Journal of Histochemistry and Cytochemistry, Vol. 45, 1247-1254, Copyright © 1997 by The Histochemical Society, Inc.


ARTICLE

ICAM-1/LFA-1 Expression in Acute Osteodestructive Joint Lesions in Collagen-induced Arthritis in Rats

Matthias F. Seidela,b, Rolf Kecka, and Hans Vetterb
a Institut für angewandte Zellkommunikationsforschung, Bonn, Germany
b Medizinische Poliklinik der Universität Bonn , Bonn, Germany

Correspondence to: Matthias F. Seidel, Medizinische Poliklinik der Universität Bonn, Wilhelmstr. 35-37, D-53111 Bonn, Germany.


  Summary
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Collagen-induced arthritis in rats is a widely used model of rheumatoid arthritis (RA). However, the joint immunohistopathology is less well characterized. The objective of this study was therefore to analyze whole ankle joints for markers known to mediate inflammatory mechanisms in RA. Indirect immunohistochemistry was performed on undecalcified cryostat sections for intercellular adhesion molecule-1 (ICAM-1, clone 1A 29) and leukocyte function-associated antigen-1 (LFA-1, clone WT.1) expression, for CD4+ lymphocytes (clone W3/25), B-cells (clone HIS 14), and macrophages (clone ED2). Acute, osteodestructive arthritis (n = 8) induced with bovine collagen Type II was verified by clinical and radiological measures. LFA-1 expression was found almost exclusively at sites associated with cartilage erosion or osteodestruction. ICAM-1 was similarly expressed in the vicinity of tissue degradation but also by blood vessels in peripheral areas of joint swelling. CD4+ lymphocytes and macrophages were more ubiquitous. B-cells were infrequent. In control animals (n = 4) ICAM-1 was expressed by synovial blood vessels. Macrophages were identified at the synovial lining. The results suggest that LFA-1 and ICAM-1 mediate important inflammatory events in this model. Similar findings in human RA synovium provide further arguments that collagen-induced arthritis in rats might be regarded as a comparable disease. (J Histochem Cytochem 45:1247-1253, 1997)

Key Words: rheumatoid, collagen, rat, arthritis, osteodestruction, immunohistopathology, ICAM-1, LFA-1, CD4


  Introduction
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Type II collagen-induced arthritis (Trentham et al. 1977 ) shares clincal, radiological, and pathological features (Stuart et al. 1988 ; Brahn 1991 ) with rheumatoid arthritis (RA) in humans. This animal model offers an exellent opportunity for detailed analysis of inflammatory mechanisms. Unlike the case in human patients, the entire collagen-induced arthritis joint pathology can be evaluated, including synovium, cartilage, and bone. However, only a few immunohistochemical studies of collagen-induced arthritis joints have been reported. These studies have focused on the CD4 and plasma cell infiltrates, which are similar to those observed in RA (Klareskog et al. 1983 ; Holmdahl et al. 1985 ). Analysis of collagen-induced arthritis joints for other tissue markers typically found in RA would therefore provide a more complete view of this experimental model and perhaps give new insights into the human disease.

Human RA synovial tissue typically demonstrates putatively autoreactive lymphocytes and macrophages infiltrating and destroying the joint tissues; B-cells are less commonly encountered (Rooney et al. 1989 ). Robust synovial fibroblast proliferation is another sequela of joint inflammation. Adhesion molecules appear partially responsible for initiating and perpetuating arthritis (reviewed by Cronstein 1994 ). These ligands stimulate transendothelial lymphocyte migration into areas of joint inflammation (El-Gabalawy et al. 1994 ). The intercellular adhesion molecule-1 (CD54; ICAM-1) and its receptor, the ß2-integrin leukocyte function-associated antigen-1 (CD11a/CD18; LFA-1) are of particular interest in RA because they are upregulated in the inflamed synovium. Furthermore, treatment of RA patients with monoclonal antibodies directed against ICAM-1 leads to a reduction of disease activity (Kavanaugh et al. 1994 ). In RA synovium, ICAM-1 is expressed by many cell types, including the vascular endothelium, lymphocytes, monocytes, and fibroblasts. ICAM-1 expressed on activated endothelium partially mediates terminal adhesion of activated mononuclear cells that bear LFA-1. This cellular receptor is found on lymphocytes and monocytes in the peripheral blood and in RA synovium (Cush and Lipsky 1988 ).

