9663 59.7543 749.7069 Tm [(LONG-TERM)-340.8(EXPRESSION)-334.8(AFTER)-337.3(PERINATAL)-338.1(GENE)-336.8(TRANSFER)-21465.7(519)]TJ ET endstream endobj 7 0 obj <> endobj 8 0 obj <>/Font<>/ProcSet[/PDF/Text/ImageC]/ExtGState<>>> endobj 9 0 obj <>stream /GS1 gs BT /F5 1 Tf 9.843 0 0 9.843 62.022 275.8109 Tm 0 0 0 1 k 0 Tc 0 Tw [(de)25.4(cr)25.5(ea)24.3(se)19.9(d)-225.7(s)0(i)26.7(g)0(n)20.8(i)16.1(c)25.6(an)25.4(tl)20.6(y)-225.7(o)0(v)26.6(e)0(r)-222.2(t)16.1(he)-216.5(ne)25.4(xt)-221.2(2)-225.7(m)0(o)25.4(n)0(t)26.5(h)0(s)-220.9(f)16.2(or)-215.3(bo)20.8(t)16.1(h)-231.5(g)16.2(r)0(o)20.9(u)16.2(ps)]TJ 0 -1.1116 TD (\()Tj /F9 1 Tf .4896 0 TD (p)Tj /F10 1 Tf .6508 0 TD (<)Tj /F5 1 Tf .7084 0 TD [(0)16.2(.)13.9(00)20.8(1)-358.2(f)16.2(or)-347.7(fe)25.5(ta)19.5(l)-358.3(a)15(nd)]TJ /F9 1 Tf 7.9369 0 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7027 0 TD [(0.)24.3(0)16.2(1)-364(f)16.2(o)0(r)-347.7(n)0(e)25.4(o)0(n)26.6(a)0(t)19.5(a)15(l\))26.5(,)-360.5(r)0(e)25.5(s)0(u)26.8(l)0(t)20.6(i)16.1(ng)-347.8(in)-347.9(a)]TJ -11.145 -1.1174 TD [(re)25.5(si)26.7(du)20.8(a)15(l)-387.1(t)16.1(r)0(a)19.7(n)16.2(sd)26.8(uc)25.4(ti)20.6(o)16.2(n)-387(l)16.1(e)0(v)25.4(e)0(l)-377.9(a)15(t)-387.1(3)-381.2(mo)25.4(nt)26.5(hs)-376.4(o)16.2(f)-387(0)16.2(.)0(3)]TJ /F10 1 Tf 18.7997 0 TD ()Tj /F5 1 Tf .7084 0 TD [(0)16.2(.)13.9(1%)-371.9(in)-376.7(t)16.1(h)0(e)]TJ -19.5081 -1.1116 TD [(fe)25.5(ta)25.3(l)-341.1(g)0(r)20.9(o)16.2(up)-330.5(c)15(o)0(m)25.4(p)0(a)25.4(r)0(e)25.5(d)-340.9(wi)25.6(th)-330.6(4.)24.3(0)]TJ /F10 1 Tf 12.3718 0 TD ()Tj /F5 1 Tf .7084 0 TD [(1)16.2(.)13.9(1%)-331.6(i)16.1(n)-340.9(th)20.7(e)-336.3(n)0(e)19.7(o)16.2(na)25.4(ta)25.3(l)-341(g)0(r)26.6(o)0(u)20.8(p)16.2(.)]TJ -13.0803 -1.1116 TD [(Al)25.6(th)26.5(ou)20.8(g)16.2(h)-415.8(th)26.5(e)-422.7(a)15(ir)26.5(wa)24.6(ys)-405.2(we)24.6(re)-406.5(ef)25.5(c)25.6(ie)25.3(nt)20.7(l)16.1(y)-415.8(tr)20.7(a)15(n)0(s)26.8(d)0(u)26.6(c)0(e)24.3(d)0(,)-407.7(g)16.2(en)25.4(e)-422.7(e)15(x-)]TJ 0 -1.1174 TD [(pr)26.6(es)25.6(si)20.9(o)16.2(n)-335.2(m)15(a)0(r)25.5(k)0(e)25.4(d)0(l)20.7(y)-329.4(de)25.4(cr)25.5(ea)24.3(se)25.6(d)-335.2(b)16.2(y)-335.2(m)15(on)26.6(th)-324.9(3)16.2(,)-331.7(pr)26.6(ov)26.6(id)20.7(i)16.1(n)0(g)-319(g)0(r)26.6(o)0(u)20.8(n)16.2(ds)]TJ 0 -1.1116 TD [(fo)26.6(r)-317.8(v)16.2(ec)24.3(to)26.5(r)-317.8(r)16.2(ea)24.3(dm)25.4(in)20.7(i)16.1(s)0(t)26.7(r)0(a)19.7(t)16.1(io)26.5(n)-317.9(t)16.1(o)-317.9(e)15(st)26.7(ab)25.4(li)20.6(s)16.4(h)-317.9(l)16.1(o)0(n)26.6(g)0(-)26.6(t)0(e)19.6(r)16.2(m)-313.3(e)0(x)25.4(p)0(r)20.9(e)15(ss)27(io)20.7(n)16.2(.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 62.022 182.6645 Tm [(Successful)-332.2(readministration)-337.8(of)-336.7(rAAV2/5)]TJ 0 -1.2266 TD [(after)-334.2(initial)-335.4(perinatal)-337.8(gene)-333.8(transfer)]TJ /F5 1 Tf 9.843 0 0 9.843 72 155.2818 Tm [(Next,)-321(we)-321(wanted)-321(to)-319.1(evaluate)-324.7(whether)-322(readministration)-328(of)]TJ -1.0137 -1.1174 TD [(an)-435.3(autologous)-438.5(vector)-441(was)-437.1(successful)-440.2(after)-439.9(initial)-435.7(perinatal)]TJ 0 -1.1116 TD [(gene)-329.3(transfer.)-331.2(In)-324.7(Fig.)-329.2(2A,)-329.1(a)-324.8(schematic)-332.6(overview)-334.7(is)-324.7(given)-329.3(of)]TJ T* [(the)-275.3(experiment)-275.2(designed)-276(to)-273(study)-270.4(the)-275.3(feasibility)-274(of)-272.9(rAAV2/5)]TJ 0 -1.1174 TD [(readministration)-500.8(to)-497.7(the)-499.9(murine)-498.7(upper)-496.2(and)-499.8(lower)-497.2(airways.)]TJ 0 -1.1116 TD [(rAAV2/5)-453.9(encoding)-454.8(fLUC)-456.7(\(1.5)]TJ /F10 1 Tf 12.8902 0 TD ()Tj /F5 1 Tf .72 0 TD (10)Tj 7.3823 0 0 6.5614 205.8519 104.8252 Tm (10)Tj 9.843 0 0 9.843 217.7007 100.4598 Tm [(GC/animal\))-454.6(was)-454.4(de-)]TJ -15.8162 -1.1116 TD [(livered)-396.2(by)-393.9(intraamniotic)-399.8(injection)-392.9(to)-394(fetal)-394(FVB/N)-399.9(mice)-394(on)]TJ 0 -1.1174 TD [(embryonic)-629.9(day)-632.3(18)-630(\(E18\))-628.7(\(term,)-633.4(E19.5;)]TJ /F9 1 Tf 17.8666 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(19\))-625.3(and)-632.3(by)]TJ -19.2547 -1.1116 TD [(snifng)-450.9(to)-445.8(neonatal)-450.3(pups)-447.7(on)-445.7(day)-448(3)-444.6(\()]TJ /F9 1 Tf 15.5339 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(22\).)-444.4(At)-446.7(3)-450.3(months)]TJ -16.922 -1.1116 TD [(postinjection,)-239.4(a)-238.4(second)-244.9(dose)-235.8(of)-238.3(the)-240.7(same)-238.2(vector)-239.4(was)-241.3(given)-237.2(by)]TJ 25.504 22.267 TD [(intratrache)-11.3(al)-452.8(instillation)-458.6(to)-457.3(approximately)-457.1(half)-460.7(of)-451.4(the)-459.6(ani-)]TJ 0 -1.1116 TD [(mals)-536.6(\(readministration)-542.2(group,)]TJ /F9 1 Tf 12.9997 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(11)-532.1(for)-533.1(the)-540.2(fetal)-532.2(group,)]TJ /F9 1 Tf -14.3877 -1.1174 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(12)-301.7(for)-297(the)-298.3(neonatal)-306.3(group\),)-298(whereas)-305.6(the)-298.3(other)-300.5(half)-299.4(of)-301.7(the)]TJ -1.3881 -1.1116 TD [(group)-402.9(was)-408.3(kept)-397.4(as)-406.3(non-readministration)-407.3(controls)-401.7(\()]TJ /F9 1 Tf 20.8386 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(8)-398.5(for)]TJ -22.2267 -1.1116 TD [(the)-471.1(fetal)-463.1(group,)]TJ /F9 1 Tf 7.0038 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(10)-468.7(for)-464(the)-465.4(neonatal)-467.6(group\).)-470.8(Gene)-462.6(ex-)]TJ -8.3861 -1.1174 TD [(pression)-303.4(levels)-302.7(were)-301.3(monitored)-306.3(by)-295.9(BLI)-299.2(at)-303(monthly)-297.2(intervals)]TJ 0 -1.1116 TD [(until)-334.2(4)-329.4(months)-336.1(after)-330.4(primary)-332.7(administration)-336.2(\(Fig.)-330.3(2B)-331.5(and)-332.8(C)]TJ T* [(and)-275.2(Fig.)-271.6(2E)-274.1(and)-275.2(F)-271.6(for)-273.9(lung)-275.2(and)-269.4(nose,)-273.8(respectivel)-11.2(y\).)-270.5(An)-273.8(adult)]TJ 0 -1.1174 TD [(control)-429.6(group)-431.7(\(23)-424.8(months)-428.2(of)-428.4(age,)]TJ /F9 1 Tf 15.004 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(6\))-428.4(was)-431.4(included)-426.1(to)]TJ -16.3864 -1.1116 TD [(control)-308.6(for)-308.5(the)-304.1(maximum)-306.4(level)-307.6(of)-301.7(gene)-306.2(expression)-312.6(expected)]TJ T* [(in)-382.5(the)-384.7(upper)-381(and)-384.6(lower)-387.8(airways)-385.2(after)-382.3(a)-382.4(single)-382.2(intratracheal)]TJ T* [(instillation)-314.6(of)-307.4(rAAV2/5-fLUC)-312.7(\(Fig.)-307.3(2D)-308.3(and)-309.7(G\).)-311.7(At)-308.5(1)-306.4(month)]TJ 0 -1.1174 TD [(postinjection,)-579.2(a)-578.2(comparable)-575.7(BLI)-575.7(signal)-578(was)-575.4(detected)-578.2(in)]TJ 0 -1.1116 TD [(the)-350.2(lungs)-351(between)-352(the)-350.2(groups)-346.2(\(Fig.)-347.6(2B)-348.8(and)-350(C\).)-346.5(The)-351.2(photon)]TJ T* [(ux,)-490.4(however,)-492.4(declined)-496.4(signicantly)-493.8(in)-486.1(both)-494.1(groups)-490.2(at)-487.3(3)]TJ 0 -1.1174 TD [(months)-318.8(postinjection)-322.4(\(5.3-fold,)]TJ /F9 1 Tf 12.8729 0 TD (p)Tj /F10 1 Tf .6624 0 TD (<)Tj /F5 1 Tf .72 0 TD [(0.001)-318.9(for)-314.3(the)-321.4(fetal)-319.1(group)]TJ -14.2553 -1.1116 TD [(and)-309.7(5.8-fold,)]TJ /F9 1 Tf 5.512 0 TD (p)Tj /F10 1 Tf .6681 0 TD (<)Tj /F5 1 Tf .72 0 TD [(0.001)-301.6(for)-308.5(the)-309.9(neonatal)-312.1(group\).)-303.7(Also)-310.5(for)-308.5(the)]TJ -6.9001 -1.1116 TD [(nose,)-354.4(a)-353.6(signicant)-353.2(decrease)-360(was)-356.5(observed)-350.7(between)-357.8(1)-352.4(and)-350(3)]TJ 0 -1.1174 TD [(months,)-356.8(resulting)-356.8(in)-353.7(a)-353.6(5.4-fold)-351(decrease)-360(for)-354.6(the)-350.2(fetal)-359.4(group)]TJ 0 -1.1116 TD (\()Tj /F9 1 Tf .5011 0 TD (p)Tj /F10 1 Tf .6681 0 TD (<)Tj /F5 1 Tf .72 0 TD [(0.001\))-510(and)-511.3(a)-509.1(7.3-fold)-512.3(decrease)-515.5(for)-510.1(neonatal)-513.7(animals)]TJ -1.8892 -1.1116 TD (\()Tj /F9 1 Tf .5011 0 TD (p)Tj /F10 1 Tf .6681 0 TD (<)Tj /F5 1 Tf .72 0 TD (0.001\).)Tj ET q 420.92 0 0 326.041 96.775 406.205 cm /Im1 Do Q BT /F7 1 Tf 9.845 0 0 9.845 62.022 389.4235 Tm [(FIG.)