Multi-component Prime-boost Chlamydia Trachomatis Vaccination Regimes Induce Antibody Thiab T Cell Responses Thiab Accelerate Clearance Of Infection in A Non-Human Primate Model

Nws yog qhov tseem ceeb thoob ntiaj teb los tsim cov tshuaj tiv thaiv kev sib deevCov kab mob chlamydia trachomatistxhawm rau tiv thaiv kev kis tus kab mob thoob ntiaj teb txuas ntxiv mus. Covkev txhaj tshuaj zoo tshaj plawslub tswv yim tseem tseem yuav tau piav qhia tag nrho. Lub hom phiaj ntawm txoj kev tshawb no yog los ntsuascov tswv yim txhaj tshuajnyob rau hauv ib tug nonhuman primate (NHP) qauv. Cynomolgus macaques (Macaqua fascicularis) tau txhaj tshuaj tom qab muaj ntau yam sib txawv.prime-boost kev txhaj tshuajteem caij thiab tom qab sib tw nrog C. trachomatis SvD nyob rau hauv qhov chaw mos. Cov tshuaj tiv thaiv antigens suav nrog cov recombinant protein antigen CTH522 adjuvanted nrog CAF01 lossis aluminium hydroxide, MOMP DNA antigen, thiab MOMP vector antigens (HuAd5 MOMP thiab MVA MOMP).

Nyem qhov no kom tau txais Herbal Cistanche Kev Tiv ThaivCov kab mob chlamydia trachomatis

Tag nrho cov antigen tsim yog heev immunogenic raising systemic C. trachomatis tshwj xeeb IgG teb. Tshwj xeeb, CTH522 cov tshuaj tiv thaiv kab mob protein tau tsa ib qho sai thiab muaj zog pecificsIgG hauv cov ntshav. Daim duab qhia ntawm B cell epitopes tshwj xeeb hauv MOMP tau pom tias txhua pab pawg txhaj tshuaj, lees paub cov epitopes nyob ze lossis nyob rau hauv qhov sib txawv (VD) ntawm MOMP, nrog cov lus teb VD4 zoo ib yam hauv txhua tus tsiaj. Tsis tas li ntawd, ntshav qab zib los ntawm txhua pab pawg txhaj tshuaj tuaj yeem ua rau hauv vitro neutralize SvD, SvE, thiab SvF. Cov lus teb ntawm cov tshuaj tiv thaiv kab mob tau cuam tshuam rau ntawm qhov chaw mos thiab lub qhov muag mucosa, uas pom tau tias muaj qib IgG. Cov tshuaj tiv thaiv kuj tau ua rau C. trachomatis-cov lus teb tshwj xeeb ntawm cov xov tooj ntawm tes, raws li qhia hauv vitro stimulation thiab intracellular cytokine staining ntawm peripheral ntshav mononuclear hlwb (PBMCs). Feem ntau, cov tshuaj tiv thaiv kab mob protein (CTH522) tau tsim muaj ntau txoj haujlwm CD4 T cell teb, thaum DNA thiab Vector cov tshuaj tiv thaiv pawg kuj tsim CD8 T cell teb. Tom qab qhov kev sib tw ntawm qhov chaw mos nrog C. trachomatis SvD, ob peb pawg tshuaj tiv thaiv tau pom.ceev tshem tawm ntawm tus kab mob, tab sis tshwj xeeb tshaj yog pab pawg DNA, txhawb nqa nrog CAF01 adjuvanted CTH522 kom ua tiavsib npaug CD4/CD8T cell teb ua ke nrog ib qhoIgG teb, ua kom pomceev tshem tawm ntawm tus kab mob.

KEYWORDS tshuaj tiv thaiv kab mob,kev tiv thaiv kab mobCov kab mob, CD4 / CD8 lymphocytes,Chlamydia trachomatis

Taw qhia

Qhov chaw mos Chlamydia trachomatis kab mob txhua xyoo ua rau kwv yees li 131 lab tus neeg mob tshiab thoob ntiaj teb (1) thiab ua rau muaj teeb meem kev noj qab haus huv thoob ntiaj teb los ntawm kev muaj peev xwm ua raumob hnyavxws li lub plabkab mob inflammatory, ectopic cev xeeb tub, thiabtsis muaj menyuam(2, 3). Cov tshuaj tua kab mob tseem muab kev kho mob zoo ntawm C. trachomatis kab mob. Txawm li cas los xij, txij li tus kab mob yog asymptomatic txog li 75 feem pua ​​​​ntawm cov neeg mob, tusFeem ntau ntawm cov kab mob no tseem tsis tau kho, thiab kis tau tus mob tsis cuam tshuam (3). Nws feem ntau pom zoo tias cov cuab yeej siv tau zoo tshaj plaws tiv thaiv kev kis tus kab mob Chlamydia txuas ntxiv yog tshuaj tiv thaiv prophylactic (4).