The objective of this study was to determine if ICAM-1 and LFA-1 expression, along with other cell markers typical of RA, occurs in the collagen-induced arthritis model. We studied the early chronic inflammatory joint lesion. Indirect immunohistology was performed on undecalcified, native cryostat sections of ankle joints using a modified protocol of a previously described tissue sectioning technique (Meacock et al. 1992 ).


  Materials and Methods
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Reagents
All reagents were purchased from Sigma (St Louis, MO) except where otherwise indicated.

Induction of Arthritis
The animal experiments were conducted with permission of the Regierungspräsidium Köln (German animal welfare act AZ 23.203.2 BN 40, 10/94). Specific pathogen-free-reared 9-10-week-old inbred male Dark Agouti rats were obtained from Harlan-Winkelman (Borchen, Germany). The rats were kept in cages with filtertops (Techniplast, Buguggiate, Italy; Becker, Castrop-Rauxel, Germany) and separate from other laboratory animals. Pathogen-free processed laboratory chow (Altromin; Lage, Germany) and water were given ad libitum. Bovine collagen Type II was obtained from Dr. Mary Griffiths and was prepared by pepsin digestion as previously described (Griffiths et al. 1992 ). The lyophilized collagen preparation was dissolved in 0.1 M acetic acid by gentle agitation overnight at 4C. The solution was then emulsified with incomplete Freund's adjuvant. In four separate experiments, eight rats were immunized with several intradermal injections on the back using 2 µg collagen/g body weight (195-210 g). Booster immunizations were given at Day 7 with 100 µg collagen. In each experiment, one rat was treated with the same volume of 0.1 M acetic acid alone.

Monitoring and Development of Arthritis
The animals were weighed two times per week and were monitored for the onset of clinical symptoms of arthritis. To quantify the intensity of the arthritis, a clinical score for each animal was calculated 6 days after the onset of arthritis. This score was composed of the sum of involvement in each limb and an element for weight loss. The scale for each limb: 0 (no swelling), 1 (faint erythema), 2 (massive erythema/faint edema), 3 (massive erythema/extensive edema). Weight loss was scored as 0 (normal weight gain), 1 (no weight gain), 2 (5% weight loss), and 3 (5-10% weight loss).

Radiography
Five to six days after the onset of arthritis, the animals were anesthetized with ether. Lateral radiographs of both hindpaws were obtained using a Phillips Rotaldex apparatus with 43 kV, 75 mAs on a standard Agfa Curix HT 1000 G film.

Processing for Whole-joint Frozen Sections of Undecalcified Tissue
After radiography, the animals were sacrificed under ether anesthesia by cervical dislocation. From some animals blood was obtained by intracardial puncture and serum aliquots were stored at -80C. For tissue embedding, the skin was removed from the hindpaws and the joints immersed in 8% gelatin. Because rapid freezing causes disruption of tissue integrity, the specimens were frozen slowly in liquid nitrogen and then stored at -80C. As control tissue for immunohistochemical staining, the spleens of some animals were removed and processed similarly. For in toto joint analysis, cryostat sections were obtained by a modification of a previously described method for unfixed joints of small animals (Meacock et al. 1992 ). In our study, tissue fixation in 95% ethanol, 100% methanol, or even acetone was accomplished by use of a novel transparent polypropylene tape (Beiersdorf; Hamburg, Germany). The frozen block was mounted laterally on the motor-driven chuck with methyl cellulose at a cabinet temperature of -30C in a Reichert-Jung Frigocut 2800. A strip of the polypropylene tape was applied to the specimen surface for each 7-µm section (using a D-knife). To prevent tissue distortion during fixation, the tape was wrapped around a microscopy slide and left to dry for 2 hr at room temperature (RT). The specimens were stored at -80C until further use. Spleen cryostat sections of diseased animals were obtained by standard methods. Some sections were stained with hematoxylin and eosin.

