Kev ua kom tiav yog tswj hwm hauv C3 Glomerulopathy Los ntawm IgG-factor H Fusion Proteins Nrog Thiab Tsis Muaj Kev Pom Zoo Targeting Domains

Hu rau:joanna.jia@wecistanche.com/ WhatsApp: 008618081934791

Alyssa C. Gilmore, Yuchun Zhang, H. Terence Cook', Deborah P. Lavin, Suresh Katti, YiWang2, Krista K. Johnson, SungKwon Kim², thiab Matthew C. Pickering

'Centre for Inflammatory Disease, Imperial College London, UK; thiab²Alexion Pharmaceuticals, New Haven, Connecticut, Tebchaws USA

C3 glomerupathiesNws yog tus cwj pwm los ntawm kev txuam nrog C3 hauv glomeruli. Ua rau muaj xws li tab sis tsis txwv rau, qhov txawv txav ntawm qhov H, qhov loj tsis zoo regulator ntawm kev ntxiv lwm txoj hauv kev. Factor H- tsis txaus (Cfh-/-) nas tsim C3glomerupathiesua ke nrog kev txo qis hauv plasma C3 qib. Siv cov qauv no, peb tau soj ntsuam qhov ua tau zoo ntawm ob lub fusion proteins uas muaj qhov ua tau H lwm txoj hauv kev tswj hwm kev tswj hwm (FH1-5)

txuas mus rau ib qho uas tsis yog-targeting nas immunoglobulin (IgG-FH1-5) lossis anti-nas properdin antibody (Anti-P- FH1-5). Ob leeg cov proteins nce ntshav plasma C3 thiab txo glomerular C3 deposition rau qhov sib npaug, tawm tswv yim tias tsis tsim nyog rau FH1-5 los hloov C3 ua kom cov ntshav los yog glomeruli. Ua raws li IgG-FH1-5 kev tswj hwm, plasma C3 qib cuam tshuam nrog kev hloov pauv hauv qib B, thaum qib plasma C5 cuam tshuam nrog kev hloov pauv hauv plasma properdin qib. Qhov tseem ceeb, qhov nce hauv plasma

C5 thiab cov qib properdin txuas ntxiv mus ntev dua li qhov nce hauv C3 thiab qhov tseem ceeb B. Hauv Cfh-/- nas IgG-FH1-5 txo qis raum raug mob thaum lub sij hawm ceev cov ntshav nephrotoxic nephritis. Yog li, peb cov ntaub ntawv qhia tau hais tias IgG-FH1-5 rov qab mus rau lwm txoj hauv kev ua haujlwm thiab txo qis glomerular C3 deposition hauv Cfh-/- nas thiab cov qib plasma properdin yog ib qho cim cim ntawm C5 convertase kev ua hauv qhov H tsis txaus. Lub immunoglobulin conjugated FH 1-5 protein, los ntawm nws qhov sib piv ntev plasma ib nrab-lub neej, tej zaum yuav muaj peev xwm kho tau C3.glomerupathies.

Cistanche tuaj yeem pab nrog Glomerulopathy

Lus Txhais Lus

C3 glomerupathies(C3G) yog kab mob raum uas tshwm sim los ntawm qhov txawv txav ntawm kev ua kom tiav C3 hauv glomeruli thiab glomerular puas. Nws yog vim uncontrolled activation ntawm complement lwm txoj kev. Fusion proteins, muaj cov haujlwm ua haujlwm ntawm lwm txoj hauv kev tswj hwm, yam H (FH), txuas nrog cov tshuaj tiv thaiv kab mob, rov qab ua kom tiav cov cai, thiab txo qis glomerular C3 hauv C3G nas qauv. Qhov kev sib txuas ntawm cov tshuaj tiv thaiv tsis yog lub hom phiaj ua rau muaj qhov sib piv ntev fusion protein plasma ib nrab-lub neej, thiab qhov no, los yog cov kev sib txuas zoo sib xws, tuaj yeem siv los txhawb FH muaj nuj nqi hauv kev kho ntawm C3G.

C3 glomerupathies(C3G) yog ib qho kev kho rau lub raum tsis zoo uas tshwm sim los ntawm qhov txawv txav ntawm C3 nyob rau hauv glomeruli.1 Nws tuaj yeem ua rau lub raum tsis ua haujlwm thiab tsis muaj kev kho mob meej.2 C3G cuam tshuam nrog kev ua kom txawv txav ntawm txoj kev ntxiv lwm txoj hauv kev (AP). Kev ua kom AP ua rau muaj kev tsim cov C3bBb (C3 convertase), ib qho enzyme uas cleaves C3. Tom qab qhov sib ntxiv ntawm lwm C3b molecule, qhov ua rau C3bBbC3b complex (C5 convertase) tuaj yeem cleave complement C5. Tsis tswj

Kev ua kom AP hauv C3G tuaj yeem cuam tshuam nrog kev txo qis hauv C3, C5, yam B (FB), thiab properdin. Ua rau tsis muaj kev tswj hwm AP ua kom muaj xws li poob ntawm kev hloov pauv hauv kev tswj hwm kev sib txuas thiab nce kev ua haujlwm hloov pauv hauv kev ua kom muaj zog ntxiv.2 Qhov tseem ceeb tsis zoo ntawm AP regulator yog yam H (FH), thiab FH tsis txaus rau tib neeg, npua, thiab nas cuam tshuam nrog C3G. 3 C3 nephritic yam, ib qho tshuaj tiv thaiv uas ua rau AP C3 hloov pauv ntau zaus hauv C3G.4 C3 nephritic yam yuav ua rau txo qis hauv C3 ib leeg (zoo- din-independent C3 nephritic factor) lossis txo qis hauv C3 thiab C5 (properdin- nyob ntawm C3 nephritic factor).5,6

Txawm hais tias properdin tsis txaus tsis tau ameliorate plasma C3 qib, C5 qib nce, qhia tias properdin yog qhov tsim nyog rau kev ua ntawm C5 tab sis tsis yog C3 convertase.8,9.

Kev tswj hwm ntawm mouse10 lossis tib neeg FH11,12 rau Cfh–/– nas txo glomerular C3 staining thiab nce circulating C3 qib. Kev tsim vaj tsev uas tsuas muaj cov kev tswj hwm thiab cov hom phiaj ntawm FH (mini-FH molecules) kuj muaj txiaj ntsig zoo hauv cov qauv no.13 Lwm txoj hauv kev suav nrog cov proteins uas lub hom phiaj ntawm kev ua kom muaj zog ntxiv. Cov no suav nrog TT30,14 cov protein uas muaj cov kev tswj hwm kev tswj hwm ntawm FH (FH1-5) txuas mus rau qhov txuas ntxiv-binding domains ntawm kev sib txuas receptor 2, thiab homodimeric FH molecules, 15 mini-FH molecules uas muaj cov khoom sib txuas-binding domains. ntawm H-related protein 1.