Txawm hais tias ntau xyoo ntawm kev tshawb fawb, tshuaj tiv thaiv tseem tsis tau muaj nyob hauv khw. Qhov no yog ib feem ntawm txoj kev ua neej nyob hauv biphasic ntawm C. trachomatis. Qhov tshwj xeeb biphasic lub neej voj voog ntawm C. trachomatis, nrog rau ob qho tib si hauv lub cev thiab cov txheej txheem ntxiv, ua rau lub cev tsis muaj zog. Nws ntseeg tau tias kev tiv thaiv kev tiv thaiv yuav tsum muaj cov lus teb ntawm tes-mediated immune (CMI), ntxiv rau serotype tshwj xeeb cov tshuaj tiv thaiv (5-13). Tsis tas li ntawd, kev tsim cov tshuaj tiv thaiv kab mob hauv qhov chaw mos ua rau muaj kev sib tw ntxiv rau cov tshuaj tiv thaiv.

Hauv kev tshawb fawb tam sim no, peb tau sim cov tswv yim tshuaj tiv thaiv sib txawv hauv tus qauv NHP chlamydia kab mob. Cov tswv yim tsom rau induce antibodies, CD4 T cells, thiab/los yog CD8 T cell teb. Tsis tas li ntawd, qee cov tshuaj tiv thaiv tsom mus tsim cov lus teb mucosal. Cov antigens uas tau xaiv rau txoj kev tshawb no yog nyob ntawm Major Outer Membrane Protein. Ua ntej, qhov kev pom zoo MOMP Antigen (Con E) raws li kwv yees li 1500 serovar E ompA sequences tau tsim los muab kev pabcuam siab epitope tiv thaiv cov kab mob C. trachomatis feem ntau (14). Thib ob, cov khoom siv hluavtaws fusion, CTH522, tau siv. CTH522 yog ib qho recombinant, engineered version ntawm MOMP, suav nrog heterologous immunorepeats ntawm VD4 cheeb tsam los ntawm plaub qhov chaw mos Ct serovars (D, E, F, thiab G) (11). Lub tswv yim tom qab cov recombinant fusion protein yog ua kom cov lus teb rau cov epitopes tiv thaiv thiab induce cross-serovar tiv thaiv (11).

Lwm qhov tseem ceeb ntawm qhov kev sim tam sim no yog qhov kev ntsuas sib npaug ntawm CTH522 hauv ob qhov sib txawv adjuvant systems (AlOH thiab CAF01), ua ke nrog kev txhawb nqa intranasal. Qhov kev sib xyaw ua ke no tau raug sim hauv cov nas thiab cov npua me uas nws tau pom tias muaj kev tiv thaiv kab mob siab heev los ntawm kev ua kom muaj cov tshuaj tiv thaiv kab mob siab, IFN-g-tsim CD4 ntxiv rau T hlwb, thiab nrawm nrawm ntawm qhov chaw mos C. trachomatis kab mob (11-13). , 15). Tsis tas li ntawd, qhov kev sib piv no tau tsom mus rau qhov kev sim tshuaj ntsuam xyuas thawj zaug hauv tib neeg tsis ntev los no yog ob qho kev tsim ua pov thawj tias yog cov tshuaj tiv thaiv kab mob, thiab qhov twg CAF01 yog qhov zoo tshaj plaws hauv kev txhawb nqa cov tshuaj tiv thaiv kab mob hauv nruab nrab thiab Th1 cell-mediated immune teb (16). Txhawm rau ua kom muaj zog tiv thaiv kab mob ntawm tes, thiab tshwj xeeb CD8 T cell teb ua ke nrog CD4 T cell teb, ib feem ntawm cov tsiaj tau txhawb nqa nrog CT522/CAF01 immunogen tom qab priming nrog Adeno vector pAL1112 (HuAd5-MOMP), ib MVA pox vector, lossis plasmid DNA pcDNA3.1 xa cov MOMP Con E antigen (14, 17). Tag nrho peb cov vectors tau ua pov thawj cov tshuaj tiv thaiv kab mob hauv NHPs, thiab nws tsis ntev los no tau pom tias qhov kev txhaj tshuaj tiv thaiv kab mob sib txuas nrog peb cov tshuaj tiv thaiv thiab kev txhaj tshuaj tiv thaiv zaum kawg nrog cov tshuaj tiv thaiv kab mob ua kom muaj zog tiv thaiv kab mob zoo tshaj plaws thiab tsis tu ncua kev tshem tawm ntawm intravaginal C. trachomatis hauv nas C. trachomatis kab mob qauv (14).

