University of Dundee
Epidermolysis bullosa simplex generalized severe induces a T helper 17 response and is improved by apremilast treatment Castela, E.; Tulic, M. K.; Rozières, A.; Bourrat, E.; Nicolas, J.-F.; Kanitakis, J.
Published in: British Journal of Dermatology
DOI: 10.1111/bjd.16897
Publication date: 2019
Document Version Peer reviewed version
Link to publication in Discovery Research Portal
Citation for published version (APA): Castela, E., Tulic, M. K., Rozières, A., Bourrat, E., Nicolas, J-F., Kanitakis, J., Vabres, P., Bessis, D., Mazereeuw, J., Morice-Picard, F., Baty, D., Berard, F., Lacour, J-P., Passeron, T., & Chiaverini, C. (2019). Epidermolysis bullosa simplex generalized severe induces a T helper 17 response and is improved by apremilast treatment. British Journal of Dermatology, 180(2), 357-364. https://doi.org/10.1111/bjd.16897
General rights Copyright and moral rights for the publications made accessible in Discovery Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.
• Users may download and print one copy of any publication from Discovery Research Portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain. • You may freely distribute the URL identifying the publication in the public portal. Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.
Download date: 30. Sep. 2021 This article Archiving. article maybeusedfornon-commercial purposesinaccordancewithWileyTerms andConditionsforSelf- Journal ofDermatology,whichhasbeenpublished infinalformathttps://doi.org/10.1111/bjd.16897.This simplex generalizedsevere inducesaTh17responseandisimprovedbyApremilast treatment', This isthepeerreviewedversion ofthefollowingarticle:Castela,E.,etal.(2018) 'Epidermolysisbullosa Lyon1, France, France, Lyon1, iis e yn France, Lyon, de Civils France, Paris, Hospital, 1 Lacour Vabres Castela E. Epidermolysis bullosa simplex generalized aninflammatory is severe disease is and response Th17 bytreatment improved Apremilast a induces severe generalized simplex bullosa Epidermolysis Article :Original Article type 0002 0000- ID: (Orcid CHIAVERINI DR CHRISTINE 0002 0000- ID : (Orcid MORICE-PICARD DR FANNY 0001-62599790) ID :0000- (Orcid MAZEREEUW-HAUTIER JULIETTE PROFESSOR 0002 0000- ID: (Orcid BESSIS DIDIER PROFESSOR 10 France, Dijon, de CHU 7 Department of Dermatology, CHU de Nice, France, Nice, de CHU Dermatology, of Department eatet f ahlg, opcs iis e yn France, Lyon, de Civils Hospices Pathology, of Department Accepted France, Toulouse, de CHU CRMRP, Article
1,13 8 , D. Bessis Bessis D. , , T. Passeron 1 is protected by copyright. ,2 , M. K. Tulic Tulic K. M. , 3 NEM 16, em 2 C3M, 12, Team U1065, INSERM 9 J Mazereeuw J. , 1 ,3 6 5 , eatet f leglg ad lncl muooy Hospices Immunology, Clinical and Allergology of Department eatet f emtlg, opcs C Hospices Dermatology, of Department C. Chiaverini 3 9 , A. Roz A. , eatet f emtlg, H d Mnplir France Montpellier, de CHU Dermatology, of Department ières ières All rightsreserved. 10 F Morice-Picard F. , 1,13 1,13 11 2 CMRMP, CHU de Bordeaux, France, France, Bordeaux, de CHU CMRMP, , E. Bourrat Bourrat E. , -6063-5409) -3815-5417) -4316-916X) -4316-916X) Nice, France, France, Nice, 2 INSERM U1111-CIRI851, Université U1111-CIRI851, INSERM 4
, J. , -F. Nicolas Nicolas -F. 11
D. , 8 eatet f Dermatology of Department Baty vl d Lyon, de ivils 4 AE, Saint-Louis MAGEC, 2 12 ,5
, J. Kanitakis J. , F Berard F. , 12 France, Scottish 5 ,
6 ,7 J. , P. , -P. British , , This article isThis article by protected copyright. All rightsreserved. articleWhat this adds to our knowledge skindisorder related to fragility. E What isalreadytopic knownonthis BULLETED STATEMENTS been never This workhas published before The authors have noconflictofinterest todeclare IRB number: 12.001 This chiaverini.c@chu de 151 route CREBHN, Christine Chiave Corresponding author: Word count Nice, Consortium Genetics Molecular Accepted (E severe generalized simplex bullosa pidermolysis Article
article
France
Department ofDerm : w
. s 25
as funded by DEBRA France and by a grant from andbyagrant as fundedbyDEBRAFrance CHU the of aint Antoineaint deGinestière 75 rini, - , nice.fr Number ofNumber references:
MD,
Ph D
, Ninewells Ninewells , atology
?
, ?
CHU Nice, de 06202 Nice Cedex2 30
Hospital ,
Table .
:
, Dundee, Scotland, Dundee, , 1, BS Hôpital Archet 2, Figures
- e sev gen
France : 7
)
s ra a is
1
3 CREBHN Nice, France. e ialn skin disabling re , CHU de de CHU
This article isThis article by protected copyright. All rightsreserved. Accepteddramatic improvement of skin. lesional CX for neutrophils of infiltration infiltrate Results: was analyzedin for processed o conducted samples Methods: EBS. ArticleObjectives: argueforadditionaldata inflammatory mechanisms mutation by Background SUMMARY after EB of skin by characterized infiltrate immune an of presence the showed We treatment with
C
L9 and L9 rm 17 from n kn ipis f lse (=7 a wl a i rbe si (n=5 skin rubbed in as well as (n=17) blister of biopsies skin in itlgcl analysis Histological
S first A - The aim of this study was to was study this of aim The 1 ptet wt severe with patients 10 n :
in gen sev gen
immunochemistry analysis and quantitative real time PCR time real quantitative and analysis immunochemistry he aut patients adult Three Epidermolysis bullosa simplex bullosa Epidermolysis KRT5 b CXCL lister
muoitceia rtopcie study retrospective immunohistochemical
EBS A
patients, and a marked improvement of skin condition condition skin of improvement marked a and patients,
premilast or fluid 10 in blistering skin blistering in 10 - e sev gen skin blistering skin blistering KRT14 and
and compared with eosin
, ananti hwd constant a showed
genes patients ophils ih EBS with - . Th17 molecule and U assess EBS in uly osdrd s mcaia disease mechanical a as considered sually A se A . . good High 7 generalized - 0% - e sev gen
e sev gen controls
the
tolerance. cond of levels of levels .
dermal
inflammation in the s the in inflammation cases .