Immunohistochemistry
Immunohistochemical staining was performed with mouse monoclonal antibodies (prepared against rat tissue) as specified in Table 1 and using an avidin-biotin-peroxidase staining kit (Vectastain Elite ABC Kit; Vector, Burlingame, CA). All incubations were carried out at 37C in a humidified chamber. The washes were done in PBS at RT for 5 min each. Antibodies were titered using spleen sections from diseased animals (Table 1). The tissue was fixed in acetone for 1 min or in 95% ethanol for 10 min at RT (see Table 1). The sections were rinsed and then blocked in 5% horse serum for 10 min. The primary antibodies (Table 1) were diluted in a mixture of PBS, 0.1% bovine serum albumin (BSA, globin-free), and 0.02% sodium azide, and were applied for 15 min. Negative controls consisted of PBS with 1% normal mouse serum or PBS alone. To efficiently quench endogenous peroxidases, the samples were then incubated in methanol containing 0.3% H2O2 for 20 min at RT (Streefkerk 1972 ). The sections were washed twice. To minimize potential crossreactions with rat tissue, the biotinylated horse anti-mouse antiserum was applied with 5% serum derived from arthritic rats; this incubation was performed for 15 min. After one wash, the avidin-peroxidase reagent and then 3,3'-diaminobenzidine tetrahydrochloride (DAB) as peroxidase substrate (Vector) were incubated for 10 min each with an intermediate wash. After color development the sections were counterstained with hematoxylin and mounted in glycerol medium. Proximal ankle joints of diseased and control animals were processed similar to spleen sections. The tape was trimmed to the size of the specimen and then fixed to the microscopy slide with an adhesive component (UHU GmbH; Brühl, Germany), which was removed before mounting the tissue samples in glycerol medium. The stained tissue sections were photographed with a Zeiss Universal microscope.


 
View this table:
[in this window]
[in a new window]
 
Table 1. Mouse monoclonal antibodies directed against rat epitopes

Quantification and Statistical Analysis of Immunostained Cells in Joints
One whole ankle joint from each animal was analyzed. Staining was scored for three locations in the inflamed joint. Areas in proximity to the joint cartilage and bone destruction (central osteodestructive lesions) were scored separately from peripheral areas of joint edema (peripheral synovial pannus areas) and from bone marrow. In the normal joint, only synovium and bone marrow were distinguished. Because individual cells could not always be delineated as positively or negatively stained, a semiquantitative score (0-4) was assigned for each joint and antigen according to percentages of stained cells: <0.5%, 0.5-5%, 5-10%, 10-25%, >25%, respectively. The mean scores derived from all joints evaluated were used to compute an average percent of positive cells in these areas. The data were analyzed with Student's t-test. Differences between the individual anatomic compartments analyzed were considered significant at p<0.05.


  Results
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Clinical, Radiographic, and Histological Characterization Confirmed the Early Nature of Arthritic Joints
All animals immunized with collagen Type II developed severe arthritis. The first symptoms of the disease were observed at the distal hindpaws between Days 14 and 17 after immunization. The interdigital erythema of distal joints spread to more proximal areas and was followed by increased swelling. In one animal, significant inflammation was observed only at the left hindpaw. By Day 20, massive erythema and edema indicated a profound inflammatory process. The manifestation of the disease at the forepaws was less constant. Other joints did not appear to be affected, as judged by clinical examination. Clinical scores ranged from 13 to 19 (16.5 ± 1.96). Compared to control animals, lateral radiographs (not shown) indicated intensive swelling of the entire surrounding joint tissue. In 15 inflamed ankle joints, bony destruction was observed radiologically (14/15 at the talocalcaneal articulation). These findings corresponded to previously described radiological changes in collagen-induced arthritis in rats (Ridge et al. 1988 ). In standard H&E stains (not shown), peripheral synovial pannus areas were dominated by a fibroblastoid cell type and edema. Cartilage erosion and mononuclear cell infiltrations were found in the central osteodestructive lesion. The joint space was mostly occupied by tissue of the proliferative inflammatory synovitis.

