Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. orsodn author: Corresponding 7 6 5 Kingdom United 4 3 Kingdom United London, Kingdom London, United London, London, College King’s 2 Medicine, and Sciences Life of 1 Santos F. clinical Alexandra and trials clinical in title: use Running for considerations by and used mechanisms be practice could test: that activation platform Basophil a and establish predict certification. to to for to ongoing assurance USA and are quality the efforts allergies and in Currently, and procedures food Europe of results. of in standardization of Clinical laboratories resolution validation, reliability biologicals. natural clinical and and such validation, the reproducibility reflects immunotherapy be procedures, analytical closely monitor ensure allergen-specific vivo can requires it as to thus in BAT As such used for and of treatments, severity. need immunomodulatory application allergen be unpredictable to the of and potentially response reactions reduces IgE can clinical BAT allergic of monitor it cause characteristics disease. can phenotype, or all allergic which EC50 patients’ account of challenges, by into the allergen diagnosis (given takes and the sensitivity BAT tests in resulting basophil intradermal tests curve test. as and sensitization bell-shaped the basophils) than the of %CD63+ From specific outcomes by release. more with (given main histamine stimulation reactivity the following with basophil are degranulation correlated patients, of similar) directly degree allergic is the in and measures BAT cytometry that from flow assay by functional controls a or is allergen (BAT) test activation basophil The Abstract 2020 16, July 3 2 1 Santos Alexandra practice in clinical use and for trials considerations clinical and mechanisms test: activation Basophil eateto eprtr iessadAlry ahsUiest optl ahs Denmark Aarhus, Hospital, University Aarhus Allergy, and Diseases Respiratory Denmark of Aarhus, Department University, Aarhus Medicine, Clinical of Department London, Hospital, U.S.A. VA, Thomas’ Fairfax, St Amerimmune, and Guy’s Hospital, Children’s Kingdom London United Evelina London, Service, Asthma, Allergy of Children’s Mechanisms Allergic in College Centre King’s UK Sciences, Asthma Microbial and Immunology of Faculty School Immunobiology, Sciences, of Course Department Life Gorer of Peter School Allergy), (Pediatric Health Children’s and Women of Department ahsUiest Hospital University Aarhus ALPAN O&O London College King’s A nalryrsac n lnclpractice clinical and research allergy in BAT 1,2,3,4 1,1 rlAlpan Oral , rlAlpan Oral , 2 5 n asJ¨urgen Hoffmann Hans and , Hans-J , re Hoffmann urgen ¨ 1 ,7 6, 3 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. fms el.Cmo ehd fietfigbspisaea SSC as are basophils identifying of methods of CD123 Common number cells. a mast through of identified be low have can Basophils assessment and HLA-DR controls. CD193 the monocytes, or allows markers: allergens and and selection with lymphocytes cytometry stimulation near-unique flow after between and using intermediate before level scatter, cells cell these side single of a state at activation the population of basophil the on focuses BAT and BAT patients allergen of for the principles comfortable to more Basic patients and of safe exposure accurate, concentration avoids process the BAT families. diagnostic measuring test, their the and than making laboratory mechanisms specific thus a more immune investigated, as be performed being explore Furthermore, assay can BAT functional to IgE. allergen, a allergen-specific technology with As BAT of stimulation disease. (Figure following state-of-the-art allergic diseases cells IgE-mediated the allergic live suspected of cover on with monitoring patients will and of of we assessment diagnosis expression review, clinical the the this 1991 in measures in In importance Knol which Edward gained and 1). by assay discovered progressively was sections laboratory has CD63 cytometry BAT basophils. all blood flow then, the of the to a surface of the of contributed is on draft (BAT) markers draft authors activation first submission. test first All to the activation and prior basophil activation. wrote the manuscript The trials, H.J.H. basophil wrote the clinical test. of of O.A. version activation in mechanisms manuscript. final basophil use the basic the the and approved the of of monitoring applications versions about immune clinical subsequent sections reactions, the the allergic about of about sections sections editing the the author: of coordinated named draft each first by the contribution of Statement Europe in laboratories Keywords: by used 190 be count: could word of that Abstract reliability certification. platform and for a USA reproducibility establish the ensure to clinical in allergen- to ongoing validation, and as assurance are analytical such quality efforts food requires Currently, treatments, and BAT of results. immunomodulatory procedures resolution of to natural of application the than response standardization Clinical monitor validation, clinical biologicals. specific to it as monitor and more used such As immunotherapy severity. and be procedures, be unpredictable the specific predict potentially vivo can of to can in of reactions thus it for and outcomes allergic phenotype, need and allergies cause patients’ main the allergen can the reduces the which BAT reflects and challenges, closely disease. are IgE allergen allergic by similar) and of of (given tests or characteristics diagnosis intradermal reactivity the EC50 all basophil in account by patients, tests histamine into (given sensitization allergic with takes sensitivity in correlated BAT BAT directly basophil from test. is and resulting and basophils) curve cytometry flow %CD63+ bell-shaped by the controls From or release. allergen with stimulation following Abstract: count: Word [email protected] address: 8640 Email 7403 20 (0) +44 number: 6424 Fax 7188 20 (0) +44 number: Telephone Kingdom 2 Allergy, United Paediatric of Department Address: Santos F. Alexandra + LD- CD203c HLADR-, + lsayoddnrtccls n D0cadFc and CD203c and cells) dendritic plasmacytoid h aohlatvto et(A)i ucinlasyta esrstedge fdegranulation of degree the measures that assay functional a is (BAT) test activation basophil The leg,aahlxs aohlatvto et D3 igoi,immunotherapy diagnosis, CD63, test, activation basophil anaphylaxis, allergy, 5043 + rCD193 or + + as xrse nSSC on expressed (also CD123 nd 2,3,4 + or ot ig tToa’Hsia,S17HLondon, 7EH SE1 Hospital, Thomas’ St Wing, South floor, 2 Fc . ε IadIE hnue sslcinmresin markers selection as used when IgE, and RI ε ...dfie h uln fterve,wrote review, the of outline the defined A.F.S. Iaeas xrse npuioetprogenitors pluripotent on expressed also are RI high low oiohl) CD123 eosinophils), CD193 + CD203c + + CD123 , as xrse on expressed (also + 1 HLA-DR n,since and, - , Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. ufc g n peuaeC6 elt o–g-eedn tmls n as fnon-responsiveness of cause phosphatase One Syk stimulus. of non–IgE-dependent a level to of low 10% well a approximately CD63 of is upregulate basophils and However, Fc through IgE allergens. stimulation surface to are respond BAT ba- not the do non-responder in population stimulants of the used enigma frequently the most The and controls response. IgE-mediated basophil and on non-IgE sophils allergen of the of and importance effect subjects The the among appreciate significantly better vary subject. to optimal surface, might same concentrations the cell point determines the allergen the factors in this on these allergens thus IgE of different activation, combination epitope-specific between The basophil the dose- of for antigen itself. different density the basophil concentration the of the the allergen antibody, affinity of factors by as IgE characteristic of such shown the intrinsic number in markers of an variability activation a results and spreading dose- surface are the basophil epitope patients There a from of IgE, different misleading. curves the seen allergen, response be of for dose be can the cell) the antigen can of impact the of can concentrations As valency to that single the different (bound with form. of tests on IgE in affinity curves, and relative epitope-specific vary response the of allergen that and curves profiles antigens the with of for dose-response for complexity basophils allergens the IgE %CD63-positive However, on of the epitopes bell-shaped. affinity for often is the curve IgE-Fc curve on antigen-specific bell-shaped response depends the a As that is patients reaction allergen. allergic of concentrations in increasing result BAT typical The C5a to and dose-response linked IL8 receptors PAF, bell-shaped like protein The substances G-coupled univalent MRGPRX2 through endogenous for activated ligands for and be receptors fMLP also by like may substances activating exogenous cells univalent the for mast of receptors origin and signalling. 