IL-38 Tiv thaiv Induction Ntawm Kev Tiv Thaiv Kev Tiv Thaiv Los Ntawm Inhibition Ntawm MTOR Signaling
Mar 06, 2023
Abstract:
Kev tiv thaiv kab mob yog qhov tau txais cov tshuaj tiv thaiv kab mob siab tshaj plaws los ntawm lub cev tiv thaiv kab mob (xws li monocytes thiab macrophages) tom qab kis kab mob lossis txhaj tshuaj tiv thaiv, lub cim xeeb de facto nonspecific nyob ntawm epigenetic thiab metabolic reprogramming ntawm cov hlwb. Peb tsis ntev los no tau qhia tias kev txhawb nqa kev tiv thaiv kev tiv thaiv yog nyob ntawm IL-1 .
Ntawm no, peb qhia tau hais tias recombinant IL-38, ib qho tshuaj tiv thaiv kab mob cytokine ntawm IL-1- tsev neeg, muaj peev xwm ua rau muaj kev hloov pauv mus sij hawm ntev thiab txo qhov induction ntawm kev tiv thaiv kab mob los ntawm -glucan hauv vivo hauv C57BL/6 nas thiab ex vivo hauv lawv cov pob txha hlwb. IL-38 thaiv mTOR signaling thiab tiv thaiv cov kev hloov pauv ntawm epigenetic thiab metabolic tshwm sim los ntawm -glucan. Hauv cov ntsiab lus noj qab haus huv, IL1F10- koom nrog ib leeg nucleotide polymorphism rs58965312 cuam tshuam nrog cov ntshav plasma ntau dua IL-38 thiab txo qis induction ntawm kev tiv thaiv kab mob los ntawm -glucan ex vivo. Cov txiaj ntsig no qhia tau hais tias IL-38 ua rau muaj kev hloov pauv mus ntev los tiv thaiv kab mob thiab tseem inhibits qhov induction ntawm kev tiv thaiv kab mob. Recombinant IL-38 tuaj yeem yog li ntawd tuaj yeem siv los ua kev kho mob rau cov kab mob uas ua rau muaj kev tiv thaiv kab mob hnyav dua.
KEYWORDS
ethanol, mob, hlawv, me RNAs
Nyem rau cistanche tubulosa pdf khoom
1 QHOV TSEEB
Ntau cov kev tshawb fawb tau pom tias monocytes thiab macrophages tuaj yeem tau txais kev hloov pauv mus sij hawm ntev, qhov tseeb ntawm lub cim xeeb uas tau raug hu ua "kev tiv thaiv kab mob." 1-3 Kev hloov kho ntawm cov hlwb myeloid yog kho los ntawm epigenetic thiab metabolic rewiring. , suav nrog ntawm lawv cov pob txha pob txha progenitors.4–7 Kev cob qhia kev tiv thaiv tuaj yeem raug ntxias los ntawm cov tshuaj tiv thaiv xws li bacillus CalmetteGuerin (BCG) uas tiv thaiv Mycobacterium tuberculosis, los yog immunomodulators xws li -glucan, ib hom fungal cell phab ntsa tivthaiv.8,9 Lub rewiring ntawm cov epigenome ntawm cov hlwb myeloid feem ntau ua rau muaj kev hloov pauv mus ntev xws li kev tiv thaiv tus tswv tsev tiv thaiv kab mob tsis sib xws.1,10 Piv txwv li, kev txhaj tshuaj BCG ntawm cov menyuam mos hauv West Africa txo qis kev tuag los ntawm kev kis kab mob, 11 thiab kev txhaj tshuaj BCG hauv cov neeg laus noj qab haus huv txo qis tom qab daj. Cov tshuaj tiv thaiv ua npaws viremia thiab kab mob parasitemia hauv cov qauv ntawm tib neeg kev sim malaria, uas cuam tshuam nrog ntau npaum li cas ntawm BCG-dependent epigenetic hloov pauv hauv cov kab mob sib kis. Lub xeev tau mus txog, uas ua rau txo cov neeg tuag los ntawm, piv txwv li, fungal thiab Staphylococcus aureus sepsis.7,9
IL-1 tsev neeg muaj 11 tus tswv cuab13,14; Ntawm cov uas IL-37 thiab IL-38 muaj cov tshuaj tiv thaiv kab mob dav dav.15,16 Yav dhau los, peb tau tshaj tawm tias IL-1 yog qhov tseem ceeb hauv kev txhawb nqa kev tiv thaiv kab mob12 thiab IL{{ 11}} inhibits qhov induction ntawm kev kawm tiv thaiv kab mob.17 Tsis tas li ntawd, IL-1 tau pom tias yog cov cytokine feem ntau nce ntxiv hauv cov pob txha ntawm -glucan-kho nas thiab ua rau induction ntawm kev cob qhia kev tiv thaiv.7 Nyob rau hauv vitro qauv ntawm kev cob qhia kev tiv thaiv kab mob, thawj tib neeg monocytes teb rau IL-1 nrog cov kev hloov pauv hauv epigenetic uas txhawb kev tsim cov cytokines xws li TNF thiab IL-6.12 Tsis tas li ntawd, A allele ntawm tib nucleotide polymorphism ( SNP) rs16944 hauv thaj tsam txhawb nqa ntawm IL1B noob tau cuam tshuam nrog kev nce ntxiv ntawm kev kawm tiv thaiv kab mob hauv tib neeg monocytes.12
Peb tau tshaj tawm yav dhau los tias IL-38 tuaj yeem cuam tshuam IL-1 , IL-6, thiab KC hauv tus qauv ntawm gouty mob caj dab.18 Raws li IL-38 tuaj yeem thaiv IL{{5 }} , peb xav tias IL-38 muaj lub luag haujlwm inhibitory hauv kev txhawb nqa kev tiv thaiv kab mob. Hauv tsab ntawv tshaj tawm no, peb tsom los txheeb xyuas lub luag haujlwm ntawm IL-38 hauv -glucan induced trained immunity. Peb qhia tau tias IL-38 inhibits induction ntawm kev cob qhia kev tiv thaiv kab mob hauv murine qauv los ntawm kev cuam tshuam lub mTOR signaling cascade, uas yog ib qho tseem ceeb rau induction ntawm kev cob qhia kev tiv thaiv kab mob.5 Hauv cov tib neeg noj qab haus huv, peb qhia tau hais tias SNP rs58965312 cuam tshuam IL{{ 12}} qib ntshav plasma thiab kev kawm tiv thaiv kab mob hauv vitro.
