Journal of Clinical Immunology, Vol. I1, No. 4, 1991
Analysis of Function-Associated Receptor Molecules on Peripheral Blood and Synovial Fluid Granulocytes from Patients with Rheumatoid and Reactive Arthritis THOMAS FELZMANN, 1 STEPHEN GADD, 1 OTTO MAJDIC, 1 DIETER MAURER, 1 PETER PETERA, 2 JOSEF SMOLEN, 2 and WALTER KNAPP 1'3
Accepted: April 1, 1991
and cells from reactive arthritis patients, suggesting a similar activation process for granulocytes in these two diseases.
In this study we report the expression pattern of 13 different function-associated surface molecules on synovial fluid and peripheral blood granulocytes from rheumatoid and reactive arthritis patients. We found increased expression of the complement receptors 1 (CD35) and 3 (CD1lb) and of the activation-associated antigens CD67, CD24, and M5 on synovial fluid granulocytes from rheumatoid and/or reactive arthritis patients compared to autologous peripheral blood granulocytes. In addition, synovial fluid granulocytes expressed IgG Fc receptor 1 (CD64) and complement receptor 4 (CDllc), neither of which can be found on peripheral blood granulocytes. Peripheral blood granulocytes from rheumatoid and reactive arthritis patients expressed higher levels of leucocyte function-associated antigen 1 (CDlla) and of the membrane proteins CD31, CD24, M5, and M6 compared to peripheral blood granulocytes from healthy controls and patients with degenerative joint disease. No significant differences in the expression of any of the molecules studied could be observed between cells from rheumatoid
KEY WORDS: Fc receptors; complementreceptors; adhesion molecules; granulocyteactivation;flowcytometry.
INTRODUCTION The expression of neutrophil granulocyte surface molecules can be modulated by a variety of agents including cytokines such as granulocyte macrophage-colony stimulating factor (GM-CSF) 4 (1), interferon-~ (IFN-~) (2, 3), tumor necrosis factor et (TNFa) (4), interleukin-8 (IL-8) (5), and complement split products (4, 6, 7) as well as by FcRs (3) and receptors for microbial wall components such as lipopolysaccharides (8). Recently we and others have shown that in vitro and in vivo stimulation of human granulocytes leads to changes in the surface expression of certain function-associated membrane molecules (3, 6, 9-13). Neutrophils are the predominant cell type in inflammatory joint effusions of patients with rheumatoid arthritis (RA) and reactive arthritis (ReA) (14). However, little is known about possible changes in the surface expression of membrane molecules on neutrophils in RA. To our knowledge, only one report has been published which described the expression of CR3 and platelet glycoprotein IIb/IIIa on synovial fluid granulocytes (SFG) and peripheral blood granulocytes (PBG) in RA (15). Here, we describe the expression pattern of 13 different surface molecules on SFG and PBG from RA and ReA patients in comparison with the expression pattern found on PBG from patients
1Institute of Immunology,Universityof Vienna, Borschkegasse 8a, A-1090, Vienna, Austria. 2Department of Medicine and LudwigBoltzmann Institute for Rheumatology, KH Lainz, Wolkersbergenstr.l, A-1130, Vienna, Austria. 3To whom correspondence should be addressed at Borschkeg. 8a, A-1090, Vienna, Austria. 4Abbreviationsused: rheumatoidarthritis(RA);reactivearthritis (ReA); degenerativejoint disease (DJD); control individuals (CO); peripheral blood granulocytes(PBG);synovialfluidgranulocytes (SFG); monoclonalantibody(mAb); IgG Fc receptor (FcR); complement receptor (CR); leukocyte functionassociated antigen (LFA); granulocyte macrophage-colony stimulating factor (GM-CSF); granulocyte-colonystimulating factor (G-CSF); interferon ~/(IFN-7); tumor necrosis factor et (TNFct); interleukin (IL); leukocyte cytoadhesion molecule (LeuCAM);mean fluorescenceintensity(MFI); phorbol myriastate acetate (PMA); formyl-methionylleucyl phenylalanine (fMLP); intercellular cytoadhesion molecule (ICAM); phosphoinositol(PI); nonsteroidalantiinflammatorydrugs (NSAID). 205
0271-9142/91/0700-0205506.50/0 © 1991 Plenum Publishing Corporation
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FELZMANN ET AL.
