DEMETRIS, AJ

DUQUESNOY, R

FUNG, J

MAKOWKA, L

MARKUS, B

SAIDMAN, S

STARZL, TE

English

PubMed

Demetris, AJ

Markus, B

Saidman, S

Fung, J

Makowka, L

Duquesnoy, R

Starzl, TE

Name not available

Establishment of primary cultures of human biliary epithelium and induction of Class II major histocompatibility complex antigens by interferon gamma

D-Scholarship@Pitt

Establishment of mrimar~ Cultures of Human Biliar, Ii Epithelium and Induction of Ctass II 'lajor 1 eistocompatibilit~ Complex Antigens ! i by Interferon Gamma • t t A J Demetrls. B Markus S Saldma:-, J ~ung L MaKIow~" R Duauesnoy ana 7: S:in", T H E BI L1A R Y erlilhelium of Inlcrmc' dl3te and ~mgf! bile duct, f~ the larfED~ of immunologic damage II' li\t:r allografl rCJCi.' Ih)r. a nd rrim;e~ bg!i;e~ eirrhosi, (PBC) I 1 Thl: Immune damage in both of the~e dl~MrD aer, I, rrygliyci~ med:;:lcd b~ T fyrnr~Dx~!<> \1;.:.1(" hipyoEeDmpPlgbilll~ c'1mpln (\f HC I anll~EKDnD arc thElu~hl 10 be the t:Hget, of the l~mrhllEDvllc :lttack In IIvcr allograft rCJe('tlor., .... here;:!' the target anllgcm In prlmar~ billie~ cirrhM~i~ ha\c \el to be defined' Cla<I~ I \1HC antlcn~ 'He norm;!ll~ eypre~Icd lln Ihe billgr~ epI:hl:k.d celk Cb~~ II \-fHe antigen, .... hJic nOI nurm;:ll~ eypre~IedI are Induced on bilt: ducI, In pathoic'gie Ie, IOn, that In\'ol\e 3('ii, \aled 1:- mrho(': Ie, such a~ PBC .ind ll\er allograft regectionKD~ Donald el alto suggeqed that the immuno, logic mediated damage and loss of the small bile duct, In human li'er allograft, i5 observed rrcferenllalh .... hen there i~ donur.reuplenl comp;l\ibilll: at the cla~~ II MHC locu~ Other grour~ haH' found that the los~ of bile duct~ In li\er allograft~ i~ as~oclated .... Ilh an incomratibilit: at the clas~ fflocu~D ~ Central 10 aorKald~onDs hypothe~l~ IS the conjecture: thai the bihar~ epithelium I' capable of actin~ as an antigen presenting cellthcreb) promot· Inf' " reacllOn directed at an incompatible cla~~ I \1 He antigen' oe~lglution of !>ome of the discrepancie~ F,,)ftl Ihr D{'T'.:J'; f>lt'n , < of Paln%gl aM Su"",.'. L II:' {'r5ll I H{'a!li: ('tlll(,'. PI/Isbu"h -4.:ia't'.I' rt!,rrll! 't'qunl.\ 16 A J lHmnrr! MD lHparlmor: of Palh%gl P'tSbllt'tOfl ("",,·trJllI Ho," pilo!. DtSulUQI O'Ho,oSI. PllIJb""h. PA fD<~/I~ C 1 VIS£; b.1 Grun" d SI'OI/Ofl. IN. 0041, J 3451881}00/·1." 453.00:0 mcntle)ned "bene m;l\ be Tn D~~ fC~ aprrl.l;'h'hec b\ :.In In \1Ir<' ~~~ft:f:K rDf~K C cuiture~ oi human billar~ epl1hl:\!L::' E~~ \\ c ha\l' rre\wu,h ~ho .... n th;;: ;D~:;D;:;DK c: ture~ of HRE. CH, b,' y!II;!bll~hcc ::"~I r, 'tt human il',er !1"UI: uDln~ i! 'L".:: , ,,. . - ~ menteC bJ'.J' mt'Clum ~Dllmb:nlDCD" .:;. _ .;I nC('tl\l' 11',Ut: ,ub;.irate' Af~oK SIr::C ~K:! ~ f~ mrhIFE:~ te, from II\er allograft bl('r,;e I natJ\c herg!e~hlm:- 'pe(,lmen;. \\I:r, PB', ( be: caIi1~ c\p:..ndl'd In culture" Ilr. t1i~ ~-e fntcrlcu~f:tI:ID epithcffg:D!