2
480 Autoantibodies 25 June 1997 - Poster presentations idiotope is decreased only in the sera of ALS patients. (This work was supported by the Ministry of Science, Belgrade, Serbia.) P.5.21.26 Synovlal tissue B-lymphocytes as source for the generatlon of human monoclonal antibodles P. von Landenberg, I. Melchers. Clinical Research Unit on Rheumato/w University Medical Center, F&burg, Germany Introduction: B-cell reactivities in the affected joints of patients with rheumatoid arthritis probably are an important factor in the chronic inflammation process. Our aim is to characterize the antibodies, being directly associated with the synovial tissue damage, in terms of specificity and V gene usage. For immortal- isation of synovial B-lymphocytes from patients with rheumatoid arthritis we use the method of electrofusion. This technique allows to produce B-cell hybrido- mas with high fusion efficiency even from mixed cell populations containing only small numbers of &lymphocytes. Materials and Methods: Synovial tissue B-lymphocytes from patients with rheumatoid arthritis (defined according to the criteria of the American Rheumatism Association) were prepared by collagenase treatment and me- chanical dissection. The viable leucocyte population in neatly all used samples contained only about 3% CD19 positive B-lymphocytes. The total number of mononuclear cells obtained after this treatment varied from 1 x IO5 to 5 x 105. This population was then immortalized by electrofusion without any in vitro stimulation or prior B-cell-selection. From this cell population we obtained B-cell hybrtdomas in a frequency of 1 in 10.000 total cells, i.e. 1 clone from 300 B-lymphocytes. Reeuh Up to now we established 10 antibody secreting hybrfdomas from synovial fluid and tissue. B-lymphocytes isolated from synovial fluid (1 patient) and from synovial tissue (2 patients) were used for electmfusion. 4 hybridomas secreted IgA and 6 hybrfdomas secreted IgG. Conclusion: Electrofusion apperantty allows to fuse B cells in a broader range of differentiation stages than conventional techniques. Synovial B-cells differ from peripheral B-cells in several aspects. Therefore the production of synovial hybtidomas is up to now difficult and rarely reported. With this technique we are now able to investigate the local B-cetl response in the synovial tissue of RA-patients. And we can expect, that antibody specificities detected with these monoclonal antibodies might reflect relevant autoantigens involved in the local inflammation process. Supported by a DFG fellowship (La 1051/l) and a DFG grant (Me 604./4). ( P.5.21.27 1 Prevalence of antlneutrophll cytoplasmlc antlbody (ANCA) In patients with autolmmune hepatltls 6. Malenica l, A. KrstuloviC l, R. OstojiC*, B. VuceliC*, D. BatlniC l. 1 Div Immunol, Dept Lab Diagnosis, Zagreb Univ School A&d, Zagreb, Zagreb, Croatia, 2Div Gastroenterolog~ Dept lntemal Med., C/in Hosp Center Zagreb, Zagreb, Croatia Introduction: ANCA autoantibodies have been described as a sensitive im- munoserologic marker of certain types of systemic vasculiis, especially We- gener’s granulomatosis, microscopic arterftis and idiopathic crescentic glomeru- lonephritis. A few studies have also shown that patients with some inflammatory bowel and hepatobiliary diseases have autoantibodies directed against neu- trophils, which am a different specificity than those associated with vasculitis. The prevalence, titer and clinical correlation of these antibodies we investigated in patients with various hepatobiliary diseases. PatIenta and Methodr: The ANCA was analyzed in 141 serum samples. The presence of ANCA was detected on ethanol-fixed cytospin preparations of peripheral blood neutmphils by indirect immunofluorescence. A perfnuclear (pANCA) and atypical (aANCA) staining pattern was considered positive. West- em blot was used for detection of autoantibodies which characterized different subtypes od autoimmune hepatitis. Reeut& Both pANCA and aANCA was detected in 20% (7/34) of patients with autoimmune hepatitis, in 7% (l/15) of patients with primary biliary cinhosis and in 4% (l/26) of patients with biliary cirrhosis. ANCA did not find in patients with chronic hepatitis (o/23) and in a group of patients with valious hepatobiliary disorders (O/43). The ANCA autoantibodies was selectively observed in patients with type 1 autoimmune hepatitis @/IO; 60%). Conclusion: These results indicate that ANCA can be used as useful clinical tool to distinguish different subtypes of autoimmune hepatitis. 