2 Cov Khoom Siv thiab Cov Txheej Txheem
2.1 Mus
Xya rau cuaj lub lis piam txiv neej C57Bl/6 nas tau txais los ntawm Jackson Laboratories (Bar Harbor, ME, USA). Cov nas tsuag tau pub kom tsis muaj menyuam hauv chav kuaj chow thiab dej ad libitum. Cov kev sim no tau pom zoo los ntawm Lub Tsev Haujlwm Saib Xyuas Tsiaj thiab Siv Pawg ntawm University of Colorado Denver (Aurora, CO, USA).
2.2 Kev cob qhia kev tiv thaiv tus qauv
Cov nas tau txais 1 ug ntawm bioactive18,19 human recombinant IL-38 (3-152) (Biotechne, Minneapolis, MN, USA) ip hauv sterile saline nyob rau hauv 200 ul ntim lossis tsuas yog saline rau peb hnub sib law liag. Thawj hnub, 1 teev tom qab txhaj tshuaj IL-38, nas tau txais ip Tag nrho ntawm 1 mg ntawm -glucan (1,3-(D)-glucan) tau ua siab zoo los ntawm xibfwb David Williams ( College of Medicine, Johnson City, TN, USA) lossis 200 ul saline raws li kev tswj xyuas tsheb. Hnub thib tsib, cov nas tau raug txi ncaj qha lossis thawj zaug nrog 5 mg / kg LPS (Escherichia coli[055:B5] Sigma-Aldrich, St. Louis, MO, USA) ip hauv 200 ul sterile saline 4 h ua ntej txi. Nas lub cev kub tau ntsuas nrog MT4 tus pas ntsuas kub (RayTek MiniTemp, Wilmington, NC, USA). Tom qab tshuaj loog nrog isoflurane, cov ntshav los ntawm orbital plexus tau sau rau hauv EDTA, thiab cov nas tau txi los ntawm lub ncauj tsev menyuam. Ntshav tau centrifuged ntawm 1000 ×g rau 10 feeb los npaj cov ntshav rau kev ntsuas cytokine.
2.3 Hauv vitro cytokine ntau lawm
Cytokine ntau lawm tau soj ntsuam ex vivo nyob rau hauv tag nrho cov ntshav, pob txha hlwb hlwb, thiab splenocytes.
Tag nrho cov ntshav tau diluted 1: 4 zaug hauv RPMI 1640. Tag nrho ntawm 200 ul ntim tau plated nyob rau hauv 96-zoo puag ncig hauv qab daim hlau thiab incubated rau 24 h tsis muaj stimulus los yog 100 ng / ml LPS (± 10 μM Nigericin [Invivogen, San Diego, CA, USA] thaum kawg teev) ntawm 37◦C hauv 5 feem pua CO2. Tom qab 24 teev, 180 ul ntawm supernatant tau sau thiab cov cell pellet tau khaws cia hauv 0.5 feem pua Triton X-100 (Sigma, St. Louis, MO, USA). Femoral pob txha pob txha raug tshem tawm hauv RPMI kab lis kev cai nruab nrab, suav rau HemaTrue cell txee (Heska, Loveland, CO, USA), thiab hloov kho rau 1 × 106 / ml. Tag nrho ntawm 200 ul ntawm lub cell ncua kev kawm tau plated nyob rau hauv 96-zoo puag ncig hauv qab daim hlau thiab incubated rau 24 teev nyob rau hauv RPMI (tsis muaj kev loj hlob yam ntxiv) nyob rau hauv tib yam mob raws li tau piav ua ntej. Spleens raug tshem tawm aseptically, homogenized los ntawm 100 μm cell strainers (Thermo Fisher Scientific, Waltham, MA, USA), thiab sau hauv RPMI. Cells raug suav thiab kho rau 5 × 106 / ml. Tag nrho ntawm 200 ul ntawm lub cell ncua kev kawm ntawv tau plated nyob rau hauv 96-zoo puag ncig hauv qab daim hlau thiab incubated rau 24 teev los yog 3 d nyob rau hauv RPMI muaj 10 feem pua FBS nyob rau hauv tib yam mob raws li tau piav ua ntej.