~ r:.rn,'_I:t Ir,:,-tJ(lr, m:.:; b~" ;-ncnJ ~k flf~! ud\ Tk fl'; .. f~ 3 rCrel~1 vn rrlmJr: culture, fKKD~ r _-:-gntrKghcr~tl~ bi\;.Jr\ erltheitJI (,:;, ::;~ : flmlnIf~:K vb";'\Juur.- un thc Ind ... f~:IK clalis II \1 He anllgem on the~e- eel;, gamma interferon. MATERIALS AND METHO;)S Cell /.Iu/ullur. and (ulrurr.1 B:ft~my e;I!!~c~:~1 CDC~f" "ere haf'"\cqec 1:.':; tv. her~leChlmy ':X';lme'1' 0: don,>r flye~ ilill"~":: .. '. trdrK~ri;grK~K:dI . )n lr- .... hrd~ue:K il..,r thr l~D1ic:~DvrK ...: .. ¥ anC ChJ'2:lerl]..JII(\n of lhc,r .:ell' are rr;-.,,:c: • : el~e .. herl· The medium u-ed for the Ir ::~D " con"5leC 0: CE'1 ~EFEyEf"llt ~dcID and S(;f,'- •. supriemenl- (S:.It: Laoo'alurle, VIe!<: \\2"""" ; William' F. "lIh 1,!!lul .. mlnl' Bo:t, mr.::,. \I.",: ., menled \O,:r 1(1'; hea: Irl.l::tI\J\ee feu: c:::' " .... er!i' "err plaice (lr, \1 A, TRIGEl EColl"t-KII~:Dy: ;;: Le\lnf1or,. "IAI C<lJlec I: .. ell pb<ll. \"··L: ., piale' Thr« hundred fif:, tC' ~lEl l,mL 0: re,,-: ,ammo Inlerferor. (Cellular Produ,,,,". BiKrK~K added to confluent monola,·er culrur~ of bd!"' :: lIal cell, (CI!!hl to J: daH after plaunfl (ani" la~ers recel"ed no anterferon suppi(mer.: IC' lhe -Both control and interferon truled mon_ffIIye~ :. ,",cre ancubaled for fi'e da~·I "Ilhout further ad: : . chanFe' wthe media The cell, .. ere ther, ha"e,:: f Hum a n B i 1'-' ss II \lajor .'''''' -\ntigens ntlgem on S AND METHODS ( ulrure,l Ir !o()(, l iml of reC{IrrOlllUI IIIj~r Product;" Bl;f:Jico! II'U ' '. \Or culture, of b!fI~ry erlthe-, iOfter piallnf I (onlrl>: mOIl(>. ~ 'on sUl'lliemen: 1(. the media, .'r. treateC mono/ale- cuhura \' "'Ilhoul further add II IOns or ~fh .. ere then hiOnesled from • l"fbr~1 1986 pp 726.730 £sID/t/rIf:",t"li (11 PnmLlrl Cuiwrl'.l and Cell "~DatlfrK :<1111/1: ~D , initial cc:1 iIEIigte~ and monolayer culture' "Slmla~ cC morphologIC (t ~ pica! cobble~tone appearanc~K Fig I) and phenotypic charactcr-, iIIIbO trrc~crgtgrK I AE-I] POSItI\C, albumin tcr211H , fgctM~ \111 related antigen nC!!;i' pel ofmtr.lher..;tll. blilgr~ epithelium, L'ltr,,· IlflIctur..!: ('\anlinZlll0n (If initial cell fsolate~ ..c mIl~KdI;ycr cultu:c, rc\calcd ePDDD~ idem!· '. "ble Intercellular JunctiOn!> (1lghl and dc~· , IIOSOmal). surface microvilli. sparse C)'lOplas· ~1fD1f11C organelles such as sm.all mitochondria. ,., rough end,)plasmic reticulin and occasional , CYtopla,mic lipid drorlcts The primary cul-. ~e~ n:mJlnec \Iable f(lr up 10 4 "eek~ rptn~ .r • conneni\'e tls~ur su bst ra te , . Fig 1 Pha.. mtCroaeoP't ~ •• '."e. oi conflu.nt lay., of prom,,'t culture oi human IIiIiaory ,plth .... 1 c~ •• Iclay 101 " .. ' Indu(//on (lr Cla,\,\ /I AnflRrn\ -ydd~:hlr of ~lEf 1\, ~lEl l ml of ~KlnIlqKI Interler,lr, !(I the cul:un: medl~ follID~ec b~ incuba\loTi f(Ir threr It I fivc dP~~ re,ulted If: morrhl'\ogi.: and rhenot~ PI{' alt{:ra:lyKm~ In the culture~ q~ED dP~~ after additIOn of intrrfer,m \0 the culture there wa, a slight enlargement and ~pindllng of the eel1~ and slight increase in indi\'idu;,1 cell di~lodgement from the pia Ie .. 