Hauv txoj kev tshawb fawb tam sim no, peb pom tau tias sib txawv ntawm kev txhaj tshuaj tiv thaiv kab mob sib txawv nrog rau cov tshuaj tiv thaiv kab mob protein, tshuaj tiv thaiv DNA, thiab cov tshuaj tiv thaiv kab mob viral vector nyob rau hauv tus qauv nonhuman primate (NHP), ua rau cov tshuaj tiv thaiv kab mob sib txawv. Tsis tas li ntawd, peb pom tias kev tswj hwm siv DNA priming thiab protein (CTH522 / CAF01) txhawb nqa cov kab mob sai sai tom qab muaj kev sib tw ntawm qhov chaw mos nrog Chlamydia trachomatis.

Cov txiaj ntsig

Txhua txoj kev txhaj tshuaj tiv thaiv tau ua pov thawj tias muaj kev nyab xeeb thiab ua rau muaj cov tshuaj tiv thaiv kab mob tseem ceeb

Tag nrho ntawm 30 cynomolgus macaques tau txhaj tshuaj tiv thaiv tom qab sib txawv ntawm cov tshuaj tiv thaiv thawj zaug nrog rau cov protein, DNA, thiab cov tshuaj tiv thaiv vector thiab cov kev tswj hwm sib txawv, raws li pom hauv daim duab 1 thiab cov duab ntxiv 6-10). Txhua tus tsiaj tau noj qab nyob zoo thaum kawm thiab tsis muaj txiaj ntsig zoo rau kev txhaj tshuaj. Tsis tas li ntawd, peb pom tsis muaj qhov hloov pauv ntawm qhov hnyav lossis qhov kub ntawm cov tsiaj (Cov duab ntxiv 1, 2).

Ntawm lub sijhawm teem sijhawm tom qab txhaj tshuaj, cov ntshav tau sau thiab qib ntawm CTH522 tshwj xeeb IgG hauv cov ntshav tau ntsuas los ntawm ELISA. Tag nrho 5 pab pawg tau txhaj tshuaj tau tsim cov tshuaj tiv thaiv tseem ceeb CTH522 IgG cov lus teb (Daim duab 2). Txawm li cas los xij, kinetics txawv me ntsis hauv 5 pawg tshuaj tiv thaiv. Txhua tus tsiaj hauv CTH522/AlOH ntxiv rau IN, CTH522/CAF01, thiab CTH522/CAF01 ntxiv rau IN pawg tau tsa ib qho tseem ceeb IgG titer tom qab tsuas yog 1 txhaj tshuaj intramuscular (IM), thiab IgG titer tseem tseem ceeb rau qhov kawg sampling point (-Figures 2. C). Cov qib ntshav IgG tau nce qeeb me ntsis hauv DNA ntxiv rau CTH522 thiab vectors ntxiv rau CTH522 pawg, qhov twg txhua tus tsiaj tau hloov pauv ntawm lub lim tiam 8 thiab lub lim tiam 4, raws li (Daim duab 2D, E).

CTH522/AlOH ntxiv rau IN, CTH522/CAF01 ntxiv rau IN, thiab CTH522/CAF01 pawg tau nce cov ntshav siab IgG ntau ntxiv tom qab 3rd IM tshuaj tiv thaiv hauv lub lis piam 16 (Figures 2A C). Cov DNA ntxiv rau CTH522 thiab vectors ntxiv rau CTH522 pawg tau nce cov ntshav hauv cov ntshav IgG qhov tseem ceeb tom qab cov protein / CTH522 / CAF01 IM booster hauv lub lis piam 20 (Figures 2D, E). Tsis muaj qhov sib txawv tseem ceeb hauv cov tshuaj tiv thaiv kab mob ntshav qab zib tom qab peb zaug txhaj tshuaj IM nrog CTH522 tsim nrog CAF01 lossis AlOH adjuvant (Figures 2A, B), thiab intranasal (IN) uas tsis yog-adjuvanted koob tshuaj tiv thaiv hauv CTH522 / AlOH ntxiv rau IN thiab CTH522/CAF01 ntxiv rau IN pab pawg tsis tau ua kom muaj kev nce ntxiv hauv cov ntshav IgG antibody titers (Figures 2A, B).

Lub kinetics ntawm cov ntshav IgG cov lus teb tau cuam tshuam txog kinetics / qib hauv cov kua hauv qhov chaw mos, qhia tias cov tshuaj tiv thaiv tau kuaj pom ntawm qhov chaw mos ntawm qhov chaw mos yuav tau muab los ntawm kev ncig. Ib yam li ntawd, cov qib ocular IgG kuj ua raws li cov ntshav IgG qib thiab, raws li tau pom hauv cov ntshav qab zib, qhia tias IM booster hauv DNA / vector pawg tau nce qhov ocular IgG titer (Cov duab ntxiv 3).

