Effect of Auranofin on Plasma Fibronectin, C Reactive Protein, and Albumin Levels in Arthritic Rats

Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

Annals of the Rheumatic Diseases, 1988; 47, 515-521

Effect of auranofin on plasma fibronectin, C reactive protein, and albumin levels in arthritic rats

KEVIN M CONNOLLY,' VERA J STECHER.2 AND DONALD J PRUDEN3 From the 'Department of Chemotheragy, Glaxo Research Divisioni, Five Moore Drive, Research Triangle Park, North Carolina 27709, USA; the Medical Division, Winthrop Pharmaceuticals, 90 Park Avenle, New York 10016, USA; and the -Department of Pharmacology, Sterling-Winithrop Research Instituite, Renlsselaer, New York 12144, USA

SUMMARY Auranofin, a member of a class of compounds with disease modifying activity, was given to arthritic rats to determine if it could reverse the abnormal plasma concentrations of fibronectin (Fn), C reactive protein (CRP), and albumin, which were unaffected by treatment with non-steroidal anti-inflammatory drugs (NSAIDs). When auranofin was orally administered for two weeks to adjuvant induced arthritic rats it significantly inhibited swelling of the injected and non-injected paws at doses of 3 and 10 mg/kg. Rocket electroimmunoassay measurement of plasma proteins in normal, arthritic, and auranofin treated arthritic rats indicated that auranofin at 10 mg/kg significantly decreased (by 77%/0) the abnormally high concentration of arthritic rat

plasma Fn, though it had no effect on Fn concentrations when administered to normal rats. CRP, copyright. which was raised approximately twofold above normal in arthritic rats, was reduced by 56% after treatment of arthritic rats with auranofin at 10 mg/kg, though CRP concentrations in normal rats were unaffected by auranofin treatment. Depressed albumin concentrations in arthritic rats were significantly enhanced (by 30%) by dosing with 10 mg/kg of auranofin. At the 3 mg/kg dose, auranofin did not significantly change plasma concentrations of Fn, CRP, and albumin in arthritic rats. At a dose of 10 mg/kg, however, auranofin, in addition to inhibiting chronic systemic paw inflammation, also altered abnormal concentrations of plasma Fn, CRP, and albumin in the http://ard.bmj.com/ adjuvant arthritic rat, thus distinguishing auranofin from standard NSAIDs we have previously tested. Key words: adjuvant arthritis, disease modifying antirheumatic drugs (DMARDs).

Certain gold compounds are classified as disease fluid"X and pannus tissue' "' of patients with RA on September 27, 2021 by guest. Protected modifying antirheumatic drugs (DMARDs) based and the plasma of lupus patients. ' It has opsonic,'2 on their ability to slow the progression of joint chemotactic,'-3 14 and adhesive'- "' properties, which destruction in rheumatoid arthritis (RA). ' Adjuvant may contribute to its suggested role in the induced arthritis in rats is a model of disease which pathophysiology of some rheumatic diseases. 7 lx shares many of the characteristics of RA2 and is Abnormally high plasma Fn concentrations have therefore useful in identifying compounds which been found not only in the arthritic rat5 but also in may be beneficial in the clinic.3 When oral gold is the MRL/MpJ (1pr) lupus mouse.'9 Agents like administered to adjuvant arthritic rats it significantly glucocorticoids and non-steroidal anti-inflammatory decreases paw inflammation.4 drugs (NSAIDs), which provide symptomatic relief Paw inflammation in the arthritic rat is accompa- without altering the progression of rheumatic nied by abnormally high concentrations of plasma disease,21 are active in decreasing paw swelling in fibronectin.5 Fibronectin (Fn) is a 440 kilodalton arthritic rats.2' Neither glucocorticoids22 nor protein found in high concentrations in the synovial NSAIDs,23 however, significantly decrease high Fn concentrations in arthritic rats as measured by Acccpted for publication 2(0 Dcccmbchr 1987. Correspondencc to Dr Kcvin M Connolly. Dcpairtmcnt of rocket electroimmunoassay, though NSAIDs have Chemotherapy. Glaxo Rcscarch Division. Fivc Moorc Drivc. been reported to lower serum concentrations of the Rescarch Trianglc Park. North Carolina 277(9. USA. acute phase protein a, acid glycoprotein.' In 515 Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