-296.4(1.)]TJ /F5 1 Tf 4.0771 0 TD [(Pe)20.1(ri)20.9(na)19.9(ta)19.7(l)-289.1(r)0(A)20.2(A)0(V)19.4(2)0(/)20.9(5)0(-)21.1(m)0(e)18.7(d)0(i)20.9(a)0(t)13.9(e)0(d)-273.8(g)0(e)19.9(n)0(e)-273.8(t)0(r)20.9(a)0(n)19.9(s)0(f)15.5(e)0(r)-273.8(t)0(o)-272.8(m)0(u)19.9(r)0(i)20.9(n)0(e)-279.6(a)0(i)19.7(r)0(w)20.2(a)0(y)19.9(s)0(.)]TJ /F7 1 Tf 24.8366 0 TD (\(A)Tj /F5 1 Tf 1.3245 0 TD [(an)19.9(d)]TJ /F7 1 Tf 1.7103 0 TD (B\))Tj /F6 1 Tf 1.2784 0 TD (b)Tj /F5 1 Tf .5413 0 TD [(-G)20.2(al)19.7(ac)18.7(to)20.9(si)15.3(da)19.9(se)-273.6(\()]TJ /F6 1 Tf 6.3805 0 TD (b)Tj /F5 1 Tf .5413 0 TD [(-G)20.2(al)13.9(\))-283.2(e)0(x)19.9(p)0(r)21.1(e)0(s)20(s)0(i)15.3(o)0(n)-272.6(i)0(n)-272.8(t)0(h)20.9(e)]TJ -40.6899 -1.0135 TD [(ai)19.7(rw)20.2(ay)14.1(s)-236.9(a)0(f)19.9(t)0(e)19.7(r)-237.1(fe)19.9(ta)19.7(l)-237.3(a)0(n)19.9(d)-237.1(ne)19.9(on)21.1(at)19.7(al)-227.9(rA)20.2(AV)13.6(2/)20.9(5)-237.1(a)0(d)19.9(m)0(i)19.7(n)0(i)20.9(s)0(t)21(r)0(a)19.9(t)0(i)14.9(o)0(n)-226.6(\()0(1)21(.)13.9(0)]TJ /F10 1 Tf 24.1225 0 TD ()Tj /F5 1 Tf .7083 0 TD (10)Tj 7.3837 0 0 6.5625 316.1196 383.8109 Tm -.0163 Tc (10)Tj 9.845 0 0 9.845 325.7574 379.4456 Tm 0 Tc [(ge)19.9(no)21.1(me)-228.9(co)19.9(pi)20.9(es)-227.6([G)20.2(C])20.1(/a)19.7(ni)15.1(m)15.1(a)0(l)13.9(\))-237.1(at)-222.2(1)-237.1(m)0(o)19.9(n)0(t)20.9(h)-237.1(po)21(st)15.3(in)20.9(je)19.7(ct)19.7(io)20.9(n.)]TJ /F7 1 Tf 21.6349 0 TD [(\(A)20.2(\))]TJ /F5 1 Tf -48.4236 -1.0077 TD [(Lo)19.9(we)19.1(r)-283.2(a)0(i)19.7(r)0(w)20.2(a)0(y)14.1(s)-283(\(l)20.9(un)21.1(g\))21.1(,)-285.5(s)0(h)21.2(o)0(w)20.2(i)0(n)20.9(g)-283.2(re)19.9(pr)21.1(es)14.3(en)19.9(ta)19.7(ti)20.7(ve)-273.8(im)19.7(ag)19.9(es)-273.6(of)-272.6(th)20.9(e)-284.3(c)0(o)19.9(n)0(d)21(u)0(c)19.9(t)0(i)14.9(n)0(g)-272.6(a)0(i)19.7(r)0(w)20.2(a)0(y)19.9(s)-283(\(t)20.9(ra)19.9(ch)14.1(ea)18.7(,)-279.8(b)0(r)21.1(o)0(n)21.1(c)0(h)14.1(i)0(,)-269.4(a)0(n)19.9(d)-283.2(br)21.1(on)21(ch)19.9(io)15.1(le)19.7(s\))21.2(.)-285.5(O)0(r)20.2(i)0(g)20.9(i)0(n)20.9(a)0(l)]TJ 0 -1.0135 TD [(ma)18.7(gn)21.1(i)21(ca)12.9(ti)20.7(on)21.1(,)]TJ /F10 1 Tf 6.0465 0 TD ()Tj /F5 1 Tf .7083 0 TD [(20)21.1(0.)]TJ /F7 1 Tf 1.9464 0 TD [(\(B)20.1(\))]TJ /F5 1 Tf 1.5433 0 TD [(Up)20.2(pe)19.9(r)-231.3(a)0(i)19.7(r)0(w)20.2(a)0(y)19.9(s)-231.2(\(n)21(os)21.2(e\))19.9(,)-228(s)0(h)15.5(o)0(w)20.2(i)0(n)20.9(g)-231.3(re)19.9(pr)21.1(es)20.1(en)19.9(ta)19.7(ti)14.9(ve)-216.2(im)19.7(ag)19.9(es)-221.8(of)-215(th)15.1(e)-226.8(p)0(r)21(o)0(x)21.1(i)0(m)19.7(a)0(l)-222.2(n)0(o)21.1(s)0(e)-221.8(r)0(e)19.9(g)0(i)20.9(o)0(n)-220.8(\()]TJ /F9 1 Tf 32.5646 0 TD [(le)19.7(ft)]TJ /F5 1 Tf 1.2496 0 TD -.0207 Tc [(\))-252(a)-20.7(nd)-246.3(a)-253.2(p)-20.7(or)-20.7(ti)-20.7(on)]TJ -44.0586 -1.0135 TD 0 Tc [(of)-266.9(th)20.9(e)-278.6(n)0(a)19.9(s)0(a)14.3(l)-271.8(ep)14.1(it)20.7(he)19.9(li)20.7(um)-268(\()]TJ /F9 1 Tf 9.6974 0 TD [(ri)21(gh)15.3(t)]TJ /F5 1 Tf 1.9003 0 TD [(\))-277.4(\()0(o)21(r)0(i)20.9(g)0(i)20.9(n)0(a)19.9(l)-277.6(ma)18.7(gn)21.1(i)15.3(ca)18.7(ti)20.7(on)21.1(:)]TJ /F9 1 Tf 10.2214 0 TD [(le)19.7(ft)]TJ /F5 1 Tf 1.2496 0 TD (,)Tj /F10 1 Tf .6968 0 TD ()Tj /F5 1 Tf .7083 0 TD [(50)21.1(;)]TJ /F9 1 Tf 1.5375 0 TD [(ri)21(gh)21(t)]TJ /F5 1 Tf 1.8946 0 TD (,)Tj /F10 1 Tf .6968 0 TD ()Tj /F5 1 Tf .7083 0 TD [(20)21(0\))21(.)-279.8(T)0(h)19.9(e)-278.6(gr)21(ap)19.9(hs)-266.7(vi)20.9(su)15.5(al)19.7(iz)19.7(e)-278.6(t)0(h)20.9(e)-278.6(av)19.9(er)19.9(ag)19.9(e)-278.6(t)0(r)20.9(a)0(n)19.9(s)0(d)15.5(u)0(c)19.9(t)0(i)20.7(o)0(n)]TJ -29.311 -1.0135 TD [(ef)19.9(c)20.1(ie)13.9(nc)19.9(y)-294.7(i)0(n)-284.3(t)0(h)20.9(e)-295.9(co)19.9(nd)21.1(uc)19.9(ti)20.7(ng)-284.2(ai)13.9(rw)20.2(ay)19.9(s)-294.5(a)0(n)19.9(d)-294.7(na)19.9(sa)20.1(l)-294.9(e)0(p)19.9(i)0(t)14.9(h)0(e)19.9(l)0(i)20.7(u)0(m)19.9(.)-291.3(Fo)15.5(r)-294.7(b)0(o)21.1(t)0(h)-284.3(t)0(h)20.9(e)-295.9(lo)20.9(we)19.1(r)-294.7(a)0(n)19.9(d)-294.7(up)21.1(pe)19.9(r)-294.7(a)0(i)13.9(r)0(w)20.2(a)0(y)19.9(s)0(,)-286.4(t)0(h)20.9(e)-295.9(re)19.9(la)19.7(ti)20.7(ve)-285.3(co)19.9(nt)20.9(ri)15.1(bu)21.1(ti)20.7(on)-284.1(of)]TJ 0 -1.0077 TD [(th)20.9(e)-272.8(d)0(i)20.9(f)0(f)21(e)0(r)19.9(e)0(n)19.9(t)-271.8(re)19.9(gi)20.9(on)21.1(s)-271.5(t)0(o)-255.5(t)0(h)20.9(e)-267.1(to)15.1(ta)19.7(l)-266.1(t)0(r)20.9(a)0(n)19.9(s)0(d)15.5(u)0(c)19.9(t)0(i)20.7(o)0(n)-255.4(e)0(f)19.9()0(c)14.3(i)0(e)19.7(n)0(c)19.9(y)-265.9(is)-261.1(vi)20.9(su)21.2(al)19.7(iz)19.7(ed)-262.3(by)-255.4(th)20.9(e)-272.8(v)0(a)19.9(r)0(i)20.9(o)0(u)21.1(s)-265.7(sh)15.5(ad)19.9(in)20.9(gs)-260.9(de)19.9(sc)20.1(ri)20.9(be)19.9(d)-271.6(i)0(n)-255.5(t)0(h)20.9(e)-267.1(le)19.7(ge)14.1(nd)21.1(s.)-257.6(Mi)20.1(xe)19.9(d-)]TJ 0 -1.0135 TD [(mo)19.9(de)19.9(l)-300.6(A)0(N)13.6(O)0(V)19.4(A)0(,)-293.1(m)0(e)18.7(a)0(n)]TJ /F10 1 Tf 9.1734 0 TD ()Tj /F5 1 Tf .7083 0 TD [(SE)20.1(M,)-293.1(**)]TJ /F9 1 Tf 3.553 0 TD (p)Tj /F10 1 Tf .6565 0 TD (<)Tj /F5 1 Tf .7083 0 TD [(0.)18.7(01)21.1(,)-302.8(*)0(*)21.1(*)]TJ /F9 1 Tf 3.7315 0 TD (p)Tj /F10 1 Tf .6565 0 TD (<)Tj /F5 1 Tf .7083 0 TD [(0.)18.7(00)21.1(1.)-292.3(Sc)20.1(al)19.7(e)-301.6(b)0(a)14.1(r)0(s)21.2(:)-300.6(10)21.1(0)]TJ /F6 1 Tf 9.0352 0 TD (l)Tj /F5 1 Tf .5471 0 TD [(m)-301.6(f)0(o)21(r)-300.4(lu)20.9(ng)-295.7(se)20(ct)19.7(io)20.9(ns)21.2(;)-306.4(2)0(0)21.1(0)-300.4(an)19.9(d)-300.4(1)0(0)15.3(0)]TJ /F6 1 Tf 13.4635 0 TD (l)Tj /F5 1 Tf .5413 0 TD [(m,)-293.5(re)19.9(sp)21.2(ec)18.7(ti)20.7(ve)19.9(ly)15.1(,)]TJ -43.4828 -1.0135 TD [(fo)21(r)-323.5(l)0(o)15.1(w)0(-)-313.8(a)0(n)19.9(d)-323.5(hi)20.9(gh)21.1(-m)19.9(ag)19.9(ni)15.1(c)20.1(at)19.7(io)20.9(n)-323.5(n)0(o)21.1(s)0(e)-313.9(s)0(e)14.3(c)0(t)19.7(i)0(o)20.9(n)0(s)21.2(.)-325.9(Co)20.1(lo)20.9(r)-323.5(i)0(m)19.7(a)0(g)19.9(e)0(s)-313.9(a)0(v)19.9(a)0(i)13.9(l)0(a)19.7(b)0(l)20.9(e)-324.6(on)21(li)20.7(ne)-314.1(at)-314.3(ww)19.4(w.)17.9(li)20.7(eb)14.1(er)19.9(tp)20.9(ub)21.1(.c)17.5(om)19.9(/h)20.9(um)]TJ /F4 1 Tf 8.9663 0 0 8.9663 62.022 749.6502 Tm (520)Tj 46.8338 0 TD [(CARLON)-336.1(ET)-338.7(AL.)]TJ ET endstream endobj 10 0 obj 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[(receiving)-359.2(a)-359.4(single)-359.2(vector)-360.4(dose)-356.7(\(Fig.)-359.1(2D)-360.2(and)-355.8(G\),)-357.8(animals)-362.7(in)]TJ 0 -1.1174 TD [(the)-269.5(fetal)-267.2(group)-264.7(reached)-271.5(comparable)-270.4(transduction)-269.1(efciencies)]TJ 0 -1.1116 TD [(as)-406.3(adult)-404.3(controls)-413.2(for)-400.7(lung)-407.7(and)-407.6(nose,)-406.2(whereas)-409.3(the)-407.8(signal)-405.3(in)]TJ T* [(neonatal)-283.3(animals)-287.8(was)-281.6(signicantly)-286.4(lower)-284.1(\(34%,)]TJ /F9 1 Tf 19.4275 0 TD (p)Tj /F10 1 Tf .6681 0 TD (<)Tj /F5 1 Tf .72 0 TD [(0.01)-277.4(and)]TJ -20.8156 -1.1174 TD (20%,)Tj /F9 1 Tf 2.4191 0 TD (p)Tj /F10 1 Tf .6624 0 TD (<)Tj /F5 1 Tf .72 0 TD [(0.001)-336.1(for)-331.5(lung)-332.8(and)-338.5(nose)-333.7(signal,)-333.8(respectively\).)]TJ -2.7877 -1.1116 TD [(Comparable)-350.9(results)-348.5(were)-353.2(obtained)-351.2(in)-347.9(an)-348.9(independent)-349.9(ex-)]TJ -1.0137 -1.1116 TD 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[(and)-275.2(B\).)-277.3(Whereas)-278.2(a)-278.7(33.3-fold)-271.5(increase)-284.1(in)-273(nose)-276.1(signal)-278.5(was)-275.8(ob-)]TJ T* [(tained)-251.1(for)-245.1(the)-246.5(fetal)-250(group)-247.4(\()]TJ /F9 1 Tf 10.7131 0 TD (p)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(0.24\),)-247.4(only)-246.4(a)-249.9(minimal)-247.9(increase)]TJ -12.0954 -1.1174 TD [(of)-393.8(4.4-fold)-391.3(was)-396.8(measured)-399.2(in)-394(the)-390.5(neonatal)-398.5(group)-391.4(\()]TJ /F9 1 Tf 20.8213 0 TD (p)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD (0.65\))Tj -22.2094 -1.1116 TD [(\(Supplementary)-302.4(Fig.)-300.4(S2D)-297.8(and)-298.2(E\).)-294.8(On)-296.8(X-Gal)-302.3(staining,)-301.6(an)-297.1(in-)]TJ T* [(crease)-385.4(in)]TJ /F6 1 Tf 4.0548 0 TD (b)Tj /F5 1 Tf .5529 0 TD [(-Gal-positive)-392.2(cells)-382.3(was)-385.3(noted)-386.9(in)-382.5(the)-379(conducting)]TJ -4.6078 -1.1116 TD 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[(du)20.8(ce)18.5(d)-312.1(a)0(f)19.7(t)0(e)19.6(r)-312.1(pe)19.7(ri)20.7(na)19.7(ta)19.6(l)-312.2(g)0(e)19.7(n)0(e)-302.9(t)0(r)20.7(a)0(n)19.7(s)0(f)21.1(e)0(r)-302.8(a)0(n)19.7(d)-312.1(af)19.7(te)19.6(r)-306.3(r)0(e)14(a)0(d)19.7(m)0(i)19.5(n)0(i)20.7(s)0(t)20.9(r)0(a)14(t)0(i)20.6(o)0(n)]TJ T* [(by)-215.3(an)19.7(al)13.8(yz)19.7(in)20.7(g)-225.7(n)0(A)20(b)-225.7(ag)19.7(ai)13.8(ns)21(t)-225.9(t)0(h)20.7(e)-226.9(ca)18.5(ps)15.3(id)-215.4(in)-215.4(se)19.9(ru)20.9(m)-226.9(a)0(n)19.7(d)-231.5(BA)18.9(LF)-216.3(\(F)21(ig)20.7(.)]TJ 0 -1.1174 TD [(3\))20.9(.)-326(A)0(s)-308.1(c)0(o)13.9(n)0(t)20.7(r)0(o)20.9(l)0(s)20.9(,)-326(we)-309.5(in)20.7(cl)13.8(ud)20.8(ed)-308.6(no)15.1(nt)20.7(re)19.7(at)19.5(ed)-314.4(an)19.7(im)19.5(al)19.5(s)-323.4(\()0(N)20(e)0(g)19.7(,)]TJ /F9 1 Tf 22.1058 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD (5\))Tj -23.4708 -1.1116 TD [(an)19.7(d)-479.1(a)0(d)19.7(u)0(l)20.7(t)-479.3(co)19.7(nt)15(ro)20.9(l)-479.3(m)0(i)19.5(c)0(e)-471.1(t)0(h)20.7(a)0(t)-470(r)0(e)19.7(c)0(e)18.5(i)0(v)20.7(e)0(d)-469.9(a)-480.3(si)20.9(ng)20.8(le)-470(ve)19.7(ct)19.5(or)-468.7(do)15.1(se)]TJ T* [(\(A)20(du)20.8(lt)-325(ct)19.6(rl)20.7(,)]TJ /F9 1 Tf 4.7806 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(6\))20.9(.)