11",0 slmil'H morphologic ch;Inge~ ~ert: !oeen in the control culture, fmmunofluore~cent stain· ing of the celb i!oolated from the gamm;: fnterfe~on pl:gte~ rc\r:J1ed poSI!iVe cvlorla~mi~D and foc;,l ~urfaee stainlnc for DR amll .. en, - . (Flf ~F 'CI similar staming ~a~ pre~ent In contwi ~rccimen<K DISCUSSION The result~ pre"ented here demonstrate that prtmgr~ culture, of blliar~ epitheliurr. can be readil~ established from .... hole or frag· mert, l)f n(lrmal human li\cr t",uc Th-: eel i • cultured displil~ phenOl) pl~ and morphologl': characteristics of differentiated biliary epithe· lium in vitro. a.nd remain viable for up to I month, Therefore. this system may be u ... eful for the stud~ of the immunologIC properties of bihar) epithelium. In fact. the prelimlnar~ resull~ presented here sug~e ... t that gamma ,'.' . - , 73C FIG 2 Immunofluo'e,cence mlCrOICOP)' of cyto'p,n prepa' .toon of human biha/)' ep'thelial call, fiva deyalltt. trNtment wnh gamma Interferon The ceU. wera "a,nad ant,-human OR antigen -Interfnor alone i~ capablc of indUCing expre,-SI(lr. of c1a~~ II MHC antlgen~ on biifar~ epIlhclial cells similar to it!> effect or: endothc-kJ. fibr"blast and epithelIal lumw cell llne~K"D: Further studle!> uSing dual markers (prekeratin and anti DR) to sho" the simulta-neous expression of phenotypic markers and immuno!electromieroscopy to confirm cell \'iabilit\ are neeessar~ to confirm these find-ing~K and are currentl~ under"Da~K 'e\enhe-les~K since the Ie\ el of contamination ,,"lib fibroblast~ or endothellill cell<. In thi~ culture i~ nq:llgiblc. :!nd ur Ii' ·W'·, t(, ~ElD~ or the ccli~ stained posltiveJ) for class II antigens after interferon treatment, we feel it safe to assume that the bile duct cells are indeed those prt> dueing the class II antigens in respon'se to ga mma- interferon REFERENCES ) [kmelrl~ AJ. iasl~ S. Van Thie' DH. ct al Tran;· plantatoon 4sK~oK:K Fn~ ~ Fennel: R Pathol Ann ~:!~~K 1 n~: 3 Ma.:Sv.ccr. o~MK Bun AD Mol A~r "1ed b.2(.9. }9~~ 4 McFariane IG. Mol Asr Med ~:~9K 19~~ 5 Ballardlnl G. Mlrallan R. Blanch. FB. cl al Lancct :100'<. }9~4 6 Donaldwn PT. lDdrad~ J. Ponmanr. B. CI ,I Lance: I :94~K 19~--' ~ Dcmelm AJ. Mar~u~ B. Burnharr . .I. e! a! qra~­plan: Pro.; 191:1 19~-~ Grone J. do~D" .... SH pEyrpem~ S. e: a! lranspian: Proc f~ f~bK 11J1lt-Cf Dcmetm AJ. Mar~u~ BH. cun~ JJ. e; al Ir. Vun (In pr~1 10 cun~ JJ. lo«\"1 4,. Starl.: TE. Cl a: Hurr. Immuno' 16 18:. 19~t- ,:o-II Pober JS. Collin' T. Glmbronc \14, 'aturr PM~ 72f 19U~ I: ~h ... am R. MombuD~ F. Molde:thaue' G. et &1 Int J Cancer }~KO4pK fnb~ 

ESTABLISHMENT OF PRIMARY CULTURES OF HUMAN BILIARY EPITHELIUM AND INDUCTION OF CLASS-II MAJOR HISTOCOMPATIBILITY COMPLEX ANTIGENS BY INTERFERON-GAMMA

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ESTABLISHMENT OF PRIMARY CULTURES OF HUMAN BILIARY EPITHELIUM AND INDUCTION OF CLASS-II MAJOR HISTOCOMPATIBILITY COMPLEX ANTIGENS BY INTERFERON-GAMMA

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