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IgE the inhibit thus can wortmannin since , 1,10,11 . 21-23 ε I h ihant g eetro lo aohl,rslsi increased in results basophils, blood on receptor IgE affinity high the RI, ε 14 I?sbnt,ado h H-oan fknssSkadLyn and Syk kinases of SH2-domains the of and subunits, RI?? osbyi obnto iheeae mut fCD45 of amounts elevated with combination in possibly , muergltdeoyoi ssSA2 n AP whereas VAMP8 and SNAP23 uses exocytosis Immune-regulated . 1 n D0ci peuae lgtyearlier slightly upregulated is CD203c and 19,20 hrfr,i speeal oicueabodrneof range broad a include to preferable is it Therefore, . 3 iue2 Figure ε Iee huhte xrs omldniiso cell- of densities normal express they though even RI 1 n atcells mast and hw xmlso aigsrtge sdin used strategies gating of examples shows 15 8 n iheooeformation exosome with and ernlto a ensuidmainly studied been has Degranulation . ε 1 Icmlxi eetraggregation receptor a is complex RI n D5cnas edtce,but detected, be also can CD45 and 14 7 ti -rnmmrn protein 4-transmembrane a is It . . 13 18 e hshrlto of phosphorylation Net . n a emodulated be may and , 16 5,6 IgE-mediated . D0aand CD107a . 9 t oein role Its . 13 The . 12 , Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. estvt a easse n hudhneb eotdfis.Torcn tde fpau leg found severity allergy symptom peanut patients of predict studies not recent could Two extract allergen first. severity wasp reported symptom to before be and reactivity measured reactivity hence be of should to relationship and needs reactivity a assessed Basophil be IgE. through can anti-Fc allergen sensitivity or to sensitisation (anti-IgE control relevant negative control biologically the positive of to IgE-mediated compared the CD63 %CD63 and express as allergen that expressed basophils be of can proportion response and the to basophil refers of and reactivity allergy, measures Basophil standardised gold-standard distinct drug more the the are being results diagnosing considered have sensitivity of best When still advantage and to is the the Reactivity which reactivity. important gives have gating, and of condition is manual developed unstimulated level it to been the compared same analysis, have in objective the analyses gating basophils change data at the fold CD63+ automated control the 2.5% During use negative and of a cells threshold control. on positive a negative CD63 set to of threshold compared percentage same the MFI respond measuring CD203c to by fluorochrome-conjugated cells in identified of the selection are up to basophils warming resilient be Activated of should surprisingly extracts method allergens is Allergen Molecular the testing determined. and concentration. Basophil be allergen used, tolerated, allergen. should major antibodies be of and allergen can protein mole of that of in dilutions concentration terms defined high log in a defined sequential methods clearly is four standard be w/v than should to 1% more than according to less Typically, solubilized response principle) (Peppys and be blood. them with to can be allergen toxic relevant brings should not the concentrations patient preparations bring can the drug patient parenteral the allergen necessary, or An if and, allergens BAT, purified the for or used recombinant extracts, standardized collection Ideally, blood of hours test 24 the to BAT before up days weeks or basophils two three day blood after steroids same fresh result the positive with on a obtained ideally stimulation, obtain fresh, basophil to used analyses. for possible best cytometry used flow material are perfor- and on, the basophils markers and be Blood key collection may of blood tested between staining being time for patient used instance: the for that antibodies BAT, medication the test BAT, of activation of results basophil stimulusmance the the a affect can of to factors results Various response to the in respond influence activated they can get whether that assess not Parameters to do important non-responders is of it degranulation IgE/Fc basophils fMLP, IgE-mediated through to Blood inhibit respond that that controls. a basophils fMLP Wortmannin, IgE-mediated to blood and tripeptide that response non-IgE Staurosporine bacterial confirming a a like The as After mounting inhibitors used valid. to often of is insensitive is capable degranulate, is test and to activation receptors control alive fMLP positive the are coupled mediated that G-protein basophils the blood allergen through i.e. relevant basophils the document, activates clinically that the to is document stimulus it to non-IgE whether important to however, time, is exposure this allergen It response. at by basophil clear decreased this not transiently modulating is is is It Syk may allergens that response. basophil against thus allergic reactions of non-responsiveness unwanted the amount basophil prevent limit to the rhinitis; of system increases, of amounts immune IgE lower incidence the with the allergen-specific of barrier associated of final was reduction a non-responsiveness IL-3 apparent of be basophil months an presence Singapore, within the and in in state Syk, basophils performed responsive culturing basophil by study a vitro large in to a experimentally revert reversed In may been also patient has a state responder transient; is phase non-responder 30 . ε I ..te onturglt D3o ees histamine. release or CD63 upregulate not do they i.e. RI, 1 ernlto hog MPocr atrta h g-eitdrsos.It response. IgE-mediated the that faster occurs fMLP through Degranulation . + aohl tagvnalre ocnrto ra h ai f%CD63 of ratio the as or concentration allergen given a at basophils 30 28 niitmnsadtpcltetet ontiflec h eutof result the influence not do treatments topical and Antihistamines . niiul en etdo A hudso ramn ihoral with treatment stop should BAT on tested being Individuals . 25,33 28,29 4 oee,i td fws eo leg,basophil allergy, venom wasp of study a in however, ; oee,ta eutmyntb h aet that to same the be not may result that however, ; ε I.I evst ouettepresence the document to serves It RI). 31 . 34 32 h atrsuystthe set study latter The . 3 . n hudb sdat used be should and , 24 steaon of amount the As . 31 24,25 ehd to Methods . h non- The . 28 tis It . + 27 26 to 3 . . . Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. n ohmxmlratvt n afmxmlratvt r eemndb tigannlna uv to curve non-linear a patient fitting the by respiratory with determined in concentration, correlates are allergen both sensitivity against reactivity level, Basophil plotted half-maximal is and concentration dose-response. allergen reactivity the histamine, each maximal of at reactivity release both of the and degree measuring The by allergen. allergen of to sensitivity basophil curve determining PGD dose-response studies the by of preceded slope reactions was the allergic on of based calculated severity be the be can can and and 100 activation by basophil plied half-maximal eliciting EC concentration as allergen expressed the to refers sensitivity Basophil ramn a enosre oteclrtalre n ytne legna ela g-eitd(but IgE-mediated which as changes, well These as basophil. allergen the bystander to intrinsic a changes and suggesting allergen to controls culprit immunotherapy positive the have non-IgE-mediated) specific to studies not allergen observed Various monitor following been treatments. to sensitivity has allergens immunomodulatory tool treatment and venom useful with insect reactivity a and or basophil respiratory be time food, in may BAT over decrease time-point, status a given allergic documented a in at of status changes classification dichotomic allergic natural the patients’ identifying beyond from response Apart allergic the of characteristics non-allergic. dose subtle versus peanut threshold more allergic the to define with challenges to sensitivity used during basophil be and reacted reactions patients milk allergic of which of forms severity at allergy all increasing to milk/egg with instance, allergic their associated For were outgrown been who egg phenotype. had children milk/whole between who patients’ fresh activation upregulation children to basophil allergic CD63 and reacting of of while of degree profiles milk/egg intermediate different aspects baked an shown tolerating have showing detailed children allergy with more egg stimulation and recognise allergen milk following of to phenotypes useful different or with be patients sensitization also IgE undetectable can of BAT immunotherapy cases stop in venom to useful bee when particularly and guide be wasp to can allergen-specific both responsiveness is and to bronchial correlation testing sensitization the this IgE double in that to component suggests value further allergic clinical which the add AHR, reflects in high BAT drop with that 20% reacting and a patients causing in challenge (PD bronchial seen allergy challenges 1s in not during food used in of allergen dose volume diagnosis the allergen expiratory the with to forced confirm correlated exposure was to stressful CD-sens use and In asthma, its risky allergic tests. supports from sensitization strongly patients specificity IgE dispense high to and Such compared 100%. sensitivity and high 96 between study retained with allergy specificity peanut CD203c immediate high a of very intensity study fluorescence its typically mean to given subjects the disease, allergic to in sample of sensitized or relevant basophils basophils patients the %CD63+ of accessible with the status in studies, an allergic 57,58 peripheral different increase the dose-dependent in in constitute reflect a allergens available showing venom to thus readily insect shown and and more has inhalant BAT are are The food, basophils anaphylaxis. cells basophils and Blood by mast reactions allergen. released allergic tissue the mediators to of than effect exposure the blood following from cells result mast anaphylaxis and and reactions reactions? syk allergic allergic by immediate about Acute regulated and us interdependent tell are BAT that can show What degranulation to crosslinking IgE from leading activation of parameters distinct rhinitis allergic in improvement asthma sensitivity allergic basophil of assess to methods developing for scene uhsuishv e oagoigfreit pligBTt h igoi fIEmdae allergic IgE-mediated of diagnosis the to BAT applying into force growing a to led have studies Such . 2 rCys-Leukotrienes or 50 36 htdcesswt nraigseverity increasing with decreases that rhinitis , 40 hc a eetyvldtdfurther validated recently was which , 36,49 3 ciaino lo aohl hudb sesda aho -2lgdilutions log 5-12 of each at assessed be should basophils blood of Activation . 37 odallergy food , n nfo allergies food in and 41,42,44,45,51,52 35 53,54 eemnto fsniiiyo aohl oalre yflwcytometry flow by allergen to basophils of sensitivity of Determination . 20 oee,sseai nlsso inligmlclsi h pathway the in molecules signalling of analyses systematic however, ; .Ti orlto a otydet ainswt o H n was and AHR low with patients to due mostly was correlation This ). 62 . 42,45,69-71 33,38-40 aohlratvt n aohlsniiiyapae obe to appeared sensitivity basophil and reactivity Basophil . 61 legnimmunotherapy allergen , A a lob aubei elcn tn challenges sting replacing in valuable be also may BAT . 25,38-40,50 nfo leg,adces nbspi eciiyduring reactivity basophil in decrease a allergy, food In . 5 63,64 33,59,65-68 42,45 35 rae rprino ciae aohl has basophils activated of proportion greater A . hthsbe hw ob sfli h diagnosis the in useful be to shown been has that , rC-eswihi h nes fEC of inverse the is which CD-sens or , n hne nsniiiyrflc h clinical the reflect sensitivity in changes and 59 hsi nte xml fhwBTcan BAT how of example another is This . h pcfiiyo A opau ranged peanut to BAT of specificity the , ´ estvt oalre tteclinical the at allergen to sensitivity s 41-45 36 25,35 nvnmalry A can BAT allergy, venom In . n niIEtherapy anti-IgE and Dsn nrae with increases CD-sens . 60 nptet with patients In . 55,56 50 multi- 46-48 . . Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. rtcl oass lnclrsos oI.Ti prahi nieyt ewl cetdb ainsand Depending challenge. patients food by oral an accepted of context well the food in be with even to allergen, diagnosed the been unlikely study to already exposure in have is of treatment required approach fearful for often be considered This are may being challenges and challenge IT. patients allergy additional as food to and practice, oral stressful clinical response an quite in clinical undergo families be challenge to assess can allergen Having allergic of to context allergy. be performance the food protocols the to in with requires known important diagnosed patient IT clinical previously particularly a food future for for been is in BAT eligibility have This moment of that treatment. the application patients allergy at key in which, for a for patients therapies, allergy, of of food eligibility outcomes of confirm clinical to of surrogate is a between trials trials. being comparable clinical to and of addition reproducible context In informative, the be the in BAT to in of aspects results and practical treatment use the are to optimal for there response order However, clinical in level. of considered cell sites. biomarker be effector given a to the as a at need both mechanisms that to trials, underlying reactions clinical possible trials in of acute clinical exploration potential of huge in risk a test the has activation BAT on basophil effect the their of ones use of the The terms i.e. activity, in specific biologicals allergen other higher higher to allergen. is are with response allergen-specific that one BAT patients sensitivity is the clinical the the intrinsic that are the result, in their IgE omalizumab a of enhances vitro to As proportion cells respond in allergen. whose the effector better the allergen these on to to the of reactivity likely receptors surface basophil most to to expression increase the surface response bound paradoxically on in can in is density reduction omalizumab progressive receptor and that a in IgE cells, to reduction omalizumab leads the effector the it of reduces on cells, dose mast and the FceRI tissue adjust circulation of and to in basophils need circulating IgE the of captures about surface decisions antibody omalizumab make with anti-IgE treatment to to the used response was the BAT monitoring in IgE-mediated. the useful study, are be treatment to shown of desensitization also effects has or reducing BAT observed tolerance therefore whether natural and confirm in basophils to allergen lacking help and of can is cells severity amount receptors mast its the ITIM-coupled of or IT reducing through through surface reaction binding signalling the allergic allergen cell on an for inhibitory inducing antibodies IgE of IgE with chance cross-link compete with the to that comparison able IgA, its is and and removed, that IgG is namely basophils IgE, surrounding containing to plasma plasma which post-treatment BAT in of blood BAT, whole effects washed suppressive the and is function adding antibodies to the prior and assess allergen antibodies pre with to which pre-incubate blocking ways to in or are approaches basophils cells primary the sensitization sensitize sensitized Passive with to immunotherapy. used strongly allergen are correlated plasma for immunotherapy post-treatment biomarker allergen diagnostic year effects of first a long-term weeks the into the during three explore treatment first to of interest effect the clinical of during cell sensitivity effector basophil an in are they following IgE, immunotherapy patients bear of and some FceRI in express basophils immunotherapy treatmentsAs oral immunomodulatory of with effect BAT in clinical Changes the to compared similar OIT treatment treatment of during is of efficacy tolerated which discontinuation allergen in transient, of difference dose be the the mirroring can of terms foods, in to SLIT increase immunotherapy with the sublingual by to measured treatment as compared on allergen, oral while the reactivity to of desensitization threshold clinical accompany in anergy, basophil of typical are al 1 Table 78 h s fseicihbtr fsgaln oeue ontemtehg-ffiiyIEreceptor IgE high-affinity the downstream molecules signalling of inhibitors specific of use The . rsnssm ftepatclise n eomnain ocruvn hmadrahan reach and them circumvent to recommendations and issues practical the of some presents 42 74 yial,ps-ramn lsacnanalre-pcfi nioiso ieetisotypes different of antibodies allergen-specific contain plasma post-treatment Typically, . 