Cov tshuaj tiv thaiv kab mob hauv cov ntshav yog qhia tawm tsam qhov sib txawv ntawm 1, 3, thiab 4 hauv MOMP ib ntus

Ntxiv elucidation ntawm qhov tshwj xeeb epitope lees paub los ntawm cov tshuaj tiv thaiv kab mob hauv cov ntshav, tom qab cov tshuaj tiv thaiv sib txawv, tau raug soj ntsuam ntxiv. Cov ntshav tau tshuaj xyuas nyob rau lub lim tiam 24, piv txwv li tom qab txhaj tshuaj tiv thaiv kab mob intranasal hauv pawg 1 thiab 2, thiab tom qab txhaj tshuaj tiv thaiv IM booster hauv pawg DNA thiab vector (pab pawg 4 thiab 5). Cov kev soj ntsuam no tau ua nrog peptide array npog CTH522, suav nrog 15 mer peptides nrog 14 amino acid sib tshooj.

FIGURE 1 Kawm tsim qauv. Daim duab qhia cov pab pawg, cov tshuaj tiv thaiv thiab cov kev tswj hwm (intramuscular, IM, lossis intranasal, IN) siv

Daim duab 2 (A–F) Serum CTH522 IgG cov lus teb. Cynomolgus macaques tau txhaj tshuaj raws li cov txheej txheem tseem ceeb sib txawv (n=5 ib pab pawg) thiab cov ntshav tau sau txhua txhua lub lim tiam thib ob. Qhov tshwj xeeb CTH522 IgG cov tshuaj tiv thaiv hauv cov ntshav tau soj ntsuam los ntawm ELISA thiab tau hais tawm ntawm no raws li titer ntawm lub log10 nplai. Grey arrowheads qhia txog kev txhaj tshuaj. Txhua kab sawv cev rau ib tug tsiaj. Kev txheeb xyuas qhov tseem ceeb yog qhia nrog asterisks *p < 0.05 thiab **p < 0.01.

Feem ntau, cov peptides lees paub tau nyob ze thiab nyob rau hauv cov chaw sib txawv (VDs 1,3,4) ntawm MOMP. Pawg CTH522/AlOH txawm pom qee qhov kev lees paub ntawm cheeb tsam VD2. Txhua tus tsiaj tau tsim cov lus teb muaj zog tiv thaiv thaj tsam VD4 uas muaj cov kev tiv thaiv nruab nrab ntawm VD4 epitope (11). Cov pab pawg DNA tau pom qhov nqaim tshaj plaws nyob rau hauv VDs (Daim duab 3), thaum pab pawg vector pom cov qauv kev paub dav me ntsis. CTH522/CAF01 thiab CTH522/AlOH pawg tau pom qhov dav tshaj plaws, tshwj xeeb tshaj yog tawm tsam SvF thiab SvG.

Cov tshuaj tiv thaiv kab mob ntshav qab zib tau tuaj yeem ua rau SvD, SvE, SvF hauv vitro

Txhawm rau ntsuas lub peev xwm ntawm cov tshuaj tiv thaiv kab mob uas ua rau inhibit qhov kis kab mob los ntawm C. trachomatis ntawm nws lub hom phiaj hlwb, ib qho kev ntsuam xyuas hauv vitro neutralization tau ua. Siv qhov kev ntsuas no, peb tau tshuaj xyuas qhov nruab nrab ntawm SvD, SvE, thiab SvF. Serum los ntawm lub lim tiam 24 tau siv rau qhov kev ntsuam xyuas, thiab cov ntshav qab zib los ntawm pab pawg neeg tsis zoo yog siv los tswj. Tag nrho tsib pawg tshuaj tiv thaiv tau pom tias muaj peev xwm ua kom tsis zoo rau tag nrho peb serovars (Daim duab 4). Tawm tsam SvD lub CTH522 / AlOH ntxiv rau IN pab pawg tau pom qhov siab tshaj 50 feem pua ​​​​ntaus qhov nruab nrab titer, thaum tawm tsam SvE pawg DNA / CTH522, ua raws li CTH522 / AlOH ntxiv rau IN pab pawg, pom qhov siab tshaj plaws neutralizing titers. Txawm hais tias tag nrho cov pab pawg muaj peev xwm ua kom tsis zoo SvF, 50 feem pua ​​​​ntawm cov neeg nruab nrab ntawm cov titers tau qis me ntsis rau txhua pab pawg tawm tsam no serovar, piv rau SvD thiab SvE. Cov vectors ntxiv rau CTH522 pab pawg tau pom qhov qis tshaj 50 feem pua ​​​​ntawm cov tshuaj tiv thaiv kab mob ntawm txhua pawg.