. Bitr li ad epidermis and fluid Blister . ee rae with treated were
multicenter severe
and an and
perivascular Th17
a performed was
is
cytokines were detected in detected were cytokines increase
a genetic disorder caused disorder genetic a
a rsetv suy was study prospective Th17 phenotype in the in phenotype Th17 . C CD4+ k ytokine ytokine in of patients with patients of in d peiat ih a with apremilast
immuno in EBS patients EBS in ) n rzn skin frozen on ,
n epidermal an lymphocytes lymphocytes expression expression staining rece ,
were nt This article isThis article by protected copyright. All rightsreserved. Accepted promising EBS phenotype show inflammation by inflammatory additional clinical the all explain to fails disease mechanical a respectively, proteins, (IF) filament epidermis the in keratinocytes basal in expressed intermediate II type and I type (K5), 5 or (K14) 14 of development Articleimprovement sev) in resulting fragility bullosa Epidermolysis INTRODUCTION approapromising newtherapeutic underline and Conclu rwn eiec fr itnt euaoy ucin o krtn i si immunity skin in keratins of functions regulatory distinct for evidence growing , -
n e sev gen blisters fo , sion
te is time, first the r new in the skin of EB of skin the in :
are Our study study Our 12
s ih age with
ains after patients - the key role key the treatment of 1 severe infections severe 5 prese , and encouraging results of topical anti topical of results encouraging and
post due to the fragility of fragility the to due nt at birth at nt i
demonstrate s a group of hereditary disorders characteriz disorders hereditary of group a s s common, is mechanisms - traumatic for the the this this
gen h ue of use the
features T , rsne of presence - disabling disorder
h 3 sev have ches disabling forthis disorder. . The disease The . 17
s blistering
patients, patients,
cells in its pathogenesis its in cells the importance of inflammation in EBS in inflammation of importance the
may be may a herpetiform pattern herpetiform a u tee s an is there but of A the disease the premilast the the an 1 and involved . result patients’ patients’ In 4 mue nitae hrceie by characterized infiltrate immune .
.
a
EBS
EB simplex simplex EB
marked improvement of skin condition in condition skin of improvement marked , s
and recent studies recent and from mutations affecting either keratin either affecting mutations from n anti an 6 - nrae neonatal increased - gen sev gen - 1
keratinocytes inflammatory treatments inflammatory 1 . 2 The later later The
and . - In addition, this study this addition, In T
generalized severe (EBS severe generalized was initially was h
involve m involve 7 molecule 17 ed by skin and mucosal and skin by ed hypothesis is supported supported is hypothesis 5 . However, this theory this However, . have suggested have ucosa. ucosa. otlt de to due mortality
- considered gen sev patients sev gen ,
suggesting 16 Progr .
We provide a essive
Th17 Th17 to be to have - that gen an d a s
This article isThis article by protected copyright. All rightsreserved. mechanicalby suctionoftheskin Controls for RNA extractio RNAlater in one and immunohistochemistry for formalin in fixed (one roofs blister 2 b) and tube), dry same the in (pooled blisters 3 least at of content liquid a) samples: biological filled investigators the patient, patients. all from obtained was 2012 study. this for eligible diagnosed Prospective established at non Retrospective Patients described previously as University Dundee of Consortium Genetic Molecular Scottish the by performed samples skin study The MATERIAL ANDMETHODS
Accepted - Article - 80° aim infected blister blister infected ) . osse of consisted l poeue were procedures All C
of of
: 17 ee enrolled were S with . this on amples
study
n). the associationofthe
study: study: and
study was to a to was study EBS
parts: 2 were design taken secondly, - e sev gen All patients All
Six obtained from 4 healthy volunteers, in whom blisters were induced were blisters whom in volunteers, healthy 4 from obtained following between 1996 and 2012 and 1996 between : first, Patients rnh center French prvd y h Hmn Et Human the by approved , nalyze and multi a
In each center, following dermatological examination of the of examination dermatological following center, each In in clinica
. an
with sadrie qetonie n otie te following the obtained and questionnaire standardized a h developed who
hi witn osn. D consent. written their immune the the le ta 1 year 1 than older l -
center EBS and skin inflammation associated with associated inflammation skin part s - immune - g histochemical retrospective study study retrospective histochemical
en sev en prospective icipated icipated for diagnosis for
oe than more - , histological who had skin biops skin had who
of hic uig h period the during g from age C s study. anss f EBS of iagnosis
2 new blisters blisters new 2 ommittee and criteria
oeua aayi was analysis Molecular whose tissue was tissue whose both sexes, who were were who sexes, both . s
. ies
EBS nomd consent Informed
( at the edge of a a of edge the at March per - - e sev gen gen sev gen using day, - October
fr stor
. were ozen was was Our ed This article isThis article by protected copyright. All rightsreserved. was doneon at at centrifuged Th analysisin Cytokine abovequantity control. housekeeping gene mRNA genes. cytokine Th17 and Th2 Th1, against directed primers Omniscr oligo with transcription reverse by prepared was cDNA USA). Scientific, MA, Waltham, Fisher (Thermo ND8000 Spectrophotometer Nanodrop a using measured was the extracted was RNA total 4°C, at hours 24 for RNAlater in roofs blister of fixation After of Analysis the gene expression roofs/ in blister 1. supplementary data (C and infiltrate), cellular the assessing S roofs ofblisters frozen. were samples skin most study, retrospective the For Histological andImmuno treatmentstart with EBS 3 to we proposed obtained, study pilot Therapeutic i scin wr peae ad tie wt hematoxylin with stained and prepared were sections kin - - bitr lis were fluids blister e