2.4 Kev ntsuas Cytokine los ntawm ELISA thiab ntsuas lactate
Supernatants thiab plasma tau khaws cia ntawm −20◦C kom txog thaum ntsuas ntxiv. ELISA tau ua raws li cov chaw tsim khoom cov lus qhia (BioTechne), tshwj tsis yog IL-38 ELISA ntawm tib neeg cov ntshav tau ua raws li tau piav qhia lwm qhov.20
Mouse plasma thiab pob txha pob txha tau txais raws li tau piav qhia thiab qib lactate tau txiav txim siab los ntawm cov khoom lag luam Lactate Fluorometric Assay (Biovision, Milpitas, CA, USA).
2.5 Western blot
Cov pob txha pob txha los ntawm nas naïve tau sau raws li tau piav ua ntej. Tag nrho ntawm 1 × 106 hlwb raug coj mus rau hauv ib qho 12-zoo tiaj tiaj-hauv qab phaj thiab incubated rau 3 d hauv RPMI uas muaj 10 feem pua serum. Lub supernatant nrog cov hlwb tsis sib txuas tau raug tshem tawm. Tom qab 4 h incubation (uas yog qhov zoo rau mTOR phosphorylation) nrog lossis tsis muaj 5 ug / ml -glucan ± 100 ng / ml IL-38 hauv RPMI, cov hlwb tau lysed hauv RIPA tsis muaj cov tshuaj tiv thaiv protease (Roche, Basel, Switzerland. ). Tag nrho cov ntsiab lus ntawm cov protein tau txiav txim los ntawm BCA assay (Thermo Fisher Scientific), thiab sib npaug ntawm cov proteins raug rau SDS-PAGE ntawm pre-casted 4-15 feem pua gels (Biorad, Hercules, CA, USA).
Cov protein sib cais tau raug xa mus rau nitrocellulose membrane (Biorad), uas tau thaiv hauv 5 feem pua BSA (Sigma) thiab incubated hmo ntuj ntawm 4◦C nrog luav polyclonal antibodies tiv thaiv (phospho) mTOR, Akt, 4EBP1, thiab S6K (Cell Signaling, Danvers, MA, USA), thiab -actin (Sigma), uas tau pom pom siv polyclonal secondary antibody (Dako, Leuven, Belgium) thiab SuperSignal West Femto Substrate (Thermo Fisher Scientific) rau cov phosphorylated proteins lossis ECL substrate rau lwm cov proteins. (Biorad).
2.6 mRNA rho tawm thiab RT-PCR
mRNA los ntawm nas pob txha pob txha raug rho tawm nrog Trizol (Life Technologies, Carlsbad, CA, USA) chloroform (Sigma) extraction, ua raws li isopropanol nag lossis daus (Sigma) thiab ob ntxuav nrog 70 feem pua ethanol (Sigma). cDNA tau tsim los siv iScript thim rov qab transcriptase (Invitrogen, Carlsbad, CA, USA). Cov txheeb ze qhia tau txiav txim siab siv SYBR Green txoj kev (Invitrogen) ntawm CFX96 Bio-Rad qPCR tshuab (Biorad), thiab cov txiaj ntsig tau nthuav tawm raws li kev sib tw nce hauv mRNA qib, txheeb ze rau cov nas tswj tsheb, nrog B2M raws li kev tu vaj tse. Primers muaj npe nyob rau hauv Kev Txhawb Cov Ntaub Ntawv Table S1.
2.7 Chromatin immunoprecipitation
Cov pob txha pob txha raug cais raws li tau piav ua ntej. Cells tau kho nyob rau hauv 1 feem pua methanol-dawb formaldehyde thiab khaws cia ntawm 4◦C. Tom qab ntawd, cov hlwb raug sonicated, thiab immunoprecipitation tau ua los ntawm kev siv tshuaj tiv thaiv H3K4me3 (Diagenode, Seraing, Belgium), raws li tau piav qhia nyob rau lwm qhov.6 DNA raug cais nrog MinElute PCR purification kit (Qiagen, Germantown, MD, USA) thiab tau ua tiav ntxiv rau qPCR tsom xam siv SYBR ntsuab txoj kev ntawm Step-one PLUS qPCR tshuab (Applied Biosciences, Foster City, CA, USA). Cov qauv raug soj ntsuam los ntawm kev sib piv Ct raws li cov chaw tsim khoom cov lus qhia. Cov epigenetics primers tau teev nyob rau hauv Kev Txhawb Cov Ntaub Ntawv Table S1.