Cov kev txhaj tshuaj sib txawv ua rau muaj qhov sib txawv T-cell profile

Txhawm rau ntsuas cov tshuaj tiv thaiv T-cell cov lus teb, PBMCs tau sau thaum lub sijhawm kawm thiab khaws cia cia. PBMCs los ntawm lub lim tiam 0 / lub hauv paus thiab lub lim tiam 22 tau rov ua dua nrog cov pas dej peptide (MOMP) thiab tshuaj xyuas rau intracellular cytokines los ntawm intracellular cytokine staining (ICS) thiab cov phooj ywg cytometry. PBMCs tau stained rau cov cim cell hauv qab no: CD3, CD4, CD8, CD154, CD137, thiab cytokines: IFN-g, IL-2, TNF-a, IL-22, IL{{14} }A. Cov tswv yim gating muaj nyob rau hauv Cov Duab Ntxiv 4, 5. Kev txhawb siab ntawm PBMCs nrog lub pas dej MOMP peptide induced ib qho tseem ceeb teb. Qhov feem pua ​​​​ntawm cov tshuaj antigen tshwj xeeb CD154 ntxiv rau CD4 ntxiv rau T hlwb qhia txog kev sib txuas ntawm IL2, IFN-g, thiab TNF-ib qho ntxiv 2 lub lis piam tom qab txhaj tshuaj tiv thaiv zaum kawg (W22) hauv txhua pab pawg txhaj tshuaj (Daim duab 5). Feem ntau ntawm cov T hlwb no tau ua ntau yam hauv kev nkag siab tias lawv tau hais ntau cytokines. Qhov feem pua ​​​​ntawm cov tshuaj antigen tshwj xeeb CD137 ntxiv rau CD8 T hlwb tsim cov cytokines tsuas yog nce hauv DNA thiab Vector Group (W22), (Daim duab 6, qhia txog kev ua haujlwm ntau ntawm CD137 ntxiv rau CD8 ntxiv rau T hlwb). Hauv

FIGURE 3 Serum epitope mapping ntawm lub lim tiam 24. Cynomolgus macaques tau txhaj tshuaj raws li cov txheej txheem tseem ceeb sib txawv (n=5 ib pab pawg) thiab cov ntshav tau sau rau lub lim tiam 24. Peptide array IgG cov lus teb rau CTH522 15mer overlapping peptides tau nthuav tawm raws li qhov nruab nrab lub teeb liab rau txhua pab pawg nrog VD1 peptides (txiv kab ntxwv), VD2 (ntsuab), VD3 (xiav) thiab VD4 los ntawm SvD, E, F thiab G (liab) tseem ceeb hauv cov xim.

FIGURE 4 Nyob rau hauv Vitro serum neutralization ntawm C. trachomatis SvD, SvE, thiab SvF. Cynomolgus macaques tau txhaj tshuaj raws li cov txheej txheem tseem ceeb sib txawv (n=5 ib pawg) thiab cov ntshav tau sau tom qab ua tiav cov sijhawm txhaj tshuaj (lub lim tiam 24). C. trachomatis SvD, SvE, thiab SvF tau incubated nrog cov ntshav los ntawm txhua tus tsiaj thiab lub peev xwm ntawm cov tshuaj tiv thaiv kab mob los tiv thaiv kab mob tau raug tshuaj xyuas los ntawm incubating cov tshuaj tiv thaiv kab mob sib xyaw rau HaK hlwb thiab staining rau inclusions. Cov kab dub qhia SvD, kab grey qhia SvE thiab tsaus grey dotted kab qhia SvF. Cov kab liab dotted qhia txhua tus serovar qhov sib cuam tshuam 50 feem pua ​​​​ntaus qhov nruab nrab.

Daim duab 5 CD4 T cell teb. Multifunctional analyses lub lim tiam 22. Cytokine ntau lawm los ntawm CD4 ntxiv rau T hlwb assayed los ntawm ICS. Multifunctional teb tom qab stimulation nrog pas dej ua ke ntawm overlapping peptides los ntawm MOMP yog sawv cev. Qhov loj ntawm txhua lub ncuav yog proportional rau feem pua ​​ntawm CD154 ntxiv rau CD4 ntxiv rau T hlwb qhia tsawg kawg ib cytokine, xws li IFN-g, TNF-a, thiab IL-2, thiab cov proportions ntawm hlwb qhia IFN-g thiab / lossis TNF-a thiab/los yog IL-2 cytokines raug tso tawm hauv txhua lub ncuav.

Kev txhaj tshuaj tiv thaiv kom nrawm nrawm ntawm qhov chaw mos C. trachomatis SvD kab mob