Accepted Article X 80°C. RNeasy - mtf ligand motif) C ipt RT kit (Qiage) and used as a template for amplification by PCR with specific specific with PCR by amplification for template a as used and (Qiage) kit RT ipt Immunoassays based on Luminex™ xMAP™ (multi xMAP™ Luminex™ on based Immunoassays
extraction Kits (Qiagen) according to the manufacturer’s protocol. RNA quantity RNA protocol. manufacturer’s the to according (Qiagen) Kits extraction the the at ha
blister’s fluidfor 2000 rpm 2000 d been conserved in Tissue Apremilast the blister fluidthe blister
s :
stor Following the promising results seen with seen results promising the Following - CXCL for histochemical analysis ed
10 minutes minutes 10 . -
t ° ad sent and 4°C at gen sev adult patients with disabling generalized blistering to generalizedblistering disabling withpatients adult sev gen
- detection detection and quantitation of and 9 immune :
at
CXCL - 4°C. The supernatant The 4°C. - labeled for CD4 and CD8 T CD8 and CD4 for labeled Tek® 10
to RNA extraction and RT and RNA extraction
. and stored at and storedat h lb ihn 4 hours. 24 within lab the eald ehiu i aalbe in available is technique Detailed
o te rsetv study, prospective the For cytokines. cytokines. - - analy phloxine - s 80°C.
were wa the z e s profiling) technology profiling)
initial initial
- expressed as relative as expressed collected
afo (P) (for (HPS) saffron - - PCR cells biological data biological
: - or chemokine or L
dT using the the using dT qis were iquids
and fr and using ozen the the
This article isThis article by protected copyright. All rightsreserved. normal rubbed, from HPS taken the biopsy of Examination skin a also had 5 included, patients Twenty andblistering rubbedskin. with Patients clinical without infection. even patients, by effective be to reported were (7/10) antibiotics topical also and (6/10) cream petrolatum or creams Emollient blisters. extensive chronic with areas, non of development the reported patients of majority from ranged healing wound of duration mean The the ofblisters. prevent onset (8/10) pre a suggesting erythema, by preceded often were blisters The steroidsof ( topical whereas (2/10) vaccines flare A are features summarized in study. prospective patients Seventeen EBS RESULTS: Accepted Article ha patients ll - - up gen sev
n i and - s w foe si bose wr aaye i te ersetv study. retrospective the in analysed were biopsies skin frozen two . improvement was induced was improvement
Interestingly n
patients haveof clinical hallmarks inflammatory disease skin t EBS d hose ,
flare teeth
Five patients participated patients Five 4 with - /10) e sev gen case -
up , inflammatory or general factors such as cutaneous infection (7/10), infection cutaneous as such factors general or inflammatory , erup , fever(3/10) EBS s s - sm ptet rpre ta tpcl plcto o seod could steroids of application topical that reported patients some ,
stained sections showed a dermal inflammatory infiltrate in all all in infiltrate inflammatory dermal a showed sections stained f hi disease their of T
in (4/10 tion
able 1 able - ae a have gen sev gen .
by protecting by
and use ofand use perivascular perivascular were included in included were )
and
and
in ee (/0 wr rpre t idc blisters induce to reported were (3/10) fever
both
c local or local omplain the skin from friction from skin the a few a
eml D+ ypoyi iflrt in infiltrate lymphocytic CD4+ dermal arms of the study the of arms - the retrospective study and 10 in the in 10 and study retrospective the days (4/10) to two weeks ( weeks two to (4/10) days
heal systemic ed
of n bitr (/0 oe friction over (8/10) blisters ing
pnaeu ad unexplained and spontaneous -
existing local inflammation local existing antibiotics ( antibiotics . Clinical and biological and Clinical
and
cold, application cold, 7 mn te 17 the Among - /10). okn skin. looking 3
/10 ).
The
This article isThis article by protected copyright. All rightsreserved. all in elevated were macrophages, and keratinocytes by produced chemoattractant Acceptedpatients Analy roofThe blister ofEBS andfluid patients had no inflammatory cells in epidermis.the Two eosinophils. of made merely was infiltrate the patients two In cases. 2 neutr in eosinophils epidermal dense a had patients Six patients. all in epidermis the of necrosis showed Immunochemistry blisters. roof 10 included study prospective The suggesting the limit Thes (2). epidermis the and/or (4) dermis the in patients these of 4 in present was infiltrate eosinophilic An possible. was blister a of non a of analysis histological study, retrospective the from patients 5 For eosinophils andneutrophilsInfiltration with can in bepresent Article ( analysis ofbiopsies taken from showed skin rubbed similar results profile mutational the to according infiltrate immune lymphocytes CD8 and ( lymphocytes T CD3+ of consisted mainly Immuno were neutrophils lymphocytic; predominantly were cells Infiltrating biopsies. 2 in (+++) dense i (++) moderate biopsies, 11 in (+) sparse was it a distribution; perivascular with dermis superficial the in localized always was infiltrate This 1a). (Figure biopsies detected i o ctkn mN epeso b Q by expression mRNA cytokine of sis
and - itceia sann fr T for staining histochemical T cells cells T results were results (
in 19/21 cases 19/21 only ( ation Fig. 2 patients eosinophils in and 1patient
1c and 1d) showed that the infiltrate mainly composed of CD4+ CD4+ of composed mainly infiltrate the that showed 1d) and 1c of technique. this compared to 4 controls 4 to compared )
( Fig. 1c). There was no difference in the type and type the in difference no was There 1c). - gen sev overexpress Th17cytokines ( gen sevoverexpress - el nies hwd ht h ifamtr infilt inflammatory the that showed antigens cell e eosinophils were not visible on the frozen sections, sections, frozen the on visible not were eosinophils e
Fig. 1b). Analysis of the lymphocytic subsets CD4 subsets lymphocytic the of Analysis 1b). - . The levels of interleukin (IL) 8, a neutrophil a 8, (IL) interleukin of levels The . C i te lse ro ws osbe o 6 for possible was roof blister the in PCR
. KRT5
lesional skin (
(Supplementary vs KRT14 pii iflrt with infiltrate ophilic Fig. - frozen skin biopsy biopsy skin frozen 9 ipis and biopsies 9 n 3 ). - Fig.