2.8 PBMC kev sim
Txoj kev tshawb no tau ua nyob rau hauv ib pawg ntawm ±200 cov neeg noj qab haus huv ntawm Western European caj ces los ntawm Human Functional Genomics Project (200FG pawg, saib www.humanfunctionalgenomics.org). PBMCs raug cais tawm ntawm kev tso cai rau cov neeg noj qab haus huv raws li tau piav qhia ua ntej.21 Kev npaj cov ntshav rau IL-38 ELISA thiab hauv vitro stimulations ntawm thawj monocyte stimulations hauv pawg no tau piav qhia lwm qhov.22 Monocytes tau kawm nrog -glucan rau 24 teev, ntxuav nrog PBS sov, thiab so hauv RPMI rau 6 d.23 Tom qab ntawd, cov hlwb raug txhawb nrog 10 ng / ml LPS. Tom qab 24 teev, supernatants tau sau thiab khaws cia ntawm −20◦C txog IL-6 thiab TNF tau ntsuas los ntawm ELISA. Plasma IL-38 qib tau txiav txim los ntawm ELISA. Cov xwm txheej ntawm IL-38 SNP rs58965312 raug soj ntsuam raws li tau piav qhia hauv cov txheej txheem ntawm Li li al.22 los ntawm cov khoom lag luam muaj SNP nti, Illumina HumanOmniExpressExome-8 v1.0.
2.9 Kev txheeb xyuas
Qhov sib txawv ntawm ntau yam xwm txheej tau txheeb xyuas nrog Wilcoxon matched-pairs kos npe rau qib xeem lossis Mann-Whitney U-test raws li qhov tsim nyog. Cov ntaub ntawv raug nthuav tawm raws li txhais tau tias ± SEM tshwj tsis yog qhia lwm yam. Cov ntaub ntawv raug tshuaj xyuas siv GraphPad Prism 8.0 (GraphPad Software, La Jolla, CA, USA). P-tus nqi hauv qab no 0.05 tau suav tias yog qhov tseem ceeb.
3 TSEEM CEEB
3.1 IL-38 inhibits LPS-induced o
Cov kev tshawb fawb no hais txog lub luag haujlwm ntawm IL-38 ntawm kev kawm tiv thaiv kab mob pib nrog kev soj ntsuam ntawm cov nyhuv ntawm recombinant tib neeg IL-38 ntawm cov kab mob hauv lub cev vim endotoxemia. Cov nas tau txais lub tsheb lossis 1 ug ntawm IL-38 ip rau hnub 4, 3, thiab 2 (Daim duab 1A). Tom qab 4 d, LPS (5 mg / kg) tau muab ip rau txhua tus nas, thiab 4 h tom qab cov nas tau txi. Raws li pom nyob rau hauv daim duab 1B, cov nas uas tau txais IL-38 pom muaj tsawg dua hypothermia piv rau cov nas uas kho lub tsheb, qhia tias txo qhov mob.24 Plasma IL-1 , TNF , thiab IL-6 concentrations tau txo qis. nyob rau hauv cov nas kho nrog recombinant IL-38 piv rau cov nas kho tsheb (Fig. 1C), whereas IL-1 concentration hauv lysates ntawm tag nrho cov ntshav tsis cuam tshuam los ntawm IL-38.
Hauv qhov sib piv, tsis muaj qhov loj IL-38- nyob ntawm kev hloov pauv ntawm cov xov tooj ntawm tes hauv cov pob txha, splenocytes, thiab tag nrho cov ntshav tau pom, sib nrug los ntawm kev txo qis hauv lymphocytes hauv cov ntshav tag nrho (cov ntaub ntawv tsis qhia). Tag nrho cov ntshav, pob txha pob txha, thiab splenocytes tau ntxiv rau hauv vitro rau 24 teev yam tsis muaj kev txhawb nqa ntxiv. Peb tau pom tias txo qis kev tso tawm ntawm TNF (75 feem pua qis) thiab IL-6 (25 feem pua qis) los ntawm tag nrho cov ntshav kab lis kev cai ntawm nas kho nrog IL-38 (Daim duab 1D saum toj). Nyob rau hauv kab lis kev cai pob txha pob txha, qhov txo qis ntawm kev tso tawm ntawm TNF (55 feem pua qis) kuj tau pom (Daim duab 1D nruab nrab). Splenocytes tsis pom qhov sib txawv hauv IL-6 lossis TNF ntau lawm (Daim duab 1D hauv qab).