-331.7(F)0(o)21(u)0(r)-324.7(m)0(o)19.6(n)0(t)20.7(h)0(s)-318.8(a)0(f)14(t)0(e)19.6(r)-329.4(pe)19.7(ri)20.7(na)19.7(ta)13.8(l)-329.5(g)0(e)19.7(n)0(e)-320.1(t)0(r)15(a)0(n)19.7(s)0(f)21.1(e)0(r)19.7(,)]TJ -6.1456 -1.1174 TD [(on)20.8(ly)-376.7(a)-382.4(w)0(e)18.8(a)0(k)-377.7(c)0(a)18.5(p)0(s)21(i)0(d)15(-)0(s)21.1(p)0(e)19.7(c)0(i)19.5()0(c)-377.6(h)0(u)20.8(m)0(o)19.6(r)0(a)19.7(l)-387.1(im)19.5(mu)19.6(ne)-372(re)19.7(sp)15.3(on)20.8(se)-377.5(wa)18.8(s)]TJ 0 -1.1116 TD [(ob)20.8(se)14.1(rv)20.9(ed)-233.7(in)-232.7(se)19.9(ru)20.9(m)-244.2(o)0(f)-232.6(t)0(h)20.7(e)-244.2(fe)14(ta)19.6(l)-243.1(g)0(r)20.9(o)0(u)20.8(p)-243(\()]TJ /F9 1 Tf 14.8255 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(8;)-232.7(in)15(hi)20.7(bi)20.7(ti)20.6(on)-232.6(of)-232.6(28)20.8(.8)18.5(,)]TJ 9.3134 12.2451 TD [(12)15.1(.)13.9(8)0(,)-223.4(a)0(n)19.7(d)-231.5(6.)18.5(7%)-222.1(at)-228.1(1:)20.7(40)20.8(,)-233.8(1)0(:)20.7(1)0(6)20.8(0)0(,)-223.4(a)0(n)19.7(d)-231.5(1:)20.7(64)15.1(0)-231.5(s)0(e)19.9(r)0(u)20.9(m)-232.7(di)20.7(lu)15(ti)20.6(on)20.8(s;)-221(Fi)15.1(g.)]TJ 0 -1.1116 TD [(3A)14.2(,)-412.4(l)0(e)19.6(f)0(t)20.7(\))0(,)-407.7(w)0(h)20(e)0(r)19.7(e)0(a)18.5(s)-415.6(in)-405.5(th)20.7(e)-417(n)0(e)19.7(o)0(n)20.8(a)0(t)19.5(a)0(l)-406.7(g)0(r)20.9(o)0(u)20.8(p)-415.8(\()]TJ /F9 1 Tf 17.3828 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD -.0207 Tc [(1)-20.7(0).1(\))-436.5(i)-20.7(nh)-20.7(i)-5.8(b)-20.7(it)-20.7(io)-20.7(n)]TJ -18.7478 -1.1174 TD 0 Tc [(re)14(ac)18.5(he)19.7(d)-271.8(9)0(9)20.8(.)0(8)18.5(,)-274.1(89)20.8(.9)18.5(,)-274.1(a)0(n)19.7(d)-271.8(56)15.1(.)13.9(9)0(%)19.8(,)-274.1(re)19.7(sp)15.3(ec)18.5(ti)20.6(ve)19.7(ly)20.7(.)-274.1(F)0(o)21(r)-277.5(co)19.7(mp)19.6(ar)19.7(is)20.9(on)15.1(,)]TJ 0 -1.1116 TD [(se)14.1(ru)20.9(m)-255.8(o)0(f)-244.1(a)0(d)19.7(u)0(l)20.7(t)-260.4(co)19.7(nt)20.7(ro)20.9(ls)-244(in)15(hi)20.7(bi)20.7(te)19.5(d)-254.5(t)0(r)20.7(a)0(n)13.9(s)0(d)21(u)0(c)19.7(t)0(i)20.6(o)0(n)-244.1(c)0(o)13.9(m)15(pl)14.9(et)19.5(el)19.5(y)-254.5(a)0(t)-245.4(a)]TJ T* [(1:)14.9(64)20.8(0)-323.6(d)0(i)20.7(l)0(u)20.7(t)0(i)20.6(o)0(n)-313.2(1)-323.6(mo)19.6(nt)20.7(h)-323.6(p)0(o)15.1(s)0(t)20.9(i)0(n)20.7(j)0(e)19.6(c)0(t)19.5(i)0(o)14.9(n)-317.9(\(9)15.1(9.)24.3(1%)-314.3(in)15(hi)20.7(bi)20.7(ti)20.6(on)20.8(\).)]TJ 1.0137 -1.1174 TD [(On)20(e)-486.1(m)0(o)19.6(n)0(t)20.7(h)-484.9(af)19.7(te)13.8(r)-484.9(v)0(e)19.7(c)0(t)19.5(o)0(r)-474.5(r)0(e)19.7(a)0(d)19.7(m)0(i)19.5(n)0(i)20.7(s)0(t)15.1(r)0(a)19.7(t)0(i)20.6(o)0(n)20.8(,)-487.2(ho)20.8(we)18.8(ve)19.7(r)-484.9(\()0(F)21(i)0(g)15(.)]TJ -1.0137 -1.1116 TD -.0204 Tc [(3)-20.4(A)-6.2(,)-363.7(r)-20.4(ig)-20.4(ht)-20.4(\))-5.4(,)-363.7(a)-368.2(c)-20.4(om)-20.4(pl)-20.4(et)-20.4(e)-358(i)-20.4(nh)-20.4(ib)-20.4(it)-20.4(io)-20.4(n)-356.7(o)-20.4(f)-356.6(t)-20.4(ra)-20.4(n)-6.5(s)-20.4(du)-20.4(ct)-20.4(io)-20.4(n)-356.7(\()]TJ /F10 1 Tf 20.3318 0 TD 0 Tc ()Tj /F5 1 Tf .7084 0 TD [(97)20.8(%)-347.7(i)0(n)20.7(-)]TJ -21.0402 -1.1116 TD [(hi)14.9(bi)20.7(ti)20.6(on)-330.5(at)-331.8(1:)14.9(64)20.8(0)-340.9(s)0(e)19.9(r)0(u)20.9(m)-342.1(di)15(lu)20.7(ti)20.6(on)20.8(\))-340.9(w)0(a)18.8(s)-346.5(me)18.4(as)19.9(ur)20.9(ed)-331.7(fo)20.9(r)-346.6(b)0(o)20.8(t)0(h)-330.6(t)0(h)20.7(e)]TJ T* [(fe)14(ta)19.6(l)-323.8(g)0(r)20.9(o)0(u)20.8(p)-323.6(\(f)15.1(et)19.5(al)19.6(-R)19.8(;)]TJ /F9 1 Tf 8.3458 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(11)15.1(\))-323.6(a)0(n)19.7(d)-323.6(ne)19.7(on)20.8(at)13.8(al)-314.5(gr)20.9(ou)20.8(p)-323.6(\()0(n)20.9(e)0(o)13.9(n)0(a)19.7(t)0(a)19.6(l)0(-)20.7(R)0(;)]TJ /F9 1 Tf -9.7109 -1.1174 TD (n)Tj /F10 1 Tf .6624 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(12)20.8(\),)-229.1(in)-227(li)20.6(ne)-233.7(wi)19.8(th)-227(ad)19.7(ul)20.7(t)-243.1(c)0(o)19.7(n)0(t)20.7(r)0(o)20.9(l)-237.4(mi)13.7(ce)18.5(.)-233.8(O)0(f)-233.4(n)0(o)20.8(t)0(e)19.6(,)-239.6(fu)20.9(rt)20.7(he)13.9(r)-237.2(s)0(e)19.9(r)0(u)20.9(m)]TJ -1.3708 -1.1116 TD [(di)14.9(lu)20.7(ti)20.6(on)20.8(s)-427.1(a)0(l)19.5(l)0(o)20.7(w)0(e)18.8(d)-427.3(rA)20(AV)19.1(2/)14.9(5-)20.9(fL)19.7(UC)-418.8(tr)20.7(an)19.7(sd)21(uc)13.9(ti)20.6(on)-411.1(to)-417(th)20.7(e)-428.5(s)0(a)19.9(m)0(e)]TJ T* [(ex)13.9(te)19.5(nt)-417(in)-417(al)19.5(l)-427.4(t)0(h)20.7(r)0(e)19.7(e)-428.5(tr)20.7(ea)12.8(tm)19.5(en)19.7(t)-427.4(g)0(r)20.9(o)0(u)20.8(p)0(s)21(,)-429.6(in)20.7(di)20.7(ca)18.5(ti)14.8(ng)-416.9(co)19.7(mp)19.6(ar)19.7(ab)19.7(le)]TJ ET q 1 i 313.625 732.245 m 551.565 732.245 l 551.565 413.745 l 313.625 413.745 l W n q 238.023 0 0 318.501 313.625 413.745 cm /Im3 Do Q Q BT /F7 1 Tf 9.845 0 0 9.845 313.0582 396.9637 Tm [(FIG.)-273.4(3.)]TJ /F5 1 Tf 4.054 0 TD [(Analysis)-194.2(of)-186.3(neutralizing)-194.6(antibody)-188(\(nAb\))-190(against)-193.7(the)]TJ -4.054 -1.0077 TD [(rAAV2/5)-291.7(capsid)-284.7(after)-289.6(perinatal)-289.4(gene)-282.7(transfer)-286.7(and)-286.3(after)-289.6(read-)]TJ 0 -1.0135 TD [(ministration.)-490.5(An)-480.8(rAAV)-484.4(transduction)-486.8(inhibition)-483(assay)-479.2(was)]TJ T* [(performed)-427(to)-416.8(analyze)-420.6(nAb)-418.4(against)-424.1(the)-418.9(rAAV2/5)-424.2(capsid)-417.1(in)]TJ T* [(serum)-179.9(and)-176.9(BALF.)-182.9(Samples)-177.3(were)-179.9(analyzed)-179.8(4)-173.7(months)-179.9(after)-180.2(fetal)]TJ 0 -1.0077 TD [(or)-618.1(neonatal)-622.2(rAAV2/5)-620(delivery,)-618.6(or)-618.1(1)-617.2(month)-616.7(after)-617.9(read-)]TJ 0 -1.0135 TD [(ministration.)-260.2(Serial)-261.8(dilutions)-257.1(of)-255.4(serum)]TJ /F7 1 Tf 15.5193 0 TD (\(A\))Tj /F5 1 Tf 1.6412 0 TD [(or)-261.1(BALF)-254.4(samples)]TJ /F7 1 Tf -17.1605 -1.0135 TD (\(B\))Tj /F5 1 Tf 1.5894 0 TD [(were)-254.7(incubated)-260.4(with)-252.8(rAAV2/5-fLUC)-259.4(and)-251.8(tested)-256.1(for)-256.3(inhi-)]TJ -1.5894 -1.0077 TD [(bition)-328.9(of)-324.5(transduction)-325.5(in)-324.6(HEK293T)-327.6(cells.)-321.8(Transduction)-326.5(was)]TJ 0 -1.0135 TD [(quantied)-374.2(by)-364.8(measuring)-375.2(relative)-369(light)-368.3(units)-367.9(\(RLU\))-370.7(per)-372.7(mi-)]TJ T* [(crogram)-332.9(of)-330.2(protein)-330.9(and)-326.6(is)-330.2(expressed)-328.8(as)-331.2(mean)-329.9(percent)-333.1(trans-)]TJ T* [(duction)-492.2(relative)-495.7(to)-491.6(no-serum)-493.8(or)-491.5(no-BALF)-493.5(control)]TJ /F10 1 Tf 21.2663 0 TD ()Tj /F5 1 Tf .7198 0 TD (SEM.)Tj -21.9861 -1.0077 TD [(The)-178.1(dashed)-180.9(line)-172.5(represents)-180.2(100%)-177.7(transduction)-175.8(as)-175.7(measured)-179.4(by)]TJ 0 -1.0135 TD [(no-serum)-332.6(or)-330.2(no-BALF)-332.2(control.)-328.5(Fetal)-329.9(or)-330.2(neonatal,)-326.1(non-read-)]TJ T* [(ministration)-204.9(controls)-205.1(4)-202.5(months)-202.9(after)-203.3(fetal)-203.4(or)-203.5(neonatal)-201.8(rAAV2/)]TJ T* [(5)-369.5(delivery;)-374.6(Fetal-R)-373.2(or)-370.5(Neonatal-R,)-370.2(fetal)-376.2(or)-364.8(neonatal)-374.6(vector)]TJ 0 -1.0077 TD [(delivery)-546.2(and)-545.4(readministration)-545.2(at)-544.6(3)-542.3(months,)-546(serum/BALF)]TJ 0 -1.0135 TD [(collection)-388.4(1)-381.1(month)-386.3(later;)-383.1(Adult)-386.2(ctrl,)-383.2(adult)-386.5(controls)-383.6(that)-379.8(re-)]TJ T* [(ceived)-245.7(a)-238.3(single)-243.4(vector)-238.8(dose,)-238.7(serum/BALF)-245.4(collection)-244.5(1)-237.1(month)]TJ 0 -1.0077 TD (postinjection.)Tj /F4 1 Tf 8.9663 0 0 8.9663 62.022 749.6502 Tm (522)Tj 46.8338 0 TD [(CARLON)-336.1(ET)-338.7(AL.)]TJ ET endstream endobj 18 0 obj <>stream AdobedC        s!1AQa"q2B#R3b$r%C4Scs5D'6Tdt& EFVU(eufv7GWgw8HXhx)9IYiy*:JZjz?7/52Kĝ:(z}Fqǿ_5yͷ^]\6Ki-xK nFܖCg8֯6&4o(XF߼gԤ.%%٢=p L1k>E $ṄyΝwwmgk5ԫ4犢 vy^Z KЮ]14M86_iq/u]J촽X$Y=1jź*yL?u0#YUz2p6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳqXɑL"mFoj kE# %ٛs;B/ݰO}ߖմǡ<\AjgB35X'TY"xVS9./hVhӴDtbɂ?{;1n9ؘC( )|yZҿ/]XҵOZkӦ%ZKZfKQz)J}ZfѯuKVO6Mk-؇_B"&Yf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf'~A^e;k CQ'g$Q"ʮ:sfwߚM'.4,:fKk~GhB{uN?-4 F4-zGLZ͹s+Qv?+i׍jZȚ7gwoM$ W.FuG̶P YR[rYUj5UR͛6D< _-3oϣ\~dbS(8j 'D|ͭySO5}Z]RTPD!2(ZT6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٰξN^73݆h.-a cb8[˄y_7aobKguNIoXcHª&|NLsf͑-Lw.'@7K-.L&Hf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛6lٳf͛!joJiUc=Ξy6р|1PM+V=uMrO-7}VZۢRo+T2Nyg|n[Tn9w@a灼kwke{aF缉cxHf"ոVkL͛!~Ӽ5P隬[D2LZNʠTwɞ`Ҽ٪7V|YCjv4w 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[(month)-410(after)-411.1(readministration,)-412.1(nAb)-407.4(increased)-417.6(in)-405.5(both)-407.7(treat-)]TJ 0 -1.1116 TD [(ment)-369.8(groups)-369.2(\(fetal-R,)]TJ /F9 1 Tf 9.2098 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(6;)-365.2(neonatal-R,)]TJ /F9 1 Tf 6.1686 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(7\),)-368.5(comparable)]TJ -18.1546 -1.1116 TD [(to)-336.4(that)-334.1(of)-330.5(adult)-335.2(controls)-338.3(\()]TJ /F9 1 Tf 10.3675 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(6\))-336.2(\(Fig.)