Vim tias qhov kev siv hluav taws xob kho mob ntawm inhibiting ntxiv C3 ua kom muaj kev tshawb nrhiav hauv C3G, 2 nws yog ib qho tseem ceeb kom nkag siab txog cov kinetics ntawm glomerular C3 deposition thiab nws txoj kev sib raug zoo rau AP ua kom nyob rau hauv ob qho tib si glomeruli thiab cov ncig. Hauv cov qauv ntawm kev tiv thaiv kab mob tsis sib haum xeeb glomerulonephritis, glomerular C3c tshem tawm tsis pub dhau 24 teev ntawm kev tiv thaiv kev ua kom muaj zog ntxiv, whereas glomerular C3d txuas ntxiv rau lub lis piam.16 Ib yam li ntawd, hauv Cfh–/– nas, exogenous FH ua rau txo qis glomerular C3c 4 teev ntawm 2. , whereas glomerular C3d tseem tsis hloov.11

Hauv txoj kev tshawb no, peb tau tshawb xyuas qhov ua tau zoo ntawm 2 cov protein fusion tshiab nrog ntxiv kev tswj hwm hauv Cfh–/– nas qauv ntawm C3G. Cov proteins muaj cov cai tswj hwm ntawm nas FH (FH1-5), uas tau txuas rau nas Ig kom ntev lub neej ib nrab. Ib qho ntawm cov fusion proteins, hu ua IgG-FH1-5, tau txuas mus rau cov tshuaj tiv thaiv kab mob uas tsis yog nas monoclonal. Lwm qhov, hu ua anti-P-FH1-5, tau txuas mus rau monoclonal antibody rau nas properdin. Qhov no tau ua tiav los ntsuas qhov kev xav tias kev tsom mus rau qhov fusion protein rau qhov chaw ntawm kev ua kom muaj zog ntxiv, los ntawm kev cuam tshuam nrog properdin deposition, yuav txhim kho cov ntaub so ntswg ntxiv kev cai. Peb cov ntaub ntawv qhia tau hais tias ob qho tib si tsis yog lub hom phiaj (IgG-FH1-5) thiab cov properdin-targeting (anti-P-FH1-5) cov proteins rov qab plasma thiab glomerular C3 kev cai hauv Cfh–/– nas. Cov kev tshawb fawb Timecourse tau pom tias kev kho dua tshiab ntawm C3 thiab FB qib mirrored qib ntawm fusion proteins hauv kev ncig. Hauv qhov sib piv, qhov nce hauv plasma C5 txuas ntxiv tom qab fusion proteins tau tshem tawm ntawm kev ncig. Peb qhia tau tias IgG-FH1-5 ameliorated glomerular raug mob thaum lub sij hawm ceev nrooj nephrotoxic nephritis hauv Cfh–/– nas.

cistanche tuaj yeem kho kab mob raum txhim kho lub raum ua haujlwm

TSEEM CEEB

Kev tsim cov fusion proteins thiab hauv vitro thiab vivo kev soj ntsuam ntawm kev ua haujlwm Fusion proteins tau tsim los ntawm kev sib txuas thawj 5 qhov kev pom zoo rov ua dua (SCR) ntawm nas FH (FH1-5) mus rau ib qho tshuaj tiv thaiv nas properdin antibody (anti -P-FH1-5) lossis cov tshuaj tiv thaiv kab mob uas tsis yog hom phiaj Ig (IgG-FH1- 5, Daim duab ntxiv S1). Cov kev ua ntawm cov proteins tau raug soj ntsuam los ntawm kev soj ntsuam AP-specific hemolytic (Daim duab 1a). Anti-P-FH1-5 thiab IgG-FH{16}} inhibited hemolysis nyob rau hauv ib koob tshuaj uas muaj zog ntau dua rau anti-P-FH1-5. Anti-properdin (anti-P) kuj txo hemolysis nyob rau hauv ib koob tshuaj, tab sis cov IgG- tswj tsis muaj kev cuam tshuam. Tom qab txhaj tshuaj sib npaug ntawm cov protein rau hauv Cfh–/– nas, IgG-FH1-5 protein tau kuaj pom txog li 11 hnub tom qab txhaj tshuaj, qhov tshuaj tiv thaiv P- FH1-5 tau kuaj pom txog li 4 hnub tom qab txhaj tshuaj (Ntxiv daim duab S2). Txhawm rau txiav txim siab seb IgG- FH1-5 thiab anti-P-FH{37}} tuaj yeem kho cov ntshav AP cov cai hauv Cfh–/– nas, thawj zaug ntsuas ntshav C3 qib tom qab txhaj tshuaj fusion proteins (Daim duab 1b) . Kev tswj hwm ntawm IgG-FH1-5 lossis anti-P-FH1-5 qhov tseem ceeb nce ntshav plasma C3 ntawm 24 teev thiab, rau qhov tsawg dua, ntawm 96 teev tom qab txhaj tshuaj (Daim duab 1b). Cov ntaub ntawv no tau ua pov thawj tias ob qho tib si IgG-FH1-5 thiab anti-P- FH1-5 tuaj yeem rov kho cov ntshav AP ib ntus hauv Cfh–/– nas. Peb tom ntej no tau ua cov ncauj lus kom ntxaws ntawm cov kev hloov pauv no los ntawm kev qhia lub sijhawm ntawm kev hloov pauv tsis yog hauv cov ntshav C3 nkaus xwb tab sis kuj ntawm plasma C5 thiab lwm txoj hauv kev proteins FB thiab properdin. Kev nce hauv C5 thiab cov qib properdin txuas ntxiv mus ntev dua li kev nce hauv C3 thiab FB tom qab kev tswj hwm ntawm IgG-FH1-5 thiab anti-P-FH1-5 hauv Cfh–/– nas.