Prevalence and Distribution of Immunological Phenotypes in Control Animals and in Collagen-induced Arthritis
Representative peripheral synovial pannus areas and central osteodestructive lesion areas of each inflamed joint were scored for the prevalence of each phenotype. We also scored normal synovium and both normal and immunized bone marrow for comparative purposes (the mean percentages for eight immunized and four control joints are shown in Figure 1). Endogenous peroxidase was not detected in control sections treated with PBS or normal mouse serum. Healthy synovium contained virtually no B-cells, whereas this cell type was readily identified in the bone marrow compartment. Vascular endothelium strongly expressed ICAM-1 (Figure 2A). LFA-1 was absent from normal synovium; approximately 7% of the bone marrow cell population was stained. No definitive CD4 staining was present in the normal synovium (Figure 3A), with subtle infiltration of the bone marrow. In contrast, many synovial cells expressed the macrophage marker and were concentrated at the synovial lining (Figure 3C), but were not significantly different from bone marrow densities. In collagen-induced arthritic joints, CD4+ cells were the predominant cell population in peripheral synovial pannus areas, mostly scattered as individual cells and infrequently as perivascular cell infiltrations. CD4+ lymphocytes were also widely distributed in central osteodestructive lesions (Figure 3B) but were less closely associated with bony destruction. Bone marrow infiltration was not significantly different from synovial infiltration, suggesting that the CD4 lymphocytes can be regarded as a general and less specific cellular response. Staining with the ED2 antibody revealed a ubiquitous distribution of macrophages in the peripheral synovial pannus area and central osteodestructive lesion (Figure 3D). Macrophages were also detected in close contact with the joint cartilage erosion. Staining for B-cells with the HIS 14 monoclonal antibody was rare in peripheral synovial pannus areas; groups of 50-100 cells were found in the vicinity of the central osteodestructive lesions (Figure 3E). Bone marrow staining demonstrated expected increases in CD4 and ED2 (macrophage)-positive cells.



View larger version (33K):
[in this window]
[in a new window]
 
Figure 1. Distribution and prevalence of immunological phenotypes in control (solvent-treated animals; n = 4) and inflamed ankle joints (collagen-immunized animals; n = 8). The mean percentages of immune phenotypes in control synovium (n = 4; SYN-Con) and bone marrow (n = 4; BM-Con) or inflamed peripheral synovial pannus areas (n = 8; PPA-CIA), central osteodestructive lesions (n = 8; COL-CIA), and bone marrow (n = 8; BM-CIA) were determined as described in Materials and Methods. Error bars show standard deviations of the scores. Nonsignificant t-test (n.s.). See text for discussion and findings.



View larger version (87K):
[in this window]
[in a new window]
 
Figure 2. Immunohistochemical staining for ICAM-1 and LFA-1 expression associated with osteodestructive lesions. In control synovium (A), ICAM-1 is expressed by synovial blood vessels (arrows). In collagen-induced arthritis (B), the same vascular pattern of ICAM-1 expression is rarely found in proximity to cartilage erosion (c). In the peripheral synovial pannus area (C), small blood vessels stain for ICAM-1 (arrows). The LFA-1 phenotype is almost exclusively associated with bone infiltration (i) in collagen-induced arthritis (D). a, articular surface; s, synovium; t, talus. Bar = 100 µm.

Figure 3. Immunohistochemical staining of other markers characteristic of rheumatoid arthritis also demonstrate inflammatory joint features. (A) Weak CD4 staining in normal synovium due to crossreaction with macrophage epitopes. (B) In collagen-induced arthritis, profound infiltration of CD4+ lymphocytes with cartilage (c) erosion. (C) Characteristic macrophage staining (arrows) of synovial cells with the ED2 antibody at the lining of normal synovium. (D) In collagen-induced arthritis, macrophage infiltration and cartilage erosion (c). Compared to the abundant macrophage presence the B-cell phenotype expression in collagen-induced arthritis (E) is quantitatively less important at the point of cartilage erosion. a, normal articular surface; s, synovium; t, talus. Bar = 100 µm.