516 Connollv, Stecher, Pruden another model of inflammation, the carrageenan volume. The systemic nature of the disease was induced pleurisy macrophage model developed by assessed by measuring non-injected (left) hind paw Ackerman et al,24 NSAIDs also failed to reduce the swelling 17 days after adjuvant injection and two high concentration of Fn in the rat pleural hours after the final dose of drug. The injected exudate.> In contrast, auranofin, as well as other (right) hind paw was also measured at this time. Paw DMARDs, sodium aurothiomalate, chloroquine, inflammation was measured by obtaining paw and hydroxychloroquine, significantly reduced con- volume with a mercury plethysmograph and record- centrations of Fn and the number of inflammatory ing with a polygraph the amount of mercury cells in carrageenan induced rat pleural exudates.>2 displaced (ml). Raised concentrations of acute phase proteins,26 particularly C reactive protein (CRP),-7 28 have D O S I N G been associated with RA. There is clinical evidence Auranofin was delivered orally in a volume of 1 indicating that abnormally high concentrations of ml/100 g of body weight. The drug was suspended in CRP in patients with RA cannot be reduced by a 1% solution of warmed (approximately 370C) gum NSAID treatment, which relieves the symptoms but tragacanth and homogenised by grinding (Eber- not the underlying progression of disease.29 bach, Ann Arbor, Michigan) before delivery at 3 or DMARDs, however, like auranofin, which are 10 mg/kg. Two days after day 1 adjuvant injection efficacious in the treatment of RA,21' also decrease daily dosing was begun and continued until the abnormally high concentrations of CRP in arthritic experiment was finished on day 17. Normal and patients. 11-32 arthritic untreated controls received vehicle alone. In contrast with the typically high concentrations Normal non-injected rats treated with auranofin of plasma CRP associated with RA,27 28 serum were given drugs orally according to the regimen albumin concentrations in patients with RA are used for arthritic rats. typically depressed.33 3 Adjuvant arthritic rats also copyright. possess abnormally low concentrations of plasma PLASMA PREPARATION albumin,35 which could not be raised by treatment Three hours after the animals received the final dose with standard NSAIDs.36 of drug and immediately after paw volume measure- As raised concentrations of plasma Fn associ- ments a 0-1 ml sample of blood was obtained by ated with adjuvant arthritis in rats5 were reduced cardiac puncture with a I ml syringe and 27 gauge after treatment with the DMARD auranofin, but needle. Blood was immediately mixed in microvials not glucocorticoids22 or NSAIDs,' reduction of with 0-012 ml sodium citrate (18.5 mg/ml), and plasma Fn in arthritic rats and dampening of the centrifuged for five minutes in a tabletop centrifuge http://ard.bmj.com/ acute phase response (typified by alteration of (Fisher Scientific, Fair Lawn, New Jersey). Whole plasma CRP and albumin concentrations) may be plasma was removed and assayed for CRP, Fn, and characteristic of a class of antirheumatic compounds albumin. Repeated freeze thawing was avoided. All distinguishable from standard NSAIDs. experiments were conducted with 10 animals in each group. Materials and methods

PURIFICATION OF PLASMA PROTEINS on September 27, 2021 by guest. Protected ANIMALS Commercially available Cohn fraction 5 purified rat Male, inbred Lewis rats (approximately 180 g) were albumin (Sigma Chemical Co, St Louis, Missouri) obtained from Charles River Laboratories. was used as the albumin standard, and antibody to rat albumin was purchased from Cooper Biomedical INDUCTION AND MEASUREMENT OF Laboratories (Malvern, Pennsylvania). It was neces- ADJUVANT ARTHRITIS sary to purify large quantities of rat Fn and CRP for Freund's complete adjuvant was prepared by adding use as antigen in obtaining appropriate antibody and 100 mg of Mycobacterium tuberculosis (Difco for calibration of a rat plasma pool standard. Laboratories, Detroit, Michigan) to 15-6 ml of Purified Fn was obtained by a modification of the squalane oil (Aldrich Chemical Co). The M tubercu- affinity chromatography procedure of Weiss and losis was then ground in a homogeniser (Eberbach Reddi37 described by us previously. 1'9 About 200 ml Corp, Ann Arbor, Michigan) followed by addition of plasma was applied to a gelatin Sepharose 4B of 1 ml of 0*15 M saline. The mixture was (Pharmacia) column after appropriate ammonium thoroughly emulsified by pulsing for 30 seconds with sulphate treatment.'9 The column was extensively a Polytron (Brinkman Instruments, Westbury, New washed with a phosphate buffered solution of 1 M York). Each rat was injected in the right hind saline (PBS), eluted with 4 M urea-trometamol footpad with 300 ,tg of M tuberculosis in a 0-05 ml (TRIS) buffer, and dialysed against PBS. The Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