3.2 IL-38 inhibits -glucan-induced cob qhia kev tiv thaiv
Tom ntej no, peb tau soj ntsuam seb IL -38 kuj tseem tuaj yeem cuam tshuam qhov induction ntawm kev tiv thaiv kev tiv thaiv los ntawm -glucan. -glucan-induced trained immunity ua rau amplified cytokine ntau lawm mus rau ib qho kev sib tw thib ob thiab tsis cuam tshuam.7,9 Raws li tau piav qhia hauv daim duab 2A, peb muab cov tshuaj recombinant IL-38 (1 ug) los yog ntsev rau cov nas 1 teev ua ntej ip -glucan (1 mg) rau hnub -4. Hnub -3 thiab -2, nas tau txais 1 ug ntawm IL-38 ib hnub. Tom qab 2 d ntawm so, tag nrho cov nas tau txais 5 mg ntawm LPS. Plaub teev tom qab kev tswj hwm LPS, lub plab phab ntsa ntawm cov nas uas kho nrog -glucan raug txo los ntawm 2◦C piv rau cov nas kho nrog lub tsheb thiab normalized los ntawm kev kho mob nrog IL-38 (Fig. 2B). Hauv cov nas uas tau kho nrog -glucan thiab tom qab ntawd sib tw nrog LPS, qhov kev hloov pauv zoo sib xws ntawm -glucan los ntawm IL-38 tau pom hauv cov ntshav TNF, IL-6, thiab IL-1 (Daim duab . 2C). Raws li kev cia siab hauv ex vivo kab lis kev cai tag nrho cov ntshav (tsis muaj kev txhawb nqa ex vivo) los ntawm -glucan-kawm nas nce hauv TNF thiab IL-6 ntau lawm tau pom, uas tau raug tshem tawm los ntawm qhov sib ntxiv ntawm IL-38 ( Fig. 2D). Qhov cytokine profile no tau tshwm sim hauv cov kab lis kev cai splenocyte thiab-txawm tias tsis yog qhov tseem ceeb hauv cov pob txha (Fig. 2D).
3.3 IL-38 tiv thaiv cov kab mob epigenetic histone hloov los ntawm -glucan
Raws li qhov nce siab ntawm cov cytokines inflammatory cytokines los ntawm -glucan yog qhov tshwm sim ntawm cov kev hloov pauv hauv epigenetic ntawm qib ntawm histones, 4,9 tom ntej no peb tau soj ntsuam qhov cuam tshuam ntawm IL-38 ntawm kev hloov pauv ntawm epigenetic ntawm cov neeg txhawb nqa ntawm cov kws tsim kev tiv thaiv kab mob ntsig txog cov noob. Cov nas raug kho nrog -glucan ± IL-38 thiab txi tom qab 5 d yam tsis muaj qhov kev sib tw LPS (Fig. 3A).2,6 Thaum tag nrho cov ntshav, pob txha pob txha, thiab splenocytes tau txhawb nqa ex vivo nrog LPS nrog lossis tsis muaj inflammasome activator nigericin, nce hauv cytokine ntau lawm tau pom nyob rau hauv -glucan cob qhia nas, uas tau tshem tawm los ntawm IL-38 (Fig. 3B).
Thaum kev qhia txog cov noob muaj feem xyuam nrog kev tiv thaiv kab mob (Tnfa, Nrlp3, Hk2, thiab Pfkp) tau txiav txim siab hauv cov pob txha ntawm LPS-chaw nas nas, nce kev qhia ntawm tag nrho cov noob tau pom uas tau tiv thaiv los ntawm IL-38 (Fig. 3 C). Txhawm rau txiav txim siab seb qhov no yog qhov tshwm sim ntawm kev hloov pauv ntawm epigenetic tshwm sim los ntawm -glucan thiab IL-38, pob txha pob txha ntawm cov nas uas tsis yog-LPS kho tau ua tiav rau kev tshuaj ntsuam epigenetic. H3K4me3 (ib qho tseem ceeb histone marker hauv kev kawm tiv thaiv kab mob 4,9) cim DNA raug cais tawm, thiab feem pua ntawm cov neeg txhawb nqa ntawm cov kab mob kev tiv thaiv kab mob uas tau txais txiaj ntsig zoo rau qhov cim histone tau txiav txim siab. H3K4me3 tau pom hauv cov nas uas tau kawm glucan, thiab qhov no tau thim rov qab thaum cov nas kuj tau txais IL38 (Daim duab 3D).
3.4 IL-38 obstructs -glucan dependent Akt/mTOR/S6K signaling
Peb tau qhia yav dhau los tias kev tiv thaiv kab mob hauv monocytes nyob ntawm qhov ua kom ntawm mTOR txoj hauv kev.5 Raws li IL-38 inhibits induction ntawm kev cob qhia kev tiv thaiv, peb xav tias inhibition ntawm mTOR txoj kev yuav siv qhov chaw, raws li kuj yog lub mechanism rau IL-37 inhibition of trained immunity.17 Raws li pom nyob rau hauv daim duab 4A, qhov induction ntawm kev cob qhia kev tiv thaiv nyob rau hauv cov pob txha pob txha los ntawm naïve nas los ntawm -glucan nyob rau hauv vitro induces phosphorylation ntawm Akt, mTOR, thiab nws downstream kinases S6K thiab 4EBP1. IL -38 txo qhov phosphorylation no, qhia tias IL38 inhibits kev ua haujlwm ntawm txoj kev no.
Raws li mTOR yog lub hauv paus tswj hwm ntawm cellular metabolism, peb txiav txim siab qhov kev ua ntawm glycolysis txoj kev, uas yog heev induced nyob rau hauv -glucan kawm monocytes thiab macrophages.5 Raws li qhov kawg khoom ntawm anaerobic glycolysis, lactate qib tau ntsuas. IL-38 kev kho mob tiv thaiv kev nce qib ntawm lactate hauv plasma 4 h tom qab kev kho mob nrog LPS thiab nyob rau hauv supernatants ntawm 72 h kab mob pob txha los ntawm cov nas uas tau kawm nrog -glucan thiab txhaj nrog LPS hauv vivo (Fig. 4B, C).