Expression of ICAM-1 and LFA-1 Staining Correlates with Intensity of Arthritic Inflammation
In collagen-induced arthritic joints, ICAM-1 stained small vascular structures in peripheral synovial pannus areas (Figure 2C), consistent with ongoing neovascularization. The same vascular staining patterns were rarely observed in central osteodestructive lesions at the cartilage surface (Figure 2B). Instead, ICAM-1 staining there was confined mostly to cells surrounding the eroded cartilage surface. The infiltration close to these bony lesions was not significantly different from the bone marrow, suggesting a more unspecific cellular response for this marker. LFA-1 staining was not observed in the peripheral synovial pannus area but was found at the cartilage surface, frequently associated with bony destruction (Figure 2D). Not all areas of osteodestruction manifested LFA-1 staining, presumably because the small groups of LFA-1+ cell infiltration were not sectioned on every plane of the slide. In two animals, intensive LFA-1 staining was observed in circular lymphoid aggregates near the joint but not in direct contact with the cartilage. Compared to control animals, the LFA-1 phenotype was greatly increased in the bone marrow in collagen-induced arthritic animals (Figure 1).


  Discussion
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Although collagen-induced arthritis in rodents is considered as perhaps the most important model of RA in humans, the joint immunopathology is not well known. Joint inflammatory mechanisms in collagen-induced arthritis may be distinct from human pathology because the autoantigen in RA is still speculative and is probably not related to collagen Type II. In this study, we therefore aimed to characterize inflammatory infiltrates in separate joint compartments for markers typically found in RA. Acute experimental arthritis was verified by clinical and radiological assessment. In toto joint analysis was accomplished by our special cryostat sectioning protocol. The novel polypropylene tape enabled us to use various tissue fixatives. Furthermore, with this procedure we did not have to consider immunohistochemical requirements for conventional bone processing, such as demineralization and paraffin-embedding. Rapid processing time (less than 6 hr for sectioning and immunostaining) and convenience can be considered as additional and substantial advantages.

Our analysis was semiquantitative because not all antibodies yielded the same quality of immunostaining. To ensure comparison to other studies, we therefore estimated the numbers of stained cells. The absence of HIS-14 (for B-cells) or LFA-1 staining in the synovium of control animals indicated a high degree of tissue specificity (although these markers were detected in the bone marrow of the same sections). Furthermore, CD4 staining in the central osteodestructive lesion but also in the peripheral synovial pannus area corresponded to previously published results in collagen-induced arthritis (Holmdahl et al. 1995 ). Weak staining with the CD4 marker in normal synovium might have been due to the presence of macrophages that are also in part detected by the antibody (Barclay 1981 ). The significance of sparse B-cell infiltration remains less well defined. Apart from production of immunoglobulins and rheumatoid factors by plasma cells, the B-cell population might be involved in antigen presentation to T-cells that trigger tissue degradation (Holmdahl et al. 1995 ). ED2+ macrophages were distributed in peripheral and central areas of joint inflammation also in direct contact with the remaining cartilage, in good agreement with their known function of tissue degradation (Sack et al. 1994 ).

LFA-1+ cells were the most sensitive indicators of osteodestructive lesions in our study. However, not all areas of osteodestruction were always associated with this phenotype, possibly owing to the small size of LFA-1-associated inflammatory infiltrates. LFA-1 expression may also change over time within the same articular surface. The important role of this molecule was demonstrated when an anti-LFA-1 antibody treatment suppressed the development of collagen-induced arthritis in DBA/1 mice (Kakimoto et al. 1992 ). Other adhesion factors may also be involved in arthritis lymphocyte migration. In the adjuvant arthritis model, complete inhibition of monocyte migration to inflamed joints was demonstrated only with a combined monoclonal antibody treatment directed against several integrin family members, i. e., LFA-1, Mac-1, and very-late-activation antigen 4 (Issekutz and Issekutz 1995 ).

