Nkag siab Kev Nkag Nkag Nkag Ntawm Adenovirus Capsids; A Chance To Change Vaccine Vector Properties Part 1
Mar 03, 2023
Abstract:
Adenovirus vector-based tshuaj tiv thaiv kab mob tau tshwm sim los ua lub tswv yim muaj zog tiv thaiv SARS-CoV-2 teebmeem kev noj qab haus huv. Qhov kev vam meej no tsis tau xav txog vim tias adenoviruses sib xyaw ua ke ntau yam kev xav tau ntawm cov tshuaj tiv thaiv caj ces. Lawv yog cov tshuaj tiv thaiv kab mob siab heev thiab muaj qhov tsawg thiab muaj txiaj ntsig zoo ntawm cov kab mob pathogenic ua ke nrog kev siv thev naus laus zis. Kev siv zog txuas ntxiv los txhim kho adenovirus-cov tshuaj tiv thaiv kab mob suav nrog kev siv cov tshuaj tsis tshua muaj neeg thiab tsis yog tib neeg adenoviruses. Hauv qhov kev tshuaj xyuas no, peb tsom mus rau tus kab mob capsid thiab qhov kev xaiv ntawm cov genotypes cuam tshuam li cas rau kev noj qab haus huv thiab kev txheeb xyuas subcellular tom ntej. Peb piav qhia txog yuav ua li cas nkag siab capsid zog, xws li kev ruaj ntseg thaum lub sij hawm nkag, tuaj yeem hloov txoj hmoo ntawm cov khoom nkag mus thiab qhov no txhais li cas rau qhov sib txawv ntawm kev tiv thaiv kab mob. Peb sib tham txog qhov kev hloov pauv ntawm daim nyias nyias lytic capsid protein VI cuam tshuam rau hom kab mob C cov kab mob tom qab nkag thiab sib tham luv luv yog tias cov txheej txheem no tuaj yeem muaj qhov cuam tshuam ntau dua hauv cov tshuaj tiv thaiv thiab / lossis kev txhim kho vector.
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1. Taw qhia
Cov kab mob kis kab mob, tsis zoo li cov kab mob kab mob uas tuaj yeem kho tau nrog tshuaj tua kab mob, tseem muaj kev hem thawj rau kev noj qab haus huv vim tias muaj qee yam tshuaj tiv thaiv kab mob. Yog li ntawd, kev txhaj tshuaj tiv thaiv yog tib lub tswv yim siv dav los tiv thaiv tib neeg (thiab tsiaj) cov pej xeem los ntawm kev kis kab mob.
Txog thaum tsis ntev los no, kev txhaj tshuaj tiv thaiv kab mob kis tau raug txiav txim siab nyob rau hauv cov ntsiab lus ntawm kev tiv thaiv kab mob thaum yau lossis raws li kev txhaj tshuaj txhua xyoo "flu" rau kev tiv thaiv kab mob khaub thuas. Nrog rau kev kis tus kabmob SARS-CoV-2 teebmeem kev noj qab haus huv hauv xyoo 2020, kev txhaj tshuaj tiv thaiv tau rov qab los rau lub hauv paus ntawm kev saib xyuas pej xeem vim tam sim no pom tias tsuas yog txoj hauv kev zoo los ntawm tus kabmob kis no. Hauv cov ntsiab lus no, adenovirus (Ad) cov tshuaj tiv thaiv vector-based tau tshwm sim los ua ib qho ntawm ob txoj kev txhaj tshuaj tiv thaiv zoo. Tom qab kev pom zoo thaum muaj xwm txheej ceev, cov tshuaj tiv thaiv Ad tam sim no tau txhaj rau hauv ib feem loj ntawm cov neeg hauv ntiaj teb hauv kev siv zog tsis tau muaj dua los txhim kho lub ntiaj teb tiv thaiv kab mob SARS-CoV-2.
Adenovirus vector vaccines tsis tau siv dav ua ntej COVID-19 tshwm sim, tab sis lawv txoj kev vam meej tsis yog qhov xav tau. Cov kab mob feem ntau yog cov tshuaj tiv thaiv zoo heev. Lawv yog cov tshuaj lom neeg ua kom zoo thiab yoog rau kev thauj mus los ntawm nucleic acid, thiab lawv elicit tiv thaiv kab mob. Adenoviruses muaj ntau yam uas ua rau lawv tshwj xeeb tshaj yog cov tshuaj tiv thaiv zoo tshaj plaws. Lawv muaj qhov tsawg thiab zoo-tus yam ntxwv pathogenic profile ua ke nrog cov kab mob siab. Lawv yog cov cuab yeej cuab tam thiab tau siv dav rau vectorization. Lawv cov khoom lag luam siab thiab kev ruaj ntseg tso cai rau kev tsim khoom hauv kev tsim khoom zoo (GMP) cov xwm txheej thiab muab cov txiaj ntsig zoo (ie, khaws cia) rau kev faib khoom hauv cheeb tsam ntawm tus nqi txo [1]. Tsis tas li ntawd, Ad cell xa mus rau cov tshuaj tiv thaiv kab mob tam sim no ua haujlwm tau zoo, thiab lawv ib txwm ua rau lub cev tsis muaj zog tiv thaiv kab mob uas ua rau kev txhawb nqa kev txhaj tshuaj zoo [2,3].
Nkag siab txog Ad biology yog qhov tseem ceeb hauv kev tsim Ad-raws li (cov tshuaj tiv thaiv) vectors. Piv txwv li, kev siv cov tshuaj tiv thaiv tsis yog tib neeg lossis tsis tshua muaj Ad serotypes hauv qee qhov kev lag luam tam sim no SARS-CoV-2 cov tshuaj tiv thaiv tau txhob txwm xaiv los tiv thaiv kev tiv thaiv kab mob ua ntej los ntawm kev txo cov tshuaj tiv thaiv. Kev siv cov kab mob gene-deleted vectors muab kev ruaj ntseg vector. Txawm li cas los xij, tseem muaj ntau yam ntawm cov kab mob hauv lub neej uas tsis tau txiav txim siab thaum tsim cov tshuaj tiv thaiv kab mob Ad-raws li, thiab kev paub txog cov genotypes tsawg lossis tsis yog tib neeg muaj kev txwv, suav nrog cov tam sim no siv los ua SARS-CoV-2 tshuaj tiv thaiv . Nrog rau qhov kev tshuaj xyuas no, peb xav kom muaj kev paub txog kev hloov kho uas tuaj yeem siv tau thaum tsim Ad vectors rau kev kho mob nrog rau kev txhaj tshuaj.
Peb tsom mus rau tus kab mob capsid thiab seb qhov kev xaiv ntawm cov genotypes cuam tshuam li cas rau kev noj qab haus huv thiab kev txheeb xyuas subcellular tom ntej. Peb piav qhia txog yuav ua li cas nkag siab txog cov khoom capsid xws li kev ruaj ntseg thaum lub sij hawm nkag tuaj yeem hloov txoj hmoo ntawm cov khoom nkag mus thiab qhov sib txawv ntawm kev tiv thaiv qhov tshwm sim tau pom. Raws li ib qho piv txwv ntawm yuav ua li cas capsid stability thiab txoj hmoo tuaj yeem hloov pauv, peb sib tham hauv kev nthuav dav yuav ua li cas hloov cov membrane lytic capsid protein VI cuam tshuam rau hom kab mob C. Thaum kawg, peb tham luv luv yog tias cov txheej txheem zoo li no muaj peev xwm raug siv los siv tshuaj tiv thaiv thiab / lossis kev txhim kho vector.
1.1. Adenoviruses tsim ib tsev neeg sib txawv ntawm cov kab mob sib kis
Keeb kwm, Ad tau raug cais tawm thawj zaug hauv xyoo 1953 los ntawm adenoids ntawm cov neeg mob uas muaj kab mob ua pa [4], thiab tsis ntev tom qab ntawd, lo lus "adenovirus" tau txais kev lees paub thoob ntiaj teb [5]. Tsev neeg ntawm Adenoviridae muaj ntau tshaj li 120 tus tswv cuab, muab faib ua tsib genera - nyob ntawm seb lawv kis tau tsiaj txhu (xws li, Mastadenoviridae), noog (xws li, Aviadenoviridae), cov tsiaj reptiles (eg, Atadenoviridae), amphibians (Siadenoviridae), los yog ntses (Ichtadenoviridae). [6,7] ib. Lub genus Mastadenoviridae kuj suav nrog ntau dua 70 hom kev tshaj tawm tib neeg [8,9], uas tau muab faib ua xya hom (A-G), raws li lawv cov khoom morphological, lom, thiab physicochemical, raws li teev nyob rau hauv Table 1 [10, 11].
Tsis tas li ntawd, Ads kuj tuaj yeem ua rau rov ua dua tshiab, yog li tsim hom tshiab [12], ib yam li AdE4, uas tsuas yog tib neeg hom E kab mob thiab zoo li yuav tshwm sim los ntawm kev sib txuas ntawm tib neeg hom B thiab simian Ad [13 ]. Txawm hais tias kev kis kab mob hauv tib neeg uas tsis yog tib neeg Ads yog ib txwm tsis tshua muaj, lawv tsis tuaj yeem ua tsis tau [14,15]. Lub peev xwm ntawm ntau yam kev tshaj tawm tsis yog tib neeg kom kis tau (lossis hloov pauv) tib neeg lub hlwb ua rau lawv txaus nyiam (cov tshuaj tiv thaiv) vector platforms. Ntawm lawv yog Ads muab los ntawm chimpanzees [16–18], gorillas [19], yaj [20], nyuj [21], dev [21,22], lossis lub ntiaj teb tshiab liab [23].
Cov kab mob Adenovirus tuaj yeem ua rau muaj kab mob sib txawv nyob ntawm lawv cov cellular lossis cov ntaub so ntswg tropism xws li ua pa, qhov muag, tso zis, lossis kab mob plab (saib Daim duab 1). Cov ntaub so ntswg tropism feem ntau yog qhov tshwm sim ntawm qhov sib txawv receptor kev siv los ntawm kev tshaj tawm sib txawv thiab kuj tseem muaj feem cuam tshuam nrog kev ruaj ntseg ntawm qhov nruab nrab [24]. Feem ntau cov kab mob nrog Ads yog me me los yog asymptomatic nyob rau hauv lub cev tiv thaiv kab mob. Kev kis kab mob hauv Primary Ad feem ntau tshwm sim thaum menyuam yaus thiab ua rau muaj kev tiv thaiv zoo. Piv txwv li, nyob rau hauv cov neeg uas muaj kev tiv thaiv kab mob, Ads tuaj yeem ua rau tsis muaj kev tswj hwm, mob hnyav, thiab ua rau muaj kev phom sij rau lub neej ua rau muaj kev phom sij loj ntawm tes, uas tuaj yeem ua rau mob ntau dhau thiab ntau lub cev tsis ua haujlwm, ua rau muaj kev tuag siab [12].