Peb piv cov kinetics ntawm cov kev hloov pauv hauv C3, C5, FB, thiab properdin los ntawm kev ntsuas cov protein ntawm lub sijhawm mus txog 27 hnub tom qab ib qho kev txhaj tshuaj ntawm IgG-FH1-5 lossis anti-P-FH1-5 ( Daim duab 2). Tom qab kev tswj hwm ntawm IgG-FH1-5, qhov siab tshaj plaws hauv C3, C5, FB, thiab properdin tau tshwm sim nyob rau hnub 1 tom qab txhaj tshuaj. Lub sijhawm rau cov kev hloov pauv no kom poob mus rau qib kev kho ua ntej sib txawv. Kev rov qab mus rau qib pretreatment tshwm sim nyob rau hnub 1 thiab 4 rau FB, nruab nrab hnub 7 thiab 11 rau C3, thiab nruab nrab hnub 14 thiab 21 rau ob qho tib si properdin thiab C5 (Daim duab 2a, c, e, thiab g). Tom qab kev tswj hwm ntawm anti-P-FH1-5, ncov C3 theem tau tshwm sim rau hnub 1 tab sis poob rau qib pretreatment ntawm hnub 4 thiab 7 (Daim duab 2b). Peak C5 theem tshwm sim rau hnub 4 thiab rov qab mus rau qib pretreatment ntawm hnub 7 thiab 11 hnub (Daim duab 2d). Qhov siab tshaj plaws hauv FB tau tshwm sim rau hnub 1 thiab poob rau qib pretreatment ntawm hnub 1 thiab 4 (Daim duab 2f). Raws li qhov xav tau, cov ntshav plasma dawb properdin tau ua tiav tom qab txhaj tshuaj tiv thaiv P-FH1-5 lossis anti-P (Daim duab 2h). Cov qib dawb hauv plasma properdin tseem qis txog li 7 hnub tom qab tiv thaiv P-FH1-5 thiab txog li 14 hnub tom qab tiv thaiv P (Daim duab 2h). Txawm hais tias ob qho tib si fusion proteins ib ntus rov qab plasma AP txoj cai, lub sijhawm ntawm kev txhim kho hauv C3, FB, thiab C5 qib tau cuam tshuam lub sijhawm ntawm lawv qhov kev tshawb pom hauv cov ntshav, uas luv dua rau kev tiv thaiv P-FH1-5 (Cov Duab Ntxiv S2) . Hauv cov ntsiab lus, kev hloov pauv hauv qib C3 thiab FB tom qab txhaj tshuaj ntawm cov protein tau luv dua li C5 thiab properdin. Cov ntaub ntawv no qhia tias plasma FB yog lub cim cim ntawm C3 convertase kev ua haujlwm hauv qhov chaw no, hos properdin yog tus cim ntawm C5 convertase kev ua. Tshwj xeeb, tom qab txhaj tshuaj tiv thaiv P, muaj qhov nce hauv C5 ntawm 24 thiab 96 teev thiab hnub 7.

Daim duab 1|(a) Ua kom tiav lwm txoj hauv kev-dependent hemolysis assay. Cov tshuaj tiv thaiv tau titrated hauv 2-fold dilutions los ntawm 60 nM txog rau 0.5 nM. Cov IgG-FH1-5, anti-P-FH1-5, thiab cov tshuaj tiv thaiv P reagents txo cov luav erythrocyte hemolysis nyob rau hauv ib koob tshuaj. Tsis muaj kev txwv tsis pub siv IgG-tswj cov protein. Kab rov tav tuav txhais tau tias qhov tseem ceeb, thiab cov kab yuam kev sawv cev rau SD. (b) Plasma ntxiv C3 hauv Cfh–/– nas tom qab txhaj tshuaj fusion proteins. Plasma C3 tau ntsuas ua ntej thiab 24 teev thiab 96 teev tom qab kev tswj hwm ntawm IgG-FH1-5 (daim duab peb sab liab, n ¼ 5) Anti-P–FH1-5 (daim duab peb sab, n ¼ 6), Anti -P (ntsuab dots, n ¼ 5), thiab IgG-tswj (xiav dots, n ¼ 5). Kab rov tav tuav txhais tau hais tias muaj nuj nqis, thiab cov nplais taw qhia SD. ***P # 0.001 piv rau pretreatment tus nqi thiab muab los ntawm 2- txoj kev ANOVA nrog Bonferroni ntau qhov kev sib piv. FH, yam H; P, properdin.

(Daim duab 2d). Raws li qhia, 8,9 qhia tias C5 convertase yog nyob ntawm properdin hauv Cfh–/– nas.

IgG-FH1-5 thiab anti-P-FH1-5 txo glomerular C3c tab sis tsis hloov C3d staining hauv Cfh–/– nas

Peb tom ntej no tau soj ntsuam qhov cuam tshuam ntawm IgG-FH1-5 thiab anti-P-FH1-5 ntawm glomerular C3b/iC3b/C3c, C3d, properdin, thiab FH- related (FHR) proteins. Thaum 96 teev tom qab txhaj tshuaj IgG- FH1-5, glomerular C3b/iC3b/C3c staining tau txo tab sis glomerular C3d (Daim duab 3a) thiab FHR staining (Daim duab 3b) tsis yog. Glomerular C3b / iC3b / C3c tseem tsis tau hloov nrog IgG-tswj lossis anti-P. Cov kab txawv txawv txawv ntawm glomerular properdin staining hauv kev kho tsis tau Cfh-/- nas9 tseem nyob 96 teev tom qab tswj IgG. Hauv Cfh–/– nas txhaj tshuaj tiv thaiv P lossis IgG-FH1-5, qib glomerular properdin yuav luag tsis pom ntawm 96 teev (Daim duab 3b). Tsis xav txog, kev txhaj tshuaj tiv thaiv-P-FH1-5 ua rau muaj cov kab mob glomerular staining qauv siv cov tshuaj tiv thaiv properdin, anti-FH/FHR, thiab cov tshuaj tiv thaiv IgG (Daim duab 3b). Qhov kev tshawb pom no tau qhia tias cov tshuaj tiv thaiv P-FH1-5 protein tau tso rau hauv glomeruli. Txhaj tshuaj tiv thaiv-P-FH1-5 cov protein rau hauv cov nas qus ua rau cov kab mob glomerular staining nrog anti-properdin, anti-FH/FHR, thiab anti-IgG tshuaj tiv thaiv ntawm 96 teev (Ntxiv daim duab S3), qhia tias qhov no reagent cuam tshuam nrog ib txwm glomeruli. Txawm li cas los xij, peb xav tias cov tshuaj tiv thaiv-P-FH1-5 cov protein kuj tuaj yeem cuam tshuam nrog cov glomerular properdin ua ntej hauv Cfh–/– nas. Thaum peb txhaj cov tshuaj reagent rau hauv Cfh–/– nas uas tau pretreated nrog anti-P kom tshem tawm glomerular properdin, muaj ib tug txo nyob rau hauv cov granular staining qauv siv anti-properdin, anti-FH / FHR, thiab anti-IgG tshuaj tiv thaiv ( Ntxiv daim duab S4). Cov kev soj ntsuam no qhia tau hais tias glomerular cuam tshuam ntawm anti-P-FH1-5 protein nyob rau hauv Cfh–/– nas yog ib feem nyob ntawm glomerular properdin.

