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In Vitro Studies in Nickel Allergy: Diagnostic Value of a Dual Parameter Analysis

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In Vitro Studies in Nickel Allergy: Diagnostic Value of a Dual Parameter Analysis In Vitro Studies in Nickel Allergy: Diagnostic Value of a Dual Parameter Analysis M ary von Blo mberg-van der Flier, M.Se., Cees K. H . van der Burg , M.D., Odette Pos, B.Se. , Ell a M . van de Pl assche-Boers, M .D ., D erk P. Bruynzeel, M.D., Ph.D., Gianni Garotta, Ph.D., and Rik J Scheper, Ph .D. Departlllellts of Patho logy (M vI3 -vd F, 0 1' , E M vd l'-B, RJ S) and Dermatology (CKH vdB, DPJ3), Free Uni ve rsity Hospital, Amsterdam, T he N etherbnds, and Centra l Research Units, F Hoffm ann- La Roche & Co., Ltd. (GG), Basel , Switze rl and A compari son was m ad e be tween the diagn osti c value o f test reacti o ns . A ccurate d ete rmination of MIF becam e fea­ assaying nickel-induced ly mphocyte pro life ra ti o n (l y m­ sible b y using cell s fro m the human monocytoid cell line phocyte trans fo rmatio n test, L TT) and mig rati o n inhibi­ U 937 as ta rget cell s in a mic ro drople t agarose assay . Using tio n facto r (MIF) pro ducti o n in nickel contact sen sitivity. this MMIT, positive reacti o ns o ccurred in 13% of the healthy Altho u g h ly mphocyte pro life ratio n was sig nificantl y in­ contro ls and fa lse-negative reacti o n s w e re fo und in 26% o f creased in the g ro up of pa ti ents w ith skin test reactivity to pati ents w ith p ositive skin test reacti vity to nickel. nickel, positive L T T w ere also frequently fo und in skin A s LTT and MMIT data appeared to be only w eak ly test-negati ve s ubjects: in 63% o f subjects with and in 30 % correlated in the individua ls tested , a dual parameter an al­ 8 of subjects w itho ut a histo r y o f m e tal a llergy. T his would ysis was perfo rmed . An excell ent correlation [p = 1.8 (1 0 - ) ] limit the value o f the L TT as a n in vi tro correlate of skin w as fo und be tween s kin test and in vitro reactivity for test reacti v ity. H o w ever, in certain pati ents positive lym­ individua ls with m atching in vitro results (60% of all in­ phocyte tra nsformati o n may re veal nicke l sen siti zatio n at dividuals teste d). In those individuals with discordant i.n a time of undetectable skin reacti vity. v itro data, skin testing will rem ain indispen sable for di­ Data o b tained w ith the m acro phage mig ra tio n inhibitio n agnosing nickel a ll erg y. J filli es! D erwa!o/ 88:362- 368, 1987 test (MMIT) sh owed a good correlatio n with nickel patch in ce nickel is the most common contact sensiti zer in theless, the individual diagnosti c va lue of this assay was limited, humans, ava il abi lity of di ag '.lOs ti c in vitro tes ts for ni ckel parti cularl y by a hi gh in cidcnce o f false-positi ve rcacti o ns. all ergy would be of g reat impo rta nce. Data o btain ed As antigen-induccd Iympho kine (LK) relcase represents a cru­ from the li terature regard in g the usc of lymphocyte cial stcp in the develo pmcnt o ftypc IV hypersensitivity reactions, transformation tes ts (LT T ) fo r this purpose, arc listed in one would ex pect assays measuring ni ckel-induced LK produc­ TSabl e I /1 - 151. Stati sti ca l differences between g ro ups o f ni ckcl­ ti on to be most suitabl e as diagnosti c tes ts in nickel all ergy. [t has sensitive pa ti ents and hea lthy contro ls were usuall y fo und. N ever- indeed been repo rted that lymphocytes from nickcl-allergic pa­ ti ents m ay produce leuk ocyte migrati o n inhibition factor (L1F) in vitro in the presencc of nickel-protcin conjugates [1 6, ·[ 7] or free ni ckel sa lts [1 8-20 1. Fro m T ablc [[ it is clear that no consistent results were o btained. This might be due to thc usc o fullseparated Man uscript received M ay 19, 1986; accepted for publi cation Septem ber bu ffy coats fo r simultaneous LlF production and mig ration (1 - 23, 1986. phase leucocyte migrati o n inhibition tes t, LM[T). In addition, This work was supported in part by Pracventicfonds, T he N etherl ands. the sho rt culture peri ods (up to 1 day) in these assays may not Reprint