62,75,76 h upesv ffcso lcigatbde nue uigtetet change A treatment. during induced antibodies blocking of effects suppressive the ; nte xeietlstpta a eue oepoeteeet fblocking of effects the explore to used be can that setup experimental Another . 73 ec odts omntrrlpeo h allergy. the of relapse monitor to test good a hence , 72 42 h eraei aohlratvt a emr ooiu in notorious more be can reactivity basophil in decrease The . swl streyaso treatment of years three as well as 6 85,86 77 vdneta lcigatbde a induce can antibodies blocking that Evidence . 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Data may be preliminary. ti motn ont htrgltosadribreetcvrg yhatcr ytm ayfrflow for vary systems healthcare by reimbursement/coverage and regulations that note to important is It oterprigo h eut.Teeaesvrlipratcmoet fteaayia aiaino a of validation analytical the of components sample important blood several the of are draw There the from results. test procedure the activation testing of basophil the reporting of the accuracy to the determines validation test Analytical validation activation clinical basophil assurance methodology, the quality the of continued of validation diagnostic and validation Analytical phenotype minimal analytical patients’ requires the against practice fulfill test clinical not the to BAT of do the who (24). of patients allergen use The to in aspergillosis, reactivity bronchopulmonary particularly clinical allergic diagnosis, is low as for data indicate such show basophils), criterion disorders, to criteria. basophils anergic allergic likely additional where rare or more cases an in non-releaser in is as value as Even finding has challenge. 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BAT food Thirdly, as is oral response. act response of that assess initiation to clinical therapies therapy and a potential on modifying asthma preventing months the allergic before immune or challenge, in improves medications for food allergy omalizumab it to oral of indication an Firstly, use an exposure the of of for reasons. prolonged confirming resolution need confirmation several require allows the natural 1) for may defer it the may groups: important Secondly, of it is main reactions. as monitoring anaphylactic allergy work-up, three 3) diagnostic of into and of confirmation categorized profile therapy - safety The specific be setting therapy. a can to clinical for which response day-to-day eligibility BAT, the 2) of allergy, in indications an applications possible different the have practice of may can clinical that BAT in conditions, test The allergic activation severe challenges. basophil with allergen of patients during considerations use outcomes for Similar The undesirable reserved treatment. of often such risk are from additional which benefit at patients biologicals, otherwise allergic be for exclude would can made that protocols be reactivity study of can of part threshold as done high challenges required, with reactivity of threshold the on • • • • noae a aacnb eoie r motn si loscmaio frslsi different in results of comparison allows it as which important in are RfB databases consistency. deposited sharing ensure assurance accurately. and would be quality and preparations perform can centers external allergen to data offer procedures, continue raw BAT to assays annotated regulation of planning validated Standardization devices are the medical (www.instand-ev.de) systems. that individualized INSTAND diagnostic assure created and have vitro to laboratories (www.rfb.bio) in 2024 testing, measures proficiency by the control established approved implemented yet quality not EU be have the bodies to regulatory 2017 has In that (IVDR), necessary. stored anticoagulant is previously chelating if control even calcium freshly, alternative prepared Testing: be Proficiency an should is lyophylised. Allergens EDTA or testing. frozen before conditions. basophils ambient all stabilises in in that response shipped concentrations. of basophil when number same even a the resolved at induce allergen been each not mostly for samples does correlations concentration of clinical allergen the of and Stability of tested importance concentration being the given allergen hence the A patients, to allergen. specific are the patient given of a in activation basophil and interferences Analytical nr-u rcso sasfrrpaso ape ttesm iepito h aedy h precision The day. same the on correlation point good time same shows BAT the at the samples for of analysis repeats for assays precision intra-run precision intra-run and Inter 90 : 91 h usino tblt ftesmlsbfr ecigtelbrtr has laboratory the reaching before samples the of stability of question The : o utie ihqaiyueo A nteciia etn,cntn quality constant setting, clinical the in BAT of use quality high sustained a For hntasotdi eai ue,smlscnsa tbeu o2 hours 24 to up stable stay can samples tubes, heparin in transported When . ie legnde o tmlt/nuebspiso o-legcpatients non-allergic of basophils stimulate/induce not does allergen given A : ne-u rcso nlsssmlsa ieettm ons whereas points, time different at samples analyses precision Inter-run : 88 . 7 92 ic A sntawdl vial sa and assay available widely a not is BAT Since . 57,87-89 . al 2 Table umrsssome summarises Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. o h ett eribre yhat aessesadisrnecmais ioosqaiyassurance quality rigorous companies, insurance ISO and in systems out care laid health as by legislation. control, reimbursed national quality by be constant required to setting, increasingly and test clinical necessary the the is For in 9001:2016, BAT test ISO and activation of ISO15189:2013 use cell 15189:2012, high-quality mast sustained the a test For non-responders, activation 10-15% basophil the the of and BAT assurance allergy blood the Quality peanut complement fresh of to clinical for study used following need be previous outcome the may a (MAT) equivocal are in with which 67% patients BAT, by in of OFC test of tests second-line number tests a sensitization the other up. as IgE all work performed to and diagnostic values compared be history routine cut-off accuracy should of for BAT diagnostic part criteria best as line. and overall BAT showed used incorporating peanut allergens when to available selection, consider BAT and patient to although (AIT) challenge by issues instance, allergen immunotherapy For influenced practical vivo allergen also is in BAT are on the the There based of years of start four success and the The three identify of after the to weeks outcomes benefit clinical alternative three with would an within correlated success patients clinics as improved treatment For which achieve used sensitivity OIT. about to be basophil as likely information can more such prognostic BAT were allergy, treatment provide responsiveness OIT, food this also starting for from may before treatments BAT most OFC for perform eligibility patients. considered. with confirm routinely is allergic correlation challenge to not strong supervised food required do its a a