Txhawm rau ntsuas qhov kev tiv thaiv kev tiv thaiv ntawm cov tshuaj tiv thaiv kab mob tiv thaiv kab mob, cov tsiaj tau tawm tsam nrog kev kis kab mob hauv qhov chaw mos nrog 5x107 IFUs C. trachomatis SvD. Tom qab inoculation, qhov chaw mos chlamydial load tau txiav txim los ntawm PCR nrhiav pom chlamydial DNA nyob rau hauv qhov chaw mos swabs (Daim duab 7). Cov kab mob no tau raug tshem tawm hauv txhua tus tsiaj los ntawm lub lim tiam 5 hauv DNA ntxiv rau CTH522/CAF01 pawg thiab Vector ntxiv rau CTH522 pawg, los ntawm lub lim tiam 6 hauv pawg CAF01/ CTH522 thiab los ntawm lub lim tiam 8 hauv CTH522/Alum ntxiv Hauv pab pawg (Daim duab 7). Hauv pawg tswj hwm, nyob rau lub lim tiam 8, 80 feem pua ​​​​ntawm cov tsiaj tau tshem tawm tus kab mob thiab 1 tus tsiaj tseem nyob PCR zoo nyob rau lub lim tiam 9. Cov IN booster hauv CTH522/CAF01 ntxiv rau IN pab pawg tsis tau nce kev tiv thaiv (Daim duab 7). Kev ntsuam xyuas hauv qib (Mantel-Cox), piv rau txhua pab pawg tau pom tus nqi P ntawm 0,0994, thiab los ntawm kev sib piv txhua pawg ntawm tus kheej tawm tsam pawg tswj hwm, tsuas yog pawg DNA-CTH522 pom tus nqi P hauv qab 0.05 (P. tus nqi =0.0257).

Kev sib tham

Txoj kev tshawb fawb tam sim no tau tsim los txhawb kev tiv thaiv kab mob ntawm tes (CD4 thiab CD8) ua ke nrog kev tiv thaiv kab mob humoral thiab kho cov phenotype ntawm kev tiv thaiv kab mob nrog MOMP-based Chlamydia tshuaj tiv thaiv. Peb ntsuas qhov kev tiv thaiv kab mob thiab kev tiv thaiv kev ua tau zoo ntawm kev sib xyaw ua ke ntawm ntau cov tshuaj tiv thaiv kab mob nrog rau cov tshuaj tiv thaiv kab mob protein (CTH522, Con E), tshuaj tiv thaiv DNA thiab cov kab mob sib kis kab mob (MVA MOMP), thiab cov tshuaj tiv thaiv kab mob adeno vector-based, hauv cov tshuaj tiv thaiv tsis yog tib neeg primate ( NHP) model.

Cov ntaub ntawv qhia tau hais tias tag nrho cov kev txhaj tshuaj tiv thaiv kab mob yog immunogenic thiab induced ob lub cellular thiab ib tug humoral teb uas muaj xws li neutralizing antibodies (Figures 2-4). Cov tshuaj tiv thaiv kab mob protein CTH522 tsim nrog CAF01 lossis AlOH tau pom muaj kev cuam tshuam sai ntawm cov kab mob antigen-specific IgG (Daim duab 2). Peb tsis tau soj ntsuam qhov sib txawv tseem ceeb ntawm cov lus teb tshwm sim los ntawm AlOH thiab CAF01 cov tshuaj tiv thaiv kab mob ntxiv, uas tau pom zoo nrog cov lus teb pom hauv tib neeg nrog cov tshuaj tiv thaiv no (16). Cov tshuaj tiv thaiv kab mob DNA thiab vector tseem ua rau muaj qhov tseem ceeb ntawm cov tshuaj tiv thaiv kab mob antigen tshwj xeeb, uas tau nce ntxiv tom qab txhaj tshuaj tiv thaiv kab mob protein (Daim duab 2). Raws li kev pom zoo nrog peb cov txiaj ntsig, kev tshawb fawb yav dhau los hauv cov nas kuj tau pom tias cov tshuaj tiv thaiv kab mob protein ua rau muaj cov tshuaj tiv thaiv kab mob tshwj xeeb ntau dua piv rau DNA thiab vector tshuaj tiv thaiv antigens thiab tseem txhawb nqa titers zoo rau cov pab pawg no (15).

MOMP muaj plaub lub VD uas tau nthuav tawm rau saum npoo ntawm daim ntawv kis kab mob ntawm Ct Cov cheeb tsam no yog lub hom phiaj tseem ceeb ntawm kev tiv thaiv kab mob-tiv thaiv kab mob, ob qho tib si los ntawm cov tshuaj tiv thaiv tsis zoo thiab Fc receptor-mediated immunity (18). Cov kev tshawb fawb yav dhau los tau pom tias kev hloov pauv ntawm cov tshuaj tiv thaiv kab mob tsis zoo hauv cov ntshav tau txo qis cov kab mob thaum ntxov tom qab tus kab mob C. trachomatis (6, 11, 19–21). Yog li, cov tshuaj tiv thaiv zoo li ua lub luag haujlwm hauv neutralizing / tswj qhov pib inoculum, tej zaum hauv kev koom tes nrog kev tiv thaiv ntawm tes. Los ntawm cov ncauj lus kom ntxaws B cell epitope daim ntawv qhia, peb pom kev paub txog VD thaj tsam 1, 3, 4 hauv txhua pawg tshuaj tiv thaiv. Cov pab pawg DNA thiab vector kuj tau lees paub VD1, 3 thiab 4 cheeb tsam. Txawm li cas los xij, cov pab pawg no tau pom qhov nqaim me ntsis hauv cov VDs piv rau CAF01 thiab AlOH pawg, tej zaum xav tias Con E antigen hauv pawg vector tsis muaj VD4 cheeb tsam ntawm SvF thiab G (Daim duab 3). Kev lees paub ntawm cheeb tsam VD4 cuam tshuam nrog lub peev xwm ntawm txhua yam tshuaj tiv thaiv cov tswv yim los txhawb cov tshuaj tiv thaiv kab mob tiv thaiv SvD, SvE, thiab SvF.