Qhov zoo tshaj plaws-kawm tib neeg Ads yog hom kab mob C hom 2 thiab 5, uas feem ntau pom nyob rau hauv cov neeg mob uas muaj kab mob ua pa sab sauv thiab plab hnyuv [25] thiab uas yuav yog qhov tseem ceeb ntawm qhov kev tshuaj xyuas no. Adenoviruses tuaj yeem ua lub luag haujlwm rau qee lub sijhawm, xws li hom B kab mob ua rau mob ntsws (xws li, Ad3, 7, thiab 14) ntawm cov neeg tiv thaiv kab mob [26] lossis Ad40/41 ntawm hom F thiab Ad12, 18, thiab 31 ntawm hom A. , uas kis kab mob hauv plab (raws li tshuaj xyuas hauv [26,27]). Cov kab mob D yog ib pawg neeg muaj ntau haiv neeg thiab muaj kev sib koom ua ke thiab yog ib qho ua rau mob qhov muag [28].
1.2. Adenovirus Capsids yog Metastable Structures
Txawm hais tias lawv muaj ntau haiv neeg, txhua qhov Ads ua raws li cov qauv tsim thiab cov koom haum zoo sib xws. Adenoviruses yog cov kab mob DNA uas tsis yog lub hnab ntim khoom uas muaj qhov loj, 26–45 kb linear ob-stranded DNA genome [29–31]. Lub capsid yog ~ 70-90 nm inch thiab muaj daim ntawv ntawm icosahedron nrog 20 facets thiab 12 vertices [32]. Lub capsid muaj peb yam tseem ceeb ntawm cov proteins (hexon, penton, thiab fiber) thiab ntau yam me me cement proteins (ie, IIIa, VI, VIII, thiab IX nyob rau hauv hom C kab mob) uas stabilize tus qauv. Cov qauv kev daws teeb meem siab ntawm ob peb Ad capsids tau txais los ntawm cryo-electron microscopy thiab X-ray crystallography [24,33–36]. Lub icosahedral capsid muaj pseudo-T25 qauv nrog 720 exons sib sau ua ke nyob rau hauv txawv trimers nyob ntawm seb lawv qhov chaw nyob rau hauv lub capsid. Lub 20 facets ntawm lub capsid yog tsim los ntawm cuaj exons sib sau ua ke (hu ua GON rau Pab Pawg ntawm cuaj), whereas ntawm 12 vertices, exons tau sib sau ua ke hauv GOS (rau Pawg Rau Rau) (saib [37,38] kom paub meej ntxiv ). Lub pentameric penton puag yog muab tso rau hauv 12 vertices thiab ua lub hauv paus rau lub protruding trimeric fiber [39,40]. Kev sib cuam tshuam ntawm cov proteins loj thiab me capsid stabilizes capsid, feem ntau los ntawm kev cuam tshuam nrog hexagons.
Piv txwv li, GON yog ruaj khov los ntawm IX [41]; IX thiab IIIa yog lawv tus kheej ruaj khov los ntawm kev sib cuam tshuam nrog VIII, uas txuas lawv nrog GOS thiab GON, feem [37]. VIII molecules qhia txog GON thiab pab kom ruaj khov hexagons ntawm sab hauv ntawm capsid. Protein VI kuj stabilizes hexon los ntawm binding lub hexon trimer central kab noj hniav raug rau lub puab kab mob lumen [42–44].
Lub capsid nyob ib puag ncig thiab tiv thaiv tus kab mob tseem ceeb ntawm cov kab mob genome, cov kab mob tseem ceeb ntawm cov proteins (piv txwv li, V, VII, Mu hauv hom kab mob C), thiab lub davhlau ya nyob twg cov protein (TP), uas yog covalently khi rau txhua 5' kawg ntawm Ad genome. Me capsid proteins cuam tshuam nrog cov proteins tseem ceeb tsim kev sib txuas ntawm capsid thiab genome; Txawm li cas los xij, lawv lub koom haum spatial ntawm cov cores proteins tseem tsis to taub. Nws tau pom tias V tswj hwm DNA hauv capsid los ntawm kev cuam tshuam nrog VI (capsid side) thiab VII (genome side) [45–47] thiab tej zaum yuav pab txhawb rau genome tso tawm ntawm lub pore nuclear [48]. VII yog lub ntsiab capsid protein txuam nrog cov kab mob DNA thiab txhawb nqa nws cov condensation hauv capsid [49-51]. VII kuj tiv thaiv kab mob genome los ntawm kev tiv thaiv kab mob los ntawm cov xov tooj ntawm tes [50,52–54]. Interestingly, capsid protein IX thiab protein V tsuas yog tam sim no nyob rau hauv Mastadenoviridae [55,56], thiab kab mob capsids tuaj yeem tsim tsis muaj IX [57] lossis protein VII [58], qhia qee qhov hloov pauv hauv cov qauv capsid.
Lub genome ntawm tib neeg Ads encodes ~ 45 cov proteins (xws li cov qauv proteins), muab tso rau hauv ib ntus tswj hwm cov ntawv sau cia [59]. Qhov kev hloov pauv hloov pauv E1A yog thawj cov protein qhia thiab yog qhov tsim nyog los pib cov kab mob kis kab mob [60]. Lub genome kuj tseem muaj qhov sib npaug ntawm 5 'encapsidation thiab inverted terminal repeats (ITR) ntawm txhua qhov kawg uas yuav tsum tau ua kom rov ua dua [59,61–64]. Adenoviruses encode ib qho adenoviral protease (AVP) uas tau ntim rau hauv cov khoom thiab ua rau ntau cov proteins (ie, IIIa, VI, VII, VIII, µ, TP, thiab 52.55K hauv hom kab mob C) [65–67]. Cov kauj ruam no yog tsim nyog rau kev tsim cov kab mob loj hlob tuaj thiab ua rau muaj cov metastable (piv txwv li, tsis ruaj khov) capsids primed rau capsid disassembly raws li kev sib txuas ntawm tes thiab nkag [68,69]
Kuj tseem muaj hom kev sib txawv ntawm cov qauv ntawm tib neeg Ads, piv txwv li, nyob rau hauv fiber ntau ntev thiab yooj, uas ncaj qha cuam tshuam rau receptor binding [70–72]. Qhov sib txawv kuj tuaj yeem pom hauv penton los ua ke. Pentons ntawm hom kab mob B tuaj yeem tsim kev sib cuag nrog penton, ua rau lwm yam thiab tsis tshua muaj cov kab mob zoo li cov kab mob uas tsis muaj genomes thiab qee zaus tsis muaj fibers. Cov khoom no hu ua dodecahedrons thiab tau kawm txog kev hloov pauv ntawm tes ntawm plasmids lossis peptides ua lwm txoj hauv kev rau Ad vectors. Dodecahedrons zoo dua nthuav tawm RGD luv peptide motif tam sim no nyob rau hauv penton ntawm feem ntau Ad hom, uas yuav tsum tau khi lub hom phiaj [73,74]. Qhov sib txawv ntawm capsid stability, raws li tau hais hauv qab no, tej zaum yuav yog ib qho khoom tseem ceeb uas txiav txim siab txoj hmoo nkag ntawm Ad hais.
1.3. Adenoviruses Ua raws li Lytic Life Cycle
Lub voj voog kab mob Ad tau raug tshuaj xyuas ntau qhov chaw [75–77]. Ntawm no, peb luv luv tsom mus rau qhov nkag mus rau hauv lub neej voj voog, thiab, tshwj tsis yog hais tias lwm yam, peb yuav xa mus rau hom kab mob C, uas yog qhov zoo tshaj plaws kawm (Daim duab 1). Adenoviruses siv capsid fiber ua lub hauv paus txuas ntawm tes molecule. Feem ntau Ads, suav nrog hom kab mob C, siv coxsackievirus thiab Ad receptor (CAR) los khi lub hom phiaj hlwb [78–81]. Hauv qhov sib piv, qee hom kab mob B nyiam khi CD46 receptor [82] lossis desmoglein 2 [83], thiab qee hom kab mob D tau pom tias cuam tshuam nrog sialic acids [84,85] lossis ncaj qha nrog v integrins (saib Table 1) [ 86].
Yog li, kev hloov pauv fiber ntau ntawm cov tsiaj yog lub tswv yim txaus nyiam los hloov tropism [87–89]. Lub luag haujlwm tseem ceeb ntawm fiber ntau molecule yog kho kom haum rau lub cev nrog lub hom phiaj hlwb, uas yog ua raws li kev sib cuam tshuam thib ob ntawm penton thiab integrins, xws li v 5 [90]. Qhov kev sib cuam tshuam no yog kho los ntawm RGD peptide motif hauv penton ib ntus [91]. Qhov tsis muaj qhov motif no, xws li hauv hom kab mob F, yog txuam nrog kev nkag ntawm tes tsis zoo [92]. Integrin binding tshwm sim hauv integrin pawg ntawm lub xov tooj ntawm tes [93] uas ua rau muaj kev cuam tshuam cov teeb meem, ua rau kev rov ua haujlwm ntawm actin cytoskeleton [94,95] thiab endocytic uptake ntawm clathrin-mediated endocytosis lossis micropinocytosis [96,97]. RGD khi kuj tseem tuaj yeem ua rau lub capsid tsis ruaj khov [98] los ntawm kev txo qis kev sib cuag ntawm penton thiab hexon, pab lub capsid kom tsis txhob coat [99]. Raws li qhov tshwm sim, Ad particles poob lawv cov fiber ntau, thiab ib tus tuaj yeem pom cov hexon dissociated thiab penton hauv endosome [100–102]. Lub endosome yog ib tug dynamic organelle primed rau mature rau hauv degradative lysosomes, thiab Ads yuav tsum tau khiav ceev ceev mus rau hauv lub cytosol. Endosome acidification tuaj yeem ua kom cov txheej txheem khiav tawm los ntawm kev ua kom tsis muaj zog ntxiv rau capsid [100,103]. Cov txheej txheem endosome nkag mus rau hauv kev tso tawm ntawm sab hauv capsid protein VI [103,104].