also Given that in before often test occur performed staff. intermediate is they clinical be safe OFC should even to important anaphylaxis, and an needs and of parents offers reactions, reactions BAT patients, diagnosis allergic allergic reactions, in the treat acute clinical anxiety for to severe significant gold-standard expertise to cause clarify The and for lead and can OFC. facilities BAT can foods requiring peanut, the It for of patients with case OFC. Even environment of but the BAT is number in detectable cases, sensitization. allergy 2 the such are h from food reduce in Ara that and allergy and instance and IgE wide for true cases is sensitive specific components, with equivocal zone allergen very and informative grey those are SPT are immunologically differentiating there this patients for IgE which in studies foods, sensitized results specific value largest most have food For and significant the of SPT cut-off. provides of i.e. majority PPV the some area, the 95% which specificity, Although grey the in their below immunologically improve and done. an to testing, were into determined prick BAT fall been skin the have performance and of diagnostic sensitivity cut-offs IgE the utility and positive about specific clinical specificity evidence as largest the the such the of is on tests, there summary which for A in cut-offs Allergology re-challenged available. of and be area are the cannot tests is patients other allergy showing Food which can no despite BAT in which still high, on allergies for but seen drug drugs are clinical lower, be life-threatening of allergies are for can of case allergies food specificity the case drug for and in the for BAT BAT sensitivity or in of of its useful sensitivity specificity determine extremely The and be to foods. sensitivity is between BAT The differences of significant validation interest. clinical of of correlates aspect essential others test An CD63, activation detecting basophil Tests”) gained the Developed has of “Laboratory as validation testing Clinical as adopted available. Basophil to allergy been commercially Italy. Many referred are has and (also America. that and South Germany kits procedures research and use Spain, house in Europe Sweden, in Eastern used Africa, use as mostly high-complexity South clinics such is including a diagnostic BAT world, countries, of the a the some capabilities throughout as Europe, acceptance in the used In test has is that clinical BAT institutions. practices a academic States, allergy as United in well the making as In decision laboratory clinical world. cytometry of the flow part of a parts as different test in assays based cytometry 40 ti airt efr knpikts rseicIEadteeoeteetsscnb sda first as used be can tests these therefore and IgE specific or test prick skin perform to easier is it , al 3 Table n a enpeiul reviewed previously been has and 93-95 76 napau I td,priiat neigtesuywt o basophil low with study the entering participants study, OIT peanut a In . ttetm fti eiwteeaesc e-p npiaeciia practice clinical private in set-ups such are there review this of time the At . 57 eoerfrigptet o F.Ti rpsdapoc reduced approach proposed This OFC. for patients referring before , 97 . 8 3,58 . 96 naohrsuy sn rs olnSCIT, pollen grass using study, another In . 40 ocruvn h ao limitations major the circumvent To . 45 . 60 . Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. ntemperature changes in and vibration by affected Recommendations be can reactivity Basophil time over reduced is reactivity Basophil issues Practical and practice clinical the 1. in Table on BAT professionals of care implementation health further Tables: of for training important as disease. are allergic research. well results for allergy treatments as BAT novel assurance and of extremely existing quality into interpretation be diagnosis trials can continuous validated clinical the and clinically for support laboratory been and thus Standardisation robust have practice A and that clinical results vitro for 4). reliable (Table in both and valuable treatments reactions consistent immunomodulatory provide allergic during can immediate which monitoring of method its surrogate and considered a diseases be as allergic can seen of basophils be can as BAT accepted The a be with can on measured two 35% the 2 is as for lab shipping 2 results high Conclusion in during lab the as between blood. processed to temperature results %CV whole is mailed The The but in B then range. 25% 1. events Donor and 2-37C Day rare below from within 1 on be is sample Lab sample processing should in blood the for locations a that a 0) 1 ensure (Day and process, Lab to collected 1) quarterly strip to (Day is temperature this mailed day it In and following day the (QA/QC). 0 the the clinical for control Day processed processed BAT offer and is is provide that assurance it A that laboratories quality where Donor states, these of from network between 6 sample a created in blood protocols. was establish laboratories harmonized AmeriBAT to and 9 counterpart, BAT place are in grade European system there its assurance quality States, to inter-laboratory well. United Similar an as with the contrast testing allergens testing in stark IgE food IgE in common time, to of is it present assurance this bring the quality and As At process external importance. this in minor of obtained of momentum improved results be the dramatically may maintain of analysis used heterogeneity of reagents the method of to the en- standardization standardizing uniquely the would surprise, that (CV test our gesting results To diagnostic the cutting-edge test. representing of a the organisation coherence external of of European establishing assurance quality the quality and the as in testing EAACI hance allergy basophil Engaging with standardizing EAACI. working of of Clinical control professionals and aim the Allergy the of under with Academy certifica- assurance force European for quality the demand task Interest laboratories, increasing a the biomedical the of launched for meet auspices Immunology To 9001/15189 the EAACI. ISO under in within year described Medicine second tion Systems ba- every and of continue Diagnosis meetings Allergy opportunities These Group discussed tests. have basophil of testing 2006 development basophil since developing testing laboratories sophil laboratories. European certified of in Representatives place in be to needs process 29 rcia susadcnieain o pia s fBTi lncltrials. clinical in BAT of use optimal for considerations and issues Practical . 29 . 98,99 n aemtrglryi h UOA etn eist teghnthe strengthen to series meeting EUROBAT the in regularly met have and , < 0 o eeto fCD63 of detection for 10% eprtr rnfro samples. of transfer temperature of stability ensure that transportation of method Ensure u o2h fbodcollection blood of 24h) hours to few (up a within BAT Perform 9 + aohl)i e uoenlaboratories European ten in basophils) eprtr oto o samples. for control temperature with system transport Prefer falst ihntesm time same the frame. within site all samples of Test samples. of timely delivery ensure to sites between system transportation Good trials clinical for Implications 100 ti motn to important is it , 31 sug- , Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. allergy venom Insect rgallergy Drug ies Examples allergy Food disease Allergic 3. Table 2. Table analyses data for adopted the method with vary can activation basophil of Quantification cytometry flow by protocol and BAT the the by identify basophils, to used markers by the influenced be can activation basophil of Measurement used anticoagulant the with vary can activation Basophil tissues the to homing and degranulation basophil induce Recommendations can infection and inflammation chronic allergen, to Exposure response basophil reduce can corticosteroids, oral including Immunosupressors, issues Practical 4,32 estvt n pcfiiyo h aohlatvto ett igoedffrn legcconditions. allergic different diagnose to test activation basophil the setting. of clinical specificity the and in Sensitivity test activation basophil the for Indications . 