4). the degree of degree the Interestingly, Interestingly,
Fig 2)
patients patients .
1 ). rate
This article isThis article by protected copyright. All rightsreserved. comparedsev skin with control significant children 4 from event initiating the be could skin sev gen chemokines cells immune for attractants potent are Keratinocytes Blistering skin patients of EBS with med inflammation especially and inflammation, of role the for argue results these together, Taken IL of levels increased showed which level, mRNA of analysis Finally, differencethe failed to reach statistical significance remained IL TNFα of levels increased significantly found We controls. healthy 4 from fluid blister the with compared was blis the of content protein cytokine The ( respectively patients, 6/6 and 4/6 in baseline above elevated were (TGFβ)] beta factor growth (IL (IFN gamma [interferon Th1 of levels The 3). (Figure controls with compared elevated significantly were which (4/6), 22 IL including response, immune Th17 of markers were as (6/6), Fig.
Accepted Article- 5 - , IL 4, iated by Th17 cells, inthepathogenesis byTh17cells, ofEBSiated
3). ( T2 cytokine Th2 a
- ) eand nhne, oee T however unchanged, remained 5) unchanged. The levels of Th1 cytokines IFN cytokines Th1 of levels The unchanged. ly 18
increase - 19
. We thus hypothes thus We . and ih h dsae and disease the with
Chemokine (C Chemokine Th17 cytokines at the protein level supported the data obtained at the the at obtained data the supported level protein the at cytokines Th17 nu d )
n IL and br of mber s -
2 ( 22 and tumor necrosis factor alpha (T alpha factor necrosis tumor and
( Fig. - ized that an increased production of chemokines in EBS in chemokines of production increased an that ized C motif) ligand 20 (CCL20) 20 ligand motif) C XL ad CXCL10 and CXCL9 Fig. - gen sevoverexpressesCXCL 5). ter fluid was analyzed in 8 EBS 8 in analyzed was fluid ter
4). for skins 4 O the immune infiltrate. Using EBS Using infiltrate. immune the - hr h ctkns sc a IL as such cytokines, Th2 ther euaoy yoie [IL cytokines regulatory
- (Supplementary gen sev. rm elh cnrl, w controls, healthy from - 17A, IL 17A,
- as they they as IL muooiie el i EBS in cells immunopositive
- - 6 and IL and 6 17F, IL 17F, , -
17 (6/6), IL (6/6), 17 two inflammatory cytokines, inflammatory two - secret 9 and 9 and NF Fig. - - 1ad IL and 21 1β were increased but increased were 1β - - 2 10 and transforming transforming and 10 lre mut of amounts large e ] and Th2 cytokines cytokines Th2 and ] gen sev patients and patients sev gen CXCL10 ) .
- 21 (6/6) and IL and (6/6) 21 e - - ad IL and 4 - gen sev skin sev gen ae found have 22 ( 22
Fig. - - gen 13, 4). - -
This article isThis article by protected copyright. All rightsreserved. skin the in lymphocytes of attractors potent three litterature cell for role a suggesting non in extent, lesser a to and, lesional in lymphocytes inf EBS with patients that confirm we study our In DISCUSSION blistering twoof thediscontinuation daysafter aft months. 7 after treatment month. one within disappeared in patients 3 The observed. was blisters of number the in decrease dramatic a respectively, days 30 and daily twice 30mg to 10mg/d from dose daily of increase starte patients The treatments. topical and measures protective despite blistering, active chronic with zones body 5 or 4 least mutation psoriasis of treatment the phosphodiesterase AMP cyclic inhibits EBS with years) 55 to 33 aged with treatment a initiated we data, these on Based Treatment of EBS er er Accepted CD3+CD4+ of infiltrate dermal a have patients Furthermore, disease. skin lammatory Article 10 and8 ,
0 11 10, mutational status was unknown for patient 3. Before treatment, all patients had at had patients all treatment, Before 3. patient for unknown was status mutational tal cmlie o ml admnl an n dare, hc progressively which diarrhea, and pain abdominal mild of complained itially . monthstreatment of
K rtncts r te an el pouig XL, XL0 n CCL20, and CXCL10 CXCL9, producing cells main the are eratinocytes - gen sev patients gen sevpatients
0 21 20, o te des eet wr rpre. hr ws o recurrence no was There reported. were events adverse other No - eitd muiy Tee eut ae n accordance in are results These immunity. mediated d Ptet 1 n 2 wer 2 and 1 Patients .