Qhov tso tawm VI yog tus cwj pwm los ntawm N-terminal amphipathic helix uas khi rau sab hauv nplooj ntawv ntawm endosome thiab induces zoo daim nyias nyias curvature, ua rau hauv zos confined membrane rupture [75,103,105,106]. Cells teb rau cov kab mob uas tshwm sim los ntawm daim nyias nyias rupture thiab qhib ib lub zos autophagy teb kom tshem tawm cov organelle puas lawm [107–109]. Txhawm rau kom tsis txhob degradation, Ads stall autophagy ntawm luv luv PPxY peptide motif nyob rau hauv protein VI kom txog thaum cov particle tau mus txog kev nyab xeeb ntawm cytosol [104,108]. Hom C kab mob tau pom tias khiav tawm ntawm cov endosomes thaum ntxov [110]. Lwm hom tsiaj suav nrog A, B, thiab D ua ntej khiav mus rau lysosome ua ntej khiav mus rau hauv cytosol [111–113], uas yuav cuam tshuam rau lub cev tiv thaiv kab mob tshwm sim los ntawm cov kab mob no [114].
Thaum Ads tau mus txog lub cytosol, lawv koom nrog dynein motors thiab siv retrograde thauj mus rau microtubules mus txog lub microtubule organizer chaw [115–117]. Lub cev muaj zog khi yog tej zaum kho los ntawm hexon [115] thiab tej zaum yuav xav tau acidic priming [118]. Nyob rau hauv ib ncig ntawm lub nucleus, capsids hloov kev thauj mus los, tej zaum los ntawm kev khi rau kinesin motor proteins thiab accumulating ntawm lub hnab ntawv nuclear [115,117,119,120]. Lawv dock ntawm lub nuclear pore complex, ces capsids yog kiag li disassembled, thiab genomes raug tso tawm thiab imported rau hauv lub nucleus [51,121–124]. Ib zaug nyob rau hauv lub nucleus, lub voj voog txuas ntxiv los ntawm kev hloov pauv ntawm cov genome thiab kev qhia ntawm cov noob E1A tam sim ntawd, uas ua haujlwm rau txhua qhov kev hloov pauv thaum ntxov [60,125–130]. Ua raws li kev qhia ntawm cov enzymes replication, kab mob genomes yog replicated thiab sib sau nyob rau hauv periphery ntawm replication chaw [131].
Tom qab ntawd, cov txheej txheem cov protein tau nthuav tawm los ntawm chav qhia lig dhau los ntawm kev tswj hwm ntawm tus txhawb nqa loj [132,133]. Kev nthuav qhia ntawm cov noob lig culminates nyob rau hauv zus tau tej cov thiab nuclear tsub zuj zuj ntawm tag nrho cov qauv proteins uas lawv sib sau ua ke rau hauv progeny thiab pob tshiab synthesized genomes. Nuclear sib sau ua ke ntawm cov tiam tom ntej ntawm Ads ces ua rau particle egress thiab cell lysis [42,134–136]. Raws li tau hais los saum toj no, kev sib sau ntawm cov capsids yog ua raws li kev loj hlob, piv txwv li, cov proteolytic cleavage ntawm ob peb capsid proteins los ntawm AVP los tsim cov kab mob sib kis [66]. Qhov tseem ceeb ntawm cov txheej txheem no yog ua piv txwv los ntawm qhov kub-sensitive mutant, Ts1 pib pom nyob rau hauv ib qho kev tshuaj ntsuam mutagenesis hauv Ad2. Ntawm qhov ntsuas kub tsis tso cai, tus mutant no tsis tuaj yeem ntim cov AVP, thiab cov khoom sib sau ua ke tsis dhau kev loj hlob, uas ua rau cov khoom tsis ruaj khov [137]. Lub luag haujlwm AVP kev hloov pauv tau raug coj mus rau hauv Ad5 khaws cov phenotype no. Ts1 capsids tseem tuaj yeem txuas mus rau cell receptors thiab yuav tsum tau endocytosed, tab sis tom qab ntawd nyob twj ywm rau hauv lub endosome vim lawv tsis ua hauj lwm kom tso cov protein VI [103,138,139].
2. Tshuaj Tiv Thaiv Kab Mob Adenovirus
Adenoviruses muaj thoob plaws, thiab feem coob ntawm cov pejxeem tau ntsib tus kabmob no, feem ntau thaum menyuam yaus. Adenoviruses yog cov tshuaj tiv thaiv kab mob siab heev, thiab txhua qhov kev sib ntsib provokes kev txhim kho tshwj xeeb los tiv thaiv Ad tiv thaiv, ob qho tib si hauv lub cev thiab kev hloov pauv, uas tuaj yeem nyob mus ib txhis. Qhov no yog qhov tseem ceeb hauv kev tawm tsam tus kab mob thiab ua rau Ads tsis tshua muaj kev phom sij rau lub cev tiv thaiv kab mob, tab sis nws tuaj yeem ua rau muaj kev cuam tshuam thaum Ads yuav tsum tau siv los ua vector hauv tshuaj tiv thaiv lossis tshuaj kho noob [140,141]. Nkag siab li cas Ad (thiab nws cov vector derivatives) ua rau lub cev tiv thaiv kab mob yuav ua rau peb txhim kho kev ua tau zoo thiab kev nyab xeeb ntawm cov tshuaj tiv thaiv. Kev soj ntsuam hauv vivo, los ntawm cov neeg mob koom nrog hauv kev sim tshuaj, qhia meej meej tias Ad-raws li vectors tuaj yeem ua rau muaj zog tiv thaiv kab mob [142,143], qee zaum ua rau tuag taus [144].
Tsis tas li ntawd, ntau yam kev tshawb fawb tau ua nyob rau hauv vitro lossis tsiaj qauv [145] qhia lub luag haujlwm tseem ceeb ntawm tus kab mob kis nws tus kheej (ie, capsid thiab incorporated DNA) hauv kev ua kom lub cev tiv thaiv kab mob. Ntawm no, peb luv luv tsom rau cov lus teb tiv thaiv kab mob cuam tshuam nrog kev nkag mus rau capsid; Yog xav paub ntxiv txog qhov tob, saib cov kev txheeb xyuas yav dhau los [146–149].
2.1. Adenoviruses ua rau Cell Intrinsic Immunity
Nrog rau txoj kev loj hlob ntawm Ads raws li vectors, nws tau pom meej tias cell-intrinsic los yog innate tiv thaiv induced los ntawm Ads yog teb rau invading virion. Qhov zoo siab, ob qho tib si cov kab mob sib txawv thiab cov kab mob tsis sib xws (piv txwv li, cov kab mob UV-inactivated, cov kab mob tsis zoo, lossis cov kab mob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khawm .
Feem ntau, Kev Tshaj Tawm tau hnov los ntawm cov hlwb ntawm ob peb kauj ruam thoob plaws tag nrho cov kab mob hauv lub neej thiab qhib txoj hauv kev tiv thaiv kab mob (Daim duab 1). Qhov kev ua kom no pib nrog Ad fiber ntau khi rau ntawm lub xovtooj ntawm tes receptor CAR, uas ua rau lub mitogen-activated protein kinase (MAPK) signaling uas ua rau kom muaj kev ua haujlwm ntawm qhov transcription factor NF-κB. Qhov no tshwm sim hauv kev tsim cov cytokines pro-inflammatory (IL-6, IL-8) [157,158]. Txawm li cas los xij, nyob ntawm Ad hom, lwm cov receptors 'kev siv yuav hloov kho lub zog ntawm cov teeb liab cascade koom nrog cytokine qhia [159]. Kev sib cuam tshuam ntawm tus kab mob penton thiab cellular integrin yog ib qho ntxiv ntawm kev tiv thaiv kab mob. Ib txoj kev tshawb fawb hauv murine macrophages tau pom tias qhov kev khi no ua rau kev qhia ntawm IL-1 ntawm RGD motif [160]. Qhov no tej zaum yuav txwv tsis pub rau lub cev tiv thaiv kab mob vim hais tias, nyob rau hauv lwm yam cell qauv (xws li, HeLa, epithelial hlwb kab), lub luag hauj lwm ntawm integrin / RGD kev sib cuam tshuam hauv chemokines thiab pro-inflammatory cytokines ntau lawm tsis tau lees paub [151,161,162]. Qhov no qhia txog lub luag haujlwm tseem ceeb ntawm cov txheej txheem internalization nws tus kheej hauv kev tiv thaiv kab mob ntau dua li kev cuam tshuam nrog integrins. Tseeb tiag, penton khi rau integrin kuj ua rau phosphoinositide 3-kinase (PI3K) signaling kom nce tus kab mob internalization [163].
Ntxiv mus, PI3K kev ua kom pom tau tshwm sim raws li kev kis kab mob los ua rau kev tsim cov cytokines pro-inflammatory, xws li TNF [164], thiab cov txheej txheem internalization tau piav qhia raws li qhov yuav tsum tau ua rau kev tiv thaiv kab mob [165]. Cov kev tshawb fawb no qhia tias kev tiv thaiv kab mob feem ntau tshwm sim tom qab-internalization kauj ruam, qhia txog qhov tseem ceeb ntawm kev txheeb xyuas tom qab nkag (saib hauv [166]).
Ib zaug nyob rau hauv endosome, Ads raug rau intraluminal pathogen-recognition receptors (PRR) xws li TLR9, uas yog ob-stranded DNA sensor txwv rau lub cev tiv thaiv kab mob. Murine macrophages tuaj yeem nkag siab Ad vectors ntawm TLR9 los txhawb kev tsim cov cytokines pro-inflammatory [167]. TLR9 kuj tau nruj me ntsis rau IFN / ntau lawm thaum murine plasmacytoid dendritic cells (pDC) tau sib tw nrog Ad [168]. Txawm li cas los xij, hauv TLR9 knock-out nas qauv, IFN / tseem tsim tawm, tawm tswv yim tias, hauv cov hlwb uas tsis yog pDC, IFN ntau lawm tshwm sim ntawm nws tus kheej los ntawm TLR9 signaling [168,169].
Viral genomes (los yog vector genomes) tuaj yeem nkag mus rau kev hnov mob thaum lossis tom qab endosomal khiav ntawm tus kab mob. Thaum lawv ncav cuag cytosol, lawv tau hnov los ntawm cytosolic ob-stranded DNA sensor cGAS [168,170]. Raws li tus kab mob genome lees paub, cGAS txhawb nqa phosphorylation thiab nuclear translocation ntawm transcription factor IRF3 los tsav IFN / kev qhia, inducing lub xeev antiviral. Endosomal passage yuav tsum muaj tus kab mob-induced endosomal membrane puas los pab txhawb lub endosomal khiav ntawm cov kab mob. Membrane rupture yog li muaj txiaj ntsig zoo rau kev tiv thaiv kab mob, hais txog qhov tseeb tias kev khiav tawm tsis zoo Ts1 mutant tsis ua kom tiav thiab ua haujlwm tiv thaiv kab mob [161,162,170,171]. Lub luag haujlwm tseem ceeb hauv cov lus teb inflammatory raws li Ad membrane nkag mus rau hauv lub tank-binding kinase, TBK1, qhia tias yog ib feem ntawm downstream cytosolic sensing ntawm tus kab mob genome ntawm cGAS / STING txoj kev [168,170].