1P.5.21.28 1 Antlbodles agalnst cardlollpln and oxLDL In lilkyear-old men predict myocardlal lnfarctlon a prospective 20 years’ study Ruihua Wu’ , Soniya Nityanand l, Lars berglund 2, Hans Lithell *, G&an Hdm ‘, Ann Karl Lefverl ‘. ‘Medicine of Dept. Karvlinska Hospital, Stc&ho/m, Sweden, *Department of Geriatrics, University of Uppsala, Uppsala, Sweden Abstract: Autoantibodies against oxidatively modified low density lipoproteins (oxLDL) and cardiolipin occur in patients with vascular diseases, including atherosclerosis. The ability of such antibodies to predict myocardial infarctton (Ml) was investigated in a prospective nested case control study in samples col- lected from 2322 males aged 50 years in Uppsata, Sweden during 197&1972. This cohort made up 82% of all 50 year old men living in Uppsala at that time. The cohort was then followed for 20 years. Autoantibodies against oxLDL and cardiolipin were measured with an enzyme-linked immunosorbent assay method. Raised levels of antibodies against oxLDL and cardidipin at 50 years of age correlated positively with the incidence of Ml and mortality related to Ml 10 to 20 years later. IgG and IgA antibodies against cardidipin were associated with Ml between 50-60 of age and IgG and IgA antibodies against oxLDL with Ml at 60-70 yean of age. Moreover, higher antibody levels were noted in those who died from acute Ml in comparison to those who survived. The predictive power of IgA and IgG antfbodies was strong and independent of that of other strong risk factors. In conclusion, raised levels of antibodies against oxLDL and cardiolipin may predict Ml and Ml-related death. LI_l P.5.21.29 The spectrum of autoantlbodles In HIV seroposltlve subjects in varlous stages of HIV infection A. Tsimyianni, M. Economou, K. Terzoglou. J. Economidou. Deperhnent of lmmunolosv-Hist~m~ocompatibiliiy, Evangelismos Hospital, Athens, Greece Intoduction: Autoimmune reactions and different types of autoantibodies (AA) have been mcognised in HIV infection. This study analyses serological aspects of organ-specific AA (PCA, AMA, SMA, ATA), non organ-specific AA (ANA, antidsDNA, anti-CL, anti-histones) and anti-neutrophil antibodies (ANCA), in individuals with different stages of HIV infection according to CDC classification (1993). Material andMethods: Sera from two groups of HIV-l seropositive subjects were investigated. Group 1 consisted of 24 asymptomatic carders (stage Al and A2) and group 2, of 23 patients with AIDS (stages A3,83, C). For the detection of AA, indirect immunofluorescence (IIF) and ELISA techniques were used. Results: The percentage of the various AA detected is shown in the table. Autoantibodies WA Al(N=lO)% AZ(N = 14)% A3,S3,C(N=32)% 0 14.3 Q-4 ANA 0 7.1 8.2 anti-DNA 0 0 0 anti-histones 50 64.3 58.4 anti-cardiolipin IgG 20 14.3 6.3 anti-cardiolipin IgM 0 0 6.3 ANCA - 30 14.3 53.1 Concluelon: In general, fewer specificities and a lower frequency of AA was found in stage Al than in stage A2 or in AIDS, with the exception of anti-histones AA which occured with equal frequency in all stages. ANCA were found to be especially increased in AIDS, they were exclusively C-ANCA and with an atypical staining pattern in IIF. These results indicate that a variety of mechanisms including apoptosis may be implicated for the immunological disregulation resulting in autoimmune reactions against a wide spectrum of autoantigens. 1P.5.21.30 ] Frequency and speclflcltles anti-neutrophll antlbodles (ANCA) In Inflammatory bowel disease M. Economou, A. Tsimyianni, K. Tetzoglou, J. Economidou. Department of Immunolcgy-Hi&compatibility, Evangelismos Hospital, Athens, Greece Introduction: Anti-neutmphil antibodies producing a p-ANCA pattern by indirect immunofluorescense (IIF) have been detected recently in sera of patients with inflammatory bowel disease (IBD). To evaluate their clinical significance, sera from patients with ulceratiie colitis (UC) and Crohn’s disease (CD) in phases with active and inactive disease have been investigated. Yaterlal and Methods: Sera from Q4 patients with IBD (69 with UC and 25 with CD) were tested for the detection of ANCA using IIF and ELISA techniques forthespecificitiesanti-PRS, anti-MPO, anti-elastase (E), anti-cathepsin G (C-G) and anti-lactoferrin (1). Reeulk The frequency of ANCA is shown in the table. No statistically significant difference was found between males and females with UC and CD