- ain 3 opand of complained 3 Patient Apremilast e e. peiati a is Apremilast sev. gen for patient 1 and 2 patient 1and for with - 4 and Th1/Th17 activation Th1/Th17 and 4
Apremilast codn t te srai rgmn (progressive regimen psoriasis the to according A premilast of patients with patients of - Apremilast
gen sev have clinical hallmarks of an an of hallmarks clinical have sev gen improves cutaneous symptoms respectively a ohr n hr agtr with daughter her and mother a e - lesional skin, and epidermal necrosis, epidermal and skin, lesional 20
.
)
persistent ml mlcl wih specifically which molecule small
in spring or summer or spring in in 3 a 3 in . Patient 3 had recurrence of . Patient 3 hadrecurrence
and has and several auto several
asa n sopd the stopped and nausea dult patients (3 women (3 patients dult been approved for approved been - . After 10, 15 10, After . immune skin immune
( Fig. with 6 K5 ) .
This article isThis article by protected copyright. All rightsreserved. IL however known, not is increase this for reason The disease. this of progress and/or development the in role crucial a play not may IgE that suggesting IL as such cytokines Th2 other Surprisingly, patient EBS of skin the in eosinophils of presence the with accordance in is patients EBS of skin in level elevated Its recruitment. and activation maturation, eosinophil in mediator the in ELISA by IL of expression increased The and increase T and cells subsequent IL as such molecules, inflammatory release and previously I and Increased cytokines. all of expression the explore to able not were we patients, material of amount small very the to due Unfortunately, IL IL of levels increased found and blisters, the of roof the and liquid the analyzed we To study, levelsofIn our the notrelated CCL20mutational were tothe status ofthepatients. shown been already has CCL20 and CCL2 are chemokines these that suggesting patients, of fluid blister the in CCL20 of level increased an and skin, control with compared skin lesional the in CXCL10 and CXCL9 of expression increased an wound healing ofphases early the inor diseases
Accepted Article- ad n bnac o T1 ctkns a bt te RA n poen levels. protein and mRNA the both at cytokines, Th17 of abundance an and 5 try to understand what understand to try
can partially explain granulocytic the infiltrate observed inthelesionalskin. L - 8 have previously been shownhavebeen 8 previously in shown that in the early stages of wound healing, keratinocytes become “activated” become keratinocytes healing, wound of stages early the in that shown IL neutrophil recruitment neutrophil - el it a oa si region skin focal a into cells - 1 and 8 and 1 likely to play an important role role important an play to likely lse fud i a itiun fnig IL finding. intriguing an is fluid, blister
is not sp not is t ype of inflammation of ype - ecific to EBS, but is more likely to be induced by the wound the by induced be to likely more is but EBS, to ecific 5, ) at both mRNA level level mRNA both at
but also but in vitro in
in the skin of K5 of skin the in )
18 CCL20 -
and in the skin of EBS patients EBS skin of the andin 4 and IL and 4 Tkn oehr tee aa ugs ta the that suggest data these together, Taken . 18, was in - 1
22
lymphocytic infiltrate. An upregulation of upregulation An infiltrate. lymphocytic and IL and -
2 induced by the the by induced ( 4 to recruit monocytes/myeloid dendritic monocytes/myeloid recruit to . Consistent. w - 13 were not increased in our patients, our in increased not were 13 on the the on
- available 8 -
- / (to initiate (to i a h ctkn ad key a and cytokine Th2 a is 5 - - blister blister
5 and 5 mice, but not in K14 in not but mice, ith these results wefoundresults these ith and the small number of number small the and cutaneous eosinophils are involved are eosinophils roof and roof innate 7, 8, 25 8, 7,
at protein level protein at levels of IL of levels CD4+ immunity
. It . It - - 8, has been has / - T cells T
IL mice
- and - 1β 1β s. 6 . , ,
This article isThis article by protected copyright. All rightsreserved. AcceptedEBS Accord approach. to up improvement sustained The treatment. topical despite blisters diffuse active had who patients adult affected im of assessment objective number limited the is study therapeutic pilot this of patients, treated 3 the In of effect the physical byinfection, orchemical stress. Th17 activation pathogenesis. of mutation psoriasis and activation Th17 in CCL20 of role critical the highlighting cytokines, Th17 by keratinocytes from released is chemokine this CCL20; by such in activated persistently is signaling Th17 the However, complete repair tissue and ablated is infection after terminated is and controlled tightly is Articleby repair tissue and CCL20, immunocytes of recruitment peptides, antimicrobial pathogens against body the protecting immunity adaptive in role a play respons immune Th17 the of involvement IL and IFNγ as such differentiation, Th17 the inhibiting IL as such cytokines effector Th17 such cells, Th17 in cells T of differentiation the inducing cytokines in increase an by characterized is EB that time, first the for show, we Here suggesting that this data casual is not auto an pemphigoid, bullous of physiopathology the in -
as gen sev psoriasis KRT5/14
Apremilast patients. 28 ing to our results, anti results, our to ing ,
rheumatoid arthritis and ichthyosis and arthritis rheumatoid
to 7 can lead to a “stressed” phenotype of keratinocytes with an uncontrolledan with keratinocytes of phenotype “stressed” a to lead can
, an approved treatment use treatment approved an , Apremilast
10
enhancing epithelial proliferation epithelial enhancing provement ots f follow of months achieved a rapid and sustained improvement. sustained and rapid a achieved 26 - .
7 IL 17, ; -
IL17 antibodies could also be of interest for treating for interest of be also could antibodies IL17 oee, hs ptet wr te ot severely most the were patients these however, e in the pathogenesis of EBS of pathogenesis the in e - 1 n IL and 21 -
p uprs h vle f hs therapeutic this of value the supports up s TGF as d in psoriasis, in psoriasis, in d 29 f patients of - via immune blistering disease of the skin, the of disease blistering immune . In psoriasis, Th17 cells are recruited are cells Th17 psoriasis, In .