Adenovirus membrane puas kuj ua rau kev tso tawm ntawm cathepsin B los ntawm endo-lysosomal compartment, ua rau oxidative kev nyuaj siab, uas activates NLRP3 inflammasome, ua rau IL-1 maturation [114]. Hom C kab mob khiav tawm ntawm cov endosomes thaum ntxov, thaum hom B kab mob khiav mus txog lig endosomes, xav tias vim qhov sib txawv ntawm kev siv receptor. Acidification nyob rau hauv lig endosome / lysosome feem ntau yuav ua kom lysosomal acid hydrolases. Ob hom induce inflammasome activation [172], tab sis, tej zaum tshuav rau lub sij hawm nyob rau hauv endosome, qhov twg ntawm lawv cov lus teb txawv. Thoob plaws ntiaj teb, hom kab mob B ua rau muaj lub cev muaj zog thiab ua kom lub cev tsis muaj zog dua li hom kab mob C [113,173,174]. Qhov no yuav piav qhia yog vim li cas hom kab mob B yog cov kab mob ntau dua, ua rau muaj kev sib kis hauv cov neeg muaj zog tiv thaiv kab mob [26]. Nws ntxiv qhia tias cov hlwb muaj peev xwm cais qhov nkag mus rau hauv qhov chaw (piv txwv li, endosome vs. lysosome, [109]) thiab kho cov kev tiv thaiv kab mob. Endosomal membrane puas thaum lub sij hawm Ad khiav txoj kev kuj yuav yog ib tug txaus ntshai teeb liab nyob rau hauv nws tus kheej. Raws li tau tham hauv cov ntsiab lus hauv qab no, kev puas tsuaj ntawm daim nyias nyias ua rau cytosolic raug ntawm intraluminal glycans, uas tau kuaj pom los ntawm lub xovtooj ntawm tes raws li kev phom sij (raws li tshuaj xyuas hauv [109]). Kev tshawb pom ntawm cov glycans raug pom los ntawm galectins tom qab ntawd ua rau autophagy, ua rau lub ceg thib ob ntawm kev tiv thaiv kab mob [108].
2.2. Adenoviruses ua rau muaj kev hloov pauv lub cev tiv thaiv kab mob thiab cuam tshuam Antiviral Autophagy thaum nkag mus hauv Cell
Innate lub cev tiv thaiv kab mob provoked los ntawm Ads los yog lawv cov vectors tom qab ntawd txhais mus rau hauv kev tiv thaiv kab mob los ntawm chemokine secretion uas nyiam cov kab mob tiv thaiv kab mob (neutrophils, natural killer cells, thiab macrophages) thiab txhawb zog antigen kev nthuav qhia [153,154]. Adenoviruses yog cov tshuaj tiv thaiv kab mob siab heev, thiab lawv cov tshuaj tiv thaiv kab mob tseem ceeb yog cov protein ntau capsid (hexon, penton, thiab fiber) uas ua rau cov kab mob tshwj xeeb tsim tawm ntawm cov tshuaj tiv thaiv kab mob los ntawm B hlwb [175-177].
Neutralizing cov tshuaj tiv thaiv feem ntau yog qhia tawm tsam hyper-variable loops ntawm hexon protein [178–180] thiab sawv cev rau qhov kev txwv loj rau kev siv Ads raws li vectors muab los ntawm tib serotype. Qhov teeb meem no tau dhau los ntawm kev siv cov seroprevalent tsawg dua lossis tsis yog tib neeg Ads [181–185]. Txawm hais tias qhov kev tiv thaiv kab mob no, thaum Ads tau siv los ua cov tshuaj tiv thaiv kab mob (xws li, txhawm rau tiv thaiv kev tiv thaiv kab mob Ebola hauv kev sim tshuaj), lawv tseem ua rau muaj zog B cell teb, tso cai rau kev tsim cov tshuaj tiv thaiv kab mob rau cov tshuaj tiv thaiv uas xav tau immunogen uas tuaj yeem kav ntev. mus txog 6 lub hlis [186]. Tsis tas li ntawd, kev tiv thaiv kab mob uas muaj nyob ua ntej tuaj yeem kov yeej nrog cov tshuaj tiv thaiv vector ntau dua, xws li nyob rau hauv rooj plaub ntawm SARS-CoV-2 tshuaj tiv thaiv vector raws li qhov siab seroprevalence AdC5 [187].
Raws li nrog rau cov tshuaj tiv thaiv tsis zoo, Ad kab mob ua rau muaj kev tiv thaiv T-cell tsis tu ncua, feem ntau los ntawm CD4 ntxiv rau kev ua kom muaj zog. Qhov kev tiv thaiv no tseem tuaj yeem pom nyob rau hauv cov neeg laus, qhia txog kev tiv thaiv ntev ntev txij li thaum yau [188–190]. Tsis tas li ntawd, kev ua kom CD8 ntxiv kuj tau piav qhia kom tswj tau cov lus teb cytotoxic [191–193]. Qhov kev ua kom T-cell no tsis tas yuav tsum muaj serotype tshwj xeeb thiab tso cai rau kev tiv thaiv kev tiv thaiv sib txawv ntawm Ad subgroups thiab serotypes [192–194].
Tsis tas li ntawd, hauv cov neeg mob stem-cell-transplanted kis los ntawm Ad, muaj zog ua kom CD4 ntxiv thiab CD8 ntxiv tau piav qhia [195]. Qhov kev ua kom no yog qhia tawm tsam hexon peptides thiab ua rau kev tshem tawm ntawm Ad viremia, hais txog lub luag haujlwm tseem ceeb ntawm CD4 ntxiv thiab CD8 ntxiv rau T-cell ua rau kev tiv thaiv kab mob. Thaum T-cell-raws li kev tiv thaiv kab mob no yog qhia tawm tsam Ad particle, Ads kuj ua rau muaj zog tiv thaiv cellular raws li Tcell teb tiv thaiv cov tshuaj tiv thaiv antigen, uas yog qhov zoo dua rau kev tiv thaiv ntev ntev thaum siv cov ad vectors recombinant raws li tshuaj tiv thaiv platforms [196,197 ]. Qhov tseem ceeb, cov ntaub ntawv los ntawm kev sim tshuaj ntsuam xyuas tau lees paub tias thaum Ad siv los ua cov tshuaj tiv thaiv kab mob (nrog rau qhov xav tau antigen li transgene), nws ua rau ob qho tib si cytotoxic CD8 ntxiv rau T-cell thiab CD4 ntxiv rau T-cell activation, ua rau lub cim xeeb tiv thaiv kab mob. qhia transgene [184,186,198–202].
T-cell ua kom tso siab rau kev nthuav tawm ntawm cov kab mob peptides los ntawm qhov loj histocompatibility complex (MHC). Luv luv, kev nthuav qhia antigen los ntawm MHC chav kawm kuv tau tshwm sim los ntawm proteasomal degradation, thiab cov peptides tau txais yuav ua rau CD8 ntxiv rau T-cell activation, tso cai rau priming thiab proliferation ntawm cytotoxic T-cell teb. MHC chav kawm II antigen kev nthuav qhia tau los ntawm lysosomal proteolysis, thiab cov txiaj ntsig peptides yuav ua rau CD4 ntxiv rau T-cell ua kom raws li kev nthuav qhia, ua rau muaj kev ua kom tom qab ntawm B hlwb [203]. Tam sim no nws yog tus cwj pwm zoo tias autophagy txhawb kev nthuav qhia peptide rau MHC chav kawm II molecules [204]. Tsis yog qhov xav tsis thoob, autophagy yog ib qho ntawm cov kev tiv thaiv qub tshaj plaws tiv thaiv kab mob thiab yog ib feem tseem ceeb ntawm cov cellular repertoire tiv thaiv cov kab mob invading, koom nrog hauv ob qho tib si hauv lub cev thiab kev tiv thaiv kab mob. Nws muaj peev xwm ncaj qha sequester tus kab mob rau degradation (xenophagy) thiab koom nrog nws kev nthuav qhia rau lub cev tiv thaiv kab mob [205–207]. Macroautophagy (feem ntau hu ua autophagy) yog ib txoj hauv kev khaws cia lysosomal degradation txoj hauv kev uas koom nrog ntau cov txheej txheem physiological xws li homeostasis thiab innate thiab adaptive immunity [208-210]. Autophagy yog tus cwj pwm los ntawm kev tsim ntawm ob daim nyias nyias vesicle hu ua "autophagosome" uas engulfs cytoplasmic cargoes (ie, organelles, aggregates, thiab pathogens) destined rau degradation (Daim duab 2). Thaum lub sij hawm tus txheej txheem no, cargo-muaj autophagosomes fuse nrog lysosomes kom degraded thiab recycled.
Antigen kev nthuav qhia ntawm autophagy tau piav qhia rau MHC chav kawm I thiab chav kawm II [204]. Tom qab induction ntawm autophagy, intracellular antigens yog engulfed nyob rau hauv autophagosomes, fusing nrog MHC chav kawm ntawv II uas muaj compartments muab lawv nrog lwm yam kab mob (Daim duab 2) [211]. Lub luag haujlwm ntawm autophagy hauv qhov tshwj xeeb MHC chav kawm II antigen kev nthuav qhia yog thawj zaug piav qhia rau Epstein-Barr tus kab mob, qhov twg inhibition ntawm autophagy ua rau txo qis CD4 ntxiv rau T-cell ua kom [212,213]. Txog niaj hnub no, piv txwv suav nrog ntau tus kab mob suav nrog tus kab mob herpes simplex [214], kab mob khaub thuas [215], thiab tib neeg kev tiv thaiv kab mob tiv thaiv kab mob hom 1 (HIV-1) [216], thiab tej zaum yuav txuas mus rau Kev Tshaj Tawm (saib hauv qab). MHC chav kawm II kev nthuav qhia yog ua raws li cov txheej txheem ntawm autophagy raws li nws yog cytosolic tiv thaiv mechanism. Autophagy kuj tseem koom nrog hauv MHC chav kawm II hla kev nthuav qhia ntawm extracellular antigens [214]. Tseeb tiag, MHC kev nthuav qhia ntawm extracellular ovalbumin yog qhov ua tau zoo dua thaum lub tshuab autophagy (thiab tshwj xeeb tshaj yog ATG5) ua haujlwm tau zoo, thiab kev siv ntawm cov khoom siv ntxiv tau pom tias muaj feem cuam tshuam rau daim ntawv uas tsis yog-canonical ntawm autophagy hu ua LC3- txuam nrog phagocytosis ( PIB) [217].
