Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. iu ugre srbo prtosta i oetrl eoealnslsnsmcs n lo healthy allow and mucosa pa- sinus for nasal all of surgery nasal remove sinus modified proportion entirely of nasal high to consist following a CRSwNP aim recurrence severe in that for disease evidence- reached operations developed of the not concepts reboot risk Despite therapy still as New high surgeries is (3). 6). particularly sinus control criteria (5, a disease CRSwNP 2020 is with of treatment, EPOS tients and There conservative level the CRSsNP under acceptable to (5). in improvement an according patients corticosteroids without CRS recommended options, nasal cases is therapy of In (3). sinuses based consist rhinosinusitis nasal (3). therapies fungal of (NSAIDs)- CRSwNP Evidence-based surgery allergic drugs in the anti-inflammatory (4). corticosteroids range and asthma non-steroidal nasal/oral a fibrosis, with are with without the cystic associated and Subtypes by determined (N-ERD), with differentiated is was phenotypes was disease (3). in and 10.9% respiratory 2020) diseases CRSsNP) of (EPOS exacerbated heterogeneous (CRSwNP/ Polyps CRS comprised Nasal polyps CRS of and The Rhinosinusitis nasal rate on (2). prevalence Paper Europe a Position (GA in socioeco- European Network 27.1% subjects, high European a to adult Asthma has 6.9 and 000 and from (1) Allergy 56 productivity Global involving work the as study In well as (1). life burden of nomic quality impairs (CRS) rhinosinusitis Chronic 29%. Introduction N-ERD impacting 19-50%, possibly CRS. asthma clusters, of endotype treatment 71-100%, CRS biologics-based NP distinct and IL- identified monitoring, (type-2-dominated), of analysis diagnosis, concentration eotaxin/CCL11 extended future high expression-based and chemokine had Our CCL18, 5-7 PARC/ (type-1-dominated), Conclusions: Clusters IP-10/CXCL10 CCL17, IL-5 13-31%). expressed TARC/ low (CRSwNP -4 (p had type-2-dominated) 5, and control Cluster-2 -3 (both CRSwNP). in Clusters eotaxin/CCL11 the (11% than CRSsNP). tissues, and chemokines-negative higher TARC/CCL17 (100% CRS inflammatory significantly MCP-3/CCL7 In and elevated was IL-5- Results: and ECP was concentration clustering. and Cluster-1 partition-based IL-5, clusters. by MIP-1 IgE, seven performed CCL7, total identified was MCP-3/ TARC, analysis CCL11, eotaxin, during The eotaxin/ of obtained CCL18, IL-5. concentration samples PARC/ and tissue inferior CCL17, for in CCL5, TARC/ Th2- n=40) scheduled determined ENA-78/ to patients CRSsNP was subjects: CXCL10, CRS-free related cytokines n=26; all of and chemokines (CRSwNP from consisted chemokines CRS of surgery (n=25) following with group profiling routine of patients control expression concentration The 66 The of included criteria. surgery. approach we 2020 turbinate study, new EPOS case-control the endotyping a this to extended and according In an CRS-patients diagnosed performed Methods: we from disease, IL-5. tissue heterogeneous cytokine nasal this type the characterize further using the to affecting CRS disease Here, inflammatory of an lining. is mucosal CRSsNP) sinus (CRSwNP/ and polyps nasal nasal without and with rhinosinusitis Chronic Background: Abstract 2020 11, May 3 2 1 Artuc F¨orster-RuhrmannUlrike the pattern on expression based chemokine rhinosinusitis chronic in clustering Endotype nvriyHsia Ghent Hospital University Berlin Universitatsmedizin Charite Charit´e Universit¨atsmedizin Berlin 1 ose Zuberbier Torsten , 1 1 rt Pierchalla Greta , lu Bachert Claus , 3 n ed Olze Heidi and , 1 gisk Szczepek Agnieszka , 1 1 2 2 oci Fluhr Joachim , E)Erpa multicentre European LEN) < .0) h analysis The 0.005). α/ C3 IP-10/ CCL3, 1 Metin , Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. neirtriaesreybtfe fCSsmtm,a e PS21 1) tris(rl aa)were nasal) (oral/ Steroids (16). 2012 study. EPOS the per undergoing in as patients participate symptoms, from to CRS recruited patients recruited of were were The free subjects Control (16) Helsinki. but consent. criteria surgery of informed 2012 turbinate Declaration written inferior EPOS the their investi- the in gave All to expressed subjects (EA2/009/07). All according principles study CRS the this with to approved diagnosed according Hospital conducted Charit´e University were the gations of Committee Ethics The population Patient uncontrolled of treatment Methods the for and biologics Patients specific to of lead choice could IL-17 the endotypes advancing CRSwNP. MPO, CRS biomarkers, severe of of parameters new understanding characterization the of additional systematic improving discovery the Therefore, performing the increasingly. and of expands CRS, influences polyps behind nasal the pathomechanisms with CRS examine severe to of therapy was the to study TNF- them the IL-22, assign of (neutrophilic), aim subsequently secondary and patients the The primarily, CRS analyzed We of IL-5 N-ERD. tissue parameter and MIP-1 the MCP-3/CCL7, asthma related eotaxin/CCL11, / comorbidities in PARC/CCL18, Th2 TARC/CCL17, associated CC-chemokines patterns the the and chemokine of CRSsNP concentration of characterize and levels CRSwNP to phenotypes ”Chronic the was the substudy study to this relation GA2LEN of in published phenotypes aim the previously with primary to patterns The immune analogous the asthma. match was to comorbid aiming study and study”, CRSsNP, present CRSwNP, cohort polyposis the monocytes, nasal activate of and rhinosinusitis mainly chemokines aim defined: stimulate CC were general chemokines e.g., families CXC The function, chemokine whereas in Following cells, differ killer families residues. natural (15). These cysteine neutrophils and