Based on these results, weexamineBased results, onthese -
- 4. These data strongly suggest the suggest strongly data These 4. - 22, and low levels of cytokines cytokines of levels low and 22, nuto o ceoie sc as such chemokines of induction , IL β, nlmaoy r eei diseases genetic or inflammatory 27 .
- The Th17 signaling pathway pathway signaling Th17 The ad IL and 6 treated EBS via -
gen sev. Th17 cells Th17 sev. gen n te bec of absence the and - -
gen sev gen 1 hg lvl of levels high 21,
n EBS In the production of production the L
patients imitation - e sev, gen 30 d d s - . .
This article isThis article by protected copyright. All rightsreserved. 8 7 6 5 4 3 2 1 References for disablingapproach this disorder. a provide also They activation. Th17 of role key the underline in inflammation of importance the demonstrate results our together, Taken Accepted Article Invest Dermatol. Invest JW. Bauer Peckl P, Peking T, Lettner V, Wally Beriault DR, Haddad O, McCuaig JV, Robinson ZJ, Russell D, Lane EB, Fudge DS. Fudge EB, Lane D, Russell ZJ, Robinson JV, McCuaig O, Haddad DR, Beriault 1991 was. PA. Coulombe E. Sprecher Epidermolysis bullosa simplex. dermatitis juvenile resembling herpetiformis. bullosa Epidermolysis RH. Meara GB, Dowling 70: 1103 classification. and diagnosis on recommendations updated Bruckner JD, Fine One. MMP V. Wally JW, Bauer CXCL8/IL S, Hainzl A, Klausegger R, Lang T, Lettner humanand in epidermolysisbullosasimplex. C Wohlenberg U, Reuter W, Roth NEB K14 mutant of behavior mechanical The
- 2013 Jul 19;8(7):e701232013 Jul 1 keratinocytes. - 26. Mol BiolMol Cell - The pathoge The 8 are potential therapeutic targets in epidermolysis bullosa simplex. bullosa epidermolysis in targets therapeutic potential are 8
Br JDermatol Discovery of keratin fun keratin of Discovery
2013 Jul;133(7):1901 - uemn , ay RA Eady L, Tuderman
PLoS One netic role of IL of role netic . 2016Sep 15;27(18):2807
1954; 66:139
. 2012;7(2):e31320.
et al. et - Schmid D Schmid - - 3 ction and role in genetic di genetic in role and ction 1β in severe epidermolysis bullosa simplex. bullosa epidermolysis severe in 1β
Cytokines as genetic modifiers in K5 in modifiers genetic as Cytokines - - Dermatol Clin R125P keratin bundles and networks networks and bundles keratin R125P 43.
Hum Mutat Hum t al. et
, Murauer EM, Hainzl S, Hi S, Hainzl EM, Murauer ,
- 10.
neie eiemlss bullosa: epidermolysis Inherited
2009; 832 30:
J Am Acad Dermatol Acad Am J rmsn nw therapeutic new promising 2009; 23 28: seases: the year that year the seases: EBS - - 41. 32. - gen sev and sev gen
ntner H, H, ntner - / - -
and 9 20
PLoS PLoS mice 14; in in J
This article isThis article by protected copyright. All rightsreserved. 16 15 14 13 1 1 10 9 2 1
Accepted Article olme ATe oeua Rvlto i Ctnos ilg: eai Gns and Genes Keratin Biology: Cutaneous in Revolution Molecular epithelia: PA.The Coulombe skin in function protein keratin epidermolysis bullosa simplex anditsaftermath. Defining CH. Lee PA, Coulombe ofregulators inflammation andimmunity inskin. f intermediate Keratin PA. Coulombe JC, Lessard RP, Hobbs Dermatol Res. simplex. bullosa epidermolysis in formation blister of mechanism proposed El Venereol. epidermo in erythema circinate migratory of A.T Cavani G, Zambruno Hac El D, Castiglia Exp Dermatol stress increased to contributes 14 keratin H Hintner A, Trost M, Wagner Rare Dis. epi for diacerein M, Wolkersdorfer W, Hitzl A, Moder F, Lagler S, Kitzmueller V, Wally 2016 May2;4(3):e1178368 apico and function barrier epithelial of regulation the roles Multiple A. Mashukova R, Forteza PJ, Salas 2017 May;137(5):e67 Challenges. and Opportunities, Diversity, Disease: Associated their - aay S, Abdel MS1, Hawary 2013 May7;8:69 2014 May;94(3):307
2013; 22: 2013; 22: 292 2015 May;307(4):371 dermolysis bullosa: a ran a bullosa: dermolysis e M Doiit A Croe , e ua , acci M, Pascucci N, Luca De T, Carbone A, Diociaiuti M, hem - e71. - - ai M, ae S, bekdr A Aoyoyi, a Apocytolysis, HA. Abdelkader SS, Sayed MR, Halim lymphocytes are directly involved in the clinical expression clinical the in involved directly are lymphocytes
- 4. -
11
et al. et .
- 7.
Imbalance of intermediate filament component filament intermediate of Imbalance
domized controlled pilot study. pilot controlled domized signaling lysis bullosa simplex patients. simplex bullosa lysis J Invest DermatolJ Invest
J Cell SciJ Cell for keratin intermediate filaments infilaments intermediate keratin for
in epidermolysis bullosa simplex. bullosa epidermolysis in - basal polarity. basal 2013; 5257 125: ilament proteins proteins ilament 2012; 13 J Invest Dermatol. Invest J Tissue Barriers. Tissue et al. al. et Orphanet J Orphanet Acta Derm Acta 2: 763 - 8.
Topical -
novel Arch - 75.