Thaum muaj nyob hauv LAP-qee, qee cov tshuaj tiv thaiv kab mob ntxiv tuaj yeem raug nthuav tawm los ntawm MHC chav kawm II kev nthuav qhia [218]. Autophagy kuj tseem koom nrog qee qhov kev nthuav qhia hauv MHC chav kawm kuv qhov kev nthuav qhia, txawm hais tias feem coob ntawm cov epitopes tau nthuav tawm los ntawm cov txheej txheem nruj me ntsis nyob ntawm tus neeg nqa khoom cuam tshuam nrog cov txheej txheem antigen (TAP) complex [219]. TAP-kev nthuav qhia ywj pheej kho los ntawm autophagy tau pom tam sim no tsuas yog nyob rau hauv TAP depletion tej yam kev mob [219,220]. Yog tias hom kev nthuav qhia no muaj tiag tiag tseem muaj teeb meem [221] thiab tau tham txog lwm qhov [204,222,223].
Txhawm rau tiv thaiv MHC kev nthuav qhia, Cov Tshaj Tawm tau muab ntau yam ntawm lawv cov genome, uas yog thaj tsam E3, txhawm rau txhawm rau MHC kev nthuav qhia hauv hlwb [224–228]. Hauv feem ntau (cov tshuaj tiv thaiv) vectors, cov noob no tsis tuaj, thiab kev ua haujlwm tsis ntev los no qhia tias kev nthuav tawm antigen los ntawm MHC tuaj yeem txhim kho Ad cov tshuaj tiv thaiv kev ua haujlwm [229,230]. Autophagy plays lub luag haujlwm tseem ceeb hauv kev ua Ad antigens rau MHC chav kawm II kev nthuav qhia hauv Ad-induced immunity [108,229].
Montespan et al. qhia tau tias Ads induce autophagy raws li endosome nkag mus thiab hais tias nkag mus rau cov khoom tshem tawm los ntawm autophagy nyob rau hauv cov txheej txheem no tau zoo nthuav tawm rau lub cev tsis muaj zog [108]. Qhov kev soj ntsuam no yog ua raws li kev tshawb fawb luam tawm los ntawm Klein li al. qhia tias nyob rau hauv cov ntsiab lus ntawm oncolytic Ads, cov kab mob antigens yog matured los ntawm JNK-dependent daim ntawv ntawm autophagy [229]. Txawm li cas los xij, autophagy nyob rau hauv cov ntsiab lus ntawm mob qog noj ntshav plays ob lub luag hauj lwm. Raws li tus neeg saib xyuas homeostasis, autophagy txwv cov qog pib [231] thaum nyob rau hauv kev mob qog noj ntshav siab heev autophagy ua cov txheej txheem ciaj sia los txhawb cov qog loj hlob [232,233]. Yog li, nyob rau hauv cov ntsiab lus ntawm oncolytic vectors, qog-specific autophagy nta yuav tsum tau ua tib zoo soj ntsuam.
3. Adenoviruses as (Vaccine) Vectors
Vectorization ntawm tus kab mob cuam tshuam nrog kev siv nws cov khoom ntuj tsim, thaum tib lub sijhawm txo cov kev pheej hmoo lom lom. Hauv cov ntsiab lus no, Kev Tshaj Tawm yog cov vector zoo tshaj plaws rau kev tsim tshuaj tiv thaiv [234]. Adenovirus vectors ruaj khov, yooj yim los tsim rau lub siab titer, thiab muaj peev xwm loj cloning. Qhov no tso cai rau kev nkag mus ntawm ib qho transgene, piv txwv li, qhia txog kev xaiv antigen. Lawv muaj cov kab mob dav dav thiab hloov cov hlwb tsis sib faib, thiab lawv cov genome tsis sib koom ua ke [1]. Raws li tau piav qhia saum toj no, lawv kuj ua rau muaj kev mob tshwm sim hauv cov kab mob antigen-tam sim no (APCs) uas ua haujlwm rau kev ua haujlwm ntxiv los txhawb lub cev tiv thaiv kab mob uas tsim nyog los txhaj tshuaj tiv thaiv zoo [2,3,142,235,236].
Ntxiv mus, Ad-derived vectors tau siv dav hauv kev sim ua ntej thiab kuaj mob los ua pov thawj lawv txoj kev nyab xeeb [237,238]. Txhawm rau tshem tawm qhov tsis xav tau ntawm lub voj voog rov ua dua, tus kab mob sib kis yuav tsum tau muab tshem tawm kom tiv thaiv cell lysis thiab kis kab mob. Adenovirus vector txoj kev loj hlob tau pib thaum xyoo 1990 thaum ntxov ua lub platform rau ntau yam kev kho mob, xws li kev kho noob, oncolytic vectors, thiab kev tsim tshuaj tiv thaiv. Feem ntau ntawm cov kab mob tshwm sim thaum ntxov tau hloov kho cov kab mob C.
3.1. Adenovirus Vector Development, Txoj Kev Tsim Nyog
Thawj tiam ntawm Ad vector yog tsim los ntawm kev rho tawm E1 thiab E3 cheeb tsam los ntawm kab mob genome, tshwj xeeb tshaj yog encoding qhov kev hloov pauv tam sim ntawd, tiv thaiv cov vectors los ntawm replicating. Cov vectors muaj lub peev xwm cloning ntawm ib ncig ntawm 8 kb los ntxig ib qho kev hloov pauv nrog cov txheej txheem tswj hwm [239–241]. Thawj tiam vectors tau tsim nyob rau hauv tib neeg lub raum lub raum (HEK) cov hlwb hloov pauv hloov pauv nrog ib feem ntawm Ad5 kab mob genome uas ntxiv rau thaj tsam E1 hauv trans [242–247]. Ib qho ntawm cov teeb meem loj ntawm cov kab ke no yog tias nws tso cai rau kev rov tsim kho dua tshiab ntawm cov genomes uas muaj peev xwm tom qab kev rov ua dua tshiab [248]. Qhov kev tshem tawm ntxiv ntawm E3, uas tsis tsim nyog rau kev tsim cov vector, nce qhov loj ntawm transgene cassette [243] thiab txhawb kev tiv thaiv kab mob zoo dua [240].
Hmoov tsis zoo, thawj tiam Ad vectors tsis tau ua tiav cov lus tsis txaus ntseeg thiab tseem ua rau lub cev muaj zog tiv thaiv kab mob cov kab mob capsid proteins uas tau qhia nyob rau theem pib [141,249,250]. Qhov kev tiv thaiv lentivector no tau pab txhawb rau vector toxicity thiab tshem tawm lub sijhawm luv luv ntawm cov hlwb hloov pauv [145]. Lub cim thib ob ntawm Ad vectors kuj tseem muaj thaj tsam E2 thiab E4 tshem tawm ntawm vector genome. E2 encodes cov replication enzymes thiab E4 ntxiv regulatory proteins [251–253]. Qhov no tso cai rau qhov chaw rau 10.5 kb ntawm cov kab mob sib kis, suav nrog txog li plaub qhov kev qhia ntawv [254]. Ib yam li ntawd, cov kev tshem tawm ntxiv tau txo qis qhov tsis xav tau rov ua dua thiab ua kom ntev transgene qhia [255]. Qhov no yog tej zaum qhov tshwm sim ntawm qhov txo qis ntawm cov kab mob kis kab mob uas txuas ntxiv txo cov kab mob tiv thaiv kab mob [141,250,251]. Adenoviral vectors ntawm tiam thib peb yog conceptually txawv. Lawv muaj cov kab mob vector genome thiab cov kab mob uas tsis tau ntim cov kab mob (tus kab mob pab) uas tsim cov kab mob vector. Cov vectors no tseem hu ua "pab-dependent", "gutless", lossis "high-capacity" (HC-Ad) vectors. Lawv muaj lub peev xwm cloning siab tshaj plaws thiab tuaj yeem nqa cov kab ke mus txog 36 kb [256-258]. Kev tshaj tawm muaj peev xwm tsis muaj kab mob kis kab mob tshwj tsis yog ITRs thiab lub teeb liab encapsidation luv luv uas tso cai rau genome ntim thiab uas tsis tuaj ntawm tus neeg pab tus kab mob [259–262]. Lub tswv yim no tau tso cai rau peb thaum kawg kov yeej cov kev tiv thaiv kab mob tiv thaiv kab mob, tso cai rau lub sijhawm ntev ntawm cov transgenes [263–267]. Txawm li cas los xij, HC-Ads tsis yog ib feem ntawm cov tshuaj tiv thaiv vector repertoire tam sim no.