Frequency and specificities anti-neutrophil antibodies (ANCA) in inflammatory bowel disease

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Page 1: Frequency and specificities anti-neutrophil antibodies (ANCA) in inflammatory bowel disease

480 Autoantibodies 25 June 1997 - Poster presentations

idiotope is decreased only in the sera of ALS patients. (This work was supported by the Ministry of Science, Belgrade, Serbia.)

P.5.21.26 Synovlal tissue B-lymphocytes as source for the generatlon of human monoclonal antibodles

P. von Landenberg, I. Melchers. Clinical Research Unit on Rheumato/w University Medical Center, F&burg, Germany

Introduction: B-cell reactivities in the affected joints of patients with rheumatoid arthritis probably are an important factor in the chronic inflammation process. Our aim is to characterize the antibodies, being directly associated with the synovial tissue damage, in terms of specificity and V gene usage. For immortal- isation of synovial B-lymphocytes from patients with rheumatoid arthritis we use the method of electrofusion. This technique allows to produce B-cell hybrido- mas with high fusion efficiency even from mixed cell populations containing only small numbers of &lymphocytes.

Materials and Methods: Synovial tissue B-lymphocytes from patients with rheumatoid arthritis (defined according to the criteria of the American Rheumatism Association) were prepared by collagenase treatment and me- chanical dissection. The viable leucocyte population in neatly all used samples contained only about 3% CD19 positive B-lymphocytes. The total number of mononuclear cells obtained after this treatment varied from 1 x IO5 to 5 x 105. This population was then immortalized by electrofusion without any in vitro stimulation or prior B-cell-selection. From this cell population we obtained B-cell hybrtdomas in a frequency of 1 in 10.000 total cells, i.e. 1 clone from 300 B-lymphocytes.

Reeuh Up to now we established 10 antibody secreting hybrfdomas from synovial fluid and tissue. B-lymphocytes isolated from synovial fluid (1 patient) and from synovial tissue (2 patients) were used for electmfusion. 4 hybridomas secreted IgA and 6 hybrfdomas secreted IgG.

Conclusion: Electrofusion apperantty allows to fuse B cells in a broader range of differentiation stages than conventional techniques. Synovial B-cells differ from peripheral B-cells in several aspects. Therefore the production of synovial hybtidomas is up to now difficult and rarely reported.

With this technique we are now able to investigate the local B-cetl response in the synovial tissue of RA-patients. And we can expect, that antibody specificities detected with these monoclonal antibodies might reflect relevant autoantigens involved in the local inflammation process.

Supported by a DFG fellowship (La 1051/l) and a DFG grant (Me 604./4).