This article isThis article by protected copyright. All rightsreserved. 23 22 21 20 19 18 17
Accepted Article M Dy M Gord RM, Day RM, future K, Papp and current applications, clinical perspectives. action, of mode diseases: mediated DP. Bogdanos A, Mavropoulos LI, Sakkas Implications inflammatory di keratinocytes: in chemokines CXCL9/10/11 of regulation Cir A, Celentano A, Marshall immunity. ge other and CCL20 upregulates injury to response M, Johnson Whalen Kennedy 10):19892004 Apr15;117(Pt mutations. keratin of spectrum a by FJ Smith HM, Horn EL, Rugg immunity. adaptive and innate linking genes other and CCL20 upregulates injury to response M, Johnson Whalen Kennedy treatmentstargeted forvitiligo. Interfe JE. Harris M, Rashighi Dermatol. Aprem of Effects the Evaluating Trial r III, phase a of Results plaque psoriasis: severe to moderate with patients in inhibitor, (PDE4) 4 phosphodiesterase
- - Reich K, Leonardi CL, Kircik L, Chimenti S, Langley RG, Hu C, Stevens C, Hu RG, Langley S, Chimenti L, Kircik CL, Leonardi K, Reich Crispin M, Billick E, Mitsui H, Gulati N, Fujita H, Gilleaudeau P, Sullivan P, Gilleaudeau H, Fujita N, Gulati H, Mitsui E, Billick M, Crispin Sullivan P, Gilleaudeau H, Fujita N, Gulati H, Mitsui E, Billick M, Crispin 2015 Jul;73(1):37 J Invest Dermatol. J Invest Dermatol. J Invest Curr Med Curr Med Chem sease. sease. - - Huang LM, Suárez LM, Huang Suárez LM, Huang PLoS One. n B Kra N, rfih CE. Griffiths NJ, Korman KB, on - 49 - . 2017 May 29. . 2017 2012 Jan;132(1):105 2012 Jan;132(1):105
99. et al. et ring with the IFN the with ring Ann Transl Med. Ann TranslMed. .
lo , culuh , otr S. Porter M, McCullough N, illo
2017 Mar 2;12(3):e0172821. 2017 Mar
andomized, controlled trial (Efficacy and Safety Safety and (Efficacy trial controlled andomized, Epidermolysis bullosa simplex in Scotland caused Scotland in simplex bullosa Epidermolysis J Invest Dermatol Invest J - - ls i Poiss ETE] 1). [ESTEEM] Psoriasis in ilast Fariñas M, Krueger JG. Humankeratinocytes'JG. Krueger Fariñas M, Fariñas M, Kru Fariñas M,
Phosphodiesterase 4 inhibitors in immune in inhibitors 4 Phosphodiesterase
2015 Dec;3(21):343. -
γ/CXCL10 pathway to develop new develop to pathway γ/CXCL10 - - 13. 13. e lnig nae n adaptive and innate linking nes
2007; 127: 574 127: 2007; eger JG. Humankeratinocytes'JG. eger
peiat a oral an Apremilast,
- Tissue 80. A Acad Am J
J C J
- for oral oral for specific ell Sci. ell - - - This article isThis article by protected copyright. All rightsreserved. 30 2 2 2 2 25 24 9 8 7 6
Accepted- Article
across the major orphan forms of ichthyosis. ichthyosis. Jan;139(1):152 of forms orphan major the across Suárez KA, Choate JG, Krueger YD, Estrada X, Zheng X, Peng R, Friedland N, Kunjravia Renert AS, Paller understanding ofpsoriasis pathogenesis. S Khalil M, Bourcier R, Vender Y, Poulin C, Lynde a Interleukin Eastham I, Kasman P, Valdez DM, Danilenko Y, Zheng migration.cell stem and leukocyte affect and patients bullosa epidermolysis in signature molecular O5. Igoucheva Salas V, Alexeev 2017 Jul;72(7):1105 D Simon HU, Simon ro a for Evidence S, Yousefi L, Borradori M, Horn C, Sitaru E, Graauw de Dermatol. Invest psor the of Kulesz CR, White JH, Raymond S, Gao JP, Lagowski Y, Liu (He of treatment disease inflammatory the chronic other and in arthritis psoriatic psoriasis, inhibition 4 Phosphodiesterase PS. Helliwell M, Wittmann canthosis. idelb). 2013 Apr27;3(1):1 - Fariñas M, Guttman M, Fariñas - iatic chemokine CCL20 by E3 ligases Trim32 and Piasy in keratinocytes. in Piasy and Trim32 ligases E3 by CCL20 chemokine iatic Nature 22, a T(H) a 22,
- J Invest Dermatol. J Invest Pro 165. - 2010 May;130(5):1384 le of eosinophils in blister f blister in eosinophils of le Alanis JC, Palisson F, Mukhtarzada L, Fortuna G, Uitto J, South A, South J, Uitto G, Fortuna L, Mukhtarzada F, Palisson JC, Alanis - - 2007;445:648 ua Y Spu M Eai H Esaki M, Suprun Y, Yuval inflammatory chemokines and cytokine and chemokines inflammatory
- 1113.
17 cytokine, mediates IL mediates cytokine, 17
- Yassky E. An IL An E. Yassky - 15. –
2017 20. Jul
651 .
- J AmAcadDermatol. 90.
ormation in bullous pemphigoid. bullous in ormation -
17 - 23
- dominant immune profile is shared is profile immune dominant , Oliva M, Huynh TN, Ungar B, B, Ungar TN, Huynh M, Oliva , - induced dermal inflammation dermal induced Alry ln Immunol. Clin Allergy J
Interleukin 17A: toward a new a toward 17A: Interleukin s dominate the blister fluid blister the dominate s - nesn , u , t al. et J, Wu J, Anderson - 2014 Jul;71(1):141 Martin MF. Regulation MF. Martin s. Dermatol Ther Dermatol Allergy. 2017 - 50. and and J .