3.2. Adenovirus ua ib qho tshuaj tiv thaiv kab mob
Ua ntej tshawb nrhiav seb Ad vectors tau siv li cas, nws yog qhov tsim nyog los hais tias kev txhaj tshuaj tiv thaiv kab mob Ad hom B hom 4 thiab hom 7 lawv tus kheej tau tsim nyob rau thaum ntxov 1970s [268]. Cov tshuaj tiv thaiv kab mob hauv qhov ncauj no yog qhov yuav tsum tau ua txij li xyoo 2011 rau cov tub rog nrhiav neeg ua haujlwm hauv Tebchaws Meskas thiab txo qis kev kis tus kabmob, mus txog zero [269–271]. Qhov no tau qhia txog qhov ua tau zoo thiab kev nyab xeeb ntawm Ads raws li kev txhaj tshuaj thiab qhib lub teb kom siv Ads ua qhov chaw txhaj tshuaj. Txog niaj hnub no, ntau dua 200 cov tshuaj tiv thaiv raws li Ad-raws li tau nkag mus rau kev sim tshuaj [272]. Ntau tus ntawm lawv tau qhia tawm tsam cov kab mob sib kis xws li HIV-1 thiab Ebola [230,273,274]. Cov tshuaj tiv thaiv kab mob Adenovirus tsis muaj qhov tsis zoo. Ib qho tshuaj tiv thaiv kab mob HIV tsis ntev los no, raws li hom kab mob C virus Ad5, tau siv cov tshuaj sib xyaw ua ke uas qhia txog tus kab mob HIV-1 gag, pol, thiab nef genes [275,276]. Cov vectors muaj kev nyab xeeb, tiv thaiv kab mob, thiab tiv taus zoo. Hmoov tsis zoo, qhov tshwm sim STEP kev sim tshuaj ntsuam xyuas yuav tsum tau nres ua ntej [277–279]. Tsis tsuas yog muaj cov tshuaj tiv thaiv no tau pom tias tsis tiv thaiv kab mob HIV, tab sis cov ntaub ntawv kab mob kis tau qhia tias kev txhaj tshuaj tiv thaiv nrog Ad nyob rau hauv rooj plaub no ua rau muaj kev pheej hmoo kis tus kabmob HIV [280–282]. Muaj kev tiv thaiv yav dhau los tiv thaiv Ad5 tau pom tias yog qhov muaj feem cuam tshuam tom qab txhaj tshuaj tiv thaiv [283,284]. Txawm li cas los xij, qhov kev soj ntsuam no tsis tau lees paub hauv cov kev tshawb fawb zoo sib xws [285,286] thiab tseem tsis paub meej txog kev siv tshuab [287,288]. Ib sab ntawm HIV, Ad-based vector tshuaj tiv thaiv tau tsim tawm hauv kev tawm tsam Ebola (Ad-EBOV), qhov kev hem thawj tshwm sim hauv Western Africa [289]. Txhua qhov tshuaj tiv thaiv Ad-EBOV siv Ebola glycoprotein (GP) ua cov tshuaj tiv thaiv antigen. Kev sim ua ntej thawj zaug hauv cov nas thiab cov tsiaj primates siv Ad5 thiab pom kev tiv thaiv ntau tshaj 3 lub hlis nrog kev tsim cov tshuaj tiv thaiv kab mob thiab CD8 ntxiv rau kev ua kom [273,290–292]. Cov tshuaj tiv thaiv no tau dhau los ntawm theem II kev sim tshuaj; Txawm li cas los xij, kev tiv thaiv yav dhau los rau Ad5 hauv cov neeg txhaj tshuaj tiv thaiv tau ua rau muaj kev hloov pauv hauv qib ntawm cov tshuaj tiv thaiv tsis zoo [293–297]. Txhawm rau kov yeej qhov teeb meem no, cov tshuaj tiv thaiv raws li hom kab mob tsawg dua D Ad26 tau tsim [186,298,299].
Txawm li cas los xij, Ad26 tsis tshua muaj tshuaj tiv thaiv kab mob hauv tib neeg, ua rau txo qis kev siv tshuaj tiv thaiv, thiab xav tau kev txhawb nqa thawj zaug siv tshuaj tiv thaiv kab mob qhua pias (MVA). Qhov kev txhaj tshuaj tiv thaiv no tau mus txog theem III kev sim tshuaj [289,300,301]. Kev siv cov ad vectors tsis yog tib neeg kuj tau txhim kho [184]. Kev siv chimpanzee ChAd3 ua lub platform txhaj tshuaj tiv thaiv EBOV tau pom muaj kev tiv thaiv hauv nas thiab chimpanzees [302,303], thiab kev sim tshuaj siv cov vectors yog nyob ntawm ntau theem ntawm kev nce qib [17,300,301,304–306].
Txog thaum tsis ntev los no, cov tshuaj tiv thaiv kab mob HIV-1 thiab EBOV yog ob qho piv txwv tseem ceeb ntawm kev siv tshuaj tiv thaiv kab mob raws li Ad vector, tab sis lawv tsis yog tib qho xwb. Kev sim sib txawv tau tsim los siv cov tshuaj tiv thaiv kab mob tiv thaiv ntau yam kab mob xws li Mycobacterium tuberculosis [307–310], dengue virus [311–313], kab mob siab B thiab C [314–320], rabies [321–324], lossis mob khaub thuas [325–328]. Txawm li cas los xij, lawv qhov kev ua tau zoo yeej ib txwm qis dua qhov kev cia siab (kom paub meej ntxiv, saib [18,142,300,329]). Tus neeg sib tw tshuaj tiv thaiv EBOV tseem tsuas yog siv tshuaj tiv thaiv Ad thiab siv nrog qee qhov kev txwv hauv tebchaws Democratic Republic of Congo. Cov tshuaj tiv thaiv no yog raws li hom Ad26 (Ad{17}}ZEBOV/MVA-BN-FILO), thiab txij li lub Tsib Hlis 2020, 20,339 tus neeg tau txais thawj koob tshuaj tiv thaiv, thiab 9560 tau txhaj tshuaj tag nrho [230,301,330].
Cov tshuaj tiv thaiv kab mob Adenovirus-vectorized tau txais kev txhawb nqa pej xeem loj hauv kev teb rau SARS-CoV-2 kis thoob qhov txhia chaw. Qhov xwm txheej ceev ntawm qhov xwm txheej tau ua rau nws muaj peev xwm los nqis peev rau lawv txoj kev loj hlob, suav nrog cov kev paub tau txais los ntawm cov tshuaj tiv thaiv kab mob coronaviruses uas ua rau yav dhau los hauv cheeb tsam SARS-CoV-1 tshwm sim [331]. Tus kab mob coronavirus spike (S) glycoprotein tau pom tias yog cov tshuaj tiv thaiv zoo tshaj plaws rau kev tiv thaiv kab mob coronavirus [332,333]. Txog niaj hnub no, plaub qhov tshuaj tiv thaiv kab mob ad vector, tag nrho cov kab mob sib kis kab mob, yog nyob rau hauv lub xeev siab heev thiab tau tso cai siv thaum muaj xwm txheej ceev hauv ntau qhov chaw hauv ntiaj teb. Cov tswv yim siv vectors tau sib txawv me ntsis, tab sis qhov tseem ceeb koom nrog kev siv cov vectors thawj tiam. Lawv sib txawv los ntawm kev siv hom C Ad5-raws li vectors (Ad{10}}nCoV), kov yeej kev tiv thaiv yav dhau los nrog cov kab mob siab (CanSino) [187,334], mus rau kev siv hom kab mob D tsawg dua. Ad26, siv qhov ruaj khov spike glycoprotein (JNJ-78436735/Ad26.COV2.S) [335]. Lwm txoj hauv kev siv ChAdOx1-S-(AZD1222), ad vector los ntawm chimpanzees [336,337]. Cov tshuaj tiv thaiv tsuas yog siv raws li lub tswv yim heterologous prime-boost lub tswv yim kom kov yeej vector tiv thaiv kab mob (Gam-COVIDVac/Sputnik V) siv thawj zaug Ad26-raws li vector thiab tom qab ntawd hauv kev txhaj tshuaj txhawb nqa siv Ad5 vector [338,339]. Nyob rau hauv txhua rooj plaub, cov ntaub ntawv soj ntsuam tau txais txiaj ntsig zoo heev, qhia txog qib siab ntawm kev tiv thaiv kev tiv thaiv, thiab kev siv thaum muaj xwm txheej ceev tau tso cai rau ntau lub tebchaws rau kev siv tshuaj tiv thaiv Ad-based. Tsis tas li ntawd, kev tshawb fawb txog kev kis kab mob ntawm cov kev txhaj tshuaj tiv thaiv tsis tu ncua hauv ntau lub tebchaws qhia tias muaj kev vam meej dhau los ntawm kev siv tshuaj tiv thaiv kab mob SARS-CoV-2.
4. Modulating Adenovirus (Vaccine) Vector Efficacy, Capsid coj txoj hauv kev
Adenovirus (cov tshuaj tiv thaiv) vector tsim yuav tsum coj mus rau hauv tus account ntau yam. Cov no suav nrog kev tswj xyuas qhov ntev thiab ntev ntawm qhov encoded antigen thiab kev ua kom lub cev tiv thaiv kab mob (adjuvant effect) uas tso cai rau kev txhais lus ntawm cov tshuaj antigen mus rau qhov kev tiv thaiv tsis tu ncua tiv thaiv kab mob tiv thaiv kab mob tab sis tsis tawm tsam vector nws tus kheej. Cov ntsiab lus ntawm kev tiv thaiv kab mob yog muab los ntawm kev tiv thaiv kab mob hauv lub cev uas cov tshuaj tiv thaiv vector elicits thaum tswj hwm. Hauv cov ntsiab lus no, qhov ua tau zoo uas lub cev tiv thaiv kab mob hauv lub cev pom cov vector yog qhov tseem ceeb, thiab yuav txiav txim siab qhov amplitude ntawm cov lus teb hloov pauv. Raws li tau piav nyob rau hauv cov nqe lus saum toj no, innate immune sensing yog feem ntau tshwm sim nyob rau hauv lub post-entry theem, concomitant nrog Ad endosome penetration. Qhov qhia tau zoo ntawm (cov tshuaj tiv thaiv) antigen nyob ntawm qhov sib npaug ntawm theem ntawm vector clearance vs. kev xa khoom zoo thiab kev qhia ntawm vector genomes rau lub nucleus. Nrog rau txoj kev loj hlob ntawm capsid zaub vectors, kev qhia ntawm transgene tsis tas yuav tsum tau ua kom lub cev tiv thaiv kab mob. Lub tswv yim no yog ua raws li kev nkag mus rau qhov epitope ntawm kev txaus siab rau hauv Ad capsid proteins, tso cai rau nws ncaj qha cuam tshuam rau lub cev tiv thaiv kab mob. Txawm hais tias txoj hauv kev no muaj qee qhov zoo, feem ntau cov tshuaj tiv thaiv kab mob vam khom cov noob caj noob ces. Yog xav nyeem ntxiv ntawm capsid zaub vectors, thov saib cov kev tshuaj xyuas no, [340,341].
Kev tiv thaiv kab mob ua ntej tawm tsam tus kab mob kab mob (xws li, 30-90 feem pua tiv thaiv kab mob C virus Ad5, nyob ntawm thaj chaw thaj chaw) tuaj yeem txo cov vector uptake thiab txo cov kab mob antigen thiab ua rau lub cev tiv thaiv kab mob [342–344]. Ntau lub tswv yim tau tsim los txo cov kev tiv thaiv kab mob tiv thaiv kab mob thiab tau raug tshuaj xyuas tsis ntev los no [249]. Luv luv, cov tshuaj tiv thaiv tsis zoo feem ntau lees paub qhov sib txawv ntawm qhov sib txawv ntawm exons thiab vim li ntawd yog serotype-specific [178-180]. Cov voj voog sib txawv no tuaj yeem hloov pauv los ntawm lwm tus, tsawg dua serotype [178,345]. Qhov no tiv thaiv cov vector los ntawm kev ua kom tsis zoo thiab tsis ua haujlwm. Raws li Ad5 yog hom seroprevalent tshaj plaws, siv cov serotypes tsis tshua muaj xws li Ad26, Ad35, lossis tsis yog tib neeg Ads kuj yuav zam qhov teeb meem no [177,346–350].