two T-lymphocytes, (14). chemokines first eosinophils, CX3C of the basophils, classification and of The C, positioning (13). CC, trafficking angiogenesis, mi- the CXC, lymphoid inflammation, directed on and recruitment, the healing, cell based regulate wound in is selectively involved inflammation, also and 2 are type cells They 1/ chemokine-receptor-positive target type cells. in the inflammatory activity of not specific status chemotactic of were activation induce gration chemokines the that elevation of influence kDa) Chemokines concentration the (8-14 and with cells. peptides presence increased small the prevalence are but asthma Chemokines (12), The SE-IgE SE-IgE. of positive far. presence phenotype and so the CRSwNP represented analyzed IL-5 the and clusters of high levels IL-5-negative proportion with IL-5 of the of group %, levels, unde- The IL-5 100 low/ increasing to IL-5. with with clusters of raised while ten concentrations phenotype, into CRSsNP high divided lymphocyte the or enterotoxin-specific was mainly Th17 CRS moderate aureus concentrations, and and Staphylococcus IL-5 IL-5 inflammation as tissue detected tectable well the 2 regarding on as type assigned parameters Based markers were (SE-IgE). 1-, included activation IgE patients neutrophil type study CRS and of The the eosinophil cytokines and pattern, comorbidity. (12). analysis, cytokines, asthma from first proinflammatory al. and tissues the et sinus remodeling, CRSwNP, followed Tomassen nasal CRSsNP, analysis of by groups markers cluster published immune second to of been This GA analysis already the the patients. have on CRS of initiative strate- focused therapy study results research case-controlled personalized FP6 First multicenter and elucidation This European specific options. Therefore, the develop therapy of to disease. framework specific aim CRS improve CRSsNP the and the with such might CRSwNP CRS, factors, within gies, in underlying of processes phenotyping the pathomechanisms range pathophysiological current the parameters, wide The of reflect remodeling a (11). sufficiently tissue by production not characterized patterns, IgE might be T-cell evidence and can and gaining eicosanoids, CRS inflammatory of are The concentration specific options CRSwNP. treatment findings, in Biological histopathological (8-10) as therapy (7). efficient mucosa an nasal as preserved the from reepithelialization C3 P1 CC1,adteCCceoieENA-78/CCL5. chemokine CXC the and /CXCL10, IP-10 CCL3, α, n IFN- and γ T1ascae)o hmkn atrs h s fbooisfor biologics of use The patterns. chemokine on (Th1-associated) 2 2 E iuii ootsuyi the in study Cohort Sinusitis LEN α Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. et h isewshmgnzd(00rm iue,4eC n etiue t10 o 0minutes 10 for containing g solution 1500 NaCl at centrifuged 0.9% and of 4degC) ml minutes, 1 5 in rpm, diluted (1000 4 homogenized was Germany). at was Mannheim, tissue Diagnostics, tissue of Roche the Cocktail, mg Inhibitor Next, Protease 100 (cOmplete cocktail protocol, inhibitor study protease the to According markers in inflammatory snap-frozen immediately of were Measurement tissues -80 The values. samples at collected. mucosal standard were or stored for CRSwNP controls source and from of nitrogen polyps case nasal liquid 25 in and the turbinates CRSsNP females, of inferior 15 case the 54]; the from in max mucosa 11 25, ethmoidal 65] from [min Samples max years 28, 39.8 [min age years (mean 47.2 material CRSsNP age Patients’ with (mean patients patients CRSwNP 40 26 and 12 included 60]; males). ) 26 were max males 65] group 18; 15 max [min CRS 25, females, years the [min 31.4 in patients years age detail, CRS 42.3 (mean In 66 age individuals from (mean 25 reactions patients obtained of of 66 consisted were males). history group of biopsies 13 control consisted the females; nasal The group 91 on males). CRS cohort, based 40 The Berlin females, made subjects. the was control from elevated diagnosis 25 patients or N-ERD and 151 results asthma pulmonologist. Of having test a as NSAIDs. prick registered by to were skin diagnosis Subjects (positive symptoms). Patients clinical surgery. allergy symptoms present upon before allergy plus weeks aeroallergens two clinical to used to IgE-levels not specific according were inhibitors allergic Leukotriene considered surgery. were before weeks four taken not ewe-lse ieecso l aaeeswr etdb sn h rsa-alsts ihmultiple with test significant. Kruskal-Wallis be the test. to U using calcu- set We Mann-Whitney were by [?]0.05 the samples. tested of using the p-values were controls in plotting the parameters and detected drawn, Overall, barely clusters all was group-comparisons. or TNF and of “cluspot” not controls SAE, differences were a and variables they Between-cluster visualization patients The because between for analysis differences additional scaling. cluster lated and the multidimensional from number plot, after optimal omitted scree dimensions The were two the (12). study on in first based subjects the following was performed clusters were of analysis cluster and analysis Component methods Statistical defined retrospectively were pg/mL). levels 151 (IL-5 IL-5 level level the IL-5 IL-5 (12), low negative protocol study follows: general as negative. the be with in to agreement presented considered are In were limits from detection detection enterotoxins of and limit of using measured the examined parameters mixture (BioCheck, below were ELISA The a toxin-1) with USA). measured syndrome against Calif., was shock City, tissue (MPO) directed toxic Foster Myeloperoxidase of IgE and Sweden). C, concentrations and Uppsala, A, (Phadia, The IgE enterotoxin UniCAP-system IFN- aureus