RNA M6 A Reader YTHDF2 Tswj NK Cell Antitumor Thiab Antiviral Immunity Part 2

YTHDF2 yuav tsum muaj rau cov tshuaj tiv thaiv kab mob ntawm NK hlwb

Txhawm rau txiav txim siab seb Ythdf2 deficiency cuam tshuam rau cov tshuaj tiv thaiv kab mob ntawm NK hlwb, peb txhaj 2.5 × 104 PFU ntawm MCMV rau Ythdf2WT nas thiab Ythdf2ΔNK nas.

Antiviral kev ua ub no thiab kev tiv thaiv kab mob yog sib cais thiab muaj kev sib raug zoo. Cov kab mob yog cov kab mob me me, tab sis lawv tuaj yeem ua rau ntau yam kab mob thiab txawm tias tuag rau qhov mob hnyav.

Antiviral kev ua ub no hais txog lub cev lub peev xwm los tua cov kab mob. Cov neeg uas muaj cov tshuaj tiv thaiv kab mob siab tuaj yeem tshem cov kab mob sai dua thiab tiv thaiv qhov pib ntawm tus kab mob. Kev tiv thaiv kab mob yog hais txog cov tshuaj tiv thaiv uas tsim los ntawm lub cev tom qab lub sijhawm hloov kho, uas tuaj yeem tshem tawm cov teeb meem ntawm lub cev.

Kev pw tsaug zog txaus, noj zaub mov zoo, thiab kev tawm dag zog yog qhov tseem ceeb heev rau kev noj qab haus huv. Cov yam ntxwv no tuaj yeem txhim kho tib neeg kev tiv thaiv kab mob thiab yog li txhim kho lub cev tiv thaiv kab mob. Tshwj xeeb tshaj yog lub sijhawm no ntawm COVID-19 kev sib kis, nws yog ib qho tseem ceeb kom noj qab haus huv.

Tsis tas li ntawd, lub siab ruaj khov kuj muaj txiaj ntsig zoo rau kev tiv thaiv kab mob. Tus cwj pwm zoo tuaj yeem ua rau tib neeg xav tias lub cev thiab lub hlwb xis nyob, yog li txhim kho kev tiv thaiv kab mob thiab tiv thaiv kab mob. Ntawm qhov tsis sib xws, kev xav tsis zoo yuav txo lub cev tiv thaiv kab mob thiab ua rau lub cev muaj kev tiv thaiv kab mob ntau dua.

Nyob rau hauv luv luv, antiviral kev ua si thiab kev tiv thaiv kab mob yog sib cuam tshuam, thiab peb yuav tsum xyuam xim rau kev txhim kho lub cev tiv thaiv kab mob thiab antiviral kev ua si nyob rau hauv ntau txoj kev los xyuas kom meej lub cev noj qab haus huv thiab noj qab nyob zoo txoj kev ua neej. Nws tuaj yeem pom tias peb yuav tsum txhim kho kev nco, thiab Cistanche deserticola tuaj yeem txhim kho kev nco, vim Cistanche deserticola muaj antioxidant, tiv thaiv kev mob, thiab tiv thaiv kev laus, uas tuaj yeem pab txo qis oxidation thiab inflammatory tshwm sim hauv lub hlwb, yog li tiv thaiv cov kev noj qab haus huv ntawm lub paj hlwb. Tsis tas li ntawd, Cistanche deserticola kuj tseem tuaj yeem txhawb kev loj hlob thiab kho cov paj hlwb, yog li txhim kho kev sib txuas thiab kev ua haujlwm ntawm neural networks. Cov teebmeem no tuaj yeem pab txhim kho kev nco, kev kawm, thiab kev xav nrawm, thiab tseem tuaj yeem tiv thaiv kev loj hlob ntawm kev paub tsis meej thiab kab mob neurodegenerative.

Nyem paub cov tshuaj ntxiv los txhawb kev nco

Cov txiaj ntsig tau pom tias Ythdf2ΔNK nas muaj qhov cuam tshuam rau MCMV kab mob ntau dua, raws li pom los ntawm qhov hnyav poob thiab nce cov kab mob kis hauv cov ntshav, tus po, thiab daim siab piv nrog Ythdf2WT nas (Fig. 3, A thiab B; thiab Fig. S2, A thiab B). .

Peb kuj tau pom qhov txo qis ntawm feem pua ​​​​thiab cov naj npawb ntawm tag nrho NKcells hauv cov poov xab thiab cov ntshav ntawm Ythdf2ΔNK nas piv nrog cov nas hauv Ythdf2WT nas tom qab kis kab mob (Fig. 3, C thiab D; thiab Fig.S2, C thiab D). Kev soj ntsuam ntxiv tau pom tias NK hlwb los ntawm Ythdf2ΔNK nas muaj qhov qhia tau qis ntawm Ki67 dua Ythdf2WT nas (Fig. 3, E–G).