Table I. Monoclonal Antibodies Used in this Study mAb
CD
Molecule
Ref. No.
32.2 IV.3 VIFcR3 To5 MHM24 VIM12 3.9 MHM23 GIOF5 1B5 AAA-6 VIM-5 VIBC5 VIMD5 VIM13
CD64 CDW32 CDt6 CD35 CD 11a CD1 lb CD1 lc CD18 CD67 CD31 ND ~ ND CD24 CD15 CD14
Fc receptor 1 Fc receptor 2 Fc receptor 3 Complement receptor 1 LFA- 1 Complement receptor 3 Complement receptor 4 13 chain of C D l l PI-linked on granulocytes Platelet GPIIa M6 M5 PI-linked on granulocy~s Lacto-N-fucopentoase III PI-linked on monocytes
20 21 Majdic et al., unpublished 22 23 24 25 23 9 13 Majdic et al., unpublished 11 26 27 26
aNot determined.
with degenerative joint disease (DJD) and from healthy control individuals. METHODS Patients and Controls. The following patients entered the study: 15 patients with RA (16), 12 of them with knee joint effusion; 7 patients with ReA, 6 of them with knee joint effusion; 11 patients with DJD; and 16 healthy CO. From these individuals, peripheral blood and, when applicable, joint fluid were subjected to the analyses detailed below. Among the RA patients three had stage I, six had stage II, two had stage M, and four had stage IV according to Steinbroker et al. (17). Twelve patients had active disease (ESR > 40 mrn/hr, >3 joint regions swollen and tender, morning stiffness > 1 hr), while three had inactive disease. Eight patients were positive for rheumatoid factor (Latex test). At the time of investigation, four patients received remission-inducing drugs (one was on gold therapy, one received D-penicillamine, two received chloroquine therapy) and two patients received low-dose corticosteroid therapy (-
Analysis of Function-Associated Receptor Molecules on Peripheral Blood and Synovial Fluid Granulocytes from Patients with Rheumatoid and Reactive Arthritis THOMAS FELZMANN, 1 STEPHEN GADD, 1 OTTO MAJDIC, 1 DIETER MAURER, 1 PETER PETERA, 2 JOSEF SMOLEN, 2 and WALTER KNAPP 1'3
Accepted: April 1, 1991
and cells from reactive arthritis patients, suggesting a similar activation process for granulocytes in these two diseases.
In this study we report the expression pattern of 13 different function-associated surface molecules on synovial fluid and peripheral blood granulocytes from rheumatoid and reactive arthritis patients. We found increased expression of the complement receptors 1 (CD35) and 3 (CD1lb) and of the activation-associated antigens CD67, CD24, and M5 on synovial fluid granulocytes from rheumatoid and/or reactive arthritis patients compared to autologous peripheral blood granulocytes. In addition, synovial fluid granulocytes expressed IgG Fc receptor 1 (CD64) and complement receptor 4 (CDllc), neither of which can be found on peripheral blood granulocytes. Peripheral blood granulocytes from rheumatoid and reactive arthritis patients expressed higher levels of leucocyte function-associated antigen 1 (CDlla) and of the membrane proteins CD31, CD24, M5, and M6 compared to peripheral blood granulocytes from healthy controls and patients with degenerative joint disease. No significant differences in the expression of any of the molecules studied could be observed between cells from rheumatoid
KEY WORDS: Fc receptors; complementreceptors; adhesion molecules; granulocyteactivation;flowcytometry.