USA). total and (S. (Minneapolis, and aureus IL-17, (ECP) Systems Staphylococcus (IL-22, protein R&D cationic cytokines from eosinophil kits additional ELISA the available of commercially concentration The USA). Diego TNF- and San IL-5- (Biolegend The Assay Chemokine Multiple LEGENDplex available USA). commercially ( using CXCL10 by IP-10/ measured MIP-1 and CCL18 CCL11, PARC/ Eotaxin/ chemokines The Germany). Darmstadt, Scientific, (ThermoFischer o .Tepoencnetainwsmaue nec apeuigCoasePu Bafr)asykit assay (Bradford) Plus Coomassie using sample each in measured was concentration protein The C. < α ocnrtoswr esrdwt h uie 0-ytm(uie,Asi,Texas, Austin, (Luminex, 100-system Luminex the with measured were concentrations 0 gm) oeaeI- ee I- 0 o11p/L rhg L5lvl(IL-5 level IL-5 high or pg/mL) 151 to 100 (IL-5 level IL-5 moderate pg/mL), 100 when o .Cnrltsuswr o sdi lse nlssbtsre sa as served but analyses cluster in used not were Control-tissues C. al 1 Table ne h ocnrto f1.8p/L(sa eeto limit), detection (assay pg/mL 12.98 of concentration the under α/ C3 C-/CL,EA7/CC5 AC CCL17, TARC/ CXCL5, ENA-78/ CCL7, MCP-3/ CCL3, eeivsiae.Teceoiecnetain were concentrations chemokine The investigated. were ) 3 γ a esrdb using by measured was ) α, L1,adIFNg, and IL-17, al 2 Table Values . > Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. lsesi lse- n oeta nfieohrcutr ncutr n ,EPwssgicnl lvtdin elevated other significantly three was ECP in 7, than and more 6 clusters in in elevated clusters significantly other were five values in not than IgE more but total and 5-7 Asthma, also cluster-5 clusters in and of clusters cluster-4. IL-5 consisted of in IL-5 concentration 31% of 4. The to to levels 1 cluster-2 high clusters with in in group 0% 23% Cluster between to group: 0 1-4 cluster in clusters levels reported in IL-5 was present low N-ERD, negative/ were the polyps in Nasal phenotypes to endotypes of Association IFN- and clusters, IL- more clusters, ( or more cluster-1 three or in two in present to than was compared higher elevated significantly significantly was was MPO 22 parameters, cases) 3 additional clusters CRS the Regarding in (9 elevated cluster-1 significantly was In and cluster-4. raised in elevated ECP clusters: significantly 2, of total were 2-4. and and Characterization IgE clusters ECP 1 total in parameters clusters and levels 2-associated in IL-5, IL-5 type low 4, significantly the and and differ Also, cluster-1 not cluster-1-3. in did in IL-5 differences IgE undetectable significant by 1-4: no categorized clusters 5): revealed was IL-5 of and group consisted 4 cluster Tables levels first (see IL-5 The association parameter low and negative/ main with the phenotypes group as clinical values Cluster IL-5 and of parameters, severity the analysis. other cluster with were chemokines, the in data of difference phenotype significant and Characterization no ( revealed cytokines ( ENA-78/CCL5 map chemokines/ chemokines, clusters of CXC heat distinct and ratios a CC- 7 and examined as Means of tabulated (indepen- identification other. only and measurements each the calculated chemokine from in on separated resulted based well patients, phenotype) were CRS clinical from of pattern. obtained data expression dent the chemokine of the analysis of Cluster clusters 7 p ( reveals controls (all analysis group vs. Cluster control cases differ concentrations the CRS ECP significantly in to and not different IL-5, compared did IgE, significantly CRS subjects N-ERD total not CRS in CCL17, or TARC/ all higher asthma, CCL11, than significantly Eotaxin/ allergies, yrs) were status, controls. 40.0 and smoking vs. cases gender, yrs CRS of 31 between (p=0.001; proportion younger The significantly were (n=66). (n=25) subjects control subjects The control and CRS of Comparison Results • • • osgicn ieecs h etohlctp -isdpteno h diinlprmtr was parameters additional the of pattern revealed 1-biased chemokines type TNF Other neutrophilic (MPO clusters. other elevated The more significantly or differences. two significant in than no cases) higher significantly CRS were other taxin/CCL11 more (13 or two cluster-4 in than In higher significantly were levels clusters 2) (type eotaxin/CCL11 and cases) TARC/CCL17) CRS (8 MIP-1 cluster-3 In pattern included (not IL-17 parameter additional present neutrophilic was The analysis) clusters. cluster other the more in or three in than cases) higher CRS (11 cluster-2 In α α/ nticue ntecutraayi)wr rsn ncutr4( cluster-4 in present were analysis) cluster the in included (not ). mxdtp 1- type (mixed C3(ye1 ye2 eeswr infiatyhge hn5o oeohrcutr.The clusters. other more or 5 than higher significantly were levels 2) type 1/ (type CCL3 etohlctp -isdpattern). 1-biased type neutrophilic > ye2iflmaoypattern). 2-inflammatory type osgicn ieecswr eetbebtenteceoielevels. chemokine the between detectable were differences significant no , > h C-/C7(xrse ntp1 ye2 ee a significantly was level 2) type type1/ in (expressed MCP-3/CCL7 the , te lses IL-22 clusters; other 3 h P1/XL0(xrse ytp ) C-/C7 and MCP-3/CCL7, 1), type by (expressed IP-10/CXCL10 the , h eeso ye2rltdceoie ACCL7adeo- and TARC/CCL17 chemokines 2-related type of levels the , mxdtp - 1 type (mixed al n al 5 Table and 4 Table al 3 Table 4 > ye2iflmaoypattern). 