ICAM-1 was found to be expressed mostly by small blood vessels in the peripheral synovial pannus area. In central osteodestructive lesions, ICAM-1 was predominantly expressed by a cell population other than the endothelium, suggesting that this adhesion molecule may not only mediate lymphocyte trafficking. Stimulated human synovial fibroblasts co-expressing HLA class II antigens and ICAM-1 have been reported to serve as potent antigen-presenting cells for staphylococcal enterotoxin B to T-lymphocytes (Origuchi et al. 1995 ). Treatment of RA patients with anti-ICAM-1 regimens may therefore perturb perpetuating T-cell activation close to the osteodestructive process. Similar T-cell activation mechanisms are likely to exist also in the collagen-induced arthritis model, although the exact identity of ICAM-1+ and LFA-1+ cells could not be determined by morphology alone. Direct correlation with mononuclear staining was technically difficult and was not always possible. The development of double-label immunohistochemistry for our tissue processing procedure will help to settle this point.

In summary, our study shows that LFA-1 and ICAM-1 expressions appear to be closely associated with osteodestruction and may therefore play an important role in tissue degradation and the chronicity of the disease. In addition, our results support further similarity to human rheumatoid arthritis (El-Gabalawy et al. 1994 ; Ishikawa et al. 1994 ; Szekanecz et al. 1994 ). Future detailed analyses and experimental manipulation of the osteodestructive mechanism in collagen-induced arthritis might therefore also be applicable to the pathogenesis of the human disease.


  Acknowledgments

We thank Dr Samuel Hunter and Dr Mary Griffiths for critically reviewing the manuscript, Dr Jörg Kriegsmann and Dr Thomas Muche for technical advice, and Ms D. Menrath for excellent assistance with taking the radiographs.

Received for publication February 24, 1997; accepted March 7, 1997.


  Literature Cited
Top
Summary
Introduction
Materials and Methods
Results
Discussion
Literature Cited

Barclay AN (1981) The localization of populations of lymphocytes defined by monoclonal antibodies in rat lymphoid tissues. Immunology 42:593-600 [Medline]

Beelen RHJ, Eestermans IL, Döpp EA, Dijkstra CD (1987) Monoclonal antibodies ED1, ED2, and ED3 against rat macrophages: expression of recognized antigens in different stages of differentiation. Transplant Proc 19:3166-3170

Brahn E (1991) Animal models of rheumatoid arthritis. Clues to etiology and treatment. Clin Orthop 265:42-53 [Medline]

Cronstein BN (1994) Adhesion molecules in the pathogenesis of rheumatoid arthritis. Curr Opin Rheum 6:300-304 [Medline]

Cush JJ, Lipsky PE (1988) Phenotypic analysis of synovial tissue and peripheral blood lymphocytes isolated from patients with rheumatoid arthritis. Arthritis Rheum 31:1230-1238 [Medline]

El-Gabalawy H, Gallatin M, Vazeux R, Peterman G, Wilkins J (1994) Expression of ICAM-R (ICAM-3), a novel counter-receptor for LFA-1, in rheumatoid and nonrheumatoid synovium. Comparison with other adhesion molecules. Arthritis Rheum 37:846-854 [Medline]

Griffiths MM, Cremer MA, Harper DS, McCall S, Cannon GW (1992) Immunogenetics of collagen-induced arthritis in rats: both MHC and non-MHC gene products determine the epitope specificity of immune response to bovine and chick type II collagens. J Immunol 149:309-316 [Abstract/Free Full Text]

Holmdahl R, Rubin K, Klareskog L, Dencker L, Gustafson G, Larsson E (1985) Appearance of different lymphoid cells in synovial tissue and in peripheral blood during the course of collagen II-induced arthritis in rats. Scand J Immunol 21:197-204 [Medline]

Holmdahl R, Vingsbo C, Mo JA, Michaëlsson E, Malmström V, Jansson L, Brunsberg U (1995) Chronicity of tissue-specific experimental autoimmune disease: a role for B cells? Immunol Rev 144:109-135[Medline]

Ishikawa H, Hirata S, Nishibayashi Y, Imura S, Kubo H, Ohno O (1994) The role of adhesion molecules in synovial pannus formation in rheumatoid arthritis. Clin Orthop 300:297-303 [Medline]

Issekutz AC, Issekutz TB (1995) Monocyte migration to arthritis in the rat utilizes both CD11/CD18 and very late activation antigen 4 integrin mechanisms. J Exp Med 181:1197-1203 [Abstract]