( P.5.21.27 1 Prevalence of antlneutrophll cytoplasmlc antlbody (ANCA) In patients with autolmmune hepatltls

6. Malenica l, A. KrstuloviC l, R. OstojiC*, B. VuceliC*, D. BatlniC l. 1 Div Immunol, Dept Lab Diagnosis, Zagreb Univ School A&d, Zagreb, Zagreb, Croatia, 2 Div Gastroenterolog~ Dept lntemal Med., C/in Hosp Center Zagreb, Zagreb, Croatia

Introduction: ANCA autoantibodies have been described as a sensitive im- munoserologic marker of certain types of systemic vasculiis, especially We- gener’s granulomatosis, microscopic arterftis and idiopathic crescentic glomeru- lonephritis. A few studies have also shown that patients with some inflammatory bowel and hepatobiliary diseases have autoantibodies directed against neu- trophils, which am a different specificity than those associated with vasculitis. The prevalence, titer and clinical correlation of these antibodies we investigated in patients with various hepatobiliary diseases.

PatIenta and Methodr: The ANCA was analyzed in 141 serum samples. The presence of ANCA was detected on ethanol-fixed cytospin preparations of peripheral blood neutmphils by indirect immunofluorescence. A perfnuclear (pANCA) and atypical (aANCA) staining pattern was considered positive. West- em blot was used for detection of autoantibodies which characterized different subtypes od autoimmune hepatitis.

Reeut& Both pANCA and aANCA was detected in 20% (7/34) of patients with autoimmune hepatitis, in 7% (l/15) of patients with primary biliary cinhosis and in 4% (l/26) of patients with biliary cirrhosis. ANCA did not find in patients with chronic hepatitis (o/23) and in a group of patients with valious hepatobiliary disorders (O/43). The ANCA autoantibodies was selectively observed in patients with type 1 autoimmune hepatitis @/IO; 60%).

Conclusion: These results indicate that ANCA can be used as useful clinical tool to distinguish different subtypes of autoimmune hepatitis.

1 P.5.21.28 1 Antlbodles agalnst cardlollpln and oxLDL In lilkyear-old men predict myocardlal lnfarctlon a prospective 20 years’ study

Ruihua Wu’ , Soniya Nityanand l, Lars berglund 2, Hans Lithell *, G&an Hdm ‘, Ann Karl Lefverl ‘. ‘Medicine of Dept. Karvlinska Hospital, Stc&ho/m, Sweden, *Department of Geriatrics, University of Uppsala, Uppsala, Sweden

Abstract: Autoantibodies against oxidatively modified low density lipoproteins (oxLDL) and cardiolipin occur in patients with vascular diseases, including atherosclerosis. The ability of such antibodies to predict myocardial infarctton (Ml) was investigated in a prospective nested case control study in samples col- lected from 2322 males aged 50 years in Uppsata, Sweden during 197&1972. This cohort made up 82% of all 50 year old men living in Uppsala at that time. The cohort was then followed for 20 years. Autoantibodies against oxLDL and cardiolipin were measured with an enzyme-linked immunosorbent assay method. Raised levels of antibodies against oxLDL and cardidipin at 50 years of age correlated positively with the incidence of Ml and mortality related to Ml 10 to 20 years later. IgG and IgA antibodies against cardidipin were associated with Ml between 50-60 of age and IgG and IgA antibodies against oxLDL with Ml at 60-70 yean of age. Moreover, higher antibody levels were noted in those who died from acute Ml in comparison to those who survived.

The predictive power of IgA and IgG antfbodies was strong and independent of that of other strong risk factors.

In conclusion, raised levels of antibodies against oxLDL and cardiolipin may predict Ml and Ml-related death.

LI_l P.5.21.29 The spectrum of autoantlbodles In HIV seroposltlve subjects in varlous stages of HIV infection

A. Tsimyianni, M. Economou, K. Terzoglou. J. Economidou. Deperhnent of lmmunolosv-Hist~m~ocompatibiliiy, Evangelismos Hospital, Athens, Greece

Intoduction: Autoimmune reactions and different types of autoantibodies (AA) have been mcognised in HIV infection. This study analyses serological aspects of organ-specific AA (PCA, AMA, SMA, ATA), non organ-specific AA (ANA, antidsDNA, anti-CL, anti-histones) and anti-neutrophil antibodies (ANCA), in individuals with different stages of HIV infection according to CDC classification (1993).