This article isThis article by protected copyright. All rightsreserved. of roof blister the infiltrate (c) neutrophils and (b) Eosinophils (a). study retrospective the Acceptedof skin the in eosinophils of infiltrate dermal andEpidermal (x200). andc) (b roof 2: Figure (HPS) stain b) Hematoxylin a) biopsies). (19/22 predominant CD4+ lymphocytic, mainly and perivascular dermal, (++), moderate as assessed was infiltrate inflammatory The x100). Fig isindicated prospective study in brackets. the in inclusion of number Their study. prospective and retrospective the both in included dominant, autosomal no NA: gene, 14 Keratin KRT14 gene, keratin KRT5: Table 1: Legends Article CCL20: (C Chemokine TGFβ:transforming factorbeta growth TNFα: tumor necrosisfactor alpha IFN interleukinIL: CXCL: (C Chemokine LC: cells Langerhans IF: intermediate filament EBS Abbreviations r 1: ure γ - : interferon gamma: interferon gen sev: Ep gen sev:
Demographic and molecular features ofEBSDemographic andmolecular features patients
P sann o a formalin a of staining HPS muotiig f foe si bos o bitr ptet 1 (magnification 11) (patient blister of biopsy skin frozen a of Immunostaining total
idermolysis bullosa simplexsevere generalized CD3
: ae F fml, : a, : ot, : er Sm ptet were patients Some year. y: month, m: day, d: female, F: male, M: - - X C motif)C ligand 20
+ lymphocytes -
C motif)C ligand
- , c) CD4, c)
ie, paraffin fixed,
+ Tcells - medd kn ipy a ad blister and (a) biopsy skin embedded , d) CD8
aalbe N: o found. not NF: available, t .
+ Tcells .
- phloxine patient 1 of1 patient - s affron AD: AD: This article isThis article by protected copyright. All rightsreserved. A 6: Figure (Parus, Abcam from bothFrance), antibodies 1:100used at dilution. CXCL10 and France) (Courtaboeuf, SAS Technologies Life from ye are chemokines both expressing Cells red. stained are cells study) 5: Figure cytokines.Th17 cells to compared patients 4: Figure or more fold. to compared median with values (IL pro of expression mRNA Figure 3: blue by with neutrophils arrows.arrows, black indicated are Eosinophils respectively. study, prospective the in 6 and 2 patients
Acceptedpremilast Article - 17, IL 17,
from 4 patients compared to control to compared patients 4 from
- Quantit 21, IL 21, itrs f patient of Pictures muotiig o CC9 n CC1 i foe si bos (retrospective biopsy skin frozen in CXCL10 and CXCL9 for Immunostaining uniaie oae y LS o te lse fud yoie o EBS of cytokines fluid blister the of ELISA by dosage Quantitative treatment (
- ative PCR roofof on the 22) and regulatory (IL regulatory and 22)
fluid of suction blisters suction of fluid b/d/f). - nlmaoy (IL inflammatory
Note the absence ofNote the absence blisters after treatment. , ad 3 and 2 1,
controls. The red line indicates when genes are increased 2 increased are genes when indicates line red The controls.
- 10, TGF 10, an . CXCL9+ cells are stained green and CXCL10+ and green stained are cells CXCL9+ . eoe (a/ before - . A .
, TNF 8, EBS
-
Cytokines reaching statistical difference. B difference. statistical reaching Cytokines - β gen sev blister. gen sev ) cytokines. Data is presented as relative as presented is Data cytokines. ) α c/e , h (IFN Th1 ),
respectively llow. CXCL9 was purchased was CXCL9 llow.
γ , h (IL Th2 ), ad . mnh after months 1.5 and )
- , IL 4, - - 5), Th17 Th17 5), e sev gen -
This article isThis article by protected copyright. All rightsreserved. Table 1 AcceptedProspective study Retrospective studyArticle 7/(8) 6 5 4 3 2/(1) 1 N° 10 9 6 5 4 17 16/(7) 15 14 13 12/(3) 11/(2) 10 9 8 - - - -
-
F M M F M F F F F F F F F M F F M M M F F F Sex
Age 28y 34y 61 y 8 y 43 y 27 y 26 y/32y 25 y 22 y 8 4y /6y 2,5y 8 m / 4 m 3 m 13 d 12 d/6y 10 d 10 d 7 d 7 d 7 d/2y 3d
de novo AD AD de novo de novo AD de novo AD AD AD de novo de novo NA de novo de novo de novo de novo de novo de novo AD de nov de novo transmission
o
NA NA (p.Asn176Ser) exon 1 K5: c.527A>G, (p.Val186leu) exon2 K5: c.556G>T, K14 K14 (p.Tyr258X) x exon K5: c.771_772delGT (p.Glu466Gly) exon 7 K5: c.XXG>A (p.Met119Val) 1 exon K14: c.355A>G NA K14 NA (p.Asn176Ser) exon1 K5: c.527A>G (p.Asn123Ser) 1 exon K14: c.368A>G Mutation K5: c.1401C>G exon 7 (p.Ile467Met) 7 K5: c.1401C>G exon del) (p.Val164_Lys185 K5: c.555+1G del) (p.Val164_Lys185 1 exon K5: c.555+1G>A 1 exon K14: c.(p.Arg125Pro) NF 6 exon K14: c.1131dupT (p.Val164_Lys185del) G.6K5: c. NA
: c.416T>C exon 1 (p.Met119Thr) exon1 c.416T>C : 56
- 751del96pb exon 1 exon 751del96pb
>A exon 1 >A exon
This article isThis article by protected copyright. All rightsreserved. Accepted Article
This article isThis article by protected copyright. All rightsreserved. Accepted Article Figure 3
This article isThis article by protected copyright. All rightsreserved. Accepted Article Figure 4
This article isThis article by protected copyright. All rightsreserved. Accepted Article