Jefferies WA, Green JR, Williams AF (1985) Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages. J Exp Med 162:117-127 [Abstract]

Kakimoto K, Nakamura T, Ishii K, Takashi T, Iigou H, Yagita H, Okumura K, Onoue K (1992) The effect of anti-adhesion molecule antibody on the development of collagen-induced arthritis. Cell Immunol 142:326-337 [Medline]

Kavanaugh AF, Davis LS, Nichols LA, Norris SH, Rothlein R, Scharschmidt LA, Lipsky PE (1994) Treatment of refractory rheumatoid arthritis with a monoclonal antibody to intercellular adhesion molecule 1. Arthritis Rheum 37:992-999 [Medline]

Klareskog L, Holmdahl R, Larsson E, Wigzell H (1983) Role of T lymphocytes in collagen II induced arthritis in rats. Clin Exp Immunol 51:117-125 [Medline]

Kroese FGM, Wubbena AS, Opstelten D, Deenen GJ, Schwander EH, De Leij L, Vos H, Poppema S, Volberda J, Nieuwenhuis P (1987) B lymphocyte differentiation in the rat: production and characterization of monoclonal antibodies to B lineage-associated antigens. Eur J Immunol 17:921-928 [Medline]

Meacock SCR, Brandon DR, Brown CP, Swann BP (1992) A novel technique for immunohistoperoxidase staining of unfixed whole joints of small animals. Histochem J 24:115-119 [Medline]

Origuchi T, Eguchi K, Kawabe Y, Mizokami A, Ida H, Nagataki S (1995) Synovial cells are potent antigen-presenting cells for superantigen, staphylococcal enterotoxin B (SEB). Clin Exp Immunol 99:345-351 [Medline]

Ridge SC, Oronsky AL, Kerwar SS (1988) Type II collagen-induced arthritis in rat. Methods Enzymol 162:355-360 [Medline]

Rooney M, Whelan A, Feighery C, Bresnihan B (1989) The immunohistologic features of synovitis, disease activity and in vitro IgM rheumatoid factor synthesis by blood mononuclear cells in rheumatoid arthritis. J Rheumatol 16:459-467 [Medline]

Sack U, Stiehl P, Geiler G (1994) Distribution of macrophages in rheumatoid synovial membrane and its association with basic activity. Rheumatol Int 13:181-186 [Medline]

Streefkerk JG (1972) Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxide following methanol. J Histochem Cytochem 20:829-831 [Medline]

Stuart JM, Watson WC, Kang AH (1988) Collagen autoimmunity and arthritis. FASEB J 22:2950-2956

Szekanecz Z, Haines GK, Lin TR, Harlow LA, Goerdt S, Rayan G, Koch AE (1994) Differential distribution of intercellular adhesion molecules (ICAM-1, ICAM-2, and ICAM-3) and the MS-1 antigen in normal and diseased human synovia. Their possible pathogenetic and clinical significance in rheumatoid arthritis. Arthritis Rheum 37:221-230 [Medline]

Tamatani T, Kotani M, Miyasaka M (1991) Characterization of the rat leukocyte integrin, CD11/CD18, by the use of LFA-1 subunit-specific monoclonal antibodies. Eur J Immunol 21:627-633 [Medline]

Tamatani T, Miyasaka M (1990) Identification of monoclonal antibodies reactive with the rat homolog of ICAM-1, and evidence for a differential involvement of ICAM-1 in the adherence of resting versus activated lymphocytes to high endothelial cells. Int Immunol 2:165-171 [Medline]

Trentham DE, Townes AS, Kang AH (1977) Autoimmunity to type II collagen: an experimental model of arthritis. J Exp Med 146:857-868 [Abstract]





This Article
Abstract
Full Text (PDF)
Alert me when this article is cited
Alert me if a correction is posted
Citation Map
Services
Similar articles in this journal
Similar articles in PubMed
Alert me to new issues of the journal
Download to citation manager
Google Scholar
Articles by Seidel, M. F.
Articles by Vetter, H.
Articles citing this Article
PubMed
PubMed Citation
Articles by Seidel, M. F.
Articles by Vetter, H.


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]