Treatment of arthritic rats with auranofin 517 dialysate was again passed over the gelatin- macia, Piscataway, New Jersey) was added to a 100 Sepharose column to purify the Fn further. When mg of purified Fn or CRP (3 mg/ml). Standard the purified Fn was assayed by sodium dodecyl procedures for protein conjugation were followed as sulphate slab gel electrophoresis it produced a described previously.1419 The protein conjugated 220 000 molecular weight double band, characteris- Sepharose was finally resuspended in degassed PBS tic of the purified Fn dimer. (0-01 M, pH 7-4) and poured into a column where it Purified CRP was obtained by a single step was allowed to equilibrate with PBS. Antibody was affinity chromatography procedure that was a syn- precipitated from the goat antiserum with a satu- thesis of the methods employed by Young and rated ammonium sulphate solution. The precipitate Williams,38 Nagpurkar and Mookerjea,9 Pontet et was resuspended with 0-01 M PBS (pH 7-2) in one al,4( and DeBeer et al.41 Approximately 100 ml of fifth the original 250 ml serum volume. The antirat serum from normal or arthritic rats was passed body solution was dialysed in 0-01 M PBS and over a 1 cm x 20 cm column of p-aminophenyl applied to the affinity column. The column was phosphorylcholine immobilised on agarose (Pierce washed with PBS (pH 7.2) and the antibody to Fn or Chemical Co, Rockford, Illinois). The column was CRP eluted with 0-2 M sodium acetate. The eluate equilibrated and extraneous proteins were eluted was immediately dialysed against PBS and stored at with a trometamol (0-02 M)/saline (0.15 M)/Ca++ - 700C. buffer, pH 7-4. The column was stripped of extraneous protein by washing with a trometamolV MEASUREMENT OF PLASMA PROTEINS saline/edetic acid (0-1 M) buffer and re-equilibrated Concentrations of Fn, CRP, and albumin were in trometamol/saline/Ca++ buffer, pH 7-4. Con- measured by rocket electroimmunoassay.19 43 4 tamination of CRP by serum amyloid protein was Plasma samples to be tested for Fn or CRP were minimised by pooling and concentrating only those diluted 1:10 in trometamol-Tricine buffer. Albumin fractions from the second half of the CRP elution measurement was made on plasma samples diluted peak. When this single step procedure was used a 1:150. A series of internal standards was run for copyright. 25% yield of CRP was obtained. The purified CRP each assay. A stock 1 mg/ml solution of rat albumin was free of serum amyloid protein contamination as was diluted 1:4, 1:5, 1:10, and 1:20 to yield measured by reduced and unreduced gradient slab standards of 250, 200, 100, and 50 [tg/ml respect- polyacrylamide gel electrophoresis gels stained with ively. The Fn and CRP standards consisted of a Coomassie brilliant blue R-250 (Biorad, Rockville sample from a rat plasma pool diluted with Center, New York) or silver stain (Biorad). Purified trometamol-Tricine buffer 1:2-5, 1:5, 1:10, and 1:20. proteins were measured spectrophotometrically The rat plasma pool was originally calibrated against http://ard.bmj.com/ (280 nm) and stored (-70°C) at a concentration -3 affinity purified rat Fn or CRP. The heights of the mg/ml. sample rocket peaks were compared with the height of the internal standards to determine Fn and CRP PREPARATION OF ANTISERA concentrations of the sample. Antibody against rat Fn and CRP was prepared by mixing the purified proteins (2 mg/ml) 1:1 with STATISTICS