-336.1(3B,)-329.2(right\).)]TJ -10.7361 -1.1116 TD [(Together,)-465.1(these)-468.5(results)-463.7(indicate)-467.7(that)-460.8(although)-465.3(only)-459.6(low)]TJ -1.0137 -1.1174 TD [(immunoreactivity)-387.1(again)-8(st)-376.5(the)-384.7(rAAV2/5)-384.8(capsid)-383.2(was)-385.3(present)]TJ 0 -1.1116 TD [(after)-382.3(perinatal)-376.6(rAAV2/5)-379(vector)-377.7(delivery)-374.2(either)-377.8(in)-376.7(serum)-376.3(or)]TJ T* [(BALF,)-414(a)-405.4(strong)-409.7(capsid-specic)-412.6(humoral)-413.4(immune)-407.8(response)]TJ 0 -1.1174 TD [(was)-465.9(induced)-465.2(on)-463(readministration)-472(to)-463.1(the)-459.6(same)-468.5(extent)-464.2(as)-463.9(a)]TJ 0 -1.1116 TD [(single)-313.1(administration)-307.4(in)-307.6(adult)-306.4(animals)-310.8(\(compare)-313(fetal-R)-305(and)]TJ T* [(neonatal-R)-315.3(with)-310.8(Adult)-311.9(ctrl;)-312.3(Fig.)-312(3A)-308.3(and)-315.5(B\),)-311.9(arguing)-308.5(against)]TJ 0 -1.1174 TD [(the)-338.7(establishment)-336(of)-336.2(tolerance)-337.3(to)-330.6(the)-338.7(viral)-335.2(vector.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 59.7543 467.603 Tm [(Increased)-336.6(immune)-336(cell)-333(inltration)-333.5(in)-336.5(retransduced)]TJ 0 -1.2203 TD [(lung)-339(after)-334.2(rAAV2/5)-330.7(readministration)]TJ /F5 1 Tf 9.843 0 0 9.843 69.7323 440.2204 Tm [(We)-466.4(then)-460.7(studied)-464(the)-459.6(local)-468.9(immune)-459.6(response)-465.7(in)-463.1(the)-459.6(re-)]TJ -1.0137 -1.1116 TD [(transduced)-377.3(lung)-378.9(in)-370.9(more)-375.4(detail.)-375.6(First,)-377.4(we)-372.9(analyzed)-376.5(relative)]TJ 0 -1.1174 TD [(differences)-414(in)-405.5(B)-405.3(cell)-410.2(and)-407.6(regulatory)-411(T)-405.5(cell)-410.2(\(Treg\))-406.3(popula-)]TJ 0 -1.1116 TD [(tions)-454.8(in)-457.3(BALF)-456.6(\(Supplementary)-457.9(Fig.)-456(S6\).)-455.8(As)-452.1(controls,)-456.9(we)]TJ T* [(included)-587.4(BALF)-583.3(of)-583.9(nontreated)-584.9(animals)-581.5(\()]TJ /F9 1 Tf 17.4519 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(5\))-578.1(and)-586.2(adult)]TJ -18.84 -1.1174 TD [(controls)-240.4(\()]TJ /F9 1 Tf 3.8014 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(6\).)-236(Four)-240.4(months)-232.4(after)-238.3(perinatal)-244.1(treatment,)-237.3(B)-238.3(cell)]TJ -5.1837 -1.1116 TD [(numbers)-482.2(\(B220)]TJ /F10 1 Tf 7.3823 0 0 6.5614 124.2141 378.8219 Tm (+)Tj /F5 1 Tf 9.843 0 0 9.843 129.5433 374.4566 Tm [(\))-479.1(i)0(n)-474.6(the)-476.9(fetal)-474.6(group)-477.8(\()]TJ /F9 1 Tf 9.181 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .7142 0 TD [(4\))-474.5(and)-476.8(neonatal)]TJ -17.6535 -1.1116 TD [(group)-512.3(\()]TJ /F9 1 Tf 3.1794 0 TD (n)Tj /F10 1 Tf .6681 0 TD (=)Tj /F5 1 Tf .72 0 TD [(5\))-509(were)-514.4(not)-510.3(different)-513.5(compared)-514.6(with)-506.6(negative)]TJ -4.5674 -1.1174 TD [(controls)-413.2(\(Supplementary)-411.8(Fig.)-409.9(S6A,)-404.9(left\).)-413.5(Likewise,)-408.4(the)-407.8(in-)]TJ 0 -1.1116 TD [(crease)-316.3(in)-307.6(B)-307.4(cells)-307.4(in)-307.6(readministration)-310.7(animals)-310.8(\(fetal-R,)]TJ /F9 1 Tf 22.1288 0 TD (n)Tj /F10 1 Tf .6624 0 TD (=)Tj /F5 1 Tf .72 0 TD (6;)Tj -23.5111 -1.1116 TD (neonatal-R,)Tj /F9 1 Tf 5.0801 0 TD (n)Tj /F10 1 Tf .6624 0 TD (=)Tj /F5 1 Tf .72 0 TD [(7\))-422.6(was)-419.8(comparable)-425.9(to)-417(adult)-421.6(controls)-424.7(\(Sup-)]TJ -6.4624 -1.1116 TD [(plementary)-511.3(Fig.)-502(S6A,)-502.8(right\).)-499.8(Treg)-502.1(\(CD4)]TJ /F10 1 Tf 7.3823 0 0 6.5614 230.57 324.0566 Tm (+)Tj /F5 1 Tf 9.843 0 0 9.843 235.8991 319.6913 Tm (Foxp3)Tj /F10 1 Tf 7.3823 0 0 6.5614 262.2614 324.0566 Tm (+)Tj /F5 1 Tf 9.843 0 0 9.843 267.5905 319.6913 Tm [(\))-502.2(levels)]TJ -21.1151 -1.1174 TD [(increased)-653.8(in)-653.2(the)-649.7(perinatal)-653.1(treatment)-648.6(groups)-651.5(after)-653(read-)]TJ 0 -1.1116 TD [(ministration)-551.7(\(Suppl)-6.5(ementary)-549.3(Fig.)-548.1(S6B,)-549(right\);)-549.4(they)-552.9(were)]TJ T* [(signicantly)-407.4(higher)-405.3(than)-403.1(in)-405.5(negative)-404.2(controls)-407.4(\()]TJ /F9 1 Tf 19.5888 0 TD (p)Tj /F10 1 Tf .6681 0 TD (<)Tj /F5 1 Tf .7142 0 TD [(0.01)-404.2(for)]TJ -20.9711 -1.1174 TD [(both)-246.4(groups\),)-247(but)-245.3(not)-245.3(different)-248.5(compared)-249.7(with)-241.7(adult)-248.8(controls.)]TJ 0 -1.1116 TD [(This)-431.8(response)-431.2(can)-426.1(be)-429.6(attributed)-428.5(to)-428.5(the)-425(fact)-433(that)-426.3(Tregs)-428.1(ac-)]TJ T* [(cumulate)-480.3(in)-474.6(regions)-475.3(undergoing)-473.1(tissue)-479.9(inammation)-477.9(\(Ka-)]TJ 0 -1.1174 TD (mikozuru)Tj /F9 1 Tf 4.1124 0 TD [(et)-274.2(al.)]TJ /F5 1 Tf 2.0332 0 TD [(,)-268.4(2009\).)-268.1(H&E)-265.8(staini)-8(ngs)-268.1(of)-267.1(lung)-269.5(sections)-270.2(\(Fig.)]TJ -6.1456 -1.1116 TD [(4A\))-401.6(demonstrated)-403.9(inltration)-405.4(of)-393.8(mononuclear)-408.6(cells)-399.6(around)]TJ T* [(conducting)-295.8(airways)-287.3(and)-292.4(blood)-287.9(vessels)-291.8(after)-290.1(readministration)]TJ 0 -1.1174 TD [(\(Fig.)-284.2(4A,)-288.7(middle)-280(and)-286.7(bottom)-284.6(rows\),)-286(a)-284.5(signal)-284.3(that)-282.3(was)-287.4(absent)]TJ 0 -1.1116 TD [(in)-382.5(non-readministered)-388.7(animals)-385.7(\(Fig.)-382.2(4A,)-380.9(top)-377.8(row\).)-383(Scoring)]TJ T* -.0215 Tc [(o)-5.3(f)-535.2(lu)-5.3(ng)-535.2(s)-5.1(e)-.7(c)-6.5(tio)-5.3(ns)-535(f)-5.3(o).4(r)-540.9(i).3(n)-5.2(amma)-6.5(tio)-5.3(n)-535.2(a)-.7(f)-5.3(t).3(er)-540.9(per)-5.3(i).3(n)-5.3(a)-.7(ta)-6.5(l)-535.3(r).5(A)-6.2(AV2)-5.3(/5)]TJ T* -.0178 Tc [(ad)4.1(min)4.1(i)4(st)4(rat)4(i)-1.7(o)4.1(n)-358.7(\()4.2(T)-2.8(ab)4.1(le)-354.1(1\))-358.7(s)4.3(h)4.1(owed)-358.7(mi)4(n)4.1(i)-1.7(mal)-358.8(i)4(n)4.3(l)4(t)-1.7(r)4.2(a)-2.8(t)4(i)4(o)-1.6(n)4.1(,)-355.3(co)4.1(mpa-)]TJ 0 -1.1174 TD -.0215 Tc [(r)-5.3(a)-.7(bl)-5.4(e)-536.4(t).3(o)-541(n).4(eg)-5.3(ati)-5.4(v).4(e)-536.4(a)-6.5(nima)-6.5(ls)-5.1(.)-531.8(R)-6.3(ead)-5.3(min)-5.3(i).3(st)-5.4(ra)-6.5(tio)-5.3(n)-535.2(r).5(e)-6.5(s).6(ul)-5.4(ted)-541(i).3(n)-535.2(i)-5.4(n-)]TJ 0 -1.1116 TD -.0217 Tc [(c)-6.7(r).3(ea)-6.7(sed)-258.9(l).1(ym)-6.7(pho)-5.5(cyt)-5.6(i).1(c)-254.3(i).1(n)-5.5().4(l)-5.6(t).1(ra)-6.7(tio)-5.5(n)-253.2(a)-.9(r)-5.5(oun)-5.5(d)-253.2(t).1(h)-5.5(e)-254.3(con)-5.5(duc)-6.7(tin)-5.5(g)-253.2(a)-.9(i)-5.6(r).3(wa)-6.7(ys)]TJ T* -.0215 Tc [(a)-6.5(n).4(d)-281.8(b).4(lo)-5.3(od)-281.8(ves)-5.1(s).6(e)-6.5(l).3(s,)-284.1(comp)-5.3(ara)-6.5(b).4(le)-282.9(to)-281.8(ad)-5.3(ult)-281.9(c)-.7(o)-5.3(n).4(tr)-5.3(ols)-5.1(.)-278.4(Mo)-5.3(re)-282.9(det)-5.4(a)-.7(il)-5.4(ed)]TJ 0 -1.1174 TD -.0387 Tc [(analysis)-298.8(de)5.1(tected)-299(CD)5.4(4)]TJ /F10 1 Tf 7.3823 0 0 6.5614 142.8094 137.707 Tm 0 Tc (+)Tj /F5 1 Tf 9.843 0 0 9.843 150.6897 133.3417 Tm -.0385 Tc [(as)-298.6(well)-298.9(a)5.3(s)-304.3(C)5.5(D)-.1(8)]TJ /F10 1 Tf 7.3823 0 0 6.5614 208.4598 137.707 Tm 0 Tc (+)Tj /F5 1 Tf 9.843 0 0 9.843 216.3401 133.3417 Tm -.0398 Tc [(T)-301.3(c)-1.7(ells)-299.9(in)-300.1(the)-301.2(l)-.7(ympho-)]TJ -15.9083 -1.1116 TD 0 Tc [(c)38.1(y)39.2(t)39.1(i)39.1(c)-296(inltrates)-336(\(Fig.)-330.3(4B)-337.3(and)-332.8(C,)-333.9(bottom)-336.4(row\).)]TJ 1.0137 -1.1116 TD [(These)-342.9(data)-340.9(demonstrate)-339.4(that)-339.9(despite)-344.2(low)-338.4(immunoreactiv-)]TJ -1.0137 -1.1174 TD [(ity)-303.1(in)-301.8(the)-304.1(transduced)-308.2(lung)-298.3(after)-307.4(perinatal)-307.5(rAAV2/5)-304.2(delivery,)]TJ 0 -1.1116 TD [(a)-359.4(strong)-363.6(local)-359.4(adaptive)-363.9(immune)-361.7(response)-362.1(is)-359.2(induced)-355.7(on)-359.3(re-)]TJ T* [(transduction)-401.6(of)-393.8(the)-396.3(lung,)-393.9(comparable)-397.1(to)-394(a)-399.7(single)-393.7(rAAV2/5)]TJ 0 -1.1174 TD [(administration)-589.6(in)-578.3(adult)-582.9(animals.)-579.2(This)-581.5(indicates)-589.6(that)-576(im-)]TJ 0 -1.1116 TD [(munological)-332.9(ignorance)-330.3(rather)-331.5(than)-328.2(tolerance)-331.6(is)-324.7(at)-326(play.)