IgG-FH1-5 ameliorated raum raug mob thaum lub sij hawm ceev nrooj nephrotoxic nephritis hauv Cfh–/– nas

Hauv cov neeg mob uas muaj C3G, mob raum raug mob tuaj yeem tsim nyob rau hauv cov ntsiab lus ntawm kev sib kis kab mob. Piv txwv li, kev poob ntawm lub raum ua haujlwm hauv FHR5 nephropathy yog txuam nrog lub sijhawm ntawm synpharyngitic macroscopic hematuria.17 Lub hauv paus ntawm C3 dysregulation hauv C3G cov neeg mob yuav ua rau muaj kev txhim kho ntxiv-kev kho lub raum raug mob tom qab qhov tshwm sim uas ua rau lub raum mob. Txhawm rau ua tus qauv no, peb tau ua kom ceev cov ntshav qab zib nephrotoxic nephritis, kev tiv thaiv kab mob

Daim duab 2|Plasma ntxiv cov sijhawm kawm hauv Cfh–/– nas txhaj nrog IgG-FH1-5 thiab anti-P-FH1-5. Plasma C3 (a, b), C5 (c, d), FB (e, f), thiab properdin (P; g, h) los ntawm kev sim pib mus txog 27 hnub tom qab txhaj ib zaug ntawm IgG-FH{{1{{21} }}}} (a, c, e, g; liab daim duab peb sab, n ¼ 5) los yog anti-P- FH1-5 (b,d,f,h; ntshav daim duab peb sab, n ¼ 6). Kev tswj hwm suav nrog cov tshuaj tiv thaiv P (ntsuab dots, n ¼ 5) thiab ib qho isotype-matched monoclonal antibody (IgG- tswj; xiav dots, n ¼ 5). Kab rov tav tuav txhais tau hais tias muaj nuj nqis, thiab cov nplais taw qhia SD. *P # 0. FB, factor B; FH, Factor H.

glomerulonephritis qauv uas suav nrog ntxiv thiab Fc receptor-mediated pathways, 18,19 hauv Cfh–/– nas. Peb yav dhau los tau pom tias Cfh–/– nas muaj qhov tsis zoo rau lub raum raug mob hauv qhov chaw no7 thiab xav tias FH1-5 pro- tein, los ntawm kev txhim kho AP cov cai, tuaj yeem kho cov mob raum mob ntxiv rau hauv cov qauv no. Vim tias peb cov ntaub ntawv qhia tias cov tshuaj tiv thaiv P-FH1-5 cov protein tau tso rau hauv ib txwm muaj glomeruli, peb siv IgG-FH1-5 thaum lub sij hawm ceev cov ntshav nephrotoxic nephritis. Nees nkaum plaub teev ua ntej txhaj tshuaj yaj nephrotoxic serum, nas tau txais kev txhaj tshuaj ip ntawm IgG-control (n ¼ 7) lossis IgG-FH1-5 (n ¼ 8). Kev sim raws tu qauv tau piav qhia hauv daim duab 4a. Cov nas raug tshem tawm 6 hnub tom qab kev tswj hwm ntawm yaj nephrotoxic serum. Tag nrho 3 qhov kev ntsuas histological ntawm glomerular pathology (qhov qhab nia hnyav, tag nrho cov xov tooj ntawm tes, thiab tus lej macrophage) tau qis dua hauv IgG-FH1-5 pawg (Daim duab 4b). Kev raug mob tubulointerstitial kuj tseem qis dua hauv pawg IgG-FH1-5 (Daim duab 4b). Serum urea tau nce qhov tseem ceeb hauv

Daim duab 3|Glomerular ntxiv kev tiv thaiv kab mob hauv Cfh–/– nas 96 teev tom qab txhaj tshuaj IgG-FH1-5 lossis anti-P-FH1-5. (a) Cov duab sawv cev ua ke nrog rau qhov ntau ntawm glomerular C3b/iC3b/C3c thiab C3d hauv 4 pawg sim: IgG-FH1-5 (daim duab peb sab liab, n ¼ 5), anti-P-FH1-5 (liab duab peb sab, n ¼ 6), anti-P (ntsuab dots, n ¼ 5), thiab IgG-tswj (xiav dots, n ¼ 5).PlasmaC3levelsatthetimeofcullareshowninFigure1. (b) Cov ntsiab lus cov ntaub ntawv sawv cev rau qhov nruab nrab qhov tseem ceeb, thiab cov tshuaj txhuam hniav qhia txog qhov sib txawv ntawm qhov sib txawv. P qhov txiaj ntsig tau los ntawm Kruskal-Wallis test nrog Dunn ntau qhov kev sib piv. (b) Cov duab sawv cev ntawm glomerularIgG, FHR, thiab, staininginthe4experimentalgroups.Bar ¼ 100 mm.AFU, arbitraryuorescent units; FH, yam H; P, properdin. Txhawm rau optimize saib cov duab no, thov mus saib hauv online version ntawm tsab xov xwm no ntawmwww.kidney-international.org.

pawg IgG-tswj, tab sis kev hloov pauv hauv cov ntshav albumin thiab cov zis albumin-rau-creatinine piv tsis txawv (Daim duab 4b). Raws li qhov xav tau, glomerular C3b / iC3b / C3c tau txo qis hauv IgG-FH1-5 pawg (Daim duab 4b), tab sis glomerular nas IgG (Daim duab 4b) thiab yaj IgG (cov ntaub ntawv tsis qhia) tsis txawv ntawm cov pab pawg. .