Txawm li cas los xij, NK cell viability wassimilar ntawm Ythdf2WT nas thiab Ythdf2ΔNK nas, raws li qhia byannexin V staining (Fig. S2 E). Cov ntaub ntawv no qhia tau hais tias qhov tsis muaj peev xwm ntawm YTHDF2 hauv NK hlwb ua rau muaj kev cuam tshuam hauv cell proliferation es tsis yog cell ciaj sia thaum kis kab mob.

NKcells inhibit MCMV kab mob los ntawm cov activating receptorsLy49H thiab Ly49D, thiab cov txheej txheem yog tus cwj pwm los ntawm perforin- los yog IFN- -mediated antiviral teb (Arase li al., 2002; Lee et al., 2009; Loh et al., 2005; Orret. al., 2010; Sumaria et al., 2009). Peb pom tias Ythdf2ΔNK nas tau txo qisLy49H+ thiab Ly49D+ NK hlwb hauv cov poov xab thiab ntshav piv nrog Ythdf2WT nas tom qab kis kab mob (Fig. 3, H–J; thiab Fig. S2, F–K).

Kev soj ntsuam ntxiv tau pom tias txawm tias ib feem pua, tsuas yog Ly49D + Ly49H + hlwb pom qhov sib txawv hauv Ythdf2ΔNK cov nas piv nrog cov Ythdf2WT nas, cov xov tooj ntawm Ly49D-Ly49H+ NK hlwb, Ly49D + Ly49H- NK9 cov hlwb, thiab LyNKH + Pleen thiab ntshav tau poob qis hauv Ythdf2ΔNK nas piv nrog Ythdf2WTmice tom qab kis kab mob (Fig. S2, L–W).

Peb cov ntaub ntawv qhia tias kev tswj hwm tus kab mob MCMV los ntawm YTHDF2 zoo li feem ntau yog kho los ntawm Ly49D + Ly49H + NK hlwb. Cov granzyme B thiab IFN- ntau lawm los ntawm NK hlwb hauv Ythdf2ΔNK nas yog piv rau cov nas Ythdf2WT (Fig. S2, X thiab Y).

Peb pom tau tias txo qis perforin ntau lawm los ntawm Ythdf2ΔNK nas piv nrog cov nas Ythdf2WT cov nas nyob rau hauv ob qho tib si poov xab thiab cov ntshav 7 d tom qab kis kab mob (Daim duab 3, K–M), qhia tias YTHDF2 feem ntau cuam tshuam rau perforin-mediated antiviral kev ua si tiv thaiv MCMV hauv NK hlwb.Cov ntaub ntawv no qhia YTHDF2 yog qhov tseem ceeb rau NK cell expansion thiab effector function thaum MCMV kab mob.

YTHDF2 tswj NK cell homeostasis thiab davhlau ya nyob twg maturation ntawm lub xeev khov kho

Cov kev tshawb pom saum toj no hais tias Ythdf2 tsis muaj peev xwm hauv NK hlwb txhim kho qog nqaij hlav metastases thiab ua rau NK cell tsis muaj peev xwm tswj tau tus kab mobMCMV txhawb kom peb tshawb xyuas seb YTHDF2 puas xav tau rau NK cell tu ntawm lub xeev khov kho.

Raws li pom hauv daim duab 4 A, qhov zaus thiab tus naj npawb ntawm NK hlwb tau txo qis hauv cov ntshav peripheral, spleen, siab, thiab ntsws tab sis tsis nyob hauv cov pob txha (BM) ntawm Ythdf2ΔNK nas piv nrog Ythdf2WT nas.

Txawm li cas los xij, tsis muaj kev hloov pauv tseem ceeb ntawm cov kab mob lymphoid progenitor, pre-NK cell progenitor, thiab refined NK cell progenitor (NKP) hauv BM (Fathman li al., 2011) ntawm Ythdf2WT thiab Ythdf2ΔNK nas (Fig. S3 A), qhia tias YTHDF2 yuav tsis cuam tshuam rau NK cell kev loj hlob ntxov hauv peb tus qauv. Txhawm rau tshawb xyuas cov txheej txheem muaj peev xwm ua lub luag haujlwm rau kev txo qis ntawm NK hlwb hauv Ythdf2ΔNK nas, peb tshawb xyuas cellproliferation, kev muaj peev xwm, thiab kev lag luam muaj peev xwm ntawm NK hlwb tom qabYthdf2 tshem tawm ntawm lub xeev khov kho.