Material and Methods: Sera from two groups of HIV-l seropositive subjects were investigated. Group 1 consisted of 24 asymptomatic carders (stage Al and A2) and group 2, of 23 patients with AIDS (stages A3,83, C). For the detection of AA, indirect immunofluorescence (IIF) and ELISA techniques were used.

Results: The percentage of the various AA detected is shown in the table.

Autoantibodies WA

Al(N=lO)% AZ (N = 14)% A3,S3,C(N=32)%

0 14.3 Q-4

ANA 0 7.1 8.2 anti-DNA 0 0 0 anti-histones 50 64.3 58.4 anti-cardiolipin IgG 20 14.3 6.3 anti-cardiolipin IgM 0 0 6.3 ANCA - 30 14.3 53.1

Concluelon: In general, fewer specificities and a lower frequency of AA was found in stage Al than in stage A2 or in AIDS, with the exception of anti-histones AA which occured with equal frequency in all stages. ANCA were found to be especially increased in AIDS, they were exclusively C-ANCA and with an atypical staining pattern in IIF. These results indicate that a variety of mechanisms including apoptosis may be implicated for the immunological disregulation resulting in autoimmune reactions against a wide spectrum of autoantigens.

1 P.5.21.30 ] Frequency and speclflcltles anti-neutrophll antlbodles (ANCA) In Inflammatory bowel disease

M. Economou, A. Tsimyianni, K. Tetzoglou, J. Economidou. Department of Immunolcgy-Hi&compatibility, Evangelismos Hospital, Athens, Greece

Introduction: Anti-neutmphil antibodies producing a p-ANCA pattern by indirect immunofluorescense (IIF) have been detected recently in sera of patients with inflammatory bowel disease (IBD). To evaluate their clinical significance, sera from patients with ulceratiie colitis (UC) and Crohn’s disease (CD) in phases with active and inactive disease have been investigated.

Yaterlal and Methods: Sera from Q4 patients with IBD (69 with UC and 25 with CD) were tested for the detection of ANCA using IIF and ELISA techniques forthespecificitiesanti-PRS, anti-MPO, anti-elastase (E), anti-cathepsin G (C-G) and anti-lactoferrin (1).

Reeulk The frequency of ANCA is shown in the table. No statistically significant difference was found between males and females with UC and CD

Page 2: Frequency and specificities anti-neutrophil antibodies (ANCA) in inflammatory bowel disease

25 June 1997 - Poster presentations Intervention in autoimmunity 481

or between active and inactive phases of the disease in the frequencies of the various antibodies tested.

Disease N Indirect IF ELISA

C-ANCA P-ANCA (%) MPO PR3 E C-G L

UC

co 69 0

2; 0

38 1551 0 0 1 3 0

7 (28) 0 0 2 0 2

Concfuslon: ANCA occur more commonly in UC than in CD, the majority remain of undefined specificity and their pattern differs from that observed in vasculitis.

1 P.5.21.31 ] Expression of recombinant proteinase-3: A comparison between different forms of pr3 and their recognition by monocionai antibodies and patient sera

Y.M. van der Geld, MC. Harmsen, P. Heerfnga, M.G. Huitema, A. Klimp, A. Tiran, C.G.M. Kallenberg. Department of Clinical /mmuno/og~ University Hospital Grvningen, The Netherlands

Introductfen: Proteinase 3 (PIN) is the major antigen for autoantibodies against cytoplasmatfc components of neutrophils (c-ANCA) in Wegener’s Granulomato- sis (WG). It has been suggested that c-ANCA recognize different, conformational epitopes and that changes in epitopespecificity of ANCA am related to disease activity.