Daim duab 4|Pretreatment nrog IgG-FH1-5 ameliorated raum raug mob thaum lub sij hawm ceev nephrotoxic nephritis hauv Cfh–/– nas. (a) Schematic ntawm ceev ceev nephrotoxic nephritis raws tu qauv. Cov nas ua ntej txhaj tshuaj nrog yaj IgG tau txais IgG-FH1-5 ( pab pawg kho mob, n ¼ 8) lossis IgG-control (tswj pab pawg, n ¼ 7) 24 teev ua ntej kev tswj hwm ntawm yaj nephrotoxic serum. (b) Lub raum muaj nuj nqi thiab histology 6 hnub tom qab induction ntawm accelerated serum nephrotoxic nephritis nyob rau hauv nas kho nrog IgG-FH1-5 (kev kho mob; liab daim duab peb sab, n ¼ 8) los yog IgG-tswj (tswj; xiav voj voog, n ¼ 7). Kab rov tav bar qhia qhov nruab nrab qhov tseem ceeb, thiab whiskers qhia txog qhov sib txawv. *P # 0.05, **P # 0.01, ***P # 0.001 tiv thaiv kev tswj thiab muab los ntawm Mann-Whitney xeem. FH, yam H; P, properdin.

Kev sib tham

Ob leeg IgG-FH1-5 thiab anti-P-FH1-5 normalized C3, FB, thiab C5 qib hauv Cfh–/– nas. Qhov kev tshawb nrhiav no yog ua raws li qhov tseeb tias cov kev cai tswj hwm ntawm nas FH nyob hauv SCR domains 1 txog 5 (a C3b-binding site thiab cofactor kev ua si rau qhov tseem ceeb I-mediated cleavage of C3b).20 Surface recognition domains, uas Kev cuam tshuam cuam tshuam rau heparin thiab endothelial hlwb thiab suav nrog qhov thib ob C3b-binding site, muaj nyob rau hauv SCR domains 18 txog

20 (FH18-20) thiab yog li ntawd tsis muaj nyob rau hauv fusion proteins.20 Txawm li cas los xij, ob qho proteins txo glomerular iC3b/C3b/C3c, qhia tias FH18-20 tsis tas yuav tsum muaj nyob rau hauv qhov chaw no. Qhov kev tshawb pom no yog ua raws li cov ntaub ntawv yav dhau los uas qhia tias Cfh–/– nas qhia txog kev hloov pauv FH protein uas muaj SCR domains 1 txog 15 (Cfh–/– .FHD16-20) tsis tau tsim qhov txawv txav glomerular C3 deposition.21,22 Txawm li cas los xij, Cfh–/–.FH D16-20 cov tsiaj tsis muaj peev xwm tswj tau C3 kom ua raws li lub raum endothelium thiab tsim thrombotic microangiopathy. Nws yog qhov ua tau tias, vim qhov tsis muaj lub hom phiaj ntawm FH 18-20 domains, kev tswj hwm ntawm fusion proteins uas muaj FH 1-5 hauv FH tsis txaus tuaj yeem ua rau muaj kev cuam tshuam rau thrombotic microangiopathy. Nws tseem muaj peev xwm hais tias qhov nce avidity rau C3b binding, nyob rau hauv lub dimeric qauv ntawm fusion proteins, yog txaus los them rau qhov tsis muaj qhov nto phiaj FH18-20 domains. Txawm li cas los xij, qhov kev lees paub saum npoo ntawm C3b kuj cuam tshuam los ntawm kev hloov pauv hauv FH.23 A Streptococcus pneumoniae FH-binding protein (PspCN, uas khi SCR domain 9), ua rau FH conformational hloov, thiab FH-PSpCN complex tau nce C3b khi thiab txhim khu kev lwj-accelerating kev ua. FH20 C3d-binding site tsis raug rau hauv FH tab sis dhau los ua rau hauv FH- PspCN complex. Qhov no yuav piav qhia yog vim li cas FH19-20 cuam tshuam nrog nto C3d tab sis FH tsis ua.24,25 Peb xav tias peb cov neeg sawv cev, los ntawm kev hloov pauv, tuaj yeem cuam tshuam nrog ob qho tib si fluid-theem thiab nto C3b. Qhov tseem ceeb, ob qho tib si IgG-FH1-5 thiab anti-P-FH1-5 tiv thaiv erythrocytes los ntawm lysis hauv AP-dependent hemolysis assay.

Tsis yog fusion protein txo glomerular C3d staining, tej zaum vim yog qhov tsis tu ncua ntawm glomerular C3d. Hauv kev sim tshuaj tiv thaiv kab mob-complex nephritis, glomerular C3c daws tsis pub dhau 24 teev tom qab tso tseg ntawm kev ua kom tiav tab sis C3d tseem nyob rau lub lis piam.16 Tsis tas li ntawd, glomerular C3d tseem nyob rau hauv lupus nephritis.26 Qhov no yuav cuam tshuam nws cov kev sib cuam tshuam nrog glomerular nto. Kev txhaj tshuaj rov qab ntawm tib neeg FH hauv Cfh–/– nas tau pom tias txo qis hauv glomerular C3d dhau 10 hnub.11 Nws zoo li qhov rov ua dua ntawm IgG-FH1-5 yuav txo qis glomerular C3d hauv cov qauv no. Kuj tsis muaj qhov hloov pauv hauv glomerular FHR staining. Me ntsis paub txog kev ua haujlwm ntawm cov nas FHR proteins, tab sis lawv tuaj yeem cuam tshuam nrog ob qho tib si C3b27,28 thiab C3d.27 Qhov tseeb, qhov pom tseeb ntawm FHR-A thiab FHR-B rau C3d muaj zog dua li ntawm FH.27. Peb xav tias glomerular FHR proteins yuav tshem tawm ntawm glomeruli tsuas yog thaum C3d raug tshem tawm. Nyob rau hauv cov ntsiab lus no, nws yog ib qho tseem ceeb hais tias nyob rau hauv tib neeg C3G, qhov H-related protein 5 associates nrog glomerular C3d.29

Cov tshuaj tiv thaiv P-FH1-5 protein depleted dawb plasma properdin ntau ntau raws li xav tau vim hais tias cov anti-properdin ib feem ntawm cov fusion protein no tuaj yeem thaiv AP kev ua si (vim properdin depletion) rau 8 hnub tom qab txhaj ib zaug.30 Txawm li cas los xij, peb cov ntaub ntawv qhia tau hais tias qhov ua tau zoo ntawm FH1-5 hauv kev nce ntshav plasma C3 qib thiab txo qis glomerular C3b / iC3b / C3c staining yog sib piv ntawm 2 fusion proteins (piv txwv li, yog ywj pheej ntawm properdin targeting). Ntxiv mus, qhov txawv txav glomerular properdin staining tau txo qis tom qab kev tswj hwm ntawm anti-P lossis IgG-FH1-5. Pom tau tias, piv rau C3d thiab FHR cov protein, glomerular properdin tau yooj yim tshem tawm tom qab kev kho dua ntawm C3 kev tswj hwm. Glomerular properdin tau hloov pauv hauv cov qauv tom qab kev tswj hwm ntawm cov tshuaj tiv thaiv P-FH{19}} cov protein, tab sis qhov no yog qhov nyuaj los ntawm kev soj ntsuam uas peb tau kuaj pom qhov tso tawm ntawm cov protein fusion no hauv ob qho tib si Cfh-/- thiab cov tsiaj qus glomeruli. Qhov no yog ib feem vim muaj kev cuam tshuam nrog glomerular properdin, tab sis qhov tso tawm hauv cov tsiaj qus-hom glomeruli qhia tias nws kuj yog ib feem ntawm-nyob ntawm glomerular ntxiv thiab yuav muaj feem cuam tshuam rau lub cev muaj zog ntawm fusion protein.