Qhov feem pua ​​​​ntawm cov hlwb proliferatingNK tau sib piv ntawm Ythdf2WT thiab Ythdf2ΔNK nas, raws li pov thawj los ntawm Ki67 staining (Fig. S3 B). Lub peev xwm ntawm NKcells kuj sib npaug ntawm Ythdf2WT thiab Ythdf2ΔNK nas, raws li qhia los ntawm annexin V staining (Fig. S3 C).

Txhawm rau txheeb xyuas seb YTHDF2 puas cuam tshuam rau kev tawm ntawm NK hlwb los ntawm BM mus rau qhov chaw, Ythdf2WT thiab Ythdf2ΔNK nas tau txhaj tshuaj iv nrog cov tshuaj tiv thaiv antiCD45 los cim cov kab mob tiv thaiv kab mob thiab txi tom qab 2 min, thiab lawv cov BM hlwb raug tshuaj xyuas. Qhov no tso cai rau peb kom muaj nuj nqis ntawm NK hlwb nyob rau hauv lub sinusoidal piv rau parenchymal cheeb tsam ntawm BM, qhov taw qhia ntawm NK cell lag luam los ntawm BM mus rau peripheral ntshav nyob rau hauv ib tug khov kho lub xeev (Leong li al., 2015).

Cov txiaj ntsig tau pom qhov txo qis hauv qhov zaus ntawm CD45+ NK hlwb hauv Ythdf2ΔNK nas piv nrog Ythdf2WT nas hauv sinusoids (Daim duab 4 B), qhia tias Ythdf2 deficiency impairs lub egress ntawm NK hlwb los ntawm BM mus rau lub circulation system nyob rau hauv vivo. .Immune cells undergo homeostatic proliferation duringlymphopenia induced by some viral infections los yog los ntawm kev kho mob (Sun et al., 2011).

Txawm hais tias peb pom tias YTHDF2 yog qhov siv tau rau NK cell proliferation ntawm qhov ruaj khov, peb tau pom qhov txo qis hauv cell proliferation thaum kis MCMV (Fig. 3, E-G). Yog li peb tau tshawb xyuas lub luag haujlwm ntawm YTHDF2 hauv kev tswj hwm NK cell homeostaticproliferation hauv qhov chaw lymphopenic hauv vivo. Peb tau hloov pauv tus lej sib npaug ntawm splenic NK hlwb los ntawm CD45.2 Ythdf2ΔNK nas lossis CD45.1 congenic nas intolymphocyte-deficient Rag2−/−Il2rg-/- nas.

Cov txiaj ntsig tau pom tias feem ntau ntawm NK hlwb tau muab los ntawm CD45.1WT tswj nas ntau dua li ntawm CD45.2 Ythdf2ΔNK nas nyob rau hnub 3 tom qab hloov pauv ntawm tes (Fig. S3 D). Kev soj ntsuam ntxiv tau pom tias qhov txo qis ntawm NK hlwb los ntawm Ythdf2ΔNK nas yog vim muaj kev cuam tshuam ntawm cell proliferation (Fig. S3 E) tab sis tsis yog cell apoptosis (Fig S3 F), qhia tias YTHDF2 tsav NK cell homeostaticproliferation hauv vivo nyob rau hauv cov mob lymphopenic.

Ntxiv qhov sib txawv ntawm murine NK hlwb tuaj yeem muab faib ua qhov tsis paub qab hau (CD11b-CD27+), nruab nrab mature (CD11b + CD27+), thiab qhov kawg paub tab (CD11b + CD27−) theem raws li CD11b thiab CD27 qib ( Chiossone li al., 2009; Geiger and Sun, 2016).

Peb pom tias Ythdf2 kev nthuav qhia tau nce nrog kev loj hlob thiab CD11b-CD27+, CD11b + CD27+, thiab CD11b + CD27- tso tawm qhov qis tshaj, nruab nrab, thiab siab tshaj plaws qhia theem ntawm Ythdf2, raws li (Fig. S3 G ), qhia tias YTHDF2 tuaj yeem koom nrog NK cell maturation. Yog li peb tau tshawb xyuas lub luag haujlwm ntawm YTHDF2 hauv NK cell maturation txhais los ntawm cell surfacemarkers CD11b thiab CD27.