complete or incomplete Freund's adjuvant. On -days An analysis of variance and Dunnett's multiple on September 27, 2021 by guest. Protected 1 and 8 a 0-5 ml aliquot of the complete adjuvant comparison were used to compare all groups (n= 10) (plus Fn or CRP) was injected subcutaneously into and to derive significance between groups. Data each of two sites along the flank of a goat. The were expressed as the mean (SEM). procedure was repeated on day 15 using Fn or CRP plus incomplete adjuvant. On day 22 the goat Results received one subcutaneous and one intramuscular injection of 0-5 ml antigen plus incomplete ad- EFFECT OF AURANOFIN ON INJECTED AND juvant. Ten days later 250-500 ml of whole blood NON-INJECTED PAW VOLUME OF ARTHRITIC was taken from the jugular vein. Goat, antirat Fn or RATS CRP antibody was obtained from the serum by Fig. 1 depicts the anti-inflammatory activity of affinity chromatography. '9 42 The antibody specific- auranofin as measured by its ability to inhibit ity against Fn or CRP was monitored by im- swelling significantly in injected and non-injected munoelectrophoresis and showed no cross reactivity rat paws 17 days after adjuvant injection. Oral drug with rat albumin or C3b or serum amyloid protein. treatment from day 3 to day 17 with 3 or 10 mg/kg of auranofin resulted in a significant (p-.001) PURIFICATION OF ANTIBODY reduction of injected and non-injected arthritic rat Ten grams of CNBr activated Sepharose 4B (Phar- paw volumes as calculated using a one tailed Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

518 ConnollY, .Stecher, Prludeni

3.00 5.0 fin treated normals 338 (7) ig/ml Fn; untreated normals 339 (13) tg/mIl Fn). g 2.75 4.5 lU -J [1EFFEC1T OF \URA\NOI IN ON P1 ASM\ CRRP IN 4.0 o 2.50 > \RTHRITIC R \TS Like the increased plasma Fn level, the CRP 2.25 3.50 C- 0 I concentration was significantly higher in arthritic 0jI ,_ -E 2.00 30 ,, E rats (718 uig/ml) than in normal controls (360 utg/ml) F- (Fig. 3). Treatment of arthritic rats with 10 but not 3 2z 1.75 2a ° mg/kg of auranofin significantly reduced the level of - z CRP by 56%0. Normal animals treatted with 10 mg/kg z 1.50 2.0 of auranofin did not have reduced CRP concentration (auranofin treated normals 451 (10) Ftg/ml 1.25L MEM____ _o _ 1.5 450 CRP). NORMAL ARTHRITIC 3 10 CRP; untreated normals (9) utg/ml AURANOFIN DOSE, mg / kg, P. 0. 800 Fig. 1 Effect ol aloanolifi oni iijl'ected oid11noi-iijectcd 750 p)aw v'oliine of arthritic rati.s. Oli ditv 1 mnale Lewi.i rats (ISO ig) were itijected iii ti/ic rig/it /)hind/it( p i it/i 00-5 oii of 700 comnplete Freutid's adi/iiatint cooi/posed of1|0 in1g of z Nlcobactcriuni tubcrculosis -.,roiunditandi iiilsiifiel inl 1560 0 650 il of squalcatic oil (and/ 1 mil ofsaliti. Fr--om daY 3 to dali 17 uJ tnitinalis icre dosel oral/l once datliY witl/ ailiratio.fin z0 600 siUspended In /l gi1on trag(acanlth. F/ic collittle of crit-ic pluts E co 550 copyright. vc/eicle wa.s calcllated so tl/ict cint ciIecceIvdI/('/1 tl of/ rulg i- p/u.s vehiclel100 g botd i'weig/ht. Rlits ii eacili of 1( hroup 500 were ble(d fion th/i liccirt oni (tciv 1 7 citi(/plcastini /)roteiti coticetitratiotis obtciniec bi rocket c'lectroonmunocisscv. Julst c) fat 450 before b/eecbing tli/c cniniacils ivcrc' ive'ig/id'cct(Icl (ciiscii'cl 0. inflammniation bi' mietsiirintg i/it' voliutic' ol it/ei'no-injcctedl 400 let pawi' cand itnjec'td(l rig/it paii.u1sing u11c'Cuirv cisplacicem'n. P/}