Peb cov ntaub ntawv kinetic tau pom cov kev tshawb pom tshiab txog kev hloov pauv ntawm lub sijhawm hauv FB, C5, thiab properdin uas nrog nce hauv C3 hauv Cfh-/- nas tom qab txhaj tshuaj fusion proteins. FB qib tau nce sai sai tom qab txhaj tshuaj

thiab rov qab mus rau theem pib ntawm lub sijhawm thaum ob qho tib si C5 thiab properdin qib tseem nce siab. Cov qib Properdin hauv Cfh–/– nas tau txo qis ntawm qhov pib thiab nce mus rau qib qub (20 mg / ml30) tom qab txhaj tshuaj IgG-FH1-5 thiab tseem nyob ntawm cov qib no tsawg kawg 14 hnub. Ib qho kev kawm zoo sib xws tau pom rau kev nce qib hauv plasma C5. Strikingly, IgG-FH1-5 feem ntau tsis tuaj ntawm kev ncig ntawm 11 hnub, qhia tau hais tias qhov tsim ntawm C5 convertase hauv Cfh–/– nas qeeb dua li ntawm C3 convertase (vim C3 qib rov qab mus rau hauv paus ntawm 7 thiab 11 hnub). Plasma C5 nce ntxiv tom qab kev kho los tiv thaiv P thiab nce ntxiv nrog cov tshuaj tiv thaiv P-FH1-5 protein. Yog li ntawd, ob qho tib si properdin lub hom phiaj thiab cov teebmeem ntawm FH1-5 tau ua rau muaj kev nce qib C5 thiab qhia tau hais tias C5 convertase yog ib feem ntawm Cfh–/– nas. Qhov kev tshawb pom no zoo ib yam nrog kev ua kom zoo ntawm C5 dysregulation hauv nas nrog kev sib xyaw ua ke ntawm FH thiab properdin.8,9 Cov qib properdin tuaj yeem txo qis hauv C3G thiab zoo li cuam tshuam nrog C5 convertase kev ua haujlwm hauv qhov sib piv rau cov ntshav C5 lossis soluble C5b{ {30}}.31,32 Peb cov ntaub ntawv txhawb kev sib txuas ntawm cov qib properdin thiab C5 convertase kev ua haujlwm thiab txhawb kev siv cov qib properdin raws li biomarker ntawm kev ua kom tsis tu ncua glomerular C5.

Qhov tshwj xeeb tshaj yog, IgG-FH1-5 cov protein tseem tuaj yeem kuaj pom hauv kev ncig mus txog 11 hnub tom qab txhaj tshuaj ib zaug. Qhov no ntev dua li qhov peb tau tshaj tawm yav dhau los rau cov nas tsis sib txuas FH1-5 cov protein, uas tau raug tshem tawm ntawm kev ncig hauv 24 teev.14 Qhov kev tshawb pom no qhia tau hais tias ntev plasma ib nrab-lub neej ntawm IgG-FH{ {9}} protein yog muab los ntawm cov tshuaj tiv thaiv kab mob. Lub plasma ib nrab-lub neej ntawm IgG-FH1-5 protein kuj tseem ntev dua li qhov peb tau pom yav dhau los rau qhov ntev FH.10,11 Piv txwv li, tib neeg FH raug tshem tawm ntawm kev ncig los ntawm 96 teev tom qab txhaj ib zaug. nyob rau hauv Cfh–/– nas.11 Ua ke nrog peb cov ntaub ntawv qhia txog kev ua tau zoo ntawm IgG-FH1-5 protein nyob rau hauv cov kev cai ntxiv, peb txiav txim siab tias conjugation ntawm FH{20}} protein kom ntev nws cov pharmacokinetic proforme muaj peev xwm kho tau. Kev siv hluav taws xob hauv C3G.

Hauv cov ntsiab lus, peb qhia tau hais tias, txawm tias tsis muaj qhov chaw lees paub qhov chaw, FH 1-5 fusion proteins tau ua haujlwm zoo hauv kev txo qis glomerular C3 ua kom thiab kho cov ntshav plasma ntxiv cov cai hauv FH tsis txaus. Tsis muaj qhov zoo tshaj plaws hauv kev sib txuas ntawm FH1-5 domains rau anti-properdin, thiab IgG-FH{5}} protein txo qis raum raug mob hauv kev sim nephritis. Nyob rau hauv sib piv rau cov kev cov nyom nyob rau hauv tsim loj-scale npaj ntawm FH rau kev siv kho mob, cov loj-scale zus tau tej cov tshuaj tiv thaiv-raws li kev kho mob yog tsim tau zoo, uas ua rau IgG-FH1-5 ib tug txaus nyiam kev kho mob rau C3G.

Txoj kev

Fusion proteins

IgG-FH1-5. Thawj 5 nas FH SCR domains (mFH1-5) txuas

mus rau qhov tsis yog hom phiaj IgG1 nas Ig (ib qho tshuaj tiv thaiv idiotypic antibody tsa tawm tsam nas monoclonal antibody).

Anti-P-FH1-5. Murine FH1-5 txuas rau nas anti-properdin antibody (anti-P), 30 muab rau Alexion los ntawm W. Song, University of Pennsylvania).

Tswj. Kev tswj hwm yog isotype-matched non-targeting nas Ig (IgG-tswj) thiab nas anti-properdin (anti-P) (Cov duab ntxiv S1). Proteins tau tsim los siv pVEK vectors thiab Expi293 hlwb (Thermo Fisher Scientific, Waltham, MA), purified siv pro- tein A (MabSelect SuRe, Cytiva, Marlborough, MA).