2-inflammatory type > ). te lses,adI-7 IFN- IL-17, and clusters), other 2 ocaatrz h lses rmthe From clusters. the characterize to ) γ nticue ntecutranalysis) cluster the in included (not < .0) h te aaeeswere parameters other The 0.005). : ye1- type iue1 Figure > ye2-inflammatory type .Teclusters The ). , γ and Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. N-8CC5ddntdffrbtentecutr.MOadI-2wr infiatyeeae ( elevated significantly were IL-22 and MPO clusters. IFN the and between clusters), differ not did in ENA-78/CXCL5 MIP-1 than and higher CCL7 significantly MCP-3/ were CCL18 cases) PARC/ CRS and (7 cluster-5 In > soitdeet fMP3CL ncutr2 ncutr3wt ie h/h namtr patterns, inflammatory Th1 elevated. Th1/Th2 significantly to were mixed pointing - with Th1- responses, typical 3 a inflammatory - cluster CXCL10 Th2 In IP-10/ and and MCP-3/CCL7 Interestingly - 2. Th2-chemokine Th1- The cluster typical concentra- CRSsNP. during a elevated in from - found expressed eotaxin/CCL11 MCP-3/CCL7 suffered we be of who levels, IL-5 to patients effects low all known associated with in is 2 (MCP-3/CCL7) cluster CCR1 chemokine In receptor single levels. a IL-5 of negative low with tions the 1 in cluster low in or the pattern negative these of were of expression levels the none IL-5 matory However, using 1-4, clusters here the (23). performed differentiation. In we for reported subjects which basis difference healthy no as endotypes, and was IL-5 CRS patients There 2-cytokine of CRS type (22). classification not between tissue the was but CRSwNP ENA-78/CCL5 provided IP-10/CXCL10 in patients of studies of infection CRS levels concentration viral in to the the increased Similarly, response in be to in (21). to increase opposed CRSsNP found as to and patients, was reported CRSwNP CRSwNP MCP-3/CCL7 into from addition, distinction obtained In providing samples CRSsNP. tissue with in patients higher MIP-1 the were ( of (20) CXC-chemokine PARC/CCL18 concentration neutrophil and the analytical mul- (19), a As that the and revealed of study“(12). CC-chemokines dataset studies cohort the vious type-2-associated polyposis in selected nasal clusters and we CRS rhinosinusitis seven targets, identified ”Chronic respectively. have GA2LEN groups, we cooperation cluster pattern, as ticenter expression high CCL11 chemokine IL-5 were eotaxin/ the in parameters as on also inflammatory well Based CRS. and 1-related as of low type CCL17 context negative/ TARC/ and the IL-5 above, and neutrophilic mentioned in in CXCL10 endotypes Additional chemokines detected cluster the IP-10/ of To biomarkers. to to as panel N-ERD. 2-related signatures such tissues this chemokines early-type and biomarkers analyzing CRS associating asthma 1-related study allow the type comorbidities first study and in current the with the disease the expression is and of the this chemokine CRSsNP results of knowledge, the The CRSwNP/ course our examined phenotypes natural of we the CRS best understand the the Here, inflammatory to with on useful choice. match based is treatment and endotypes endotypes a into into make subdivided CRS subjects. be to of could of division CRS The 50% that 29%. to markers. demonstrated to 43% (12) 19 in research of present Previous proportion was a asthma with only, and 6 100%, and Discussion to 5 clusters 71 group: in from cluster diagnosed levels ranged was IL-5 formation N-ERD high polyp the with in CRS phenotypes to endotypes of Association te lsesi lse- n n infiatyeeae in elevated significantly and 7 and cluster-5 in clusters other 5 • • h lse nlss a infiatyeeae,adTNF and in elevated, significantly elevated was significantly analysis) also cluster the was SE-IgE differences. significant cases) ( cluster-6 CRS eotaxin/CCL11 in (2 present was cluster-7 analysis) In cluster the in included ( elevated ( levels cases) TARC/CCL17 CRS (16 cluster-6 In pattern noyegop.Crooaigterslso oasne l 1) edtce o inflammatory low a detected we (12), al. et Tomassen of results the Corroborating groups. endotype , ). ncutr3 h eeso oai/CL1adTR/C1 eesgicnl ihrta in than higher significantly were TARC/CCL17 and CCL11 eotaxin/ of levels the cluster-3, in > γ te lses.MOwssgicnl lvtd( elevated significantly was MPO clusters). other 5 nticue ntecutraayi)wspeeti cluster-5 in present was analysis) cluster the in included (not > ie infiatyhge hnohrcutr.Alohrceoie eeldno revealed chemokines other All clusters. other than higher significantly times 3 α/ > C3wr infiatyhge than higher significantly were CCL3 te lses eesgicnl lvtd EIEwsas significantly also was SE-IgE elevated. significantly were clusters) other 3 h eeso ye2rltdceoie oai/CL1 AC CCL17, TARC/ CCL11, eotaxin/ chemokines related type-2 of levels the , h hmknsTR/C1,PR/C1 were PARC/CCL18 TARC/CCL17, chemokines the , h oai/CL1( CCL11 eotaxin/ the , 5 > α/ te lses lo h h/T2chemokines Th2 Th1/ the Also, clusters. other 5 C3(7,etxn C1 1) TARC/CCL12 (18), CCL11 eotaxin/ (17), CCL3 > α inflammatory hnfieohrcutr) ACCL8 and PARC/CCL18, clusters), other five than a rsn ncutr7( cluster-7 in present was ye2iflmaoypattern 2-inflammatory type > > hni he te lsesi cluster-6. in clusters other three in than te lses P1/XL0and IP-10/CXCL10 clusters. other 5 > > te lses P icue in (included MPO clusters. other 5 te lses,adIFN- and clusters), other 2 rlwmxdT1 Th2 Th1/ mixed low or srn h/h pattern). Th1/Th2 (strong ye2-inflammatory type al 1 Table ). > ie and times 5 > inflam- .Pre- ). other 5 γ (not Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. otetuprrsiaoytatcniin ul utfisteidvda,ptetdrce igotcapproach and diagnostic patient-directed diagnostic individual, the used the drugs augment justifies biologics to fully of conditions CRS, expansion tract recent about respiratory The knowledge upper approach. general treat therapeutic to Additional the the tune-up IgE. increase and and to observations possibilities, IL-5 Our monitoring expanded of CRSwNP. and further upregulation CRSsNP without be in patients characterization Weshould in and endotype CRSwNP, markers to in contribute early (12). biomarkers biomarkers as type-2 disease neutrophilic serve with may