30 . e venom Bee venom Wasp agents blocking muscular Neuro- lactams Beta- g allergy Egg entallergy Peanut 101 4 . . 104 105 103 29 . 105 102 40 econsidered. be can analyses data analyses. Automated data of step for each defined be should Criteria conditions. optimised and method validated with a performed be should BAT condition. inflammatory chronic or infection during or exposure allergen after BAT performing Avoid BAT. for collection blood before immunosupressors Avoid olce nohprno EDTA. or heparin blood into in collected performed be can BAT ug/ml 1 – 0.0001 venom, Bee ug/ml 1 – 0.0001 venom, Wasp ouoim4 CD63+ 4% Rocuronium aiu %CD63+ 5% Various 10ug/mL Ovalbumin 0ng/mL 10 extract Peanut stimulation Allergen 10 basophils CD63+ 10% basophils CD63+ 10% basophils basophils basophils CD63+ 5% u-ffSniiiySpecificity Sensitivity basophils CD63+ 8.11% cut-off Optimal lncltrial. clinical the throughout and between centers used be to needs methodology same exact The standardized. be should cytometers flow and trial a clinical throughout used be protocol should and reagents same The studies BAT. of using criteria exclusion an be should conditions inflammatory infections Active SPT). not but challenge (namely exposure allergen to be prior to collected needs BAT for Blood using BAT. studies of criteria exclusion an be should with immunosupressors treatment continue to Need trials clinical for Implications sites. between and studies during methodology and material same the using be collected should BAT for Blood 1 93% 91% 5 83% 85% 0 96% 80% 5 80% 55% 7 100% 77% 8 96% 98% Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. A. 2: Figure 1. Figure IgE. circulating and density receptor IgE in decrease the of consequence a as allergen to response basophil decreases Omalizumab threshold. allergen clinical in change the reflecting in important particularly be to seem CD-sens) (or EC50 and immunotherapy specific allergen of course the over decrease sensitivity basophil and reactivity legends: Basophil figure allergy. and IgE-mediated Figures of diagnosis the confirms cut-off positive the above test activation basophil A messages clinical Key 4. Table Examples allergy Respiratory disease Allergic Bspiswr dnie sSCo D0c D2+HLA-DR- CD123+ CD203c+ SSClow on as gate identified monocyte were – Basophils Lymphocyte 2. 1. cells: CD193+ CD123c+ SSClow as identified were Basophils 1. eaiecnrladbspi ciaini esrdaoetegt o h te tmlto conditions, stimulation controls. other is positive both the or for but on gate allergen 5%, the Gate with above (was measured either is % 3. activation 2.5 basophil SSC-H, and population. to control vs positive negative threshold SSC-H the negative Assess then CD63 5. FSC-A, Set 2.5%), vs 4. at FSC-H universal CD193, more exclusion x Doublet CD203c simultaneously 2. markers plot, FSC/SSC a lnclapiain ftebspi ciaintest. activation basophil the of applications Clinical xmlso aigsrtge o basophils: assurance. for quality strategies EQA, gating (BAT). of test Examples activation basophil the of time-line Historical Aspergillus pollen Grass 106 36 rrs 1 f rAsp or ul) (10 extract fumigatus A stimulation Allergen SQU/ml 0.0001 – 100 extract, pollen Grass 11 DN ND ND Specificity ND Sensitivity cut-off Optimal basophils CD63+ 2.5% 4 h D3gt ssto the on set is gate CD63 The . DND ND Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 12 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 7 rnaA aog ,AiaA ta.I ir vlaino g-eitdhprestvt ecin to reactions hypersensitivity IgE-mediated of evaluation vitro In al. et A, Ariza C, quinolones. Exocytosis. Mayorga Compound A, on Aranda Focus 17. Cells: Mast of cells. Degranulation Res. mast Anaphylactic Immunol R. human J Sagi-Eisenberg mature O, but of Klein VAMP-8, degranulation and 16. (VAMP)-7 activation-induced protein for membrane mediators. Immunol. associated required cell Vesicle J are mast al. Eur VAMP-3, et for or S, VAMP-2 pathways Bolat SP, not exocytic Frank LE, Diverse Sander S. 15. Sugita mast NR, fine-tuning Bin in H, CD45 2018;46(2):235-247. for Xu Requirement M. systems. 14. Huber ligand-receptor A, different Weiss by M, mediated Kuhny responses (BAT). ML, Testing cell Hermiston Basophil Clinical G, of Grochowy Cons differential 13. and to Pros relationship al. et basophils: M, Rep. Ferrer human EF, Asthma in Knol HJ, CD63 Hoffmann and processes. 12. CD203c degranulation anaphylactic of and Expression piecemeal of Jr. histamine basophils. regulation D, of Analyzing MacGlashan patterns LS. degranulation Clerck the 11. De study WJ, to Stevens instrument Methods. novel MM, TGFbeta-1. a Immunol Hagendorens surface J (HistaFlow): CH, exosome cytometry Mertens via flow CH, by signalling Bridts release Endosomal DG, Ebo al. 10. et SC, Jang Y, cells Yin Vesicles. mast GV, Shelke Recruitment human Cell Immune 9. in Cultured Tetraspanins of FcepsilonRI. Roles R. Varied and receptor Many Dahl Migration. The MD. and IgE Wright PO, MJ, Hickey high-affinity L, Schiotz Yeung the 8. LH, of Christensen expression PM, Frandsen for munol. into HJ, heterogeneous Insight Hoffmann are apigenin: versus Apigenin-7-O-glucoside actions. 7. al. cytotoxic et and anticandidal D, of Stojkovic modes D, basophil- the Stanisavljevic novel M, as Smiljkovic CD164 and 6. CD107a, upregulation. IgE-dependent CD13, of of kinetics with Identification time in decrease al. patterns Res. response et can two A, of CD123 dissection Jakob and of S, markers activation Florian expression F, The test. Hennersdorf activation G. 5. basophil Lack the V, of Turcanu strategy gating A, the 2016;6:11. Stephens for diagnosis N, implications in Becares activation: testing activation basophil AF, basophil Santos of utility clinical 4. The disease. al. allergic et CCR3 of C, of Mayorga monitoring expression AF, and Santos Robust cytometry. HJ, CA. flow Hoffmann Dahinden in WJ, 3. marker Pichler mono- selection S, CD63 basophil Ducrest via single M, activation Fux a basophil T, as human Gentinetta OV, Monitoring Hausmann D. Roos 2. J, Calafat H, 435. Jansen antibody FP, clonal Mul EF, Knol 1. References: B. 2005;15(5):325-335. 2012;157(3):246-250. 2019;8(1):1650458. Allergy. 2016;16(8):56. rn Immunol. Front 2019;2019:9542656. 2008;38(3):855-863. 2011;66(2):247-254. leg lnImmunol. Clin Allergy J 2012;375(1-2):30-38. 2018;9:1644. Allergy. 2015;70(11):1393-1405. 918( t1):328-338. Pt 1991;88(3 XL J. EXCLI 13 Allergy. 2017;16:795-807. elSignal. Cell 2011;66(1):85-91. lnEpAllergy. Exp Clin 2009;21(8):1277-1286. n rhAlryIm- Allergy Arch Int 2010;40(9):1365-1377. ice o Trans. Soc Biochem lnTas Allergy. Transl Clin urAllergy Curr Extracell J Cell Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 6 alnB opA oaso G dad ,SeigrM ddynJ aohlalre threshold allergen Basophil J. Adedoyin asthma. M, in Skedinger sensitivity transfusion. M, allergen blood Eduards of measure SG, by activa- a Johansson IgE-sensitization is A, Passive basophil CD-sens, Nopp sensitivity, The al. B, Dahlen et immunotherapy. HF. M, 36. specific Merk Hage van monitoring I, A, and Sauer Nopp 2005;60(9):1192-1199. specificity SG, S, Johansson sensitivity, Moll-Slodowy 35. R, the allergy: reflect Kwiecien venom test B, activation wasp basophil lergy. Sachs in the SM, of test parameters Erdmann tion Distinct peanut. al. to et 34. reactions A, allergic basophil Douiri of of G, threshold analysis Toit and to Du severity approach AF, programmatic Santos Data-driven external 33. al. and et standardization M, test: Schneider activation A, tests. basophil activation Calatroni basophil the SU, in The Patil confidence W. force Building 32. task Aberer al. EAACI et an A, M, - Groselj-Strele C factors. assurance N, critical A, quality A and P, Heinemann issues M technical EM, 31. allergy: Sturm of diagnosis B, mass the and Kranzelbinder in cytometry test flow GJ, activation using Sturm by analysis. activation basophil before 30. Assessing hours 24 al. stored et S, blood Gupta in N, cytometry activation Gaudenzio basophil K, Mukai blood whole 29. the of standardisation nonreleasing peanut. Improved anti-IgE to AF. in test Santos events G, Intracellular Lack D. M, autocrine Kwok Roos of AJ, 28. evidence Verhoeven TW, IL-3: Kuijpers FP, secrete basophils. Mul human basophils reactions EF, disease. allergic Human Knol allergic of AP. in 27. threshold responses Bieneman functional and KL, and severity phenotypic Chichester of for Biomarkers JT, anergy.priming Schroeder basophil al. et of 26. C, characterization O’Rourke Systematic challenges. G, peanut Toit oral Du al. during AF, et Santos B, 25. Lee AK, Andiappan KJ, 2017;72(3):373-384. and Puan Syk of 24. loss the re-capitulates desensitization. basophils of human disease. methods of 1070. other allergic desensitization by basophils characterized Subthreshold in expression human mechanisms Jr. FcepsilonRI in D, basic signalling MacGlashan IgE-mediated down-regulation: 23. of Syk Regulation in U. role Muchhal its 2014;44(5):713-723. G, and Moore CD32b Jr., eosinophils. by D, and Macglashan basophils, testing. activation 22. cells, Basophil mast Jr. DW, IgE, MacGlashan 21. 