Peb pom tias poob ntawm Ythdf2 hauv NKcells ua rau muaj qhov txo qis hauv qhov zaus ntawm qhov kawg ntawm NK hlwb thiab / lossis nce hauv cov hlwb tsis paub tab thiab nruab nrab ntawm cov laus NK hlwb hauv tus po, siab, ntsws, thiab ntshav tab sis tsis nyob hauv BM (Fig. 4 C thiab Fig. S3 H), qhia tias YTHDF2 zoo tswj lub davhlau ya nyob twg NK cell maturation.Ua raws li cov ntaub ntawv no, cov qib ntawm KLRG1, uas yog atherminal NK cell maturation marker, tau txo qis hauv Ythdf2ΔNK nas nyob rau hauv tus po, siab, thiab ntsws tab sis tsis BM piv nrog uas ntawm Ythdf2WT nas nyob rau hauv cov kab mob sib xws los yog cov ntaub so ntswg compartments (Fig. 4 D).

Txhawm rau txiav txim siab seb qhov txo qis ntawm cov laus NK hlwb los ntawm Ythdf2 deficiency iscell intrinsic, peb tsim chimeras hauv Rag2−/−Il2rg−/− nas los ntawm txhaj BM hlwb los ntawm CD45.1 WT thiab CD45.2 Ythdf2ΔNK nas, sib xyaw ntawm 1: 1 piv.

Raws li tau pom los ntawm kev ntws cytometry ntawm 8 wk tom qab hloov pauv, qhov txo qis ntawm lub davhlau ya nyob twg NK hlwb tau muab los ntawm CD45.2 Ythdf2ΔNK BM hlwb dua li ntawm CD45.1 WT tswj hlwb (Daim duab 4 E), qhia tias NK cellterminal maturation tswj los ntawm YTHDF2. yog cell intrinsic.T-box transcription yam Eomes thiab Tbet yog qhov tseem ceeb rau NKcell maturation (Daussy li al., 2014; Gordon li al., 2012).

Intracellular staining qhia qhov txo qis ntawm cov protein ntau ntawm Eomes hauv NK hlwb los ntawm Ythdf2ΔNK nas piv nrog Ythdf2WT nas (Fig. S3 I). Tsis tas li ntawd, peb pom tias qhov txo qis ntawm cov protein thiab mRNA qib ntawm Eomes tshwj xeeb tshwm sim nyob rau hauv cov laus laus (CD11b + CD27−) NK hlwb (Fig. S3,J–L).

Txawm li cas los xij, nyob rau hauv sib piv rau Eomes, qhov kev qhia ntawm Tbet yog sib npaug hauv NK hlwb ntawm Ythdf2ΔNK thiab Ythdf2WT nas (Fig. S3, I thiab K), qhia tias YTHDF2 tejzaum nws tswj NKcell davhlau ya nyob twg maturation los ntawm lub hom phiaj Eomes.

YTHDF2 txhawb nqa NK cell effector muaj nuj nqi

Peb qhov kev tshawb pom tias Ythdf2 tsis muaj peev xwm cuam tshuam NK cell homeostasisand maturation txhawb peb los txiav txim siab seb YTHDF2 tseem cuam tshuam NK cell effector function. Lub tshuab tseem ceeb uas tswj hwm NK cell muaj nuj nqi yog qhov txheeb ze ntawm inhibitory thiab activating receptors (Chan li al., 2014).

Peb tau txheeb xyuas cov theem qhia ntawm cov molecules cuam tshuam nrog NKcell ua kom lossis inhibition. Peb pom tias qhov kev qhia theem ntawm cov activating receptors CD226 thiab NKG2D, tab sis tsis yog CD69, tau txo qis hauv splenic NK hlwb los ntawm Ythdf2ΔNK nas piv nrog cov los ntawm Ythdf2WT nas (Fig. 5 A), whereas theinhibitory receptors NKG2A, TIGIT, thiab PD{ 7}} tau zoo sib xws ntawm Ythdf2ΔNK nas thiab Ythdf2WT nas (Fig. 5 A thiab Fig.S3 M).

Tsis tas li ntawd, Ythdf2ΔNK NK hlwb tsim tawm tsis zoo IFN- dua li Ythdf2WT NK hlwb thaum txhawb nrog IL-12plus IL-18 lossis nrog YAC-1 nas lymphoma hlwb (Fig. 5, B, thiab C. ).

Peb tom qab ntawd kuaj lub peev xwm cytotoxic ntawm Ythdf2ΔNK NK hlwb. Peb pom tias Ythdf2ΔNK NK hlwb tau txo qis cytotoxicity tawm tsam MHC chav kawm I-tsis muaj nas lymphoma celllines RMA-S (Fig. 5 D) tab sis pom zoo li cytotoxicity tawm tsamMHC chav kawm I- txaus RMA hlwb (Fig. S3 N) raws li qhia los ntawm 51Crassay. . Ua ke, cov kev tshawb pom no qhia tau tias YTHDF2 yog qhov tseem ceeb rau NK cell effector function.

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