requests to: M ary von Blo mberg- va n der Fli er, Depa rtment o f have becn suffi cient fo r adequate ni ckcl prescntatio n by accessory Pathology, Free University Hospi ta l, De Boelela an 11 17, 1007 ME Am­ ce ll s, Iympho kinc release by effector cc ll s, and inhibition o f mi­ sterdam, T he Netherlands. g ratin g ta rget cell s. A bbreviatioll s: Con A: concanavalin A [n th e present study, therefo re, w e adaptcd a 2-phase protocol DTH : delayed-type hypersensiti vi ty in w hi ch peripheral bl ood lymphocytes (PBL) w ere precultured FCS: feta l ca lf serUIll to all ow fo r m acro phage mig rati o n inhibitio n facto r (MIF) pro­ L1 F: leukocyte migration inhi bition factor ducti on. Subsequentl y, MIF activity was measurcd on a ho m og­ LK: Iymphokine enous populatio n o f ta rgct cell s. In preliminary experimcnts 3 LMIT: leukocyte migrati on inhibition test(s) presently ava ilable hum.an m yelo/monocyti c ccll lin es (KG-l (22], LTT: lym phocyte transfo rmatio n tes t(s) HL60 r231, and U 937 r24,25]) w ere screened fo r M[F sensitivity. MI : m igration inhibitio n U937 ce ll s, found to provide o ptimal targets, w ere then used to MI F: macrophage migration inhibition fac tor eva lu ate the diagnosti c value o f a 2-phase macrophage mig rati on MMIT: macro phage migration inh ib ition test(s) PEL: peri pheral blood Iymphocyte(s) inhibition tes t (MM IT ) in ni ckel all ergy. The MMIT data w ere PBS: phosphate-buffe red saline compared w ith lymphocyte transfo rmatio n data as to their cor­ PM N : polymorphonuclea r relati on w ith skin test reactivity. [n addition, considering that SI: stimulatio n index both in vitro parameters (MIF producti on and lymphocyte pro- 0022-202X/87/S03.S0 Copyrig ht © 1987 by T he Society for In vestigative Dermatology. In c. 362 VOL. 88. NO. 4 APRIL 1987 IN V ITRO TESTS IN NICKEL ALLERGY 363 Table I. Lymphocyte Transformati on in the Diagnosis of Nickel All ergy-Review of Literature Results Patients Control Group In Vitro Test % False % False Year First Author Reference Antigen Concentration" N i> N egative' N i> Positived Si gn' 1962 Aspegren [1] NiCIz 100 /L M 15 0 15 100 1970 Pappas [21 NiAc 112 '/LM 3 0 8 100 1970 M acLeod [3] NiSO. 50 /L M 12 17 14 0 + 1972 Form an 14] NiSO. 18 II 12 66 + 1972 Hutchinson 151 NiSO./ NiAc 50/LM 8 25 7 14 + 1973 Mill ikan [6 1 NiCL2 40 /L M 8 0 6 0 + 1975 G imenez 17] NiSO. 0.65/65 /L M 25 0 10 0 + 1976 Kim 181 NiSO./ NiAc 40 /L M 21 0 17 < 10 + 1978 Svejgaard 19] NiSO. 20 /L M 8 13 9 11 + 1979 Veien 110 1 NiSO. 36/72 /L M 8 13 4 25 + 1980 S i Iv en noincn-Kassi nen 111 J NiSO" 8/40 /L M 35 II 49 20 + 1981 AI-Tawil 11 2] NiSO, 24/LM 13 15 9 22 + 1982 MacLeod 11 3J NiSO., 50 /L M 6 33 6 33 1984 Nordlind 11 4] NiSO" 20 /L M 10 0 10 40 + 1985 AI -Tawil [1 5] NiSO" 80 /LM 41 15 34 32 + 1987 Blomberg IThis pa pcrl NiSO., 7- 80 /L M 27 7 23 39 + "Nickel concentrations showin g :111 optimal difference between patient and co ntrol g roups. bNull1ber of individuals tested . 'Stimulati on ind ex (5 1) :s 2 in nickel-allergic patients. "51 > 2 in health y controls. 'Statisti cal differences between groups of nickel-a ll ergic pa tients and health y cont rols : +. liferation) relate to different phases of the cell-mediated immune Skin Tests Patch rests were performed w ith 5% NiS0 4 in response (representing effector and memory functions, respec­ petrolatum. Patches were removed after 48 h and the reactions tively), a dual parameter analysis was performed. were read 15 min later and graded according to Fregert [26]: negati ve (-), weak (nonvesicular) reacti ons (+), and strong MATERIALS AND METHODS (edematous and vesicular) reactions (+ +), or even bullous re­ Donors Twenty-seven ni ckel-all ergic indi viduals showin g pos­ actions (+ + +). itive ni ckel patch tests (13 + and 14 + + or + + +) were ran­ Lymphocyte Transformation Tests Mononuclea r cell s were domly selected from paticnts visiting our Allergy C linic, and from isolated fro l11 heparinized blood by Fi coll-isopaque centrifugati on junior hairdressers w ho had been skin tested in a parall el study prior to or 2-3 weeks after skin testing and cryopreserved.
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