5. DD. a Mini-Primer):S450-456. Metcalfe markers: activation Suppl C, basophil 2006;117(2 human Prussin of response 20. Differential al. of et Occupation approach. R, Off-Target cytometry Ortolani Does flow A, multi-parameter Vella Hypersensitivity: S, Rocuronium Chirumbolo al. 19. et Role? CH, a Play Bridts Receptor J, MRGPRX2 Elst the AL, Gasse Van 18. 2004;59(10):1102-1109. lnTas Allergy. Transl Clin yoer lnCytom. Clin B Cytometry leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J 078(up 2)(26):15-16. (Suppl 2017;8 leg lnImnlPract. Immunol Clin Allergy J Allergy. lnMlAllergy. Mol Clin 2017. 1992;90(1):92-103. 2020. leg lnImmunol. Clin Allergy J 14 leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J 2020. 2008;6:12. lnEpAllergy. Exp Clin Immunol. J 2019;7(3):998-1003. lnEpAllergy. Exp Clin 07193:8-9 e811. 2017;139(3):889-899 Allergy. 2015;135(1):179-186. 2013;132(4):777-787. 2009;182(4):2432-2438. leg lnImmunol. Clin Allergy J 2011;41(8):1091-1097. 2009;64(9):1319-1326. lnEpAllergy. Exp Clin 2012;42(7):1060- Allergy. Allergy. Al- Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 4 aGahnD,J.Rlaaiiyo ua aohl:clua estvt n aia histamine maximal and as sensitivity basophils cellular allergies: basophils: on human focus of Releasability Series; variables. independent Review Jr. are Clinic DW, release the MacGlashan in 54. subcutaneous Immunology modulation. pollen WG. immune grass Shreffler assessing of for SU, biomarkers effect Patil controlled Sustained P. nonrandomized 53. real-life, Korosec a N, response; Bajrovic basophil M, allergen-specific Silar study. of M, suppression Kosnik on asso- immunotherapy M, are Zidarn pollen fraction. grass E by 52. immunoglobulin caused grass-specific symptoms larger Rhinitis a P. component- and Korosec Allergy. sensitivity and A, Exp allergen Grahek sensitivity basophile M, elevated threshold Silar with allergen M, ciated Kosnik Basophil M, Zidarn al. 51. et diagnosis. SG, allergy Johansson hazelnut improve A, associated diagnostics Nopp a is resolved J, (CD-sens) Xolair; sensitivity Brandstrom with allergen basophil years 50. High children. 6 in al. After asthma et H. OB, allergic Nilsson severe Oman B, with M, Nordlund Palmqvist JR, Konradsen J, 49. Ankerst with- J, during Adedoyin changes clinical SG, follow-up. withdrawal and Johansson 3-year CD-sens A, H. Nopp treatment. Oman to of M, 48. approach years Palmqvist useful 6 J, after a Ankerst Xolair sensitivity: SG, of threshold Johansson drawal allergen A, Basophil Nopp al. 47. et J, evaluation. of Ankerst efficacy outcome SG, treatment clinical Johansson anti-IgE long-term A, reflects Nopp allergen sensitivity Basophil basophil 46. patients. al. in pollen-allergic et grass changes P, in G-dependent Siddhuraj immunotherapy PA, Immunoglobulin subcutaneous Wurtzen P. JM, Korosec Schmid M, 45. immunotherapy. Silar pollen in birch M, side-effects during Kosnik predicts sensitivity basophils N, threshold of Lalek sensitivity High P. 44. Korosec predicts E, sensitivity Music basophil N, Bajrovic in immunotherapy. M, improvement venom Silar Early M, Kosnik HJ. Hoffmann 43. immunotherapy. R, pollen grass Dahl changes with PA, immunological relief and Wurtzen of symptom measure JM, allergy biological Schmid between a discriminates CD-sens: 42. H. test Oman activation SG, Basophil ASIT. Johansson by LO, stimulated Cardell al. A, et Nopp N, 41. Becares basophil children. A, of peanut-sensitized Douiri in analyses AF, tolerance Combining Santos A. outcome. Nopp challenge timothy MP, 40. and wheat gliadin Borres of omega-5 SG, prediction wheat, hydrolyzed the Johansson to enhances G, antibodies grass IgE Hedlin and CD-sens, C, sensitivity, threshold Nilsson allergen N, Nilsson threshold the allergen in 39. Basophil C. complement children. Nilsson peanut-sensitized easy-to-use MP, Borres in and M, DBPCFC reliable 247. Rudengren and SG, a IgE-sensitization Johansson CD-sens, be A, sensitivity, Nopp can S, CD-sens Glaumann SG. 38. Johansson rhinitis. LO, allergic Cardell of A, diagnosis Nopp 37. Allergy. 2012;42(1):49-57. 2015;70(5):547-555. Allergy. Allergy. Allergy. 2009;64(5):811-814. n rhAlryImmunol. Allergy Arch Int 2005;60(11):1401-1406. leg lnImmunol. Clin Allergy J 2010;65(1):56-60. Allergy. leg lnImmunol. Clin Allergy J eit leg Immunol. Allergy Pediatr Allergy. lnEpImmunol. Exp Clin 2006;61(3):298-302. leg lnImmunol. Clin Allergy J 2007;62(10):1175-1181. 15 lnEpAllergy. Exp Clin 2013;161(1):87-90. lnEpAllergy. Exp Clin 1993;91(2):605-615. n rhAlryImmunol. Allergy Arch Int Allergy. 2014;134(3):645-652. 2012;23(4):376-384. 2012;167(1):59-66. 2020. 2015;45(9):1412-1418. 04143:4-4 e745. 2014;134(3):741-744 2010;40(8):1186-1193. Allergy. 2013;162(1):50-57. 2012;67(2):242- Clin Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 2 haaaa ,JnsS,CltoiA ta.Eiec fptwyseicbspi nryidcdby induced anergy basophil pathway-specific of Evidence children. al. peanut-allergic et in A, immunotherapy Calatroni biomarker oral SM, novel peanut Jones a A, oxidase: Thyagarajan diamine of have 72. expression who Basophil subjects al. response. et in immunotherapy GW, peanut allergen Scadding to of JA, Layhadi unresponsiveness MH, Sustained Shamji 71. al. et children. M, immunotherapy. in oral Kulis allergy peanut AM, egg completed Scurlock of BP, treatment Vickery for immunotherapy 70. Oral children al. in et threshold Med. RA, reactivity Wood J SM, Predicting Engl allergic Jones of AW, al. Burks severity et 69. predicting KC, tool Carlsen peanut. a Lodrup to MM, of anaphylaxis basophil Michelsen Development with the T, al. Reier-Nilsen of et Benefit 68. S, A. Andorf Bernard challenge. N, peanut M, children. Purington during allergic Albertini reaction RS, in B, milk Chinthrajah cow’s Saggio reintroduce with 67. T, to IgE Bourrier when allergen-specific deciding M, activation, in Vivinus-Nebot basophil test A, between activation Correlations Rubio al. challenges. 66. et food oral N, of Leung severity J, and Wang outcome Y, egg-specific Song not but 65. activation allergy. basophil egg and clinical IgE Egg-specific of phenotypes al. e148. with et M, correlate Masilamani counts A, T-cell Grishin MC, as- Berin suppression allergy. 64. basophil milk with Allergen-specific patients WG. in Shreffler e720. tolerance HA, clinical Sampson with A, Nowak-Wegrzyn sociated N, pa- Wanich venom-allergic insect 63. between differences subjects. Immunological sensitized asymptomatically al. and et 1231. immunotherapy C, without Schrautzer and with D, tients Bokanovic data L, improved including Arzt venom: test wasp 62. activation and basophil bee tool? and to determinants. research positivity testing carbohydrate Double specialist IgE cross-reactive J. allergen-based or Ring about recombinant M, tube Ollert by test U, procedure Darsow a diagnostic L, in Krischan B, challenge Eberlein food 61. test: activation Basophil during Allergy. events G. Transl adverse activation. severe Lack Clin basophil with AF, assessing children Santos allergic by Identifying studies 60. al. LEAP et the C, in Rourke Applications challenges ’ O New peanut G, and oral Toit Old Du AF, Test: Santos Activation food 59. Basophil in AF. test Santos activation R, basophil McKendry Allergy. the M, in of Kwok O, application Hemmings clinical the 58. for study. map population Road a WG. Shreffler basophils: allergy. in AF, profiles Santos expression 57. protein signal Early population. Biol. Jr. Leukoc D, general phos- J MacGlashan 5’ the S, phosphatidylinositol Ishmael in and 56. basophils kinase human tyrosine from spleen secretion between munol. Relationship and expression Jr. phatase DW, MacGlashan 55. 