inflammatory associated levels eotaxin CCL18 Increased heterogeneous PARC/ and markers. and TARC a has inflammatory CCL11, of eotaxin/ pattern being diversity CCL17, wide expression TARC/ CRS a of chemokine by of characterized on endotypes findings based CRS detailed rhinosinusitis previous identified chronic extended of and clustering confirmed consistent endotype endotypes, the IFN- type-2 conclusion, of in In relevance only The N-ERD diagnosed al. We et patterns (37). non-type-2 Tomassen inflammatory cluster-7. before also of type-2-biased in the Additionally, results 50% before to the contrast of consider biologicals. with in prevalence to 2-related confirms cluster-2), asthma importance in observation type an of (9% This of with is subjects which use CRSwNP cluster-7. profile, in regarding in exists IL-5 decision asthma negative 100% a therapeutic to with with the positively cluster-1 subjects correlated making in confirm CRSwNP CRSwNP 11% to about of from phenotype data follow the recent increasing should study, concentrations, studies our IL-5 in More also the (12), shortcoming. findings. results a the previous IL-17 to considered Corroborating al., weight be et statistical to Tomassen add to has and contrast study results in our IL-22and however, our (12); cytokines in al. cluster related et identified type-1 Tomassen the of results Similarly, TNF the and also to 36). and comparable asthma, (35, are neutrophilic asthma clusters, results comorbid all eosinophilic with in with elevated CRSwNP significantly CRSwNP IFN- not-Th2 was the Th2 - in the MPO neutrophiles – in of inflammation impact neutrophilic the of marker stressing typical a In interest, Of (35). CCR3 and 34). CCR1 CCL7 (33, via eosinophils MCP-3/ eosinophils expressing and 34). neutrophils, basophils, MIP-1 (33, cells, environment chemokines contrast, NK CC inflammatory lymphocytes, Both type-2 cells, chemokines. the dendritic these in of MIP-1 eosinophils effects chemoattract and inflammatory to 3 type-2 of known phase additional levels recent are at MIP-1 the in pointing and serum group”, CCL7 al. in IL-5 MCP-3/ et reduced nasal of Jonstam also concentration in the and of significantly increased Further, (31) study An decreased dupilumab a CCL26) (29). with In (CCL24/ dupilumab therapy dermatitis in -3 of atopic (30). studies and weeks in immune polyps 16 eotaxin-2 described type-2 nasal following eotaxin-group as of in secretions the regulation cells, described and considered the T were are for CCL18 memory cells PARC/ essential CCL17 of TARC/ are dendritic trafficking and and of for CCL18 cells number as Elevated PARC/ dendritic well (28). by (20). as sputum produced tissues response in chemokines asthma CRSwNP eosinophils typical was of the of sputum CCL18 be number in inflammatory PARC/ the to the found of in type-2 with were CCL18 concentration the PARC/ correlates levels the markedly upregulate PARC with also cytokines CCL18 association PARC/ CCL11, Type-2 and eotaxin/ in clusters. patients, and IL-5 these CCL17 in of TARC/ higher concentrations to significantly in elevated addition higher observed In was we isolated concentration profile. (27). fibroblasts 5-7, its patients in CRSsNP addition, clusters elevated to In In be compared to (26). CRSwNP found the IL-4 was 25). from with (18, and secretions stimulation controls nasal (19) following to type-2-profile patients, compared a CRSwNP patients with inflammation. CRSwNP from a related associated for suggesting type-2 (24) is difference, of chemokine biomarkers significant CCL17 eosinophilic TARC/ early no as as showed serves CCL17 concentrations TARC/ CCL11 IgE and Eotaxin/ and CCL11 eotaxin/ IL-5 using time, for same possibility the At clusters. other γ eeoeepesdi o-adhg-L5cutr cutr ,4 n ;IFN- 5; and 4, 1, (clusters clusters high-IL-5 and low- in overexpressed were α α/ eeol rsn ntelwI- lse ru.Yt h eaieysalnme fptet per patients of number small relatively the Yet, group. cluster IL-5 low the in present only were C3peoiatyatattp-/ye2clswt C-/CL trcigmonocytes, attracting CCL7 MCP-3/ with cells type-1/type-2 attract predominantly CCL3 α/ C3atat IFN- attracts CCL3 versus lcb-rae RwPptet (8). patients CRSwNP placebo-treated γ atvtdnurpisa ela ml upplto fCCR1- of subpopulation small a as well as neutrophils -activated 6 α/ γ C3(2 a infiatygetri h “high the in greater significantly was (32) CCL3 en rsn nNEDptet a described was N-ERD-patients in present being γ loi lse ) These 6). cluster in also Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. 7 oi ,KeesC ac ,Ncl S ekrS rmrM,e l yoiepten nnslse- nasal in patterns European Cytokine Allergy al. polys. 2012: et nasal MF, without EPOS or Kramer 2016;12:19. with S, Immunol. rhinosinusitis Becker al. chronic Clin MS, between Asthma et Nicolo distinguish patients M, F, Rhinology. non-atopic Haack of C, Baroody otorhinolaryngologists. cretion Klemens I, for K, summary Konig Alobid A 17. C, 2012. Bachert polyps J, nasal Mullol and 2012;50(1):1-12. rhinosinusitis VJ, 2015;13(3):469-73. on Lund paper Paulo). Immunity. WJ, (Sao position Einstein immunity. Fokkens immunity. in and role 16. Chemokines their LC. and Marti system DC, Palomino classification new 15. 