Txawm li cas los xij, qhov teeb meem loj ntawm kev siv cov hom tsis tshua muaj yog lawv cov tshuaj tiv thaiv qis dua uas ua rau lawv tsis tshua muaj txiaj ntsig txog kev tsim tshuaj tiv thaiv [298]. Yog li ntawd, cov tswv yim los txo cov kev tiv thaiv kab mob tiv thaiv kab mob yuav tsum tau tsim nyob rau hauv tib lub sijhawm rau cov tswv yim los txhim kho vector immunogenicity kom tau txais vector zoo tshaj. Txawm hais tias thawj theem ntawm neutralizing antibodies tiv thaiv vector yog tsawg, kev txhaj tshuaj nrog ib hom vector yuav ua rau cov lus teb uas tiv thaiv nws txoj kev siv yav tom ntej. Priming ntawm cov lus teb los tiv thaiv vector kuj tshwm sim nyob rau theem tom qab nkag, thaum cov vectors raug txheeb xyuas thiab ua tiav raws li antigens lawv tus kheej. Kev tshem tawm cov ntsiab lus ntawm kev tiv thaiv kab mob zoo rau kev txhaj tshuaj tiv thaiv zoo tuaj yeem los ntawm kev siv nyiaj ntau ntxiv rau kev tiv thaiv kab mob vector. Nkag siab txog cov kev txwv uas txiav txim siab txoj hmoo ntawm Ad vectors tom qab uptake yuav muab lub sijhawm rau kev tsim vector.
4.1. Stability dag nyob rau hauv hom
Receptor-mediated uptake thiab khiav tawm ntawm endo-lysosomal compartment yog cov cim ntawm Ad kab mob thiab vector transduction. Qhov pib uptake yog txiav txim los ntawm kev sib cuam tshuam ntawm tus kab mob fiber ntau thiab penton molecules nrog cell receptors. Fiber thiab penton muaj ntau rau tag nrho cov Ads, tab sis, raws li lawv hom, lawv koom nrog sib txawv cellular receptors (Table 1). Cov kab mob C siv CAR, thiab nkag mus rau cov khoom tawm ntawm qhov chaw endosomal thaum ntxov [79]. Yog tias hom kab mob C (Ad5) raug tsim kom muaj fiber ntau molecule los ntawm hom kab mob B (Ad16), qhov tshwm sim particle (Ad5F16) ces khiav tawm ntawm lub lysosomal compartment (raws li qhia los ntawm staining nrog lysosomal marker LAMP1) es tsis txhob thaum ntxov. endosome [351]. Qhov kev ncua qeeb no los ntawm lysosomes yog ib qho kev qhia nrog txhua hom kab mob B kuaj [111,352,353].
Tsis tas li ntawd, hybrid Ad5F16 nrog rau hom B kab mob elicit muaj zog pro-inflammatory teb dua li hom kab mob C [159,351]. Qhov no yog ntaus nqi los ntawm kev tshawb pom ntxiv ntawm intraluminal TLR9 hauv lub cev tiv thaiv kab mob [354]. Tsis tas li ntawd, lysosomal membrane puas, tsis zoo li thaum ntxov endosome puas, ua rau muaj zog oxidative kev nyuaj siab. Lysosome rupture co-tso tawm lysosomal hydrolases xws li cathepsin B uas tuaj yeem ua rau mitochondria puas, yog li ua kom muaj kev puas tsuaj los ntawm kev ua kom inflammasome [355-357]. Cov txheej txheem Ad uptake destabilizes lub capsid, tso cai rau tso tawm cov kab mob protein VI [103,106]. Tso tawm cov protein VI ces nkag mus rau hauv daim nyias nyias los ntawm hauv lub endo-lysosomal compartment kom tus kab mob nkag mus rau cytosol. Sib piv hom C thiab B qhia tau hais tias ob tus kab mob no sib npaug ntawm cytosol nkag thiab genome xa, tab sis txawv ntawm lawv txoj kev khiav tawm [111]. Qhov tseeb tias kev sib pauv fiber ntau yooj yim tuaj yeem hloov pauv qhov nkag thiab cov khoom tiv thaiv kab mob los ntawm hom B mus rau hom kab mob C yog qhov zoo kawg li. Nws muab cov ntaub ntawv pov thawj ntawm lub tswv yim tias hybrid vectors nrog cov khoom tshwj xeeb tuaj yeem tsim tau. Qhov sib txawv hauv qab no zoo li tias cov protein VI tso tawm qeeb hauv hom kab mob B (lossis tus kab mob hybrid) thiab tej zaum yuav xav tau acidification lossis lwm yam, tsis paub txog kev cuam tshuam.
Txij li thaum cov protein VI tso tawm qeeb nyob rau hauv rooj plaub no, qhov no tuaj yeem cuam tshuam txog kev ruaj ntseg capsid. Tseeb tiag, hom kab mob C Ad5 tau pom tias tso tawm cov fiber ntau molecule ntawm kev khi ntawm tes, ib kauj ruam xav tias yuav ua rau lub capsid tsis muaj zog thiab ua rau cov txheej txheem disassembly rau cov protein VI tso tawm. Nyob rau hauv cov ntsiab lus no, fiberless Ad5 tau pom tias tsis tshua muaj kev ruaj ntseg dua li ib txwm hais [358]. Nyob rau hauv sib piv, tsis-matured, hyper ruaj khov Ts1 Ad2/5 tsis tso fiber ntau los yog protein VI thiab yog cov tsis zoo inducers ntawm o, vim lawv tsis ua rau TLR9 los yog ua rau cov membrane puas [114,138,359,360].
Txawm hais tias fiber ntau swapping nrog hom B tiv thaiv kev tso tawm fiber ntau thiab ua rau hom C capsid ruaj khov dua tsis tau tshawb xyuas tab sis yog qhov tshwm sim. Capsid stability tej zaum yuav yog ib qho tseem ceeb ntawm kev txiav txim siab tom qab nkag capsid txoj hmoo thiab, los ntawm kev txuas ntxiv, ib qho uas tuaj yeem siv rau kev tsim vector. Cov kab mob B hom yuav ruaj khov dua vim yog siv cov receptor sib txawv lossis los ntawm kev sib cuam tshuam fiber ntau-penton. Hauv qhov sib piv, hom F kab mob Ad40/41 kis tau rau hauv plab hnyuv thiab tau nce capsid stability vim lawv cov tropism thiab lawv hloov mus rau lub plab hnyav ib puag ncig [24,361]. Lub plab muaj qhov kub ntawm qhov nruab nrab uas yog ob peb degrees siab tshaj qhov ua pa sab sauv tsom los ntawm hom kab mob C (37 ◦C vs. 33 ◦C) [362]. Yog li, Ad40/41 yog intrinsically ntau thermostable, txhawb kev kis tus kab mob [24], ua rau lawv kuj nyuaj rau propagate [24,361,363].
Tsis tas li ntawd, lawv tsis muaj RGD motif nyob rau hauv lub penton molecule, uas ua rau ib tug ntxiv txo cell nkag [92]. Txawm hais tias cov khoom tshwj xeeb capsid no txhais tau rau hauv kev khiav tawm ntawm lysosomal compartment los yog ib nrab disassembly / protein VI tso tawm qhov tsis xws luag thaum nkag tsis paub. Hauv qhov sib piv, cov khoom no tuaj yeem muab cov ntsiab lus rau kev tsim cov tshuaj tiv thaiv qhov ncauj. Tsis zoo li hom B fibers, hom F fiber ntau-harboring vectors muaj lub cev tsis muaj zog [364] thaum siv rau cov qauv ib puag ncig. Yog hais tias ib tug tib neeg Ad hom yog tam sim no coj tawm ntawm cov ntsiab lus thiab ua hauj lwm raws li ib tug tshuaj tiv thaiv vector, nws yuav tsim nyog los saib lub ntuj tropism ntawm no vector mus nrhiav cov lus qhia los kwv yees nws tus cwj pwm. Cov tshuaj tiv thaiv kab mob tam sim no siv raws li Ad26 yog hom kab mob D, thiab nws cov tropism ntuj yog qhov muag thiab lub plab zom mov [365]. Cov kab mob D nrog rau Ad26 yog cov tshuaj tiv thaiv tsawg dua li Ad5 [366] thiab tau pom tias muaj kev khiav mus rau qhov kawg ntawm cov kab mob endosomal, tej zaum cuam tshuam nrog kev tso tawm protein VI qeeb.
Yog li ntawd, thaiv cov acidification ntawm lub endosomal compartment lig ntxiv txo nws cov immunogenicity [113]. Nyob rau hauv sib piv rau tib neeg Ads, kev paub ntawm tsis yog tib neeg Ads kuj siv nyob rau hauv kev txhaj tshuaj thiab vectorization yog heev txwv. Chimpanzee-derived Ads suav nrog ChAd3 yog ze ze rau hom D kab mob [367,368]. Interestingly, Ad5 vectors yog immunogenic ntau dua li chimpanzee-derived Ad vectors thiab muaj peev xwm hloov tau ntau dua. Qhov kev soj ntsuam no tsis tau txuas nrog qhov sib txawv ntawm penton RGD motif lossis fiber ntau, thiab nws cov laj thawj tseem ceeb tseem tsis paub [369]. Muaj peev xwm sib txawv ntawm capsid stability tsis tau tshawb xyuas. Lwm qhov tsis yog tib neeg Ad exploited li vector yog canine Ad hom 2 (CAV-2). Cov vector no muaj cov cuab yeej zoo tshaj plaws los hloov cov neurons xaiv, tab sis tsis nyob ib puag ncig glial hlwb, tsis zoo li Ad5 tswj vector, txawm tias ob leeg vectors siv tib receptor [370–372]. Qhov kev muaj peev xwm no tuaj yeem txuas rau kev nce capsid stability, tso cai rau tsis muaj coating tsuas yog hauv cov hlwb neuronal. Cov dev muaj lub cev kub uas yog ~ 2 ◦C siab dua tib neeg, uas yuav ua rau kom siab CAV-2 capsid stability [373], piv rau hom F kab mob. CAV-2 tsis zoo elicits ib qho kev tiv thaiv kab mob hauv lub cev, zoo ib yam nrog qhov tsis muaj kev cuam tshuam zoo hauv cov phiaj hlwb [372]. Yog li, CAV2 vectors yuav yog lwm qhov piv txwv uas cov cuab yeej biophysical txuas nrog nws cov tropism lossis tus tswv tsev muaj peev xwm muab cov khoom siv vector tshiab.