-331.7(The)]TJ 25.5097 67.9184 TD [(increase)-358.9(in)-353.7(Tregs)-359(after)-359.2(readministration)-362.6(is)-353.5(likely)-359.5(secondary)]TJ 0 -1.1174 TD [(to)-359.4(induced)-367.3(tissue)-359(inammation)-368.5(and)-361.6(does)-362.5(not)-360.5(play)-362.8(a)-365.1(role)-362.8(in)]TJ 0 -1.1116 TD [(the)-315.6(induction)-316.7(and)-309.7(maintenance)-324.7(of)-313.2(immunological)-319.1(tolerance.)]TJ /F11 1 Tf 8.9663 0 0 8.9663 310.8472 681.3352 Tm [(Second)-331.2(vector)-336(readministration)-337.8(does)-328.9(not)-334.8(increase)]TJ 0 -1.2266 TD [(gene)-333.8(expression)-329.5(after)-334.2(perinatal)-337.8(gene)-333.8(transfer)]TJ /F5 1 Tf 9.843 0 0 9.843 320.8251 653.8959 Tm [(In)-336.2(a)-342.1(n)0(e)13.9(x)0(t)-336.4(s)0(t)20.9(e)0(p)19.7(,)-343.3(we)-338.3(ev)13.9(al)19.6(ua)19.7(te)19.6(d)-346.7(a)-347.8(se)19.9(co)19.7(nd)-336.3(re)19.7(ad)19.7(mi)19.5(ni)14.9(st)20.9(ra)19.7(ti)20.6(on)-336.3(of)]TJ -1.0137 -1.1116 TD [(rA)20(AV)19.1(2/)20.7(5-)15.1(fL)19.7(UC)-343.9(\(1)20.9(.5)]TJ /F10 1 Tf 8.3919 0 TD ()Tj /F5 1 Tf .7084 0 TD (10)Tj 7.3823 0 0 6.5614 410.0597 647.3195 Tm -.0162 Tc (10)Tj 9.843 0 0 9.843 420.8314 642.9542 Tm 0 Tc [(GC)18.9(/a)19.5(ni)20.7(ma)18.4(l\))-342.1(6)-352.4(m)0(o)19.6(n)0(t)20.7(h)0(s)-347.6(a)0(f)19.7(t)0(e)19.5(r)-352.4(pe)19.7(ri)20.7(-)]TJ -11.1738 -1.1116 TD [(na)19.7(ta)19.6(l)-433.2(t)0(r)20.7(e)0(a)18.5(t)0(m)19.5(e)0(n)19.7(t)-433.2(or)-422.6(3)-433.1(m)0(o)19.6(n)0(t)20.7(h)0(s)-422.5(a)0(f)14(t)0(e)19.6(r)-433(th)20.7(e)-434.2()0(r)21(s)0(t)-422.6(r)0(e)19.7(a)0(d)19.7(m)0(i)19.5(n)0(i)15(s)0(t)20.9(r)0(a)19.7(t)0(i)20.6(o)0(n)]TJ 0 -1.1174 TD (\()Tj /F9 1 Tf .3225 0 TD (n)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(5)-392.8(f)0(o)15.1(r)-392.7(th)20.7(e)-399.7(f)0(e)19.7(t)0(a)19.6(l)-392.9(an)13.9(d)-392.8(n)0(e)19.7(o)0(n)20.8(a)0(t)19.5(a)0(l)-389.4(g)0(r)20.9(o)0(u)20.8(p)0(\))-388.1(\()0(F)21(i)0(g)20.7(.)-395.1(5A)20(\).)-390.4(Af)20(te)19.6(r)-392.7(a)0(n)]TJ -1.6876 -1.1116 TD [(in)20.7(cr)19.7(ea)18.5(se)-573.4(in)-566.8(BL)18.6(I)-582.8(s)0(i)20.9(g)0(n)20.8(a)0(l)-573.7(m)0(e)18.4(a)0(s)19.9(u)0(r)20.9(e)0(d)-573.6(1)-577.1(mo)19.6(nt)14.9(h)-577.1(a)0(f)19.7(t)0(e)19.5(r)-582.8(pr)20.9(im)19.5(ar)19.7(y)]TJ T* [(re)19.7(ad)19.7(mi)19.5(ni)20.7(st)15.1(ra)19.7(ti)20.6(on)-382.3(\(d)20.9(es)19.9(cr)14(ib)20.7(ed)-377.7(pr)15.1(ev)19.7(io)20.7(us)21(ly)-382.5(in)-382.5(Fi)20.9(g.)-384.7(2,)-378.9(bu)20.8(t)-392.9(s)0(h)21(o)0(w)20(n)]TJ 0 -1.1174 TD [(ag)19.7(ai)19.5(n)-352.4(i)0(n)-347.9(F)0(i)20.9(g)0(.)-344.4(5)-352.4(as)-343(gr)15.1(ay)-343.2(do)20.8(ts)20.9(\),)-344.3(on)20.8(ly)-347.9(a)-353.6(m)0(i)19.5(n)0(o)20.8(r)-352.4(de)19.7(cr)14(ea)18.5(se)-343(in)-342.2(BL)18.6(I)]TJ 0 -1.1116 TD [(si)20.9(gn)20.8(al)-280(wa)13(s)-288.9(n)0(o)20.8(t)0(e)19.6(d)-289.1(fo)20.9(r)-294.8(t)0(h)20.7(e)-290.2(fe)19.7(ta)19.6(l)-289.2(a)0(n)19.7(d)-294.8(ne)19.7(on)20.8(at)19.5(al)-280(gr)20.9(ou)15.1(ps)-278.5(be)19.7(tw)19.8(ee)18.5(n)-289.1(4)]TJ T* [(an)19.7(d)-300.6(6)-300.6(m)0(o)19.6(n)0(t)20.7(h)0(s)-290(\()0(1)15.1(.)13.8(4-)15.1(fo)20.9(ld)20.7(,)]TJ /F9 1 Tf 9.7224 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)24.3(47)-295.9([f)20.9(et)19.5(al)19.5(])-300.6(a)0(n)19.7(d)-300.6(1.)18.5(2-)20.9(fo)15.1(ld)20.7(,)]TJ /F9 1 Tf 10.1198 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)24.3(77)]TJ -22.5723 -1.1116 TD [([n)20.9(eo)19.7(na)19.7(ta)19.6(l])-307.6(fo)20.9(r)-317.8(l)0(u)20.7(n)0(g)-307.5([)0(F)21(i)0(g)20.7(.)-314.4(5B)-308.5(an)19.7(d)-312.1(C)0(])19.8(;)-318(1.)18.5(9-)20.9(fo)20.9(ld)20.7(,)]TJ /F9 1 Tf 18.4887 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(80)-307.5([f)20.9(et)19.5(al)19.5(])]TJ -19.8537 -1.1174 TD [(an)19.7(d)-404.3(3)0(.)24.3(1)0(-)15.1(f)0(o)20.9(l)0(d)20.7(,)]TJ /F9 1 Tf 5.5984 0 TD (p)Tj /F10 1 Tf .6624 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)24.3(50)-393.9([n)20.9(eo)13.9(na)19.7(ta)19.6(l])-394(fo)20.9(r)-404.2(n)0(o)20.8(s)0(e)-394.8([)0(F)21(i)0(g)20.7(.)-406.6(5E)-395.1(an)19.7(d)-398.5(F)0(])15.3(\))0(.)]TJ -6.9692 -1.1116 TD [(Af)20(te)19.6(r)-381.2(t)0(h)15(e)-382.4(se)19.9(co)19.7(nd)-376.6(re)19.7(ad)19.7(mi)19.5(ni)20.7(st)15.1(ra)19.7(ti)20.6(on)-370.8(\(o)15.1(ra)19.7(ng)20.8(e)-382.4(a)0(r)19.7(r)0(o)15.1(w)-382.1(in)-371(Fi)20.9(g.)-373.2(5B)]TJ T* [(an)19.7(d)-381.2(C)-382.3(an)19.7(d)-381.2(F)0(i)20.9(g)0(.)-373.2(5)0(E)-372(a)0(n)19.7(d)-381.2(F\))21(,)-383.6(l)0(u)20.7(n)0(g)-370.8(a)0(n)19.7(d)-381.2(no)20.8(se)-371.8(si)15.1(gn)20.8(al)19.5(s)-381(r)0(e)19.7(m)0(a)18.5(i)0(n)20.7(e)0(d)]TJ 0 -1.1174 TD [(un)20.8(ch)19.7(an)19.7(ge)19.7(d)-461.9(w)0(h)20(e)0(n)-446.9(a)0(n)13.9(a)0(l)19.5(y)0(z)19.7(e)0(d)-446.9(1)-461.9(mo)19.6(nt)20.7(h)-456.1(l)0(a)13.8(t)0(e)19.5(r)-456.1(bo)20.8(th)-451.6(in)-451.6(th)20.7(e)-457.3(f)0(e)19.7(t)0(a)19.6(l)]TJ 0 -1.1116 TD [(gr)20.9(ou)20.8(p)-352.4(\()]TJ /F9 1 Tf 3.1333 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(79)-336.3(fo)20.9(r)-352.4(l)0(u)20.7(n)0(g)20.8(;)]TJ /F9 1 Tf 5.9383 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(79)-336.3(fo)20.9(r)-352.4(n)0(o)20.8(s)0(e)19.9(\))-352.4(\(F)21(ig)20.7(.)-349(5)0(B)-343(a)0(n)19.7(d)-346.7(E\))]TJ -11.8016 -1.1116 TD [(an)19.7(d)-473.4(i)0(n)-468.9(t)0(h)20.7(e)-474.6(ne)19.7(on)20.8(at)19.5(al)-470(gr)20.9(ou)20.8(p)-473.4(\()]TJ /F9 1 Tf 11.9053 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7084 0 TD [(0.)24.3(08)-468.7(fo)20.9(r)-473.4(l)0(u)20.7(n)0(g)15.1(;)]TJ /F9 1 Tf 6.3126 0 TD (p)Tj /F10 1 Tf .6566 0 TD (=)Tj /F5 1 Tf .7085 0 TD [(0.)18.5(13)-463(fo)20.9(r)]TJ -20.9481 -1.1174 TD [(no)20.8(se)19.9(\))-502.2(\()0(F)21(i)0(g)20.7(.)-504.5(5C)-487(an)13.9(d)-496.4(F)0(\))21(.)-504.5(Co)19.8(mp)19.6(ar)19.7(ed)-492.9(wi)19.8(th)-491.9(ad)19.7(ul)20.7(t)-502.3(c)0(o)19.7(n)0(t)20.7(r)0(o)20.9(l)0(s)-491.7(r)0(e)19.7(-)]TJ 0 -1.1116 TD [(ce)18.5(iv)20.7(in)20.7(g)-306.4(a)-301.8(si)15.1(ng)20.8(le)-297.2(ve)19.7(ct)19.5(or)-295.9(do)20.8(se)19.9(,)-302.9(t)0(h)15(e)-301.8(lu)20.7(ng)-295.9(si)20.9(gn)15.1(al)-291.5(af)19.7(te)13.8(r)-300.6(t)0(h)20.7(e)-307.5(se)19.9(co)19.7(nd)]TJ T* [(re)19.7(ad)19.7(mi)19.5(ni)20.7(st)15.1(ra)19.7(ti)20.6(on)-353.5(in)-347.9(th)20.7(e)-365.1(f)0(e)19.7(t)0(a)19.6(l)-364.1(gr)20.9(ou)20.8(p)-358.2(w)0(a)18.8(s)-363.8(on)20.8(ly)-353.7(32)20.8(%)-359.3(\()]TJ /F9 1 Tf 20.9308 0 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(01)20.8(;)]TJ -22.2958 -1.1116 TD [(Fi)20.9(g.)-332.8(5D)20(\))-346.6(a)0(n)19.7(d)-340.9(th)20.7(e)-347.8(n)0(o)20.8(s)0(e)-331.5(s)0(i)15.1(g)0(n)20.8(a)0(l)-331.8(w)0(a)18.8(s)-346.5(on)20.8(ly)-336.4(16)20.8(%)-342(\()]TJ /F9 1 Tf 18.5462 0 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(00)20.8(1;)-336.4(Fi)20.9(g.)]TJ -19.9113 -1.1174 TD [(5G)20(\).)-263.7(Fo)15.2(r)-277.5(t)0(h)20.7(e)-273(ne)13.9(on)20.8(at)19.5(al)-268.4(gr)20.9(ou)20.8(p,)-269.5(31)-267.1(an)19.7(d)-271.8(5)0(%)-268.2(o)0(f)-267.1(t)0(h)20.7(e)-278.7(lu)20.7(ng)-267.1(an)19.7(d)-277.6(n)0(o)20.8(s)0(e)]TJ 0 -1.1116 TD [(si)20.9(gn)20.8(al)19.5(s)-473.2(o)0(f)-457.2(a)0(d)19.7(u)0(l)20.7(t)-473.5(co)19.7(nt)20.7(ro)20.9(l)-473.5(m)15(ic)13.8(e)-468.8(w)0(e)18.8(r)0(e)-458.3(o)0(b)20.8(t)0(a)13.8(i)0(n)20.7(e)0(d)-458.4(\()]TJ /F9 1 Tf 19.3065 0 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(01)-457.2(an)19.7(d)]TJ /F9 1 Tf -20.6716 -1.1116 TD (p)Tj /F10 1 Tf .6566 0 TD (<)Tj /F5 1 Tf .7084 0 TD [(0.)18.5(00)20.8(01)20.8(,)-320.2(r)0(e)14(s)0(p)21(e)0(c)18.5(t)0(i)20.6(v)0(e)19.7(l)0(y)15(;)-323.8(Fi)20.9(g.)-309.8(5D)-314.1(an)19.7(d)-323.6(G)0(\))14.2(.)]TJ -.3513 -1.1174 TD [(The)-478(failure)-476.8(to)-480.4(retransduce)-483.2(the)-476.9(airways)-483.1(efciently)-481.2(by)-480.3(a)]TJ -1.0137 -1.1116 TD [(second)-498.3(readministration)-500.8(\(Fig.)-497.3(5\))-497.5(can)-495.2(be)-498.7(attributed)-497.6(to)-497.7(the)]TJ T* [(presence)-479.8(of)-474.5(capsid-specic)-481.7(nAb,)-479.9(measured)-479.8(both)-476.8(1)-473.4(month)]TJ 0 -1.1174 TD [(\(described)-313.9(previously)-306(in)-307.6(Fig.)-312(3\))-307.4(and)-309.7(3)-306.4(months)-307.3(after)-313.2(primary)]TJ 0 -1.1116 TD [(readministration)-495(\(i.e.,)-492.9(before)-492.7(the)-494.2(second)-492.6(readministration;)]TJ T* [(Supplementary)-451.1(Fig.)-450.2(S7\),)-444.3(supporting)-448.8(the)-453.8(inhibitory)-449.2(role)-449.2(of)]TJ 0 -1.1174 TD [(nAb)-545.6(in)-549.5(preventing)-555(retransduction)-554.7(after)-549.3(the)-551.8(second)-550.2(read-)]TJ 0 -1.1116 TD [(ministration)-338.6(\(Fig.)-330.3(5\).)]TJ 1.0137 -1.1116 TD [(In)-313.2(co)19.7(nc)19.7(lu)20.7(si)15.1(on)20.8(,)-326(o)0(u)20.8(r)-323.6(re)13.9(su)21(lt)20.6(s)-329.2(d)0(e)19.7(m)0(o)19.6(n)0(s)21(t)0(r)20.7(a)0(t)19.5(e)-330.6(lo)20.7(ng)20.8(-t)20.7(er)14(m)-324.9(g)0(e)19.7(n)0(e)-314.4(e)0(x)19.7(-)]TJ -1.0137 -1.1116 TD [(pr)20.9(es)19.9(si)15.1(on)-370.8(up)-376.6(to)-376.7(7)-387(m)0(o)19.6(n)0(t)20.7(h)0(s)-376.4(a)0(f)19.7(t)0(e)19.5(r)-387(in)20.7(it)20.6(ia)13.8(l)-381.4(p)0(e)19.7(r)0(i)15(n)0(a)19.7(t)0(a)19.5(l)-387.1(ge)19.7(ne)-377.7(tr)20.7(an)19.7(sf)21.1(er)]TJ 0 -1.1174 TD [(fo)20.9(ll)20.6(ow)20(ed)-216.5(by)-215.3(a)-226.9(s)0(i)15.1(n)0(g)20.8(l)0(e)-216.6(r)0(e)19.7(a)0(d)19.7(m)0(i)19.5(n)0(i)20.7(s)0(t)15.1(r)0(a)19.7(t)0(i)20.6(o)0(n)-215.3(i)0(n)-215.4(a)0(d)19.7(u)0(l)14.9(t)-225.9(li)20.6(fe)19.7(.)