3.5 SNP thiab IL-38 plasma qib kwv yees lub zog ntawm kev cob qhia kev tiv thaiv induction
Txhawm rau txheeb xyuas qhov cuam tshuam ntawm IL-38 ntawm kev kawm tiv thaiv kab mob hauv tib neeg, peb tau tshawb xyuas cov txiaj ntsig ntawm 11 SNPs cuam tshuam nrog IL-38 (IL1F10) thiab cov nyob ib sab IL-1Ra (IL{{6 }}RN) ntawm qhov induction ntawm kev cob qhia kev tiv thaiv kab mob hauv tib neeg PBMC los ntawm 118 yam kev noj qab haus huv ntawm 200FG pawg.25 Peb ntsuas cov ntshav IL-38 concentrations, thiab peb ntsuas lub peev xwm ntawm lawv cov monocytes kom tau txais kev tiv thaiv kab mob. Raws li pom hauv daim duab 5A, qhov induction ntawm IL-6 thiab TNF tom qab kev cob qhia hauv vitro nrog -glucan muaj feem xyuam rau SNP rs5896312. Tsis tas li ntawd, qib plasma ntawm IL-38 yog nyob ntawm qhov SNP no, raws li cov neeg nqa khoom ntawm ob qho kev xaiv alleles muaj qib siab dua piv rau cov ntsiab lus nqa ob qho kev siv alleles (Fig. 5B). Tsis muaj qhov sib txawv hauv plasma IL-1RA qib ntawm cov tib neeg no (cov ntaub ntawv tsis qhia). Hauv cov ntsiab lus, kev txwv tsis pub muaj kev tiv thaiv kab mob hauv cov neeg ua haujlwm noj qab haus huv tau cuam tshuam nrog ntau dua ntawm cov ntshav IL -38, uas nyob rau hauv lem yog nyob ntawm SNP rs5896312.
4 Kev sib tham
Hauv tsab ntawv tshaj tawm no, peb pom tau tias IL-38 txo qis qhov induction ntawm kev tiv thaiv kab mob los ntawm -glucan. Qhov kev soj ntsuam no zoo ib yam nrog cov tshuaj tiv thaiv kab mob ntawm IL-38 ua ib qho inhibitor ntawm innate thiab adaptive immunity.16,25 Thaum nas tau kawm nrog -glucan, kev hloov pauv ntawm epigenetic tshwm sim hauv cov hlwb ntawm lub cev tsis muaj zog, ua rau txhim kho cov lus teb proinflammatory ntawm lub cev tsis muaj zog tiv thaiv kab mob raws li qhov kev sib tw thib ob. Txhim kho kev tsim cov proinflammatory cytokines kuj ua rau muaj kev hloov pauv hauv cov kab mob hauv cov pob txha, 7 thiab txo cov neeg tuag hauv cov qauv ntawm cov kab mob C. albicans lossis S. aureus infections.8,9 Ntxiv mus, IL-1 tau pom tias ua si loj. Lub luag haujlwm hauv kev txhawb nqa kev tiv thaiv kev tiv thaiv, 7,12 uas tseem ceeb los ntawm Van der Meer li al., uas tau pom tias kev kho cov nas nrog cov tshuaj me me ntawm cov tib neeg recombinant IL-1 txo qis kev tuag ntawm cov nas granulocytopenia los ntawm cov kab mob Pseudomonas tom qab, ib lub tswv yim nyob rau lub sijhawm ntawd hu ua nonspecific resistance to infection.27 Lwm tus tswv cuab ntawm IL-1 tsev neeg, IL-37, tau pom tias inhibit NLRP3 inflammasome thiab tso tawm ntawm IL{18}} , thiab induction ntawm kev cob qhia kev tiv thaiv kab mob.17 Muab cov tshuaj tiv thaiv kab mob ntawm IL-38,28 xws li nws lub peev xwm los thaiv IL-1 upregulation hauv nas qauv ntawm gouty mob caj dab,18 peb tshawb xyuas seb IL{{ 25}} tuaj yeem thaiv qhov induction ntawm kev cob qhia kev tiv thaiv.