INTRODUCTION The expression of neutrophil granulocyte surface molecules can be modulated by a variety of agents including cytokines such as granulocyte macrophage-colony stimulating factor (GM-CSF) 4 (1), interferon-~ (IFN-~) (2, 3), tumor necrosis factor et (TNFa) (4), interleukin-8 (IL-8) (5), and complement split products (4, 6, 7) as well as by FcRs (3) and receptors for microbial wall components such as lipopolysaccharides (8). Recently we and others have shown that in vitro and in vivo stimulation of human granulocytes leads to changes in the surface expression of certain function-associated membrane molecules (3, 6, 9-13). Neutrophils are the predominant cell type in inflammatory joint effusions of patients with rheumatoid arthritis (RA) and reactive arthritis (ReA) (14). However, little is known about possible changes in the surface expression of membrane molecules on neutrophils in RA. To our knowledge, only one report has been published which described the expression of CR3 and platelet glycoprotein IIb/IIIa on synovial fluid granulocytes (SFG) and peripheral blood granulocytes (PBG) in RA (15). Here, we describe the expression pattern of 13 different surface molecules on SFG and PBG from RA and ReA patients in comparison with the expression pattern found on PBG from patients
1Institute of Immunology,Universityof Vienna, Borschkegasse 8a, A-1090, Vienna, Austria. 2Department of Medicine and LudwigBoltzmann Institute for Rheumatology, KH Lainz, Wolkersbergenstr.l, A-1130, Vienna, Austria. 3To whom correspondence should be addressed at Borschkeg. 8a, A-1090, Vienna, Austria. 4Abbreviationsused: rheumatoidarthritis(RA);reactivearthritis (ReA); degenerativejoint disease (DJD); control individuals (CO); peripheral blood granulocytes(PBG);synovialfluidgranulocytes (SFG); monoclonalantibody(mAb); IgG Fc receptor (FcR); complement receptor (CR); leukocyte functionassociated antigen (LFA); granulocyte macrophage-colony stimulating factor (GM-CSF); granulocyte-colonystimulating factor (G-CSF); interferon ~/(IFN-7); tumor necrosis factor et (TNFct); interleukin (IL); leukocyte cytoadhesion molecule (LeuCAM);mean fluorescenceintensity(MFI); phorbol myriastate acetate (PMA); formyl-methionylleucyl phenylalanine (fMLP); intercellular cytoadhesion molecule (ICAM); phosphoinositol(PI); nonsteroidalantiinflammatorydrugs (NSAID). 205
0271-9142/91/0700-0205506.50/0 © 1991 Plenum Publishing Corporation
206
FELZMANN ET AL.
Table I. Monoclonal Antibodies Used in this Study mAb
CD
Molecule
Ref. No.
32.2 IV.3 VIFcR3 To5 MHM24 VIM12 3.9 MHM23 GIOF5 1B5 AAA-6 VIM-5 VIBC5 VIMD5 VIM13
CD64 CDW32 CDt6 CD35 CD 11a CD1 lb CD1 lc CD18 CD67 CD31 ND ~ ND CD24 CD15 CD14
Fc receptor 1 Fc receptor 2 Fc receptor 3 Complement receptor 1 LFA- 1 Complement receptor 3 Complement receptor 4 13 chain of C D l l PI-linked on granulocytes Platelet GPIIa M6 M5 PI-linked on granulocy~s Lacto-N-fucopentoase III PI-linked on monocytes
20 21 Majdic et al., unpublished 22 23 24 25 23 9 13 Majdic et al., unpublished 11 26 27 26
aNot determined.
with degenerative joint disease (DJD) and from healthy control individuals. METHODS Patients and Controls. The following patients entered the study: 15 patients with RA (16), 12 of them with knee joint effusion; 7 patients with ReA, 6 of them with knee joint effusion; 11 patients with DJD; and 16 healthy CO. From these individuals, peripheral blood and, when applicable, joint fluid were subjected to the analyses detailed below. Among the RA patients three had stage I, six had stage II, two had stage M, and four had stage IV according to Steinbroker et al. (17). Twelve patients had active disease (ESR > 40 mrn/hr, >3 joint regions swollen and tender, morning stiffness > 1 hr), while three had inactive disease. Eight patients were positive for rheumatoid factor (Latex test). At the time of investigation, four patients received remission-inducing drugs (one was on gold therapy, one received D-penicillamine, two received chloroquine therapy) and two patients received low-dose corticosteroid therapy (-