-228.1(A)0(n)20(a)0(l)19.5(y)0(s)21(i)0(s)-215.2(o)0(f)]TJ 0 -1.1116 TD [(ca)18.5(ps)21(id)20.7(-s)15.3(pe)19.7(ci)19.5(c)-371.8(nA)20(b)-375.5(i)0(n)15(d)0(i)20.7(c)0(a)18.5(t)0(e)19.6(d)-381.2(lo)20.7(w)-376.3(i)0(m)19.5(m)0(u)19.6(n)0(o)15.1(r)0(e)19.7(a)0(c)18.5(t)0(i)20.6(v)0(i)20.7(t)0(y)-370.9(a)0(g)19.7(a)0(i)19.5(n)0(s)21(t)]TJ T* [(th)20.7(e)-226.9(r)0(A)20(A)0(V)-217.1(v)0(e)19.7(c)0(t)19.5(o)0(r)-215.3(a)0(f)19.7(t)0(e)19.6(r)-225.7(pe)19.7(ri)15(na)19.7(ta)19.5(l)-225.9(g)0(e)19.7(n)0(e)-216.5(t)0(r)20.7(a)0(n)19.7(s)0(f)21.1(e)0(r)19.7(,)-228.1(bu)20.8(t)-225.9(a)-226.9(si)20.9(gn)20.8(i)20.9(ca)18.5(nt)]TJ 0 -1.1174 TD [(in)20.7(cr)19.7(ea)18.5(se)-498.5(in)-497.7(nA)20(b)-507.9(l)0(e)19.6(v)0(e)19.7(l)0(s)-497.5(a)0(f)19.7(t)0(e)19.5(r)-507.9(re)19.7(ad)19.7(mi)19.5(ni)20.7(st)15.1(ra)19.7(ti)20.6(on)20.8(.)-504.5(I)0(n)-497.5(a)0(d)19.7(d)0(i)20.7(t)0(i)14.8(o)0(n)20.8(,)]TJ 0 -1.1116 TD [(mu)19.6(lt)20.6(ip)20.7(le)-412.4(re)19.7(ad)19.7(mi)19.5(ni)15(st)20.9(ra)19.7(ti)20.6(on)20.8(s)-421.4(w)0(e)18.8(r)0(e)-412.3(n)0(o)20.8(t)-421.7(fe)19.7(as)14.1(ib)20.7(le)19.6(,)-418.1(d)0(e)13.9(m)15(on)15.1(st)20.9(ra)19.7(ti)20.6(ng)]TJ T* [(th)20.7(at)-383.6(im)19.5(mu)19.6(no)20.8(lo)20.7(gi)20.7(ca)18.5(l)-392.9(i)0(g)20.7(n)0(o)15.1(r)0(a)19.7(n)0(c)19.7(e)-393.9(ra)19.7(th)20.7(er)-383.5(th)20.7(an)-383.5(to)20.7(le)19.5(ra)19.7(nc)13.9(e)-388.2(w)0(a)18.8(s)-392.6(in)20.7(-)]TJ 0 -1.1174 TD [(du)20.8(ce)18.5(d)-323.6(a)0(g)19.7(a)0(i)19.5(n)0(s)15.3(t)-318(rA)14.2(AV)19.1(2/)20.7(5)-323.6(c)0(a)18.5(p)0(s)21(i)0(d)-313.4(a)0(f)19.7(t)0(e)13.8(r)-323.6(pe)19.7(ri)20.7(na)19.7(ta)19.5(l)-323.8(g)0(e)19.7(n)0(e)-314.4(t)0(r)20.7(a)0(n)13.9(s)0(f)21.1(e)0(r)19.7(.)]TJ /F4 1 Tf 8.9663 0 0 8.9663 310.8472 204.6047 Tm (Discussion)Tj /F5 1 Tf 9.843 0 0 9.843 320.8251 188.1637 Tm [(Transduction)-218.4(with)-212.9(a)-215.4(nonintegrat)-9(ing)-210.8(rAAV)-220.1(vector)-216.4(results)-221.8(in)]TJ -1.0137 -1.1116 TD [(transient)-369.5(gene)-369.6(expression)-370.2(in)-365.2(actively)-369.8(dividing)-372.1(cells)-370.7(and)-367.3(re-)]TJ 0 -1.1174 TD [(mains)-157.6(a)-152(challenge)-158.8(in)-152.1(many)-156.6(gene)-156.5(therapy)-159.9(studies)-152.7(\(Flageul)]TJ /F9 1 Tf 22.1288 0 TD [(et)-153.2(al.)]TJ /F5 1 Tf 1.9122 0 TD (,)Tj -24.041 -1.1116 TD [(2009;)-178.4(Wang)]TJ /F9 1 Tf 5.034 0 TD [(et)-182(al.)]TJ /F5 1 Tf 1.941 0 TD [(,)-176.2(2012\).)-181.7(At)-181.8(present,)-180.4(many)-179.6(methods)-182.8(are)-178.4(being)]TJ -6.975 -1.1116 TD [(evaluated)-308.6(for)-302.7(rAAV)-306.5(readministration,)-308.4(for)-302.7(example,)-309.7(by)-301.7(sero-)]TJ 0 -1.1174 TD [(type)-328.2(switching)-334.7(\(Weinstein)]TJ /F9 1 Tf 11.0241 0 TD [(et)-331.8(al.)]TJ /F5 1 Tf 2.0908 0 TD [(,)-331.7(2010;)-328.2(Wang)]TJ /F9 1 Tf 5.921 0 TD [(et)-331.8(al.)]TJ /F5 1 Tf 2.0908 0 TD [(,)-331.7(2012\),)]TJ -21.1266 -1.1116 TD [(capsid)-302.6(engineering)-298.1(\(Vandenberghe)]TJ /F9 1 Tf 14.3762 0 TD [(et)-297.2(al.)]TJ /F5 1 Tf 2.0562 0 TD [(,)-302.9(2009\),)-296.9(or)-301.7(immune)]TJ -16.4324 -1.1116 TD [(modulation)-370.9(and)-373.1(tolerance)-371.9(induction)-374.3(to)-370.9(prevent)-373(immune)-373.2(re-)]TJ 0 -1.1174 TD [(sponses)-325.1(to)-324.9(the)-321.4(vecto)-8(r)-323.6(and/or)-324.7(transgene)-324.3(\(Goudy)]TJ /F9 1 Tf 19.3526 0 TD [(et)-320.3(al.)]TJ /F5 1 Tf 2.085 0 TD [(,)-320.2(2011;)]TJ -21.4376 -1.1116 TD (Wang)Tj /F9 1 Tf 2.6495 0 TD [(et)-256.9(al.)]TJ /F5 1 Tf 2.0159 0 TD [(,)-251.1(2011\).)-256.6(Alternatively,)-258.9(the)-252.3(viral)-260.3(vector)-256.7(can)-253.3(be)-256.8(ad-)]TJ -4.6654 -1.1116 TD [(ministered)-250.9(to)-244.2(an)-251(immature)-250(immune)-246.5(system)-250.6(by)-249.9(perinatal)-249.9(gene)]TJ 0 -1.1174 TD [(therapy)-344.2(\(Sabatino)]TJ /F9 1 Tf 7.5452 0 TD [(et)-343.3(al.)]TJ /F5 1 Tf 2.1023 0 TD [(,)-343.3(2007;)-345.5(Sinn)]TJ /F9 1 Tf 5.3968 0 TD [(et)-349.1(al.)]TJ /F5 1 Tf 2.1081 0 TD [(,)-343.3(2008;)-345.5(Hu)]TJ /F9 1 Tf 4.7806 0 TD [(et)-349.1(al.)]TJ /F5 1 Tf 2.108 0 TD (,)Tj -24.041 -1.1116 TD [(2011\).)-567.6(We)-570.1(report)-566.5(here)-571.1(for)-567.7(the)-563.3(rst)-568.6(time)-571.5(the)-569(successful)]TJ /F4 1 Tf 8.9663 0 0 8.9663 59.7543 749.7069 Tm [(LONG-TERM)-340.8(EXPRESSION)-334.8(AFTER)-337.3(PERINATAL)-338.1(GENE)-336.8(TRANSFER)-21465.7(523)]TJ ET endstream endobj 22 0 obj <> endobj 23 0 obj <>/Font<>/ProcSet[/PDF/Text/ImageC]/ExtGState<>>> endobj 24 0 obj <>stream /GS1 gs BT /F5 1 Tf 9.843 0 0 9.843 62.022 199.0488 Tm 0 0 0 1 k 0 Tc 0 Tw [(readministration)-287.7(of)-278.6(rAAV2/5)-281.1(to)-284.5(the)-281.1(upper)-277.3(and)-280.9(lower)-284.1(murine)]TJ 0 -1.1174 TD [(airways)-252.7(after)-255.6(initial)-251.4(perinatal)-255.7(gene)-254.4(transfer.)]TJ 1.0137 -1.1116 TD [(In)-359.3(an)-354.7(ef)19.7(fo)15.1(rt)-353.6(to)-359.4(be)19.7(tt)20.6(er)-360.4(un)20.8(de)19.7(rs)15.3(ta)19.6(nd)-353.5(th)15(e)-365.1(u)0(n)20.8(d)0(e)19.7(r)0(l)15(y)0(i)20.7(n)0(g)-353.5(i)0(m)13.7(m)15(un)15.1(o-)]TJ -1.0137 -1.1116 TD [(lo)20.7(gi)20.7(ca)12.8(l)-266.2(m)0(e)18.4(c)0(h)19.7(a)0(n)19.7(i)0(s)15.1(m)0(,)-259.2(w)0(e)-257.7(d)0(e)13.9(t)0(e)19.6(c)0(t)19.5(e)0(d)-262.5(a)-267.2(si)20.9(gn)15.1(i)20.9(ca)18.5(nt)20.7(ly)-261.5(lo)20.7(we)18.8(r)-271.8(l)0(e)19.6(v)0(e)19.7(l)-271.9(of)]TJ T* [(ca)18.5(ps)21(id)15(-s)21.1(pe)19.7(ci)19.5(c)-354.5(nA)20(b)-358.2(a)0(f)19.7(t)0(e)19.5(r)-358.2(pe)13.9(ri)20.7(na)19.7(ta)19.6(l)-358.3(t)0(r)15(e)0(a)18.5(t)0(m)19.5(e)0(n)19.7(t)-358.3(co)19.7(mp)19.6(ar)19.7(ed)-348.9(wi)14.1(th)]TJ 0 -1.1174 TD 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[(in)15(du)20.8(ct)19.5(io)20.7(n)-254.5(o)0(f)-249.8(t)0(o)20.7(l)0(e)19.6(r)0(a)19.7(n)0(c)19.7(e)-255.7(ag)19.7(ai)13.8(ns)21(t)-254.7(f)0(o)20.9(r)0(e)19.7(i)0(g)15(n)-254.5(pr)20.9(ot)20.7(ei)19.5(ns)21(,)-256.9(s)0(u)21(c)0(h)-251(a)0(s)-245.1(v)0(e)19.7(c)0(t)19.5(o)0(r)]TJ T* [(or)-290.1(tr)20.7(an)13.9(sg)21(en)19.7(ic)-291.5(pr)20.9(ot)20.7(ei)19.5(ns)21(,)-302.9(d)0(e)19.7(s)0(p)21(i)0(t)20.6(e)-301.8(th)20.7(e)-301.8(i)0(m)19.5(m)15(at)13.8(ur)20.9(it)20.6(y)-300.6(o)0(f)-284.4(t)0(h)14.9(e)-296(im)19.5(mu)19.6(ne)]TJ 0 -1.1174 TD [(sy)15.3(st)20.9(em)18.4(.)]TJ 1.0137 -1.1116 TD [(We)-339.7(demonstrated)-340.5(that)-339.9(readministration)-345.3(to)-336.4(the)-338.7(mouse)-341.8(air-)]TJ -1.0137 -1.1116 TD [(ways)-444(was)-442.9(feasible)-444.3(after)-439.9(both)-442.3(fetal)-445.8(and)-436.4(neonatal)-444.5(rAAV2/5)]TJ ET q 1 i 97.002 732.245 m 517.153 732.245 l 517.153 335.905 l 97.002 335.905 l W n q 420.151 0 0 396.34 97.002 335.905 cm /Im4 Do Q Q BT /F7 1 Tf 9.845 0 0 9.845 62.022 319.1243 Tm [(FIG.)-400.1(4.)]TJ /F5 1 Tf 4.1807 0 TD [(Evaluation)-399.8(of)-393.6(lung)-401.5(inammation)-396(after)-404.8(perinatal)-398.8(rAAV2/5)-401.1(administration)-398.1(or)-399.3(readministration)-401.3(in)-399.5(adult)-398.1(life.)]TJ /F7 1 Tf 44.2199 0 TD (\(A\))Tj /F5 1 Tf -48.4006 -1.0135 TD [(Inltration)-312(of)-312.9(immune)-314.8(cells)-312.7(in)-313.1(lung)-309.3(parenchyma)-315.2(was)-310(evaluated)-319.1(by)-307.2(hematoxylineosin)-315.7(\(H&E\))-313.3(staining)-314.6(4)-312(months)-312.3(after)-312.7(fetal)]TJ T* [(and)-320.9(neonatal)-322.7(rAAV2/5)-320.5(administration)-323.3(\(1.0)]TJ /F10 1 Tf 17.4541 0 TD ()Tj /F5 1 Tf .7198 0 TD (10)Tj 7.3837 0 0 6.5625 250.8094 303.5338 Tm (10)Tj 9.845 0 0 9.845 261.3542 299.1685 Tm [(GC)-320.5(of)-318.7(rAAV2/5-)]TJ /F6 1 Tf 6.9621 0 TD (b)Tj /F5 1 Tf .5471 0 TD [(-Gal)-322.9(per)-320.9(animal)-326.5(and)-315.1(1.5)]TJ /F10 1 Tf 9.945 0 TD ()Tj /F5 1 Tf .7198 0 TD (10)Tj 7.3837 0 0 6.5625 450.1416 303.5338 Tm (10)Tj 9.845 0 0 9.845 460.6865 299.1685 Tm [(GC)-320.5(of)-318.7(rAAV2/5-fLUC)]TJ -40.4941 -1.0135 TD [(per)-361.2(animal\))-362.1(and)-355.4(1)-358(month)-363.3(after)-358.7(readministration)-361(at)-360.4(3)-358(months.)-361.7(Samples)-361.5(of)-359(negative)-363.1(and)-355.4(adult)-363.5(control)-359.7(mice)-358.9(that)-356.8(received)-364.1(a)]TJ 0 -1.0077 TD [(single)-456.5(vector)-451.9(dose)-454.1(were)-456.3(analyzed)-456.2(in)-451.3(parallel.)