IH((1ct!lllCll f a itiic r(t1% cit/il (11111(,1110fi 5 19

800r Fi 1: (-I () I1 \ lK\N0RN () F I N () N P1 A S \1 750 F \1 Bi x1IN IN' \R I IIRITIIC R xS To dceteriline xx hctheli reduction otf aithritic Ilat 700 K plhsima Fni and CRP by arl-anofill \V\as dIueC to cneralisled inihibitioni of livcir priotein sv ihesis. 650 k plasma albLmnIiIl concenitratioins xere meastiured (Fi-. a. - 600 4). In contirast x ith the ralised lcels of Fii and C'RP C.) 'E secen in the unl1treated artliritic rats. plasma albuLIIIIIn - 550 concentrations in airthritic rats cire depressecd V) compared wxith normals (6 4 mg/nil I 18 4 mi/mil). !5 500 Treatilmenit ot airthritic rats xith aLuranioftiln did 11ot reduLICe the of 450 alireadx lox concentrations plasmia albinLI1ill a-s xxoUld be expectedi if airanofin xxere 400 CaluilSI 'ceniaLlised inhibitiOnl of lixver protein sxyn- thesis. In the experimncit shown in Fig. 4 aRuranofin 350 at 1() tmg/kle sieificaiitlx enhanced plasma albIinIi lev'xlsbv!( 300 NORMAL ARTHRITIC 3 10 Discussion AURANOFIN DOSE, mg / kg, P. 0. In this study aIdljuLxIlt induLCe1d arthritic raits had abnorniallx high Plasma concentrations of F Fic} 3 ft1 ot auratioo ooi oin / m ( i,, and p CRP1\i rthrii 0t. CRP ( R P w's 0n1?i llrc'd h1 Act c/ O c. and depressed concentrations of plassma

1 c albumnin. Ti eitment of arthritic rats wxith auranofin Ol't/lli/i(' il7ll.' ¢t'.$'{t% 1'f'//'('6('1 1/1('c171'll111/. '.1 (}cIed 11 l,Iltl rpic Sk1)0 reduLced pax inflammini'ation and significantly altered copyright. arthriic7(18'1(sanlinalsl.Rcuh the abnormal plasma concentrations of Frn CRP. itald albt1umnill. 20 Fibronectin is a1 plasma.t pirotein norilialix Svii- tthesisedi in the liver. Thic high plasmia F-l conicelitltra- tions seen in arthiritic rats' mayx he a iCsLIlt of increased liver sxnthesis but additionally probably 15 reflect increased in situ pi oductionI in the) inflamned z Joints. aS VsxnoxiaIl fllids from patients xith RA http://ard.bmj.com/ contain a hiehei- concentration of FtI than the 10 plaIsIlmI. S stemllic x asIClitis. chara'icteristic of sex ere airthritis.4 mav also contribUte totohe 11h'JI1 COnICCIe- E trations of plasma Fn in airthritic rats as larue .imoilIts of crioprecipitates associated with Fn ha Ve 5 been detectedi in the) seruLIm of patients xith RA and Vasculitis.4" In the case of the rat model. wxith its on September 27, 2021 by guest. Protected severe and xxidelx disseminated dliselse and rela- tixclx small blood xVolImIe. high plasma Fn coiiceni- 0 NORMAL tratioiis max be par tiallx at i-CsIlt ot sxnithesis from ARTHRITIC 3 10 the In AURANOFIN inflamde tissuC. human111sartiS-itis is rarelx alloxxwed to progress to the deurce of sexeritx seen in DOSE, mg / kg, P. 0. the arthritic riat so thiat plasma Fn conceiltrCations in Fig. 4 1 1(C/oftouraioOii oilp/)/(1(0ti8'1 (1//)1/ill i'il (O1't/il'i/1 hiUMMim s xWxOUld he X\pcCtedI to be norial' in aIll but ro((,,P 0cc/dlc lr i ieCUN11101,11" pj .s p1()tcI II the most sevxrce cases ot RA. One of the cffects of zi I'0ilCC/1i((' )11 ()tl. 1(v(ce' c 1110.0(/1c(1(111(1o11( illcIc/)e(/ M 2 DMA\ RD sucih aIs alil-aofill mx he its abilitx to itt/lli [i/c f(l'o/ iii (1(ad()11 .'ol l '( 1p1101(' li (l fp(l. / 1 tici damipen the overactixitx of thie ifltllimedi joilnt and x ascI.Ilar tissuLe. resuIltine in at iedUced conicenitaitionl trfOf lnetooio f/ ic i,( 1)1//,')l,aIiea odl1p (s1d olli/ol (1/0(1 (1o of plasma Fni in rats. and a reduced sxyno tgo oil oUc.1 0/1h 1111c )1/11c (lc/i ll s 1f/lldItilr w .si"l'iii possiblx ial fluid Fn concentration in (/lilct(/ 1:20. 1:10. 1. (111(1 1:4 wit/ill roinc11l'tnOi / I1l(ICl(' huLnillIs. /ttt .71-1 allilbod-v l 1-t,aolb/lui7lill Wt'¢.s s'I(l }ipplic bY '0/(/ I.Tnlike Fn the ICLtC phase pi-oteill CRP is Biotiedlic'O/ LlhOrOtOli /:)0-.01 (comipare/ Ivil/i produced onlx in the lixver.4 Although there is no utr(SE l db 1t0 r/itic anlinaIci . Rl(cm(11/i.lcr actixve sxvthesis alt the sitc of infliamimiaitioni CRP is ( S E.\1 *1(ot 0710 /}(2Z(>tl1 pcr gru.(/ aenerallx accepted as a uLseftll Clin l measCiSUrement Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