AP-kev ntsuam xyuas hemolytic

Nees nkaum feem pua ​​​​ntawm cov nas ib txwm muaj titrated los ntawm 60 nM mus rau 0.5 nM tau incubated nrog luav erythrocytes (1.5 106 hlwb / ml) ntawm 37 C rau 30 feeb. Heme tso tawm yog quantified spectrophotometrically (optical ntom 415 nm); 100 feem pua ​​​​lysis yog cov ntshav tsis muaj inhibitor.

Plasma FB thiab FH

Plasma tau txais tom qab centrifugation ntawm cov ntshav sau rau hauv cov hlab uas muaj ethylenediamine tetraacetic acid (Sarstedt, Nordrhein-Westfalen, Lub teb chaws Yelemees). Proteins raug ntsuas los ntawm capillary electrophoresis immunoassay (WES, ProteinSimple, San Jose, CA). Cov tshuaj tiv thaiv uas siv tau yog polyclonal anti-nas FH antibody (Alexion Pharmaceuticals, Boston, MA) thiab anti-nas FB antibody (Abcam, Cambridge, UK); WES anti-luv nrhiav module (ProteinSimple, San Jose, CA) tau siv. Chemiluminescent signals tau soj ntsuam nrog Compass rau SW software (ProteinSimple, version 5.0.1), quantified li peak area, and normalized to the assay control.

Cov nas

Txhua tus nas tau nyob hauv qhov tshwj xeeb tsis muaj kab mob; cov txheej txheem tau ua raws li lub koom haum cov lus qhia thiab pom zoo los ntawm United Kingdom Home Office. C57BL/6 nas qus tau yuav los ntawm Jackson Laboratory (Bar Harbor, ME), thiab Cfh–/– nas raug tsim tawm raws li tau piav qhia yav dhau los.7 Cov nas tau sib phim rau hnub nyoog thiab poj niam txiv neej; equimolar koob tshuaj ntawm cov protein tau muab los ntawm kev txhaj tshuaj ip (1 mg rau anti-properdin thiab IgG-tswj; 1.4 mg rau IgG-FH1-5 thiab anti-P-FH1-5). Accelerated serum nephrotoxic nephritis tau tshwm sim los ntawm iv txhaj tshuaj yaj nephrotoxic ntshav rau cov nas ua ntej txhaj tshuaj nrog yaj IgG.7 Kev tswj hwm ntawm fusion proteins tau ua 24 teev ua ntej yaj nephrotoxic serum induction (Daim duab 4a).

C3, C5, thiab dawb properdin Plasma C3 tau ntsuas los ntawm enzyme-linked immunosorbent assay.14 Plasma "dawb" properdin thiab C5 qib tau ntsuas siv electrochemiluminescence immunoassays (Meso Scale Discovery [MSD], Meso Scale anti Diagnostics, Rockville, MD) - properdin los yog kev cai los tiv thaiv murine C5 coated ntawm high-binding MSD daim hlau. Bound "dawb" properdin thiab C5 tau kuaj pom nrog anti-properdin (los ntawm W. Song) lossis biotinylated monoclonal anti-C5 (Alexion) ua ke nrog streptavidin-SULFO (MSD, Meso Scale Diagnostics). Lub teeb liab chemiluminescent tau txais los ntawm kev siv SECTOR S 6000 imager (MSD, Meso Scale Diagnostics) thiab tau txheeb xyuas nrog MSD Discovery Workbench software (Meso Scale

Diagnostics, version 4.0.12.1). Recombinant murine properdin thiab C5 tau siv ua qauv.

Lub raum ua haujlwm thiab histology

Hematuria thiab proteinuria raug soj ntsuam siv Hema-Combistix (Bayer, Nyeem Ntawv, UK); plasma urea raug ntsuas thiab lub raum cov ntaub so ntswg ua tiav raws li tau piav qhia dhau los.9 Cov zis creatinine tau txiav txim siab ntawm cov zis tso zis siv Creatinine Companion Protocol assay (#1012; Excell, Philadelphia, PA), thiab cov zis tso zis / plasma albumin tau ntsuas los ntawm enzyme-linked immunosorbent assay (#E99-134; Bethyl Laboratories, Montgomery, TX). Periodic acid-Schiff-stained raum seem raug soj ntsuam nyob rau hauv ib tug dig muag zam thiab ranked raws li qhov hnyav ntawm glomerular thiab tubulointerstitial raug mob (0 [tsis muaj], 1 [mob], 2 [moderate]). Kaum glomeruli ib ntu tau raug soj ntsuam los txiav txim siab tus lej glomerular cell. Immunofluorescence staining tau ua rau ntawm 5-mm cryosections mounted siv Vectashield nruab nrab nrog DAPI (Vector Laboratories, Burlingame, CA). Cov tshuaj tiv thaiv uas siv tau yog fluorescein isothiocyanate (FITC)-conjugated polyclonal tshis anti-nas C3b/C3c/ iC3b (1:200; #55500; MP Biomedical, Santa Ana, CA)9; Fig-chain-spe-cific FITC-conjugated polyclonal tshis anti-nas IgG (1:400; #F5387; Sigma-Aldrich, St. Louis, MO); FITC-conjugated monoclonal nas anti-tshis / yaj IgG (1:100; #F5137; Sigma-Aldrich); los yog FITC-conjugated tshis los tiv thaiv tib neeg properdin (1:50; #GAHu/PPD/FITC, Nordic Immu- tsis muaj logic Laboratories, Copenhagen, Denmark).9 Biotinylated tshis anti-nas C3d (1:10; # BAF2655; R&D Systems , Minneapolis, MN) tau siv rau ntawm biotin-blocked seem (Biotin Blocking System; Agilent Dako, Santa Clara, CA), nrog streptavidin AF488 Secondary antibody (1:200; #S-32354; Thermo Fisher Scientific). FH tau pom nyob rau hauv puri-rat anti-nas CD16/CD32- ntu thaiv (1:100; BD Biosciences, Franklin Lakes, NJ) siv tshis tiv thaiv tib neeg FH (1: 1000; #A312; Quidel, San Diego , CA) thiab monoclonal anti-tshis IgG-FITC Secondary antibody (1:100; clone GT-34; F4891; Sigma-Aldrich). Glomerular macrophages tau txheeb xyuas siv FITC-conjugated nas anti-nas CD68 (FA-11 clone GTX43518; GeneTex, Irvine, CA). Kev soj ntsuam ntau ntawm cov tshuaj tiv thaiv kab mob tau ua tiav siv Leica DM4B optical microscope txuas nrog Leica DFC700T cov koob yees duab digital (Leica Microsystems, Wetzlar, Germany) thiab Image-Pro Plus software (Media Cybernetics, Rockville, MD, version 7). Kaum glomeruli raug tshuaj xyuas rau ib ntu, thiab qhov ntsuas qhov hnyav tau qhia nyob rau hauv cov fluores- science units.