2000;18:217-42. a Immunol. Rev Chemokines: Annu O. receptors. Yoshie 2000;12(2):121-7. their A, and chemokines Zlotnik of biology 14. The of A. endotypes Zlotnik 2016;137(5):1449-56 Inflammatory D, Immunol. Rossi al. Clin 13. et Allergy H, J Olze biomarkers. J, of Arebro analysis LO, e4. cluster Cardell on Differentiation T, based Zele al. 2006;61(11):1280-9. rhinosinusitis Van et chronic Allergy. G, P, Vandeplas Cauwenberge mediators. P, Van Tomassen inflammatory G, of Holtappels 12. measurement G, by Maele in Van diseases P, effective sinus Gevaert is 2013;131(1):110-6 chronic S, Omalizumab Claeys of Immunol. T, al. Clin Zele et Allergy Van W, J 11. Bauters asthma. N, and Ruyck polyps De nasal K, in with Blomme e1. surgery patients T, for nonallergic Zele need Van and L, Reduced 2017;140(4):1024-31 allergic Calus Immunol. al. P, Gevaert Clin et Allergy S, 10. J Nasser P, trial. Gevaert Randomized GK, mepolizumab: Scadding e14. with VJ, Lund polyposis AR, nasal severe Sousa LIBERTY C, and parallel-group Bachert dupilumab SINUS-24 of placebo-controlled, 9. safety NP 2019. double-blind, and (LIBERTY randomised, Efficacy Lancet. polyps multicentre, al. trials. two nasal et 3 SE, from with phase for Lee results rhinosinusitis N, Surgery chronic Amin SINUS-52): Sinus PW, severe NP Endoscopic Hellings with C. M, patients Desrosiers Bachert JK, in G, 2019;129(6):1286-92. Han Holtappels C, Laryngoscope. P, Bachert Study. Gevaert 8. Retrospective T, Endotype-Based Zele An van 2014;28(3):192-8. wNP: K, recurrent Allergy. CRS between Jonstam Rhinol Type-2 immunoprofiles S, J initial Am Alsharif in Differences polyps. 7. C. nasal Bachert with P, rhinosinusitis showing Gevaert 2017;72(2):282-90. chronic study G, Real-life nonrecurrent Allergy. Holtappels and al. centre. T, et referral Zele WJ, tertiary Van Fokkens M, a 6. Jorissen association in P, its surgery Levie and M, sinus adults Timmermans after in SF, rhinosinusitis 2012;67(1):91-8. Asthma Seys uncontrolled J, Allergy. al. Veen et der Europe. van T, in Keil 5. survey P, Tomassen EPOS GA2LEN D, of the Hastan summary rhinosinusitis: J, Executive Lotvall chronic R, al. with Newson et D, S, Jarvis Reitsma 2020. R, 4. Rhinology. Kern pathways. PW, care Hellings integrated C, Hopkins including VJ, 2020 2011;66(9):1216-23. Lund rhinosinusitis Allergy. WJ, Chronic Fokkens study. al. GA(2)LEN 3. et A, A Bockelbrink J, disease. Bislimovska underestimated RB, surgery Newson Europe–an C, sinus 2015;125(1):25-32. Bachert in endoscopic WJ, Fokkens of Laryngoscope. D, evaluation Hastan rhinosinusitis. Economic chronic TL. 2. refractory Smith RJ, for Schlosser therapy JC, medical Mace continued ZM, versus Soler L, Rudmik 1. CRS. of monitoring References: and treatment successful a warrant to 7 Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. 4 arrM o tbtE arpaeiflmaoypoen1 n ice elBiol. Cell Biochem J Int protein-1. inflammatory Macrophage E. Stebut macrophage and von RANTES M, CA. Dahinden Maurer 2004;36(10):1882-6. TJ, granulocytes. Schall eosinophil SC, human 34. normal Bischoff of Exp activation 1992;176(6):1489-95. T, J and Med. migration Brunner Exp Th2s. the M, J induce and alpha Krieger (Th1s) 1 protein cells A, inflammatory helper Rot expression T Differential 1 33. al. type et of A, Borsatti responsiveness R, 1998;187(1):129-34. chemotactic Lang and Med. D, D’Ambrosio receptors PP, 2019;74(4):743-52. chemokine Bordignon type Allergy. G, local of Bianchi reduces polyposis. Dupilumab R, nasal Bonecchi al. with et 32. Y, rhinosinusitis Wang chronic N, ex- Tian in The LO, biomarkers Cardell LP, al. pro-inflammatory Mannent 2 et BN, Swanson Immunobiology. G, K, Vos altered. Jonstam De is 31. K, polyps nasal Crombruggen with Van rhinosinusitis N, chronic Ruyck severe in De 2014;219(9):729-36. subsets G, cell Venereol. Holtappels dendritic Derm of CA, Cells pression Perez-Novo Acta Dendritic R, Blood Chemokines. Pezato CD1c+ Disease-specific al. 30. et Produce K, Can Gaspar and Z, Bacso Premature QM, 2017;97(3):325-31. are Doan-Xuan asthma: Dermatitis G, with Atopic Nagy patients G, in in Beke analysis 2009;135(2):295-302. A, microarray Chest. Kapitany Protein H. sputum. 29. Kita in T, PARC/CCL18 Kobayashi chemokine K, the the Iijima of CK, in elevation Kim Asthma CCL17 HB, Allergy Kim of rhinitis expression 28. chronic alpha-defensins. in Decreased and assessment Biomarker neopterin, al. TARC/CCL17, E. 2016;37(1):35-42. 2018;13(5):e0197355. et Magen Endothelin-1, Proc. EV, One. CY, Fefelova PLoS rhinosinusitis: BI, Kang chronic Kuznik IL-5. HK, EV, and and Egorova Kim IL-4 NN, DY, by Tsybikov regulation Lee 27. its NR, and Im epithelium HJ, polyp nasal Lee disrupted Laryngoscope. B, patients. Kim the rhinosinusitis immunoreactivity of chronic -3 26. and activator eosinophilic -2, of Eotaxin-1, potent polyps al. a et nasal K, represents the Kawano 2009;119(6):1053-9. T, in eotaxin Saito concentration H, Yokoi Human protein A, Koyanagi and A. Y, 1996;26(8):1919-25. Kojima Kapp Immunol. T, Yao J D, 25. Eur Kimmig eosinophils. R, human 2006;36(6):748-59. of Hochstetter burst Allergy. J, respiratory Exp Clin Elsner rhinosinusitis. chemotac- chronic granulocyte 24. in and CXCL10 oncogene-alpha chemoattractants growth-related Increased neutrophil of role as Primary al. protein-2 J. Alberty tic et Int. F, Allergol Sachse K, C, Rudack asthma. Matsumoto and 23. chemotactic N, rhinosinusitis Monocyte Okada chronic refractory D, with al. Asaka patients et T, from 2013;62(4):495-502. D, Yoshimura fibroblasts nasal Miotto K, 1998;27(5):281-7. in Wada A, Otolaryngol. expression M, J Luster Yoshikawa sinusitis. K, chronic al-Ghamdi 22. non-allergy-associated O, and Ghaffar allergy of Immunol. in expression S, Clin expression Frenkiel Increased Allergy protein J ED, al. Wright polyps. et nasal LA, 21. with Suh rhinosinusitis AT, chronic Peters with RT, by patients e1-9. Chustz recognition in 2012;129(1):119-27 DR, 18 CCL17 Nagarkar ligand chemokine JA, chemokine Poposki into CC S, insights Peterson Structural of 2018;13:27-31. 20. GL. source Rep. rich Gilliland Biophys a Biochem G, are Obmolova M116. polyps antibody nasal A, Teplyakov Eosinophilic EO. Luger 2006;44(2):145-50. 