Ntawm no, peb qhia tau hais tias ntxiv rau kev tiv thaiv LPS-induced o nyob rau hauv nas, 29 IL -38 abrogated induction ntawm kev cob qhia kev tiv thaiv thiab cov lus teb tom qab rau ib tug thib ob stimulus nrog LPS. IL -38 kev tswj hwm inhibited -glucan-induced phosphorylation ntawm Akt/mTOR signaling pathway. Ib yam li ntawd, IL-38 tiv thaiv -glucan nyob ntawm proinflammatory epigenetic reprogramming, raws li qhia los ntawm kev txo qis hauv H3K4me3 ntawm promotors ntawm Tnfa, Nrlp3, Hk2, thiab Pfkp hauv nas pob txha. Hauv vivo kev kho mob nrog -glucan thiab IL-38 thiab kev tswj hwm LPS tom qab tau ua rau qis qis ntawm lactate hauv cov ntshav thiab hauv ex vivo cultured pob txha pob txha supernatants. Kev txo qis ntawm lactate qhia tias IL-38 thaiv glycolysis, feem ntau yuav yog los ntawm inhibition ntawm mTOR txoj hauv kev. Cov kev soj ntsuam no tau lees paub los ntawm kev txheeb xyuas ntawm pawg neeg noj qab haus huv uas SNP rs5896312 nyob ib sab ntawm cov noob encoding IL-38 cuam tshuam rau ob theem plasma IL-38 thiab inducibility ntawm kev kawm tiv thaiv kab mob hauv monocytes ex vivo. Ua ke, peb qhia tau tias qhov txo qis ntawm IL{19}} ua rau muaj kev nce ntxiv ntawm kev tiv thaiv kev tiv thaiv, thaum kev tswj hwm ntawm recombinant IL -38 inhibits cov lus teb lom neeg no.
IL-38 khi rau IL-36R, thiab IL-1R9 yog nws cov coreceptor putative.16,30 Nws tau raug tshaj tawm tias N-terminally truncated IL-38 attenuates lub JNK/AP1 txoj hauv kev uas ua rau txo qis IL-6 ntau lawm los ntawm macrophages.30 Ntawm no peb qhia tau tias IL-38 kuj inhibits txoj kev Akt/mTOR, uas yog ib qho tseem ceeb hauv induction ntawm kev cob qhia kev tiv thaiv kab mob.5 Txaus siab, inhibition ntawm mTOR txoj hauv kev kuj tau pom zoo rau IL37,31,32 uas qhia txog ib feem ntawm kev ua haujlwm zoo sib xws ntawm cov kev tiv thaiv dav dav IL-1 cov neeg hauv tsev neeg.
Dhau li lub luag haujlwm zoo ntawm kev tiv thaiv kev tiv thaiv kab mob hauv kev tiv thaiv kev kis kab mob, cov nyhuv tshem tawm tau raug pom zoo hauv cov kab mob auto-immune.3 Yog li ntawd, IL-38 yuav raug siv kho kom npau taws rau kev tiv thaiv kab mob hauv cov kab mob no. Kev cob qhia kev tiv thaiv kab mob tshwj xeeb tau qhia tshwj xeeb los ua lub luag haujlwm pathologic hauv lub cev lupus erythematosus (SLE), Sjögren's syndrome, thiab rheumatoid mob caj dab.3 Lub luag haujlwm tiv thaiv ntawm IL-38 tau npaj rau hauv SLE, 26 hauv cov nas raug kuaj mob caj dab, 33 nyob rau hauv Sjögren tus kab mob IL-38 tau upregulated raws li ib tug counterbalance rau IL-36-nyob inflammatory.34 Li no, ib tug tshiab hypothesis yog hais tias recombinant IL-38 yuav siv tau los inhibit o nyob rau hauv auto-immune kab mob los ntawm inhibiting kev kawm tiv thaiv kab mob.
Kev cob qhia kev tiv thaiv kuj tau pom zoo los ua lub luag haujlwm hauv cov kab mob autoinflammatory. Peb tau tshaj tawm yav dhau los hais tias nyob rau hauv cov neeg mob hyper IgD syndrome, qhov tsis txaus ntawm mevalonate kinase ua rau muaj kev sib txuam ntawm mevalonate, thiab kev ua haujlwm dhau los ntawm kev tiv thaiv kab mob phenotype tshwm sim raws li kev tawm tsam ntawm sterile o. Txog niaj hnub no, muaj cov ntaub ntawv txwv txog lub luag haujlwm ntawm IL-38 hauv IL-1 lossis NLRP3- ntsig txog auto-inflammatory syndromes. Qhov inhibition ntawm IL-1 los ntawm IL-38 thiab homology nrog IL-1Ra tau lees paub ntxiv kev tshawb nrhiav txog qhov cuam tshuam ntawm IL-38 hauv cov kab mob no thiab tshwj xeeb ntawm kev sib sau ua ke ntawm cov inflammasome thiab IL-1 ua, raws li ib tug anti-inflammatory luag hauj lwm rau IL-38 nyob rau hauv cov IL{11}}cov kab mob uas tau tsav, tuaj yeem xav txog. Peb tam sim no tsis paub tias txoj kev receptor twg raug thaiv los ntawm IL-38 uas ua rau txo qis ntawm IL-1 kev taw qhia. Peb xav tias IL{14}} tuaj yeem cuam tshuam nrog IL-36R signaling, uas tuaj yeem khi IL{16}} cytokines, thiab induces IL17 thiab IL{18}} . Tsis tas li ntawd, IL-38 tuaj yeem cuam tshuam nrog IL-1R9 uas tseem koom nrog inducing IL-17.35 IL-1 nws tus kheej tuaj yeem ntxias IL-36 cytokines thiab IL -17, ua rau lub voj tsis zoo uas tuaj yeem tawg los ntawm IL38. Txawm li cas los xij, qhov tseem ceeb tshaj plaws peb qhia tau tias IL-38 inhibits txoj kev mTOR, elucidating ib qho ntawm cov txheej txheem uas IL-38 siv los inhibit IL-1 ntau lawm.