Parenteral txhaj tshuaj nrog CTH522 nyob rau hauv CAF01 thiab AlOH induced tseem ceeb theem ntawm mucosal antibodies (IgG) nyob rau hauv qhov chaw mos thiab qhov muag thiab theem correlated nrog serum titers (Ntxiv daim duab 3, lub lim tiam 18). Cov tshuaj tiv thaiv DNA thiab vector ib leeg tsis ua rau cov mucosal IgG, uas qhia txog cov ntshav qab zib qis dua piv rau cov protein ib leeg. Txawm li cas los xij, tom qab CTH522/CAF01 IM booster, cov pab pawg no tseem ceeb nce hauv cov ntshav qab zib Ab. theem thiab ntawm qhov chaw mos thiab qhov muag IgG, ntawm qhov sib npaug nrog cov protein ib leeg (Cov duab ntxiv 3, lub lis piam 18). Qhov kev ntsuam xyuas sib npaug ntawm CTH522 hauv qhov sib txawv priming adjuvant systems (AlOH thiab CAF01) ua raws li ob zaug intranasal boosting nrog antigen tsis muaj boosting nyhuv ntawm mucosal antibody qib (Ntxiv daim duab 3, lub lim tiam 22). Yog li tag nrho cov ntshav ntshav IgG qib tau cuam tshuam rau ntawm cov mucosa rau txhua pawg. Ib qho kev sim tib neeg tsis ntev los no-Kuv kuj tau soj ntsuam qhov sib npaug ntawm CTH522 hauv AlOH thiab CAF01, ua raws li ob zaug kev txhawb nqa intranasal. Ntawm no mucosal IgG titers kuj sib raug zoo nrog cov ntshav IgG concentrations (16). Txawm li cas los xij, qhov sib txawv ntawm qhov kev tshawb pom hauv NHP qauv, tib neeg kev sim tau pom tias muaj txiaj ntsig ntawm kev txhawb nqa hauv pawg CAF01, ua rau nce qib ntawm IgG thiab IgA. Hmoov tsis zoo, kev ntsuam xyuas IgA tsis tuaj yeem ua tau hauv txoj kev tshawb fawb tam sim no, vim muaj cov khoom siv tsawg thiab cov cim tsis zoo uas tshwm sim los ntawm tus ntoo khaub lig reacting cov tshuaj tiv thaiv kab mob thib ob siv hauv kev ntsuam xyuas. Ob qhov kev tshawb fawb siv VaxINator ntaus ntawv, uas yog tsim los rau atomize qhov ntswg tshuaj rau mucosal adsorption. Txawm li cas los xij, qhov ntswg kab noj hniav ntawm tib neeg yog qhov dav ntau dua li hauv NHP thiab nws tuaj yeem xav tias qhov no tuaj yeem cuam tshuam rau kev faib tawm thiab kev nce ntxiv.

Ntau qhov kev tshawb fawb hauv tib neeg thiab nas tau pom qhov tseem ceeb ntawm Th1 T cell tiv thaiv kab mob thiab cov nyhuv cytokine IFN-g (8, 22–27). Hauv qhov sib piv, tsawg dua paub txog lub luag haujlwm tiv thaiv CD8 T hlwb (28–33). Txhua pawg tshuaj tiv thaiv tau ntxias CD4 T hlwb, tab sis tsuas yog cov DNA thiab Vector pawg tau pom CD8 T cell teb (Daim duab 6 thiab 7). Raws li cov cytokines ntsuas, CD8 T hlwb tau tsim los ntawm ib tug tsawg tus naj npawb ntawm subsets piv rau CD4 T hlwb, uas yuav muab faib ua 6 los yog ntau subsets, feem ntau ntawm cov uas tsim tau ob peb cytokines (Daim duab 7). CD4 T hlwb tau pom ntau dua IL-2-qhia subsets dua CD8 T hlwb, uas feem ntau suav nrog tsuas yog ib qho IL-2-qhia subset, ib qho uas qhia IL-2 ua ke nrog IFN-g thiab TNF- a (Daim duab 6). CD8 T hlwb tau tswj hwm los ntawm cov pej xeem loj uas tsim tau IFN-g nkaus xwb, thaum lwm qhov tseem ceeb CD8 T cell subset tsim ob qho tib si IFN-g thiab TNF-a. Cov qauv cytokine no ntawm CD4 T hlwb thiab CD8 T hlwb tsis zoo li qhov tau pom hauv nas, thaum kuaj cov tswv yim tshuaj tiv thaiv tuaj yeem ua rau CD4 thiab CD8 T hlwb (34).