Kev txheeb cais

GraphPad Prism, version 8.0 (GraphPad, San Diego, CA) tau siv los txheeb xyuas cov lej. Rov ua dua-kev ntsuas 2-txoj kev tsom xam ntawm qhov sib txawv nrog Bonferroni ntau qhov kev sib piv (qhov tseem ceeb yog lub sijhawm thiab kev kho mob) tau siv rau kev tshuaj xyuas lub sijhawm. Kruskal- Wallis nrog Dunn ntau qhov kev sib piv tau siv thaum sib piv ntau pab pawg, thiab kev sim Mann-Whitney tau siv rau 2 pawg.

Qhia tawm

MCP tau txais kev pab tswv yim los ntawm Alexion, ChemoCentryx, Novartis, Gyroscope, thiab Achillion Pharmaceuticals. HTC tau txais kev pab tswv yim los ntawm Alexion, Novartis, Aurinia, thiab Achillion Pharmaceuticals. YZ, YW, KKJ, thiab SK-K yog cov neeg ua haujlwm ntawm Alexion Pharmaceuticals. SK yog ib tus neeg ua haujlwm qub ntawm Alexion Pharmaceuticals thiab tam sim no ua haujlwm los ntawm Gemini Pharmaceuticals. Tag nrho lwm tus kws sau ntawv tshaj tawm tsis muaj kev sib tw nyiam.

TXOJ CAI

MCP yog Wellcome Trust Senior Fellow hauv Kev Tshawb Fawb Kev Tshawb Fawb (piv txwv li 212252/Z/18/Z), thiab ACG thiab DPL tau txais kev txhawb nqa los ntawm qhov kev sib raug zoo no. YZ, SK, YW, KKJ, thiab SK-K tau txais nyiaj los ntawm Alexion Pharmaceuticals.

SUPPLEMENTARY KHOOM

Cov ntaub ntawv ntxiv (PDF)

Daim duab S1. Fusionproteinsusedinthisstudy.IgG-FH1-5 muaj cov thawj 5 thawj ntawm nas yam H (FH1-5) txuas mus rau tus nas uas tsis yog hom phiaj Ig. Anti-P-FH1-5 muaj mFH1-5 txuas mus rau ib qho tshuaj tiv thaiv nas anti-nas properdin antibody (anti-P; saib Miwa et al.30). Tswj cov proteins suav nrog cov isotype-matched non-targeting nas Ig (IgG-tswj) thiab cov nas tsis muaj zog tiv thaiv nas properdin antibody (Anti-P). Daim duab S2. WES blot tsom xam txhawm rau txheeb xyuas FH1-5 hauv cov qauv ntshav plasma los ntawm FH- cov nas tsis muaj peev xwm txhaj nrog (A) IgG-FH1-5 lossis (B) Anti-P-FH1-5. Lub 94-kDaIgheavychainlinkedtomouseFH1-5 (HC-FH1-5) tuaj yeem kuaj pom tau txog li 11 hnub tom qab txhaj tshuaj ntawmIgG-FH1-5 thiab txog li 4 hnub tom qab txhaj tshuaj anti-P-FH1-5 protein (cov thawv liab ).Controlsincludedplasmafromwild-type nass qhov twg kuaj pom H protein tag nrho (FH, lub thawv dub, kab C1) thiab cov nas uas tsis txhaj tshuaj FH-tsis muaj peev xwm, qhov twg tsis muaj FH pov thawj (cov thawv dub, kab C2,). Raws li qhov xav tau, tsis yog FHnorFH1-5 tsis tuaj yeem kuaj pom hauv cov ntshav plasma los ntawm nas txhaj nrog Anti-P lossis IgG-control.

Daim duab S3. Glomerular ntxiv kev tiv thaiv kab mob 96 teev tom qab txhaj tshuaj IgG-tswj lossis anti-P-FH1-5 hauv cov nas qus. Cov duab sawv cev glomerular pom (bar ¼ 100 mm).

Daim duab S4. Lub luag haujlwm ntawm properdin ntawm kev tso tawm ntawm anti-P-FH1-5 hauv glomeruli ntawm FH-tsis muaj cov nas. Cfh–/– nas tau txhaj nrog IgG- tswj lossis tiv thaiv P (kev kho 1) ua raws 24- teev tom qab los ntawm IgG- tswj lossis tiv thaiv P-FH1-5 (kev kho 2). Tsiaj txhu raug tshem tawm 120 teev tom qab kev kho mob ib qho thiab glomerular staining rau IgG, properdin, thiab FH / FHR raug soj ntsuam. Cov duab sawv cev tau qhia. Anti-P txhaj tshuaj txo qis properdin (kab 1, kem 2) tab sis tsis yog FH / FHR (tseem hloov pauv) lossis IgG (tseem tsis muaj) staining. Kev tswj hwm ntawm cov tshuaj tiv thaiv-P- FH1-5 24- teev tom qab IgG-tswj tau ua rau cov tsos ntawm granular staining nrog anti-IgG, anti-FH / FHR, thiab anti-P tshuaj tiv thaiv raws li pom tom qab txhaj tshuaj tiv thaiv P-FH 1-5 ib leeg (saib daim duab 3b). Qhov no granular staining tau txo qis thaum txhaj tshuaj tiv thaiv P-FH1-5 ua ntej los ntawm kev txhaj tshuaj tiv thaiv P, uas depletes ob qho tib si ncig (saib daim duab 1h) thiab glomerular (saib daim duab 3b). Glomerular staining nrog anti-IgG tsuas yog tshwm sim hauv cov nas uas tau txhaj tshuaj tiv thaiv P-FH1-5 (kab 3, kab 3 thiab 4) thiab tau qis qis hauv cov nas uas tau txais anti-P-FH1-5 ua ntej los ntawm anti-P. Peb xaus lus tias qhov tso tawm ntawm anti-P-FH1-5 nyob rau hauv glomeruli ntawm Cfh–/– nas yog ib feem nyob ntawm properdin. Kab rov tav bar qhia qhov nruab nrab qhov tseem ceeb, thiab whiskers qhia txog qhov sib txawv ntawm qhov sib txawv. P-tus nqi tau muab los ntawm Mann-Whitney xeem. Bar ¼ 100 mm.

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