Txoj kev tshawb no nthuav tawm cov pov thawj tias IL-38 inhibits qhov induction ntawm kev tiv thaiv kev tiv thaiv los ntawm kev cuam tshuam nrog mTOR txoj hauv kev thiab nrog IL1 teeb liab. Sib nrug los ntawm kev cuam tshuam nrog cov kev taw qhia tseem ceeb no hauv kev txhawb nqa kev tiv thaiv kev tiv thaiv los ntawm -glucan, IL-38 nws tus kheej kuj ua rau muaj kev hloov pauv ntawm epigenetic, uas qhia tias cais los ntawm inhibiting kev tiv thaiv kev tiv thaiv IL-38 yuav muaj sijhawm ntev. immunosuppressive nyhuv. Lub luag haujlwm ntawm IL-38 nyob rau hauv tib neeg cov kab mob uas nyob ntawm qhov kev tiv thaiv kev tiv thaiv yuav tsum muaj tus cwj pwm ntxiv, piv txwv li los ntawm kev soj ntsuam ntawm tib neeg cov qauv los txiav txim siab qhov cuam tshuam ntawm IL-38 ntawm qhov mob hauv cov kab mob no. Cov teebmeem metabolic ntawm IL-38 thiab nws cov teebmeem ntawm kev tiv thaiv kev tiv thaiv tau dhau los tshwj xeeb hauv kev teeb tsa ntawm cov kab mob plawv. Kev cob qhia kev tiv thaiv kab mob yog xav tias yuav pab txhawb rau qhov mob tsis tu ncua uas ua rau atherosclerosis.36 Ib qho cim ntawm qhov mob tsis tu ncua no yog C-reactive protein (CRP) thiab hauv kev tshuaj ntsuam meta ntau dua 80,000 cov ntsiab lus, muaj cov kab mob polymorphisms pom. tsuas yog tsib lub cev tiv thaiv kab mob uas cuam tshuam nrog kev ncig CRP, uas suav nrog CRP nws tus kheej, NRLP3, IL-6R, thiab tawm tsam IL-1F10 (IL-38).37 Peb tau tshaj tawm tsis ntev los no tias IL-38 plasma concentrations sib cuam tshuam nrog IL-6 thiab CRP hauv cov neeg rog rog, thiab feem ntau txo qis hauv cov neeg mob metabolic syndrome.20 Cov no thiab peb cov kev soj ntsuam tam sim no qhia tias IL-38 yuav yog ib qho Tus neeg ua haujlwm tseem ceeb hauv kev txwv txoj hauv kev metabolic thiab o uas yog tus tsav tsheb tseem ceeb ntawm cov kab mob plawv.
Hauv kev xaus, hauv txoj kev tshawb no, peb qhia tau hais tias IL-38 yog ib qho tseem ceeb ntawm cov tshuaj tiv thaiv kab mob uas cuam tshuam qhov induction ntawm kev tiv thaiv kab mob. IL-38 inhibits induction ntawm epigenetic thiab metabolic kev hloov pauv tshwm sim los ntawm -glucan, thiab ib qho SNP uas cuam tshuam nrog ntau dua IL-38 concentrations hauv cov neeg ua haujlwm noj qab haus huv ua rau txo qis ntawm kev tiv thaiv kev tiv thaiv los ntawm -glucan ex vivo. Cov ntaub ntawv no txhawb nqa qhov kev xav tias kev hloov kho ntawm IL-38 tuaj yeem yog kev siv tshuaj kho mob hauv cov kab mob uas tau txais kev tiv thaiv kab mob tsis zoo thiab ua rau muaj kab mob pathology thiab MGN pab kuaj kuaj mob. LABJ, MGN, thiab CAD pab txhawb cov khoom siv / cov khoom siv. DMG thiab RJWA tau sau / kos duab thiab ua tiav daim ntawv. Txhua tus kws sau ntawv tau nyeem thiab pom zoo cov ntawv sau kawg.
TXOJ CAI
FLV tau txais kev txhawb nqa los ntawm VIDI nyiaj pab los ntawm Netherlands Lub Koom Haum rau Kev Tshawb Fawb Txog Kev Tshawb Fawb. MGN tau txais kev txhawb nqa los ntawm ERC Advanced Grant (#833247) thiab Spinoza Grant ntawm Netherlands Lub Koom Haum rau Kev Tshawb Fawb Txog Kev Tshawb Fawb. CAD tau txais kev txhawb nqa los ntawm NIH grant AI 15614. RJWA tau txais kev txhawb nqa los ntawm VENI nyiaj pab los ntawm Netherlands Lub Koom Haum rau Kev Tshawb Fawb Txog Kev Tshawb Fawb (#09150161810007)
Qhia tawm
Cov kws sau ntawv tshaj tawm tsis muaj kev sib cav txog kev txaus siab.
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KEV PAB CUAM
Cov ntaub ntawv txhawb nqa ntxiv tuaj yeem pom hauv online hauv Cov Lus Qhia Txhawb Nqa ntawm qhov kawg ntawm tsab xov xwm.
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