520 Connolly, Stecher, Pruden of disease activitv. The present study supports Wc are zraitct'Lfl t'or tthc tchilincal assistarinc ot Tcrcsia La B3r-IC John Rithrilannn Shirlv Allin RLIssell Bcckei Ic Bohilt use as a marker of arthritic disease, as the of CRP Charles FILIIIo. and Phyllis Spchioht The atithot-s also \\ ish to the results indicate that plasma CRP concentrations acknowsledge Joseph Deleskicssics and Jo Anin MaLz of the are significantly raised in arthritic rats and can be "rraphics dcpairtmcnt. is \\cll as Virgzinia Dixon. Wends lanklc. reduced by treatment with auranofin. Darlcne DcFraiiccsco. anid Doniia Sparrosw for tlicii cxperIt t\psll( ahilitv Atirainofin (lot No KFK-8933 -101) sas a -f( ft' toill the As the liver accounts for all or part of the Smith. Klinec and Ficnch Lahoratories synthesis of CRP and Fn it was important to establish that the auranofin mediated reduction of References plasma Fn and CRP was not due to generalised Empire Rhcumatismii C'ouncil sLhIcomilmittcc. Gold therap\ ill Clinical rhcumatoid arthritis, final rcport of a muilticcntrc conti-ollcd inhibition of protein synthesis by the liver. trial. Ann R/ieumt I)i 1961 20: 315-33. results indicate that a subnormal concentration of 2 Billinhamliar M E. Models of arthritis aind the scact t'ot plasma albumin is associated with RA.'3 Data anticirthritic drugs Phlarmnacol Tlhe, 1983: 21: 389-428 from this laboratory35 36 and others48 indicate that 3 Rainsford K D Adjusant polvarthritis in rats: Is this a satisfactorv modcl for screcninig antilrthritis drugs? A4geuts arthritic rats also possess abnormally low concen- Actions 1982: 12: 452-8. trations of plasma albumin. Although these low 4 Walz D T. Mechanisms of action of gold salts in rhCeInumatoid concentrations may be due to increased catabolism arthritis Advances int Infltaininatioo Researcht 1984. 7: 239-47. of albumin,4" there is evidence that a low plasma 5 Stcchcr V J. Kaplan J E, Connolls K M. Miclcns A. Salccns J K. Fibroncctin in acute aind chronic inflammiation. Arth/ritir albumin concentration is an indicator of liver Rlieito 1986: 29: 394-9 dysfunction.11- If auranofin reduced plasma Fn 6 Carsons S. Moscsson M W. Diamond 11 S. Detction iaid and CRP merely by inhibiting protein synthesis in quantitation otf fibroncctin in ssnosvial fluid from patients w ith the liver then plasma albumin concentrations in rheumattic discasc. Artluritios Rlietiti 1981: 24: 1261-7. 7 M. lamnmartino A J. Schmid F R. et arthritic rats treated with auranofin would also be Lu-Stcffcs ah. Fibroncetin in rhCUmratoid and non-rhcumnatoid arthritis ss nos ial fltuids ilaid expected to be low. Auranofin treatment. however, in svnovial fluid crsoprotcins. Allul Clio lab .Sci 1982. 12: reversed the low albumin concentrations seen in 178-85 arthritic rats. In view of the inverse relation between 8 Scott D L. Farr M. Crockson A P. Wialton K W. Ss\nosvia fltLidcopyright. and plasma t'ibronectin lscls in rhcumratoid airthritis. (/liot S the ability of auranofin to lower plasma CRP and Fn 1981: 62: 71-6. concentration while at the same time raising plasma 9 Vartio .1. Vahcrt A. Von Esscn R. lsomiaki II. Stcnmain S. albumin concentration it is unlikely that auranofin is Fibroncctin in synov'ial flulid and tissuc in rhcumaitoid airthritis. causing generalised inhibition of protein synthesis in fLur J C(hi /Iin't 198X 11: 