-458.7(Representative)-458.7(images)-452.9(at)-458.3(various)-451.5(time)-455.8(points)-455.3(depict)-452(peribronchial,)-457.9(peri-)]TJ 0 -1.0135 TD [(bronchiolar,)-276.1(and)-274.8(perivascular)-275.9(regions)-273(in)-272.8(which)-272(inltration)-277.7(of)-272.6(immune)-268.7(cells)-278.1(was)-275.4(assessed.)]TJ /F7 1 Tf 36.0658 0 TD (\(B\))Tj /F5 1 Tf 1.6066 0 TD [(Anti-CD4Alexa)-279.3(488)-267.9(staining)]TJ -37.6724 -1.0135 TD [(for)-250.6(identication)-247.4(of)-249.6(CD4)]TJ /F10 1 Tf 7.3837 0 0 6.5625 161.0078 263.6787 Tm (+)Tj /F5 1 Tf 9.845 0 0 9.845 168.7747 259.3133 Tm [(T)-249.8(cells)-249.3(in)-249.8(lymphocytic)-245.3(inltrates.)]TJ /F7 1 Tf 13.5383 0 TD (\(C\))Tj /F5 1 Tf 1.6354 0 TD [(Anti-CD8Alexa)-250.5(555)-250.6(staining)-245.5(for)-250.6(identication)-253.2(of)-243.8(CD8)]TJ /F10 1 Tf 7.3837 0 0 6.5625 538.4125 263.6787 Tm (+)Tj /F5 1 Tf 9.845 0 0 9.845 546.1227 259.3133 Tm (T)Tj -49.1722 -1.0135 TD [(cells)-370.3(in)-364.9(lymphocytic)-371.9(inltrates.)-367.2(4)]TJ /F10 1 Tf 13.6535 0 TD ()Tj /F5 1 Tf .2534 0 TD [(,6-Diamidino-2-phenylindole)-369.6(\(DAPI\))-370.2(was)-367.6(used)-367.7(for)-371.5(nuclear)-367.6(staining.)-369.8(Scale)-365.5(bars:)-368.9(50)]TJ /F6 1 Tf 34.2979 0 TD (l)Tj /F5 1 Tf .5528 0 TD (m,)Tj -48.7576 -1.0077 TD [(except)-280.3(scale)-279.1(bar)-274.8(H&E)-282.5(bottom)-272.4(row:)-281.4(200)]TJ /F6 1 Tf 16.0318 0 TD (l)Tj /F5 1 Tf .5528 0 TD (m.)Tj /F9 1 Tf 1.3072 0 TD (Abbreviations)Tj /F5 1 Tf 5.6261 0 TD [(:)-277.6(readmin,)-278.9(readministration)-280.4(at)-274(3)-277.4(months)-277.8(and)-274.8(tissue)-277.8(collection)-284.8(at)-274(4)]TJ -23.5179 -1.0135 TD [(months;)-336.5(non-readmin,)-334.7(non-readministered)-340.7(controls)-331.7(4)-335(months)-335.3(after)-335.7(perinatal)-335.4(gene)-334.5(transfer.)]TJ /F4 1 Tf 8.9663 0 0 8.9663 62.022 749.6502 Tm (524)Tj 46.8338 0 TD [(CARLON)-336.1(ET)-338.7(AL.)]TJ ET endstream endobj 25 0 obj <>stream Adobed       :k  s!1AQa"q2B#R3b$r%C4Scs5D'6Tdt& EFVU(eufv7GWgw8HXhx)9IYiy*:JZjzm!1AQa"q2#BRbr3$4CS%cs5DT &6E'dtU7()󄔤euFVfvGWgw8HXhx9IYiy*:JZjz?N*f{mmq;PƽY܅U2sb?4?4)\]M܍28f-d< 9@8*~͊n͊6*ثb͊6*ثb͊6*ثb͊6*ثb͊6*ثb͊6*ثbZk$y-R ,ZD$P"1˛dV~bMWYoRYuq#=QzR~Kjިմ 9nY-/&3P5ٱTU])mRɱ]43A%ԧQl XTz6V 0kkAn1 ͊,ثb͊6*ثb͊6*ثb͊6*ثb͊6*:Հ;s?0f_BK4z\) 6^b^MIlU8}/ZGe o MqQKW>˛Yn[ϴZւ^,_Ԛe𲵲4#6*s^za{lHV14T̾bC_,ǙDih2/* 6lUCshsy7>b\㰳͒7Gש͊:IU71"X+ٗ6ҫ[j0:-SyҲԖw+%%ǔ͊ʳqްڳմޱ54N˰ڰ/MrⲵϮ7u+[ݳ(֎lu-ޥɳ5]N-X *@DKUm9T,Էݭ7&kۨ-k» ;Ei\تf^iCk]+ 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[(regulatory/suppressor)-429.2(T)-422.9(cells)-427.9(in)]TJ -11.6556 -1.1284 TD [(human)-333.9(fetuses.)-338.8(Blood)-334.3(105,)-336(47154721.)]TJ -.8844 -1.1223 TD [(Deprest,)-268.1(J.A.,)-263.5(Devlieger,)-266.8(R.,)-261.2(Srisupundit,)-266.6(K.,)]TJ /F9 1 Tf 18.255 0 TD [(et)-260.1(al.)]TJ /F5 1 Tf 2.2811 0 TD [(\(2009\).)-261.8(Fetal)]TJ -19.6517 -1.1284 TD [(surgery)-356.6(is)-346.5(a)-352(clinical)-353(reality.)-357.8(Semin.)-348.5(Fetal)-352.4(Neonatal)-355.8(Med.)-351.6(15,)]TJ 0 -1.1223 TD (5867.)Tj -.8844 -1.1284 TD [(Dismuke,)-578.3(D.J.,)-574.5(Tenenbaum,)-570.5(L.,)-575.5(and)-571.9(Samulski,)-573.1(R.J.)-577.7(\(2013\).)]TJ .8844 -1.1223 TD [(Biosafety)-636.9(of)-624.7(recombinant)-631(adeno-associated)-636.9(virus)-629.8(vectors.)]TJ 0 -1.1284 TD [(Curr.)-339.4(Gene)-333.8(Ther.)-332.3(13,)-335.8(434452.)]TJ -.8844 -1.1284 TD [(Flageul,)-432.1(M.,)-426.5(Aubert,)-423.2(D.,)-428.8(Pichard,)-428.9(V.,)]TJ /F9 1 Tf 15.98 0 TD [(et)-424.8(al.)]TJ /F5 1 Tf 2.6105 0 TD [(\(2009\).)-426.4(Transient)]TJ -17.7061 -1.1223 TD [(expression)-363.6(of)-362.5(genes)-362(delivered)-362.8(to)-359.6(newborn)-362.9(rat)-360.1(liver)-359.9(using)-359.1(re-)]TJ 0 -1.1284 TD [(combinant)-361.9(adeno-associated)-362.4(virus)-355.3(2/8)-353.6(vectors.)-358(J.)-353.1(Gene)-358.2(Med.)]TJ 0 -1.1223 TD [(11,)-335.8(689696.)]TJ -.8844 -1.1284 TD [(Flotte,)-286.9(T.R.,)-287.7(Zeitlin,)-285.5(P.L.,)-285.9(Reynolds,)-283.2(T.C.,)]TJ /F9 1 Tf 17.2486 0 TD [(et)-284.5(al.)]TJ /F5 1 Tf 2.3238 0 TD [(\(2003\).)-286.2(Phase)-285.6(I)]TJ -18.6881 -1.1223 TD [(trial)-389.8(of)-386.9(intranasal)-391.9(and)-388.9(endobronchial)-387.7(administration)-386.7(of)-386.9(a)-388.6(re-)]TJ 0 -1.1284 TD [(combinant)-276.5(adeno-associated)-277(virus)-276(serotype)-272.7(2)-274.6(\(rAAV2\)-CFTR)]TJ 0 -1.1223 TD [(vector)-474.6(in)-469.3(adult)-473.4(cystic)-476.9(brosis)-473.5(patients:)-470.1(A)-471.3(two-part)-474(clinical)]TJ 0 -1.1284 TD [(study.)-334.8(Hum.)-337.1(Gene)-333.8(Ther.)-338.4(14,)-329.7(10791088.)]TJ -.8844 -1.1223 TD [(Fontenot,)-428(J.D.,)-422(Dooley,)-420.9(J.L.,)-422.4(Farr,)-421(A.G.,)-424.5(and)-419.4(Rudensky,)-424.1(A.Y.)]TJ .8844 -1.1284 TD [(\(2005\).)-243.5(Developmental)-243.8(regulation)-242.2(of)-240.5(Foxp3)-235.4(expression)-241.6(during)]TJ 0 -1.1223 TD [(ontogeny.)-334.2(J.)-334.8(Exp.)-337.9(Med.)-333.3(202,)-329.9(901906.)]TJ -.8844 -1.1284 TD [(Gao,)-579.6(G.,)-581.3(Vandenberghe,)-583.7(L.H.,)-577.3(Alvira,)-581.2(M.R.,)]TJ /F9 1 Tf 19.8957 0 TD [(et)-577.2(al.)]TJ /F5 1 Tf 2.9154 0 TD (\(2004\).)Tj -21.9268 -1.1223 TD [(Clades)-403(of)-399.1(adeno-associated)-405.1(viruses)-401.6(are)-401.5(widely)-402(disseminated)]TJ 0 -1.1284 TD [(in)-335.2(human)-333.9(tissues.)-336.9(J.)-334.8(Virol.)-332.9(78,)-335.8(63816388.)]TJ -.8844 -1.1223 TD [(Gao,)-311.3(G.P.,)-303.8(Alvira,)-306.8(M.R.,)-309.5(Wang,)-308.1(L.,)]TJ /F9 1 Tf 14.6504 0 TD [(et)-302.8(al.)]TJ /F5 1 Tf 2.3726 0 TD [(\(2002\).)-304.5(Novel)-304.8(adeno-)]TJ -16.1386 -1.1284 TD [(associated)-277.6(viruses)-267.5(from)-267(rhesus)-274(monkeys)-266.2(as)-272(vectors)-266.5(for)-267.3(human)]TJ 0 -1.1223 TD [(gene)-253(therapy.)-249.1(Proc.)-247.9(Natl.)-255.4(Acad.)-248.5(Sci.)-251(U.S.A.)-250.7(99,)-244.3(1185411859.)]TJ -.8844 -1.1284 TD [(Goudy,)-490.3(K.S.,)-492.9(Annoni,)-489.9(A.,)-489.8(Naldini,)-487(L.,)-490.1(and)-492.6(Roncarolo,)-492.1(M.G.)]TJ .8844 -1.1223 TD [(\(2011\).)-597.2(Manipulating)-598.7(immune)-595.3(tolerance)-598.7(with)-598.7(micro-RNA)]TJ 0 -1.1284 TD [(regulated)-338.4(gene)-338.3(therapy.)-334.5(Front.)-334.8(Microbiol.)-334.8(2,)-329.6(221.)]TJ -.8844 -1.1223 TD [(Grubb,)-492.6(B.R.,)-491.9(and)-492.6(Boucher,)-492.3(R.C.)-491.8(\(1999\).)-493.5(Pathophysiology)-488.5(of)]TJ .8844 -1.1284 TD [(gene-targeted)-300.9(mouse)-296.4(models)-289.9(for)-297.8(cystic)-293.9(brosis.)-290.6(Physiol.)-294.7(Rev.)]TJ 0 -1.1223 TD [(79,)-335.8(S193S214.)]TJ -.8844 -1.1284 TD [(Halbert,)-555.1(C.L.,)-550(Standaert,)-554.8(T.A.,)-552.9(Aitken,)-554.5(M.L.,)]TJ /F9 1 Tf 19.9506 0 TD [(et)-546.8(al.)]TJ /F5 1 Tf 2.8605 0 TD (\(1997\).)Tj -21.9268 -1.1223 TD [(Transduction)-353.9(by)-347.9(adeno-associated)-356.3(virus)-355.3(vectors)-351.8(in)-347.4(the)-351.7(rabbit)]TJ 0 -1.1284 TD [(airway:)-250(Efciency,)-246.2(persistence,)-249.7(and)-248.6(readministration.)-247.2(J.)-243.3(Virol.)]TJ 0 -1.1223 TD [(71,)-335.8(59325941.)]TJ -.8844 -1.1284 TD [(Halbert,)-286.7(C.L.,)-275.5(Rutledge,)-281.7(E.A.,)-278.4(Allen,)-279.9(J.M.,)]TJ /F9 1 Tf 17.4194 0 TD [(et)-284.5(al.)]TJ /F5 1 Tf 2.3177 0 TD [(\(2000\).)-280.1(Repeat)]TJ -18.8527 -1.1223 TD [(transduction)-497.8(in)-493.7(the)-498.1(mouse)-491.6(lung)-494(by)-494.3(using)-493.3(adeno-associated)]TJ 0 -1.1284 TD [(virus)-495.6(vectors)-492.1(with)-488.9(different)-497.4(serotypes.)-491.6(J.)-493.4(Virol.)-491.5(74,)-488.3(1524)]TJ 0 -1.1223 TD (1532.)Tj -.8844 -1.1284 TD [(Harvey,)-342.6(B.G.,)-336.2(Leopold,)-336(P.L.,)-340.8(Hackett,)-337.4(N.R.,)]TJ /F9 1 Tf 18.3709 0 TD [(et)-339.4(al.)]TJ /F5 1 Tf 2.4336 0 TD [(\(1999\).)-341(Air-)]TJ -19.9201 -1.1284 TD [(way)-530.7(epithelial)-529.6(CFTR)-533.4(mRNA)-526.5(expression)-534.4(in)-530.3(cystic)-525.7(brosis)]TJ 27.0135 70.9036 TD [(patients)-385.1(after)-385.3(repetitive)-386.1(administration)-386.7(of)-386.9(a)-382.5(recombinant)-387(ad-)]TJ 0 -1.1223 TD [(enovirus.)-335.7(J.)-328.7(Clin.Invest.)-339.7(104,)-329.9(12451255.)]TJ -.8844 -1.1284 TD [(Hu,)-331.3(C.,)-328.3(Cela,)-330.4(R.G.,)-330.1(Suzuki,)-330.7(M.,)]TJ /F9 1 Tf 13.2597 0 TD [(et)-327.2(al.)]TJ /F5 1 Tf 2.4153 0 TD [(\(2011\).)-334.9(Neonatal)-331.4(helper-)]TJ -14.7906 -1.1223 TD [(dependent)-239(adenoviral)-234.8(vector)-242.8(gene)-234.7(therapy)-236.8(mediates)-243.1(correction)]TJ 0 -1.1283 TD [(of)-411.3(hemophilia)-416.4(A)-416.5(and)-413.3(tolerance)-421.9(to)-414.5(human)-413.2(factor)-415.9(VIII.)-417.8(Proc.)]TJ 0 -1.1223 TD [(Natl.)-334.6(Acad.)-333.9(Sci.)-336.4(U.S.A.)-336.1(108,)-329.9(20822087.)]TJ -.8844 -1.1284 TD [(Kamikozuru,)-415.4(K.,)-416.6(Fukunaga,)-415.1(K.,)-410.5(Hirota,)-416.5(S.,)]TJ /F9 1 Tf 18.2428 0 TD [(et)-412.6(al.)]TJ /F5 1 Tf 2.5861 0 TD [(\(2009\).)