hbterfals and Methods: Using PCR with tailored EcoRl restriction sites and coding infomation for a C-terminal 6 histidine tag, we amplified the open reading frame of Pr3. The insert was cloned and expressed in a prokaryotic expression vector. Recombinant E. co/i Pr3 (rcP13) was induced with IPTG and was re- tained in the cytoplasm. The P&encoding insert was further subcloned into a Pichia Pastoris recombination vector, which could recombine with the methanol oxktase gene, yielding mut- (methanol utflisation miuns) recombinants. Recom- binant Pichia Pt3 (rpPr3) was induced with methanol and was secreted into the medium as detected with antigen-specific capture-ELISA. The secreted rpPr3 and rcPr3 were isolated through IMAC (immobilised metal chelate chromatogra- phy) and concentrated. Western blot analysis and ELISA were used for protein detection. Enzymatic activity was determined by proteolytic cleavage of a spe- cific substrate. Antigen specific ELISA was used to study recognition of rPr3 by 11 monoclonal antibodies and sera from 22 patients with WG.

Reeuftez On SDS-PAGE the rcP13 had the full length size of native Pr3 (approx. 28 kDa) and reacted with a polyclonal rabbi antiserum. RpPr3 revealed a prominent band at approx. 32kDa which also reacted with polyclonal and monoclonal antibodies.

RpPr3 was recognised by all monoclonals in a catching ELISA while 4 moabs also recognized rpPr3 in a direct ELISA. In a catching ELISA 4 monoclonals recognized rcPA while none recognized rcPr3 in a direct ELISA. RpPr3 was recognized by 70% and the rcPt3 by 40% of the anti-Pr3 positive sera in a catching ELISA. However, none of the WG sera reacted with rcPr3 in a direct ELISA, whereas only 2 sera reacted with rpPt3 by direct ELISA. Furthermore, rpPt3 but not rcPr3 harboured protease activity which could be inhibited with PMSF and al-antftrypsin.

Conclueion: As 46-70% of the WG sera recognized the rPt3 products, these products presently can’t be used in a diagnostic test. The results further suggest that there are differences between WG sera in the epitopes recognized.

P.5.21.32 Anti-neutrophii cytopiasmic antibodies (ANCA) in primary scterosing cholangitis

In antigen-specific ELlSAs, 55% of the serum samples recognized one or more antigens. Forty-six percent contained antibodies to bacterfci- daf/peneabBty-increasing protein, 23% to cathepsin G, and 22% to lactoferrin. Frequently, antibodies to combinations of those antigens were simultaneously present in a serum sample. Only 4% of the patients had antibodies to proteinase 3, whereas antibodies to myeloperoxidase or elastase were not detected. The presence of ANCA was not related to the duration of PSC. Also, ANCA as de- tected by indirect immunofluorescence were not associated with the presence of cirrhosis nor with the coexistence of inflammatory bowel disease. However, antibodies to bacterfcidal/permeability-increasing protein and cathepsin G were both associated with the presence of cirrhosis, and antibodies to lactoferdn were more frequently detected in patients with PSC in conjunction with ulcerative coi- itis.

Conclusion: We conclude from this study that bactedcidal/permeabitii-in- creasing protein is a major ANCA antigen in PSC, often in combination with cathepsin G or lactoferdn. Antibodies to bacterfcidal/penneability-increasing protein and cathepsin G are related to cfrrhosis, and antibodies to lactofentn in PSC are associated with the concurrent presence of ulcerative colitis.

09:00-l 8:30/ 12:00-l 4:00 Forum lounges

P.5.22 Intervention in autoimmunity

P.5.22.01 MC 1288 -A vitamin D anaiogue with immunosuppressive effects which suppresses collagen-induced arthritis in rats

L. Ma&son I, L. Svelander ‘, A. Bucht ‘, C. Johnsson 2, Lars Klareskoo ‘, Per Larsson ‘. ’ Dept of Rheumatology, Kamlinska Hospital, Stoddmlm~ Uppsala, Sweden, 2 Dept of Tmnsplantation Surgery, University Hospital, ti~psala, Sweden

Introduction: Cholecalciferol has been shown to have immunomodulatory and antiinflammatory effects but also serious side effects such as hypercalcemia. Here we show that the vitamin-D analogue. MC 1288 (provided by dr Lisa Binderup, Leo Pharmaceutical Products, Denmark) have immunomodulatory effect on collagen-induced arthritis in rats at doses that do not induce hypercal- cemia.