19. L, Rhinology. Morawietz eotaxin-3. T, and Zuberbier eotaxin-2 U, eotaxin, Forster H, Olze 18. 8 Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. C4 C8receptor CCR8 CCR4, chemokine activation-regulated chemokine TARC/CCL17 IFN-gamma-inducible Chemokines al. of et model A, mouse Mathew a in RN, 1 inflammation Smith Table airway JA, and 2002;168(10):5278-86. Maclean hyperreactivity Immunol. airway AM, J to Tager asthma. contributes 2017;168(1):e54-e8. A, (CXCL10) Ter. Sauty 10 Clin BD, protein chemokines. CXCR3 Medoff and rhinitis 49. Allergic P. Fallahi 1992;21(6):589-96. V, neutrophil-activating Invest. Mazzi novel a Immunol 48. of ELISA. detection sensitive The A. a Walz PM, using Lincoln (ENA-78) MD, Burdick peptide J SL, Kunkel inflammation. RM, airway Strieter allergic 47. produce during cells chemoattractant Mast neutrophil al. 1998;63(6):746-51. potent et a HL, Biol. Evanoff as Leukoc MD, chemokine Burdick function 1997;62(5):604-11. SW, CXC can Chensue Biol. the which SL, Leukoc Kunkel ENA-78, of CM, J function Hogaboam NW, disease. and Lukacs in Regulation 46. role S. its Schnyder-Candrian synthesis, and C, Chemical monocytes Mueller biologically in J. P, into ENA-78 Schmutz Damme MCP-3 A, Van and Walz G, MCP-2 45. proteins Opdenakker chemotactic R, 1995;7(2):97-104. monocyte Ebberink Cytokine. human A, chemokines. the Wuyts active of P, folding Leuven and Van Cytokine purification P, chemokines. CC Proost of subfamily 44. MCP/eotaxin in The approach G. 1999;10(1):61-86. Opdenakker therapeutic Rev. J, novel Factor Damme Van Growth a E, antagonists: Coillie asthma? Van receptor for 43. marker Chemokine diagnostic U. potential 2004;59(12):1243-58. Forssmann a Allergy. as SE, diseases. CCL11 Escher allergic al. J, et Elsner M, Huang 42. W, Gao L, Allergy Li J 2014;51(8):847-54. H, reactivity. Asthma. Bi airway J, J allergic Zhou D, of Wu production alveolar inhibition Enhanced 41. and with al. fluid associated et BAL O, is Tureci 2012;130(6):1384-93. serum, cells U, Immunol. in Luxemburger dendritic Clin CCL18 J, tolerogenic Maxeiner by al. H, et CCL18 Martin CCL18 2013;107(9):1444-52. S, J, of a Reuter Med. Guzman I, Respir as R, Bellinghausen Sarria diseases. CCR8 40. SU, lung human interstitial Sixt of in X, Identification supernatant He culture AD. F, macrophage Bonella Luster M, WG, patients Cai Shreffler in 39. J, IFN-gamma 2013;210(10):1889-98. of Leung Med. role MF, Exp Prominent J Ling e1-3. L. receptor. SA, 2013;132(4):856-65 Borish Islam Immunol. SC, Payne Clin 38. Allergy J, J Negri Dis. P, disease. Huyett Respir respiratory L, Adv aspirin-exacerbated Different Liu Ther with JW, asthma. Steinke al. noneosinophilic 37. of et management N, the DeRuyck to G, approaches Immunol. Novel 2016;10(3):211-34. Clin Holtappels NC. Allergy N, Thomson J Bruaene disease. 36. sinus Van chronic C, in Perez-Novo inflammation mucosal T, orchestrate 2008;122(5):961-8. Zele cells T-effector Van of N, types Zhang 35. rgnadfntoso nlzdchemokines. analyzed of functions and Origin hmsand thymus ) ( CC rgnFunction (19) cells epithelial and dendritic monocytes, from secreted Origin 9 trcsTlmhcts(Th2) lymphocytes T attracts Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. Ctffvle codn oTmse ta.21 (12) 2016 3 al. Table et Tomassen to according values *Cutoff categorical. as analyzed 2 Table receptor CXCR3 10 protein chemokine) /CXCL10 IP-10 receptor CXCR2 peptide neutrophil-activating cell-derived chemokine) ENA-78/CCL5 receptors CCR5 protein-1 inflammatory chemokine) ΜΙΠ1α/῝῝Λ3 receptor CCR3 CCR1, protein-3 chemotactic chemokine) MCP-3/CCL7 receptor CCR3 protein chemotactic chemokine) Eotaxin-1/CCL11 (38) receptor CCR8 chemokine regulated activation- chemokine PARC/CCL18 ecitv ttsiso R ainsadcnrlsubjects. control and patients CRS of statistics Descriptive akr sdfrtecmoetadcutraayi,tecto au o akr htwere that markers for value cut-off the analysis, cluster and component the for used Markers L2 ye1activity Type activity Neutrophilic marker immunity Adaptive activity activity 2 2 Type Type NA pg/ml* 12.98 NA activity kU 2 3.85 Type activityTNF- 2 Type NA IL-22 activity 2 Type MPO activity IL-5 inflammation 2 Neutrophilic type and SE-IgE 1 Type IgE activity NA 2 ECP Type NA CXCL10 IP-10/ NA concentrations PARC/CCL18 increased of Interpretation NA TARC/CCL17 values* Cutoff ENA-78/CXCL5 NA MIP-1 NA CCL7 MCP-3/ CCL11 Eotaxin/ Marker F- 58p/l ye1activity 1 Type activity TH17 pg/ml* 85.8 pg/ml* 25.06 IFN-g IL-17 umnr and pulmonary ) IFN epithelial the macrophage monocyte eosinophil α ( CC ( α/ ( γ- CC ( CC CXC ( C3N ye1adtp activity 2 type and 1 Type NA CCL3 inducible CC ( CC α CCR1, 89 gm*Tp 1activity Type pg/ml* 38.94 oteT1ctkn IFN cytokine Th1 the to response in (48) keratinocytes fibroblasts, T-lymphocytes, monocytes, from secreted 46) (45, fibroblasts keratinocytes, cells, mast cells, endothelial monocytes, from secreted (34) cells and NK cells mast cells, dendritic neutrophils, B-lymphocytes, and T- from secreted (43) cells mononuclear from (42), secreted eosinophils attracts (41) eosinophils from secreted (39) neutrophils and eosinophils cells, mast cells, antigen-presenting from secreted A L akro ueatgneeto oa mucosa local on effect superantigen or Marker /L* 10 γ trcsnurpis(47) neutrophils attracts IFN attracts (44) neutrophils and basophils eosinophils, cells, killer natural lymphocytes, cells, dendritic monocytes, attracts (24) receptor CCR3 on selective 40) (38, immature cells and dendritic cells, B cells, T CD4 naive attracts ypoye (49) lymphocytes Th1 activated attracts (34) eosinophils expressing CCR1 of subpopulations small only and neutrophils γ- activated + n CD8 and + Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. n