Ib lub tswv yim zoo sib xws nrog kev txhaj tshuaj DNA ua raws li kev txhawb nqa nrog kev tiv thaiv kab mob (ISCOM) ntawm MOMP protein (35) ua rau muaj zog tiv thaiv kab mob Chlamydia muridarum ntsws. Hauv txoj kev tshawb no, DNA MOMP (los yog MOMP / ISCOMS) ua rau muaj kev tiv thaiv tsawg dua (35). Lwm txoj kev tshawb fawb tau pom tias kev txhaj tshuaj tiv thaiv nas nrog DNA MOMP plasmids, tsis muaj cov tshuaj tiv thaiv subunit, ua tsis tiav los tiv thaiv qhov chaw mos C. trachomatis challenge (36), qhia tias cov tswv yim DNA tau txais txiaj ntsig los ntawm kev ntxiv nrog cov tshuaj tiv thaiv subunit inducing CD4 T hlwb thiab cov tshuaj tiv thaiv. . Nws yuav tsum raug sau tseg tias cov tshuaj tiv thaiv DNA tau pom tias muaj kev tiv thaiv kab mob tsawg tsawg hauv tib neeg kev sim tshuaj, uas yuav tsum tau hais txog hauv kev tshawb fawb yav tom ntej (37).

Txawm hais tias cov koob tshuaj Ct uas siv rau hauv txoj kev tshawb no yog siab dua li cov koob tshuaj uas tau ntsib los ntawm cov kab mob hauv tib neeg, nws tau raug xaiv los xyuas kom meej tias txhua tus tsiaj tau kis tus kab mob, uas yog qhov tseem ceeb rau kev txiav txim siab tsawg tus tsiaj hauv txhua pab pawg. Cov koob tshuaj yog nyob rau hauv txoj kab nrog cov koob tshuaj siv hauv kev tshawb fawb yav dhau los, uas nyob ntawm 106 txog 108 (38, 39). Kev txo cov koob tshuaj hauv peb tus qauv ua rau muaj kev hloov pauv ntau ntxiv txog kev kis kab mob (cov ntaub ntawv tsis qhia). Cov koob tshuaj qis dua tau siv ntau zaus (40, 41). Txawm li cas los xij, qhov no tsis yog ib qho kev pom zoo thaum lub hom phiaj yog los ntsuas kev tiv thaiv kev tiv thaiv kev ua haujlwm, tshwj xeeb tshaj yog tias kev tiv thaiv ntuj yog kev kho kom haum xeeb kev tiv thaiv ua ntej kev tiv thaiv kab mob ua tiav. Nws tuaj yeem xav tias kev siv koob tshuaj ntau yuav xav tau lub cev tiv thaiv kab mob uas muaj tag nrho cov caj npab ntawm lub cev tiv thaiv kab mob, xws li antigen tshwj xeeb CD4 T hlwb, CD8 T hlwb nrog rau cov tshuaj tiv thaiv tsis zoo, txhawm rau txo cov kab mob. qib. Qhov ntawd kuj tseem yuav piav qhia tias vim li cas cov tshuaj tiv thaiv kab mob tsis zoo hauv txhua pab pawg tsis ua rau muaj kev tiv thaiv tseem ceeb hauv txhua pab pawg [txawm tias peb tau pom muaj kev tiv thaiv ntxov hauv ntau pab pawg, qhov twg cov tshuaj tiv thaiv tau xav ua lub luag haujlwm (Daim duab 7)] . Tam sim no peb tab tom sim tsim ib qho qauv ntawm lub cev ntau dua.

Ua ke, peb cov ntaub ntawv tau pom tias CTH522 antigen tsim nyob rau hauv adjuvant CAF01 los yog AlOH tuaj yeem ua rau ob qho tib si T-cell multifunctional thiab neutralizing antibodies hauv NHPs. Ntxiv mus, peb cov txiaj ntsig tau qhia tias cov tshuaj tiv thaiv kab mob heterologous muaj xws li cov tshuaj tiv thaiv DNA thiab CTH522 / CAF01 subunit vaccine sawv cev rau cov tshuaj tiv thaiv zoo rau induction ntawm ntau lub cev tiv thaiv kab mob nrog rau qib siab ntawm neutralizing Abs ua ke nrog CD4 thiab CD8 T cell teb muaj zog. Nws tuaj yeem ua kom nrawm nrawm ntawm tus kab mob C. trachomatis.

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