2'(7-12. the liver. Although 30 mg/kg of auranofin has been 1ff Scott D L. Wainwright A C. Walton K W. Williamson N. Significancc otf fibroncctin in rhcumatoid airthritis and ostco- cited as the minimum lethal dose in rats,'- the 10 airthrosis. Atiii Rhliltoi DOi 1981: 40: 142-53. mg/kg dose used in our study resulted in no lethality 11 Carsons S. Parcnti D. Lasvictes B B. Diamond H S. Singcr A. http://ard.bmj.com/ or weight loss. Results of a three month toxicology Boxcr M. Plasnmia fibronectin in systemic lupus crythcmilatosus: indicated that the side effect of rclationship to clinical actisity. DNA binding and acute phase study only dosing protcins. J Rliewlmto(otol 1981X: 12: 1(088-92. with 12 mg/kg of auranofin was a less than average 12 llvncs R O. Yamadia K M. Fibroncctins: multifunctional weight gain.4 This supports our data showing that modular gIvcoproteins. J Cell Biol 1982: 95: 369-77. treatment of normal animals with 10 mg/kg of 13 Norris D A. Clark R A F. Swigart M. Huff J C. Wcston W L. Howcil S E. Fibroncctin are chcmotactic for human not or CRP concen- fragments auranofin did alter plasma Fn pcripheral hlood monocytcs. J lhnmnaol 1982: 129: 1612-8 trations. Thus it does not appear that reduction of 14 Moscsson M W. Amrani D L. The structurc aind hiologic on September 27, 2021 by guest. Protected Fn and CRP in arthritic rats is due to non-specific activities of plasma fibroncctin. Blood 198ff. 56: 145-58. toxicity. The alteration of Fn,1 CRP,1' and 15Akivama S K. Yamada K M. Havashi M. The structure of fibronectin aind its rolc in ccilular adhcsion. Jolurtt(ti of albumin57 18 production may be under regulation by Supranole ulair Striuclure t(d Cellildat- Biochlemistr 1981: 16: one or more cytokines. Hepatocyte stimulatory 345-58. factor159 has been reportedly involved in the induc- 16 livncs R 0. Fibroncctins. Sci Amtl 1986: 254 42-51. of tion of CRP in human hepatoma cell lines,?6 while 17 Scott D L. Delamerc J P. Walton K W. The distribution fibronectin in the pannus in rhcumatoid arthritis. Br J L xp we and others have evidence that high concen- Pathol 1981: 62: 362-8. trations of splenic interleukin-l in adjuvant arthritic 18 Wcissman G. Activation of ncutrophils and the lesions of rats36 can be significantly reduced after treatment rheumatoid arthritis. J Lab Clitt Med 1982: 100: 322-33. with auranofin. 19 Connolly K. Stechcr V J, Saclens J K. Kaplan J E. The levels and autoimmunc abnormal rclationship hctwecn plasma fibroncctin We have also recently shown that disease activity in MRU1 micc. Proc Soc Erp Biot Med 1985: concentrations of plasma Fn, CRP, and albumin in 180: 149-54. the adjuvant arthritic rat are not altered by NSAID 2(0 Stecher V J. Carlson J A. Connolly K M. Bailey D M. Discasc- treatment.36 Thus measurement of these acute modifying antirheumatic drugs. Med Res Rev, 1985; 5: 371-9ff. 21 Piliero S J. Gracme M L. E B. Chinca G. Colomho C. in the arthritic rat be Sigg phase proteins adjuvant may Action of anti-inflammatory agents upon blood and histopatho- useful in differentiating potential antirheumatic logic changes induced hv pcriarthritis in rats. Life Sci 1966; 5: compounds from standard NSAIDs. 11157-69. Ann Rheum Dis: first published as 10.1136/ard.47.6.515 on 1 June 1988. Downloaded from

Treatmnent of arthritic rats with auranlofin 521

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