RsP shaadNED n lse ihngtv L5 lseswt o L5ad3clusters 3 and IL-5 low with clusters 3 CRSwNP of IL-5, proportion negative given. the with are as values cluster well IL-5 One as high N-ERD. chemokines, with and characteristic their asthma and CRSsNP, clusters and the of schema Simplified 5 Table 1 Cluster underlined. are differences TNF-alpha, cluster a (SAE, CRS in limit the used detection between were the values parameters under elevated Some measurements Significantly of columns. IFN-gamma). rate in and high given the IL-17 are to analysis according cluster manner in categorical used Variables CRS. with patients 4 Table aoaoyFindings Laboratory Clinic soito fteedtpswt h lnclpeoye:Smaygraph. Summary phenotypes: clinical the with endotypes the of Association noyepols oie etmpo lseigo niiulcss osdfiecutr of clusters define Rows cases. individual of clustering of map heat Modified profiles: Endotype 7 6 5 4 3 2 .3 50.43.71 .9632231 407313.3 65541825282524 234 .8110500 positive Ratio 0 0.285714 0.125 0 0.166667 0.076923 0 positive Ratio 0.090909 0 0 0 0153846 0 positive Ratio 0.5 0 0 0 0.076923 0 positive Ratio 0 1 0.181818 0.538462 0.25 positive Ratio 0 1 0.857143 1 0 0 0 0.266667 MV 0.142857 0 6.18 1.459333 MV 1.241429 0 0 0 0 1233.43 312.8451 372.8626 931.37 MV 530.6479 640.023 693.7425 2046 524.4044 265.3933 17.185 66.88688 142.835 MV 42.74111 298.265 346.2869 178.0429 30.69 17.7375 83.46462 MV 6.49 15812.5 9846.44375 11930.28571 2673.592308 1294.98875 373.86 686.4611111 MV 26.53524 54.08828 140.0067 21.36967 18.46822 9.585244 6.566953 MV 1739.438 742.6069 881.4973 155.4964 168.232 127.0965 80.5841 MV 14.00733 9.569145 23.5649 6.773338 6.324046 3.164666 3.334867 2.263917 MV 2.65226 5.415359 2.647437 6.601246 3.019662 1.363239 3.494613 3.10241 MV 7.15083 2.613834 8.489828 2.909686 1.260987 analysis cluster in analyzed not Variables analysis cluster 33.57911 in analyzed not Variables analysis 87.71971 cluster in analyzed 173.8526 not Variables 264.4073 analysis MV 1110.102 cluster in analyzed 346.3294 not Variables analysis 332.7771 cluster in analyzed not Variables analysis cluster in analyzed not Variables analysis cluster in analyzed Variables 35603.54 analysis cluster in analyzed Variables 2795.157 analysis 5507.627 cluster in analyzed 4034.732 Variables 2426.655 analysis cluster in 1982.33 analyzed MV Variables 2866.261 analysis cluster in analyzed Variables analysis cluster in analyzed Variables analysis cluster in analyzed Variables analysis cluster 7.239 in analyzed Variables 10.73267 analysis cluster 74.70387 in analyzed Variables 45.04413 analysis 82.2664 cluster in analyzed 53.37202 Variables 32.91463 analysis cluster in analyzed MV Variables ENA analysis cluster in analyzed Variables 8/CC5MOI-0/CC1 I1 C3MP3/CL AC/CL7PR C1 oai/CL1EPI- oa-g L2 A A ai ai L N-lh-aeoilI-7ctgra IFN-gamma-categorial IL-17-categorial TNF-alpha-categorial % IL5 Ratio ratio SAE SAE IL-22 Total-IgE IL-5 ECP CCL11 / Eotaxi CCL18 / PARC CCL17 / TARC CCL7 / MCP-3 CCL3 / MIP1a CXCL10 / IP-10 MPO CXCL5 / 78 A UlM .4M .0p=0.073 p=0.283 0.00 MV 484.64 MV 2.91 629.97 MV 8.34 MV MV 194.37 412.83 MV MV 5122.77 MV 30.00 MV TNF- 337.34 MV pg/ml IL-22 p=0.287 kU/l SAE p=0.038 274.25 MV pg/ml IL-5 119.03 5.86 MV MV pg/ml 424.12 ECP MV p=0.063 kU/l 383.61 IgE MV Total 113.19 MV pg/ml 8.42 IP-10/CXCL10 MV CCL18pg/ml PARC/ 43.97 pg/ml MV CCL17 TARC/ CXCL5pg/ml ENA-78/ L1 gm V1.6M 44 p=0.067 14.49 MV 14.56 MV IFN- pg/ml IL-17 p=0.334 p=0,.088 p=0.415 0% 26.4% p=0.226 16.0% 5.73 MV p=0.232 8.3% p=0.626 p=0.474 2% 11.68 MV 3.98 26.4% 25 MV 48.0% 30.96 64.0% MIP-1 MV 37.98 56.0% MV pg/ml CCL7 MCP-3/ pg/ml CCL11 40.04 Eotaxin/ MV 33.3% % N-ERD 69.4% 66 65.3% % Asthma % Allergy % current Smoker % former Smoker % male Gender MV Age Number γ α α/ gm V5.3M 6.1p=0.227 163.51 MV 55.23 MV pg/ml gm V1.9M 00 p=0.715 10.06 MV 13.29 MV pg/ml C3g lM .8M .4p=0.334 5.94 MV 4.18 MV ml CCL3pg/ 11 ainsCnrl Significance Controls Patients p=0.000 p=0.000 p=0.002 p=0.000 p=0.002 p=0.001 Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. 7 6 5 4 3 2 1 h ce ltwsue odtrieteotmlnme fcutr ob eandi ute nlss as analysis, further in retained be to clusters of number optimal the determine (12). to elsewhere used described was plot scree The 1 Figure Legends: Figure ClusterNumber 100 100 2 53.8 25.0 16 7 13 8 11 9 cases of ce plot: Scree high 5 IL- low 5 IL- tive a- Neg- IL-5 5 IL- 85.7 18.2 0.0 % tio Ra- tive i- pos- 5 IL- 5 IL- > > > > 3 5 5 2 > > > > > C Total ECP 5 5 3 2 2 > IgE +SE > IgE +SE > > IgE 3 5 5 2 12 > (CCL18) PARC > > 5 3 5 > > > > > (CCL17) TARC 5 3 2 5 2 > > > > > (CCL11) Eotaxin 2 5 2 3 5 > > > (CCL7) MCP3 5 5 0 9 0 3 > > (CCL3) ΜΙΠ1α 5 5 (CCL5) 78 ENA- > > > > > P IP- MPO 0 00 50 19 100 19 81 5 2 14 29 43 71 5 3 2 > (CXCL10) 10 5

> > > 22 IL- 300 0 13 2

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12 0 22 11 3 % NP % Asthma % ERD N- Posted on Authorea 11 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158921683.38986270 — This a preprint and has not been peer reviewed. Data may be preliminary. Osuycnetaddsg,suyla,dt nlssaditrrtto,mnsrp rf n nlversion final and draft manuscript interpretation, and analysis data lead, study manuscript design, the and to concept study input HO critical design, and concept study CB manuscript the to input manuscript critical the TZ to input critical measurements, laboratory MA manuscript the to input version critical final JWF and draft manuscript interpretation, manuscript data the draft AJS to manuscript input data, critical interpreted collection, and data analyzed GP measurements, laboratory and collection data UFR interest. Contributions: of conflict no declare authors The interest: of Conflict 13