-414.2(The)]TJ -19.9445 -1.1223 TD [(expression)-522.2(prole)-522.1(of)-527.1(functional)-522.8(regulatory)-525.7(T)-520.5(cells,)-525.6(CD4)]TJ /F10 1 Tf 6.9712 0 0 6.196 544.9321 656.0502 Tm (+)Tj /F5 1 Tf 9.2951 0 0 9.2951 319.0676 641.4802 Tm (CD25high)Tj /F10 1 Tf 6.9712 0 0 6.196 358.9227 645.6187 Tm (+)Tj /F5 1 Tf 9.2951 0 0 9.2951 363.9684 641.4802 Tm [(/forkhead)-440.7(box)-433.5(protein)-439.4(P3)]TJ /F10 1 Tf 6.9712 0 0 6.196 463.181 645.6187 Tm (+)Tj /F5 1 Tf 9.2951 0 0 9.2951 468.17 641.4802 Tm [(,)-439.2(i)0(n)-432.8(patients)-440(with)-440.1(ul-)]TJ -16.041 -1.1223 TD [(cerative)-244.8(colitis)-246(during)-246.6(active)-244.1(and)-242.5(quiescent)-249.8(disease.)-243.1(Clin.)-248.2(Exp.)]TJ 0 -1.1284 TD [(Immunol.)-331.3(156,)-336(320327.)]TJ -.8844 -1.1223 TD .0119 Tc [(Li,)-518.8(H)4.1(.,)-518.8(M)11.9(u)-5.9(rph)5.7(y)-.4(,)-518.8(S)2.8(.L.,)-512.7(Gil)11.9(e)-4.2(s-Davi)11.9(s,)-518.8(W.,)]TJ /F9 1 Tf 17.6695 0 TD .0112 Tc [(et)-519(al.)]TJ /F5 1 Tf 2.8483 0 TD .0125 Tc [(\(2)6.3(00)6.3(7\).)-518.2(P)3.4(re-)]TJ -19.6334 -1.1284 TD .0105 Tc [(existing)-599.6(A)-3.4(AV)-601.1(capsi)10.5(d)-7.5(-)1.9(specic)-597.7(CD8)]TJ /F10 1 Tf 6.9712 0 0 6.196 462.0471 603.7794 Tm 0 Tc (+)Tj /F5 1 Tf 9.2951 0 0 9.2951 472.7621 599.6407 Tm .607 Tc [(Tc)596.6(e)590.5(l)607(l)589.6(s)-2.2(a)596.6(r)592.3(eu)594.7(n)600.8(a)596.6(b)594.7(l)595.2(e)]TJ -16.535 -1.1284 TD .0099 Tc [(to)-447.7(eli)9.9(m)-7.7(inate)-451.9(A)2.1(AV-t)9.9(r)-10.4(a)-.5(nsduce)-6.6(d)-447.7(h)3.7(epatoc)-6.6(ytes.)-447.6(M)-1.7(ol.)-447.6(T)-4.3(he)-6.6(r.)-447.6(15,)]TJ 0 -1.1223 TD .0123 Tc [(7)6.1(92)6.1(80)6.1(0.)]TJ -.8844 -1.1284 TD 0 Tc [(Limberis,)-385.3(M.,)-383.8(Bell,)-386.6(P.,)-387.4(and)-388.9(Wilson,)-381.4(J.M.)-389.2(\(2007\).)-383.7(Detection)-391.9(of)]TJ .8844 -1.1223 TD [(reporter)-442.9(gene)-442(expression)-442.9(in)-438.9(murine)-439.4(airways.)-445(Methods)-442.2(Mol.)]TJ 0 -1.1284 TD [(Biol.)-333.6(411,)-329.9(2534.)]TJ -.8844 -1.1223 TD [(Limberis,)-543.9(M.P.,)-545.5(and)-541.4(Wilson,)-546.1(J.M.)-541.6(\(2006\).)-542.3(Adeno-associated)]TJ .8844 -1.1284 TD [(virus)-324.8(serotype)-327.6(9)-329.5(vectors)-327.4(transduce)-332(murine)-329.7(alveolar)-326(and)-327.9(nasal)]TJ 0 -1.1223 TD [(epithelia)-426.4(and)-425.5(can)-429.7(be)-425.3(readministered.)-429.2(Proc.)-430.9(Natl.)-426.1(Acad.)-425.4(Sci.)]TJ 0 -1.1284 TD [(U.S.A.)-336.1(103,)-329.9(1299312998.)]TJ -.8844 -1.1223 TD [(Limberis,)-360.9(M.P.,)-362.5(Vandenberghe,)-364.1(L.H.,)-357.7(Zhang,)-366.8(L.,)]TJ /F9 1 Tf 20.3227 0 TD [(et)-363.8(al.)]TJ /F5 1 Tf 2.4824 0 TD (\(2009\).)Tj -21.9206 -1.1284 TD [(Transduction)-494.2(efciencies)-501.6(of)-496.6(novel)-498.4(AAV)-493.1(vectors)-498.2(in)-499.8(mouse)]TJ 0 -1.1223 TD [(airway)-335.9(epithelium)]TJ /F9 1 Tf 7.7033 0 TD [(in)-335.2(vivo)]TJ /F5 1 Tf 3.1289 0 TD [(and)-334(human)-333.9(ciliated)-340.8(airway)-342(epithe-)]TJ -10.8322 -1.1284 TD (lium)Tj /F9 1 Tf 2.1774 0 TD [(in)-329.1(vitro)]TJ /F5 1 Tf 3.0069 0 TD [(.)-335.5(Mol.)-334.7(Ther.)-332.3(17,)-335.8(294301.)]TJ -6.0687 -1.1223 TD [(Lipshutz,)-718.9(G.S.,)-718.6(Flebbe-Rehwaldt,)-724.3(L.,)-715.8(and)-724.3(Gaensler,)-717.7(K.M.)]TJ .8844 -1.1284 TD [(\(2000\).)-420.3(Reexpression)-427.8(following)-424.3(readministration)-430.1(of)-423.4(an)-425.3(ade-)]TJ 0 -1.1223 TD [(noviral)-378.4(vector)-383.1(in)-377.9(adult)-375.8(mice)-380(after)-385.3(initial)]TJ /F9 1 Tf 16.9437 0 TD [(in)-384(utero)]TJ /F5 1 Tf 3.6534 0 TD (adenoviral)Tj -20.5971 -1.1284 TD [(administration.)-338(Mol.)-334.7(Ther.)-332.3(2,)-335.7(374380.)]TJ -.8844 -1.1223 TD [(Liqun)-416.8(Wang,)-417.9(R.,)-419.8(McLaughlin,)-418.6(T.,)-416.9(Cossette,)-420.2(T.,)]TJ /F9 1 Tf 20.2129 0 TD [(et)-418.7(al.)]TJ /F5 1 Tf 2.5922 0 TD (\(2009\).)Tj -21.9206 -1.1284 TD [(Recombinant)-413.9(AAV)-407.7(serotype)-413(and)-413.3(capsid)-410.4(mutant)-406.1(comparison)]TJ 0 -1.1223 TD [(for)-297.8(pulmonary)-293.5(gene)-295.7(transfer)-301.7(of)]TJ /F6 1 Tf 12.7779 0 TD (a)Tj /F5 1 Tf .5001 0 TD [(-1-antitrypsin)-296.7(using)-298.1(invasive)]TJ -13.278 -1.1284 TD [(and)-334(noninvasive)-337.3(delivery.)-334.1(Mol.)-334.7(Ther.)-332.3(17,)-335.8(8187.)]TJ -.8844 -1.1223 TD [(Liston,)-403.3(A.,)-404.4(and)-407.2(Rudensky,)-405.8(A.Y.)-406.1(\(2007\).)-402(Thymic)-408.5(development)]TJ .8844 -1.1284 TD [(and)-303.5(peripheral)-304.2(homeostasis)-310.7(of)-301.5(regulatory)-306.1(T)-300.9(cells.)-306(Curr.)-302.8(Opin.)]TJ 0 -1.1223 TD [(Immunol.)-331.3(19,)-335.8(176185.)]TJ -.8844 -1.1284 TD [(M)12.8(a)0(n)19.9(n)0(o)18(,)-244(C.)19.4(S.)21.3(,)-244(P)0(i)21.8(e)0(r)17.6(c)0(e)15.7(,)-237.9(G.)16.5(F.)21.3(,)-244(A)0(r)20.2(r)0(u)15.7(d)0(a)19.9(,)-237.9(V.)16.5(R.)19.4(,)]TJ /F9 1 Tf 16.2179 0 TD [(et)-229.6(al)18.6(.)]TJ /F5 1 Tf 2.214 0 TD [(\(2)21.8(00)18(6\))21.8(.)-244(S)0(u)21.2(c)0(c)15.7(e)0(s)20.8(s)0(f)16.6(u)0(l)]TJ -17.5475 -1.1223 TD [(transduction)-418.5(of)-423.5(liver)-420.9(in)-414.5(hemophilia)-422.5(by)-421.1(AAV-Factor)-424.1(IX)-418.9(and)]TJ 0 -1.1284 TD [(limitations)-306(imposed)-302.2(by)-305.2(the)-302.9(host)-304.1(immune)-302.5(response.)-309.1(Nat.)-304.6(Med.)]TJ 0 -1.1223 TD [(12,)-329.7(342347.)]TJ -.8844 -1.1284 TD [(Mingozzi,)-318.8(F.,)-320.3(and)-321.8(High,)-318.8(K.A.)-320.8(\(2011\).)-316.6(Therapeutic)]TJ /F9 1 Tf 20.7313 0 TD [(in)-316.9(vivo)]TJ /F5 1 Tf 3.0923 0 TD (gene)Tj -22.9392 -1.1223 TD [(transfer)-417.6(for)-413.7(genetic)-415(disease)-419.9(using)-414(AAV:)-413.4(Progress)-413.7(and)-419.4(chal-)]TJ 0 -1.1284 TD [(lenges.)-337.3(Nat.)-335.1(Rev.)-332.8(Genet.)-333.5(12,)-335.8(341355.)]TJ -.8844 -1.1223 TD [(Mingozzi,)-251.7(F.,)-253.3(Maus,)-253.3(M.V.,)-251.4(Hui,)-251.6(D.J.,)]TJ /F9 1 Tf 15.0834 0 TD [(et)-254(al.)]TJ /F5 1 Tf 2.2628 0 TD [(\(2007\).)-249.6(CD8)]TJ /F10 1 Tf 6.9712 0 0 6.196 520.214 269.0078 Tm (+)Tj /F5 1 Tf 9.2951 0 0 9.2951 527.5841 264.9259 Tm (T-cell)Tj -22.433 -1.1284 TD [(responses)-418(to)-414.5(adeno-associated)-423.4(virus)-416.3(capsid)-422.6(in)-414.5(humans.)-418.6(Nat.)]TJ T* [(Med.)-333.3(13,)-335.8(419422.)]TJ -.8844 -1.1223 TD [(Moss,)-278.7(R.B.,)-278.4(Rodman,)-277.9(D.,)-276.3(Spencer,)-281.2(L.T.,)]TJ /F9 1 Tf 16.474 0 TD [(et)-278.4(al.)]TJ /F5 1 Tf 2.3177 0 TD [(\(2004\).)-274(Repeated)]TJ -17.9074 -1.1284 TD [(adeno-associated)-569.8(virus)-568.8(serotype)-571.6(2)-561.3(aerosol-mediated)-573.9(cystic)]TJ 0 -1.1223 TD [(brosis)-266.2(transmembrane)-271.1(regulator)-269.4(gene)-265.2(transfer)-265.1(to)-268.1(the)-266.3(lungs)-267.6(of)]TJ 0 -1.1284 TD [(patients)-574.2(with)-574.3(cystic)-580.6(brosis:)-576.8(A)-575(multicenter,)-575(double-blind,)]TJ 0 -1.1223 TD [(placebo-controlled)-337.1(trial.)-335(Chest)-337.6(125,)-329.9(509521.)]TJ -.8844 -1.1284 TD [(Moss,)-486.1(R.B.,)-485.8(Milla,)-484.4(C.,)-486.9(Colombo,)-486.7(J.,)]TJ /F9 1 Tf 15.8519 0 TD [(et)-485.8(al.)]TJ /F5 1 Tf 2.7325 0 TD [(\(2007\).)-481.3(Repeated)]TJ -17.7 -1.1223 TD [(aerosolized)-457.9(AAV-CFTR)-461.7(for)-456.4(treatment)-459.3(of)-460(cystic)-458.6(brosis:)-454.8(A)]TJ 0 -1.1284 TD [(randomized)-485(placebo-controlled)-483.4(phase)-484(2B)-480.8(trial.)-481.3(Hum.)-483.5(Gene)]TJ 0 -1.1223 TD [(Ther.)-332.3(18,)-335.8(726732.)]TJ -.8844 -1.1284 TD [(Mueller,)-235.6(C.,)-242.9(and)-236.4(Flotte,)-238.1(T.R.)-238.9(\(2008\).)-237.4(Clinical)-237.7(gene)-240.8(therapy)-236.8(using)]TJ .8844 -1.1223 TD [(recombinant)-387(adeno-associated)-392.9(virus)-385.8(vectors.)-388.5(Gene)-382.6(Ther.)-387.2(15,)]TJ 0 -1.1284 TD (858863.)Tj -.8844 -1.1223 TD [(Nathwani,)-670.4(A.C.,)-671.7(Tuddenham,)-670(E.G.,)-668.8(Rangarajan,)-677.5(S.,)]TJ /F9 1 Tf 23.293 0 TD [(et)-668.7(al.)]TJ /F5 1 Tf -22.4086 -1.1284 TD [(\(2011\).)-591.1(Adenovirus-associated)-599.3(virus)-593.2(vector-mediated)-599(gene)]TJ 0 -1.1223 TD [(transfer)-338.3(in)-329.1(hemophilia)-337.1(B.)-334.4(N.)-331.2(Engl.)-337.4(J.)-334.8(Med.)-333.3(365,)-329.9(23572365.)]TJ -.8844 -1.1284 TD [(Parsons,)-319.8(D.W.,)-317.5(Grubb,)-315.8(B.R.,)-321.1(Johnson,)-316.5(L.G.,)-321.1(and)-315.7(Boucher,)-321.6(R.C.)]TJ .8844 -1.1223 TD [(\(1998\).)-389.8(Enhanced)]TJ /F9 1 Tf 7.6545 0 TD [(in)-396.2(vivo)]TJ /F5 1 Tf 3.2448 0 TD [(airway)-390.8(gene)-393.2(transfer)-399.3(via)-388.3(transient)]TJ /F4 1 Tf 8.9663 0 0 8.9663 59.7543 749.7069 Tm [(LONG-TERM)-340.8(EXPRESSION)-334.8(AFTER)-337.3(PERINATAL)-338.1(GENE)-336.8(TRANSFER)-21465.7(527)]TJ ET endstream endobj 36 0 obj <> endobj 37 0 obj <>/ProcSet[/PDF/Text]/ExtGState<>>> endobj 38 0 obj <>stream /GS1 gs BT /F5 1 Tf 9.2951 0 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