2007;119(3):626-633. lnEpAllergy. Exp Clin urAlryAtm Rep. Asthma Allergy Curr 2012;367(3):233-243. 2009;86(2):313-325. 2016;6:10. 2017;47(9):1115-1124. lnEpAllergy. Exp Clin n leg shaImmunol. Asthma Allergy Ann leg lnImmunol. Clin Allergy J 2018;18(12):77. leg lnImmunol. Clin Allergy J n leg shaImmunol. Asthma Allergy Ann 2018;48(4):415-423. leg lnImmunol. Clin Allergy J 16 lnEpAllergy. Exp Clin 05154:1-2 e919. 2015;135(4):913-921 leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J 2014;133(2):468-475. 08111:97 e62. 2018;121(1):69-76 2012;130(1):155-161. 2012;42(8):1197-1205. Allergy. 2015;114(4):319-326. Allergy. Allergy. 2019;74(S106):73. 2011;66(1):92-100. 2018;142(1):149-158 2009;123(4):789-794 leg lnIm- Clin Allergy J 2018;73(6):1223- N Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 0 yedM lsmyrM lxne ,e l mrvdpnl o lnclimn phenotyping: immune clinical for panels Improved al. et C, Alexander laser. M, violet Plassmeyer the of reconsideration. M, Utilization a Ryherd for time tests: 90. activation Basophil al. et Immunol. MA, Clin Faber V, Rev Sabato AP, and Implementation Uyttebroek test: 89. instruments. activation Basophil between basophil C. and of Lambert systems analysis M, between Jeraiby to standardization K, approach Sidi-Yahya programmatic AE, Depince-Berger Data-driven 88. al. relation et in M, fraction Schneider antibody A, IgE tests. relevant activation Calatroni disease treatment. SU, the (Xolair) Patil of anti-IgE size 87. The of efficacy using al. the et responses predicts H, cell Oman mast ’total-IgE’ A, and to Nopp basophil SG, on Johansson omalizumab of 86. to Effects basophils challenge. al. allergen human et cat D, of Kelly intranasal sensitivity PM, an Sterba intrinsic JA, the Eckman cell, increases 85. mast Omalizumab SS. of Saini Kinetics allergy. stimulation. Jr., RA. peanut IgE-mediated DW, Wood with Macglashan adults SS, omalizumab-treated Saini 84. in DW, responses Macglashan challenge PM, Immunol. food Sterba Clin oral JP, and Courneya oral peanut basophil, JH, facilitates Savage omalizumab dosed Individually 83. al. serum adolescents. et low allergic AC, a peanut with Sundqvist to in allergics M, immunotherapy Vetander in response effective J, is Brandstrom basophil Xolair 82. H. the Oman J, of factors. Adedoyin SG, Suppression competing Johansson level. A, IgE SS. 2 Nopp J, Saini on Ankerst RA, dependent 81. Wood is omalizumab JH, Savage with as munol. treatment Jr., basophils during DW, in allergen release MacGlashan histamine IgE-mediated 80. and expression food receptors. omalizumab. Syk to of FcgRII SS. efficacy responses e1610. and clinical Saini cell the FcgRII predicting Jr., mast for the DW, biomarkers and via MacGlashan Basophil mediated S. not 79. AF are LK, patients James tolerant O, but lergy. Hemmings sensitised M, in Kwok allergens RT, McKendry 78. activation cell mast allergens. and major basophil peanut-induced peanut inhibits to IgG4 1256. precedes sensitized al. 2 children et h HT, peanut-tolerant Bahnson Ara in LK, to James AF, sensitivity Santos basophil 77. in decrease Early al. et immunotherapy.e1314. A, oral peanut Calatroni after does J, unresponsiveness but sustained Steinbrecher antibodies SU, blocking induces Patil immunotherapy IgE. oral 76. peanut-specific Peanut of al. characteristics et functional M, Kwok change LK, not immunotherapy. James allergen AF, in Santos perspectives 75. and approaches Novel al. during et peanut O, to Pfaar Allergy. E, response Valovirta immunologic HJ, the Hoffmann of 74. Suppression al. et transient. AL, often Guerrerio is SD, immunotherapy Narisety M, Gorelik 73. 2019;74(S106):97. 02105:1013 e1135. 2012;130(5):1130-1135 2017;72(7):1022-1034. n rhAlryImmunol. Allergy Arch Int 02105:1312 e1122. 2012;130(5):1123-1129 yoer lnCytom. Clin B Cytometry 2014;10(10):1325-1335. leg lnImmunol. Clin Allergy J yoer lnCytom. Clin B Cytometry leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J 2010;152(1):71-74. 2018;94(4):667-673. lnEpAllergy. Exp Clin 03124:0-1 e901-904. 2013;132(4):906-911 17 2018;94(5):671-679. 2015;135(5):1283-1292. yoer A. Cytometry leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J leg lnImmunol. Clin Allergy J 00154:8-9 e887. 2010;125(4):889-895 leg lnImmunol. Clin Allergy J 2019;49(10):1328-1341. Allergy. 2009;64(10):1472-1477. 2017;91(3):261-269. 2019. 2019;144(5):1310-1319 2017;139(5):1680-1682 leg lnIm- Clin Allergy J 2015;135(5):1249- Allergy J Expert Al- Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159493194.42634850 — This a preprint and has not been peer reviewed. Data may be preliminary. 0.Gre ,Wtr ,Mroi ,e l lo aohlatvto sarlal imre fallergic of biomarker reliable a is activation basophil Blood al. et B, fibrosis. cystic Mirkovic in J, aspergillosis Waters activationbronchopulmonary Y, basophil CD63 Gernez study. The prospective W. 106. a Aberer A, miti- allergy: Heinemann not venom I, Weiglhofer Hymenoptera probably M, in is Trummer test E, test anaphylaxis activation Bohm experiment. Rocuronium-induced GJ, basophil vitro Sturm DG. in new Ebo 105. an A LS, from J. Clerck evidence Ring De sugammadex: H, CH, by Bridts Behrendt gated in J, F, allergy antibiotics. Leysen Rueff food to U, 104. of allergy Darsow diagnosis in I, CCR3 the Suarez and for human CD63 Leon tests in using activation B, CCL2 Basophil Eberlein and al. 103. receptors, et L, IgE Hanssens high-affinity S, Basophils, Mulier children. A, al. Ocmant et 102. M, proficiency Silar sIgE V. PJ, Turner e. INSTAND P, anaphylaxis. of Korosec years diagnostics. Six allergy 101. vitro I. Schellenberg in potentials of K, more evaluation Kabrodt basophils: An S, human testing: Goseberg on based N, tests Wojtalewicz Diagnostic 100. al. et pitfalls. PM, than Gamboa ML, perspectives tests Sanz and Diagnostic AL, Weck BF. de Gibbs 99. LK, perspectives. Poulsen cell and Jr., mast pitfalls DW, human potentials, MacGlashan novel 90. EF, basophils: A Knol human G. S, on Lack Erdmann based HT, J, Bahnson Kleine-Tebbe M, 98. Kwok allergy. N, peanut Becares for N, test Couto-Francisco activation AF, Santos immunother- oral 97. peanut successful Sustained SJ. IgE. Galli peanut-specific e886. and KC, considerations. 896 activation Nadeau basophil fluorescence postanalytic low RS, with cell-based Chinthrajah - associated K, of apy IV Mukai Validation part M, CA. Tsai - Hanson 96. ICCS T, and issues. Oldaker ICSH J, preanalytical as- the De Cytom. - fluorescence from P, Clin II Gambell cell-based guidelines R, practice part of Louzao Validation - assays: D, A. ICCS Barnett Estrellado and 95. JY, ICSH Han the S, from Kussick guidelines Cytom. A, practice aims. Dasgupta and BH, rationale says: - Davis I guidelines practice part 94. assays: - fluorescence ICCS cell-based of and Validation D. ICSH Barnett the T, Oldaker from B, Wood BH, mass Davis and 2020. 93. cytometry July flow 6th using Accessed by analysis. activation https://eur-lex.europa.eu/eli/reg/2017/746/oj. basophil before 92. Assessing hours 24 al. stored et S, blood Gupta in N, cytometry Gaudenzio K, Mukai 91. 2013;84(5):286-290. lnEpAllergy. Exp Clin 2013;84(5):309-314. leg lnImmunol. Clin Allergy J 2009;39(8):1234-1245. n rhAlryImmunol. Allergy Arch Int leg lnImmunol. Clin Allergy J 07103:5-5 e715. 2017;140(3):750-758 lnEpAllergy. Exp Clin u eprJ. Respir Eur leg lnImmunol. Clin Allergy J 18 leg Int. J Allergo yoer lnCytom. Clin B Cytometry 2008;146(3):177-189. Allergy. 08122:8-9 e689. 2018;142(2):689-691 Anaesthesia. 2016;47(1):177-185. n rhAlryImmunol. Allergy Arch Int 2004;59(10):1110-1117. 2010;40(3):411-418. leg lnImmunol. Clin Allergy J 2017;26(2):43-52. 2011;66(6):526-527. 2017;139(3):889-899.e811. 2013;84(5):282-285. yoer Clin B Cytometry 2020;145(3):885- 2006;141(1):79- yoer B Cytometry