Materials and Mothode: Arthritis was induced by immunisation of DA rats with rat type II collagen in FIA. lmmunised rats were treated, either before arthritis development or after arthritis development, with MC 1288 intraperttoneally at doses ranging from 0.1 fig&g to 0.025 &kg daily or with the vehicle. The incidence, the severity of arthritis as well as the cytokfne production of lymph node cells and the serum levels of anti-Cll antibody were evaluated.

ReeulB: 1) Treatment with the vitamin D analogue before arthritis had developed inhibited joint inflammation. Only 50% of the rats developed arthritis as compared 100% arthritis in the control rats. 2) Treatment initiated after the onset of the disease did not affect the incidence. However, the severity of the joint lesions was significantly suppressed. 3) Treatment with low levels of the analogue which did not induce hypercalcemia did diminished arthritis severity equally well as higher doses of MC 1288.4) Rats treated with MC 1288 developed significantly lower titers of IgGPa antibodies to CII. Titers of other IgG isotypes were not effected.

C. Roozendaal ‘, A.W.M. van Milligen de Wit2, E.B. Haagsma’, G. Horst ‘, C. Schwarze3, H.H. Peter3, J.W. Cohen Tewaert’, P.C. Limburg ‘, C.G.M. Kallenberg ‘. ’ Dept. htemal Medicine, University Hospital, Grcningen, The Nethedands, 2 Dept. Gastmenterobgy and HepatofogK Academic Medical Centre, Amsterdam, The Netherlands, 3Dept. Rheumatology and Clinical

Conclusion: It has previously been shown that cholecaldfeml has im- munomodulatory properties. Here we demonstrate a strong suppressive effect on experimental arthritfs in rats of the vitamin D analogue MC 1288. This may be explained by a decrease in the autoantibody response to collagen II and effects on cytokines.

Immunol~, University Hospital, Freibug, Germany

Introduction: Autoantibodies directed against cytoplasmic constituents of neu- troohil aranulocvtes IANCA) have been detected in serum samoles from oatients with pn’mary &ero&ng chblangitis (PSC). The antigenic specificities and the clinical correlates of those ANCA have, however, not been fully elucidated. Aim of this study was to determine the antigenic specificities and the clinical corre- lates of anti-neutrophil cytoplasmic antibodies (ANCA) in serum samples from 69 patients wfth PSC.

Methods: The presence of serum IgG ANCA was analyzed by indirect immunofluorescence on ethanol fixed granulocytes. Antibodies directed against bactericidav~~ability-increasing protein, cathepsin G, lactofentn, proteinase 3, myefopemxidase, and elastase were identified by ELISA.

Reeults: IgG ANCA were detected by indirect immunofluorescence in 67% of the PSC patients. All serum samples produced a pednuclear fluorescence pattern (PANCA).

P.5.22.02 MHC Wpeptide llpoeomes as artificial antigen oresentina cells

Annemiek J.M.L. van Rensen ‘, L.S. Taams’ , M.C.J.T. Grosfeld-Stulemeyer 2, A. Besselings, W. van Eden2, D.J.A. Cmmmelin’, M.H.M. Wauben*. ’ Gnmingen Utmcht institute for Drug Exp/oration, Dept. of Pha~aceutics, faculty of Pharmacy; Utrecht University, TB Utrecht, The Netherlands. 2 institute of Infectious Diseases and Immunolcgy, Facu/ty of Veterinary Medicine, Utrecht UniversiQ The Netherlands

Intmductlon: Interference in the responses of self-reactive T cells has been the focus of extensive research, since these cells play a major role in certain autoimmune diseases. Our goal is to modulate T cell responses via antigen presentation in the absence of costimulatory signals. In this way, we aim at the induction of antigen-specific T cell unresponsiveness or apoptosis. A way to