4.2. Fine-Tuning lub Capsid Structure, piv txwv ntawm Protein VI hauv Hom C Ad2/5
Yog tias capsid stability thiab protein VI tso tawm txiav txim siab downstream txoj hmoo ntawm tus kab mob capsid, cov lus nug tshwm sim seb qhov no puas tuaj yeem hloov kho hauv Ad vectors, piv txwv li, los muab cov khoom tshiab rau cov kab mob viral (Daim duab 3). Feem ntau ntawm peb txoj kev nkag siab txog yuav ua li cas Ads nkag mus rau endo-lysosomal compartment los ntawm hom C kab mob Ad2/5. Qhov kev tshawb pom ntawm cov protein VI raws li cov txheej txheem membrane lytic yog qhov tseem ceeb kom nkag siab txog cov txheej txheem no [103,106]. Protein VI encodes N-terminal amphipathic helix uas yog qhov tseeb membrane lytic ib feem ntawm cov protein. Thaum lub sij hawm tsim cov kab mob, cov protein VI tau qhia tias yog cov protein ua ntej uas koom nrog hexon trimers ntawm amphipathic helix [42,374]. Lub complex yog imported mus rau hauv lub nucleus los ntawm kev thauj cov teeb liab encoded nyob rau hauv lub N- thiab C-terminus ntawm protein VI, uas raug tshem tawm thaum lub sij hawm tus kab mob sib sau ua ke thiab maturation los ntawm tus kab mob protease. Lub C-terminal 11 amino acid peptide kuj ua rau cov protease, muab txoj hauv kev ntse los txuas cov kab mob sib dhos nrog kev tiv thaiv ntawm amphipathic helix. Tus kab mob Ts1 Ad2/5 mutant muaj lub ntim tsis xws luag rau cov protease, thiab tsis muaj cov capsid proteins, suav nrog cov protein VI, ua tiav. Cov khoom ua tau zoo yog qhov ruaj khov, khaws cov fibers, thiab tsis txhob siv cov protein VI.
Raws li qhov tshwm sim, Ts1 particles raug txheeb rau hauv lysosomes raws li receptor-mediated endocytosis. Qhov tseeb qhov chaw ntawm cov protein VI nyob rau hauv tus kab mob mature yog tsis meej, tab sis lub cleaved N-terminus localizes mus rau sab hauv hexon nto ntawm peripersonal exons [37,104,375]. Ob lub ntsiab lus hloov pauv hauv cov protein VI, G33A, thiab S28C, tau pom tias ua kom Ad5 capsid stability (Daim duab 3A) [376,377]. Tus thawj mutant ua ntej N-terminal protease cleavage site nyob rau hauv cov protein VI thiab impairs kev ua. Cov txiaj ntsig tshwm sim yog qee qhov tsis paub qab hau thiab yog tus cwj pwm los ntawm qhov tsis zoo ntawm endosome lysis, qhia tias N-terminus ntawm protein VI stabilizes intra-virion protein-protein kev sib cuam tshuam [376]. Qhov thib ob mutant raug xaiv rau nws lub peev xwm los tsim intra-molecular disulfide txuas hniav [377].
Tus kab mob no muaj cov proteins uas tsis tau ua tiav ib nrab, uas muaj feem cuam tshuam nrog rau cov protein VII, tab sis xav tsis thoob tias tsis muaj kev cuam tshuam txog kev kis kab mob, txawm tias nws muaj kev ruaj ntseg ntxiv. Txawm hais tias ib qho ntawm cov kab mob muaj qhov hloov pauv lub cev tiv thaiv kab mob tsis paub. Lwm qhov kev hloov pauv hauv cov protein VI uas cuam tshuam rau tus kab mob txoj hmoo yog cov protein VI L40Q kev hloov pauv, ncaj qha nyob rau hauv amphipathic helix (Daim duab 3A) [378,379]. Cov kab mob nrog qhov kev hloov pauv no ib txwm muaj tab sis qee qhov tsis zoo rau qib endosome lysis. Nyob rau hauv sib piv rau cov saum toj no-hais mutants, L40Q virions tsis tshua muaj ruaj khov dua lawv cov tsiaj qus-hom counterparts. Es tsis txhob ncua sijhawm, protein VI thiab penton raug tso tawm ntxov los ntawm L40Q capsid. L40Q virions tsis tshua muaj kab mob thiab raug rau ib nrab lysosome sorting. Txawm li cas los xij, kev hloov pauv L40Q tsis tas tshem tawm cov txheej txheem membrane, thiab qee cov kab mob khiav tawm mus rau cytosol, ua rau tag nrho ~ plaub-fold txo qis ntawm L40Q vector [379,380]. Cov kev soj ntsuam pom tau hais tias cov amphipathic helix pab nrog nws cov kev cuam tshuam rau cov particle stability. Nws tsis paub yog tias thiab yuav ua li cas L40Q kev hloov pauv hloov cov cuab yeej immunogenic ntawm Ad5 vectors. Cov kev soj ntsuam tsis ntev los no qhia tias kev tso tawm ntawm cov protein VI los ntawm capsid yog tshwm sim los ntawm kev sib tw sab hauv nrog cov protein VII rau tib yam lossis sib tshooj binding qhov chaw hauv exons [375].
Raws li txoj cai, tus kab mob mutant uas tsis muaj protein ntau VII tsis khiav tawm ntawm endosome thiab muaj qhov tsis xws luag hauv endosome lysis [58]. Kev nkag siab ntxiv ntawm cov txheej txheem thiab cov koom haum muaj zog ntawm sab hauv capsid (piv txwv li, protein VI) thiab cov proteins tseem ceeb (piv txwv li, protein V thiab VII) hauv cov particle thiab thaum lub sij hawm ntawm tes, kev nkag yuav zoo heev pab kom nkag siab txog dab tsi txiav txim siab (reversible) capsid stability.
Kev tswj hwm kev ua haujlwm ntawm kev tiv thaiv kab mob ntawm Ad vectors yog qhov kev txaus siab loj rau kev tsim cov tshuaj tiv thaiv, thiab nthuav dav MHC kev nthuav qhia ntawm antigens yuav txhim kho kev tsim tshuaj tiv thaiv [381]. Raws li tau hais dhau los, autophagy tswj kev tiv thaiv kab mob thiab pub peptides rau MHC. Autophagy yog ib qho ntawm cov txheej txheem tiv thaiv cellular tshaj plaws tiv thaiv kab mob, tsis tau Ads, raws li kev yoog rau tib neeg cov kab mob, tsis tau hloov kho cov txheej txheem kom tsis txhob autophagy thaum nkag. Hloov chaw, lawv zoo li puag autophagy activation thiab hloov lub luag haujlwm los txhawb kev kis kab mob. Protein VI plays lub luag haujlwm tseem ceeb hauv kev tswj cov lus teb autophagy ntawm tes. Adenovirus membrane lysis los ntawm cov protein VI ua rau kev nrhiav neeg ua haujlwm ntawm galectin 3 thiab galectin 8, uas tau lees paub los ntawm lub xovtooj ntawm tes raws li kev phom sij (Daim duab 3B) [108,109,382]. Kev tshawb pom ntawm qhov chaw nkag mus los ntawm galectins qhib kev xaiv autophagy hauv cov kab mob ntawm tes, raws li qhia los ntawm kev nrhiav neeg ua haujlwm ntawm autophagy receptors (xws li, NDP52 thiab p62) thiab LC3 punctae tsim [108].
Interestingly, Ad5 vector hais tswj kom khiav tawm ntawm lub ruptured endosome thiab yuav khiav mus rau lub nucleus. Inhibiting autophagy tsis cuam tshuam rau kev kis tus kab mob, tab sis ncua lawv cov genome xa mus rau lub nucleus, qhia tias Ads hijack lub cellular machinery rau lawv cov txiaj ntsig [108]. Kev khiav tawm ntawm endosome, yog li ntawd, tso cai rau tus kab mob kom tsis txhob degradation los ntawm autophagy ntawm tus txheej txheem nquag. Cov txheej txheem no tau tshawb pom los ntawm lwm cov protein VI mutant, M1 mutant. Txhua tus tib neeg Ads (thiab ntau yam tsis yog tib neeg Ads) encode ib qho kev txuag PPxY peptide motif hauv protein VI. Hauv M1 mutant, qhov motif no tau hloov mus rau PGAA (Daim duab 3) [104]. Ob hom tsiaj qus thiab M1 mutant zoo tso tawm cov protein VI thaum nkag thiab ua rau autophagy tom qab tawg ntawm endosomal membrane. Txawm li cas los xij, tus kab mob M1 mutant tsis muaj peev xwm khiav tau zoo rau hauv cytosol, thiab EM cov duab tau pom tias cov kab mob M1 mutant tau daig hauv cov kab mob ruptured endosomes, tsom los ntawm autophagy.
Yog li ntawd, tus kab mob M1 mutant tau pom tias muaj cov kab mob sib kis tau zoo, uas tau rov qab los ntawm kev siv tshuaj thiab tshuaj tua kab mob ntawm autophagy. Txoj kev tshawb no qhia tias cov kab mob qus siv lawv cov PPxY motif los nrhiav cov ubiquitin ligase NEDD4.2 thiab txwv qhov loj hlob ntawm autophagosomes ntev txaus kom khiav mus rau cytosol. Tsis tas li ntawd, cov cuab yeej no tso cai rau tus kab mob no txwv nws qhov kev nthuav qhia antigenic [108]. Qhov tseem ceeb, M1 mutant vector txhaj rau hauv nas ua rau muaj zog tiv thaiv kab mob tiv thaiv kab mob thiab txo CD8 ntxiv rau cov lus teb rau cov hloov pauv hloov pauv, qhia tias kev hloov pauv yooj yim tuaj yeem muaj kev cuam tshuam loj heev rau kev tiv thaiv kab mob ntawm Ad5 capsid. Txawm hais tias ib qho ntawm lwm cov protein VI kev hloov pauv cuam tshuam rau immunogenicity tsis paub. Txawm li cas los xij, qhov muaj PPxY motif nyob rau hauv tus kab mob L40Q tuaj yeem piav qhia tias vim li cas cov vector khaws cov kab mob sib kis tau zoo txawm tias muaj cov kab mob lysis tsis raug [379,380]. Vim tias qhov endosome lysis tsis tshua muaj txiaj ntsig, thiab L40Q vectors xaus rau hauv ib feem ntawm lysosomal compartment, nws yog qhov pom tau tias cov tshuaj tiv thaiv kab mob tiv thaiv kab mob tseem txawv ntawm hom tsiaj qus, Ts1, lossis M1 Ad5 capsids.
