Collagen-induced arthritis (CIA) is an experimental animal model of human rheumatoid arthritis being characterized by synovitis and progressive destruction of cartilage and bone. CIA is induced by injection of heterologous or homologous collagen type II in a susceptible murine strain. DBA/1J mice deficient of complement factors C3 (C3−/−) and factor B (FB−/−) were generated to elucidate the role of the complement system in CIA. When immunized with bovine collagen type II emulsified in CFA, control mice developed severe arthritis and high CII-specific IgG Ab titers. In contrast, the C3−/− and FB−/− were highly resistant to CIA and displayed decreased CII-specific IgG Ab response. A repeated bovine collagen type II exposure 3 wk after the initial immunization led to an increase in the Ab response in all mice and triggered arthritis also in the complement-deficient mice. Although the arthritic score of the C3−/− mice was low, the arthritis in FB−/− mice ranked intermediate with regard to C3−/− and control mice. We conclude that complement activation by both the classical and the alternative pathway plays a deleterious role in CIA.

Rheumatoid arthritis (RA)3 is a common chronic inflammatory disease affecting joints and giving rise to cartilage and bone destruction. Collagen-induced arthritis (CIA) is an experimental animal model for human RA being characterized by inflammation of multiple joints, accompanied by synovial hyperplasia, infiltration of mononuclear cells, pannus formation, and destruction of cartilage and subchondral bone. The disease is elicited by immunization of genetically susceptible mice with type II collagen (CII) in CFA (1). CIA pathology is dependent on both humoral and cell-mediated immunity (2). Indeed, the arthritic severity in murine arthritis is positively correlated with the IgG autoantibody response to CII (3). In addition, passive transfer of CII-specific Abs elicits synovitis in naive recipients (4, 5, 6) and mice lacking B cells are resistant to induction of CIA (7).

Complement activation has previously been implicated in the pathogenesis of RA (8, 9). Complement components and activation products have been demonstrated both in the synovial fluid and in the synovial membrane of the affected joints (10, 11). Animals depleted of complement by cobra venom factor were found to be refractory to arthritis until their complement levels were recovered (12). However, as this depletion is achieved through an enormous activation of the complement system, the effects of which are both unknown and difficult to control, this approach has obvious drawbacks. Systemic administration of anti-C5 Ab prevented the onset of CIA and ameliorated established disease in this animal model (13). C5-deficient mice were partly resistant to CIA despite normal cellular and Ab responses to immunization with CII and substantial intra-articular deposition of complement-fixing Abs (14). These experiments could not elucidate the role of the earlier components of the complement activation cascade. Soluble complement receptor 1, a potent inhibitor of the classical and alternative complement pathways, prevented the progression of arthritis in both rats (15) and mice (16).

In this study, we have investigated the role of some of the early components of the complement cascade, C3 and factor B (FB), respectively, in the pathogenesis of CIA. The mice deficient for C3 or FB, previously generated in our laboratory (17, 18), were immunized with bovine CII and arthritis was evaluated both clinically and histopathologically. We show that the Ab response in C3−/− and FB−/− mice is impaired but can be boosted by repeated immunization. Regardless of the specific anti-CII Ab levels, the complement-deficient mice display reduction or almost complete absence of clinically or histologically verifiable arthritis. These results show that the classical pathway and possibly also the alternative pathway are involved in the pathogenesis of CIA.

Mice carrying the C3 (17) or FB null allele (18), respectively, were backcrossed to the CIA-susceptible DBA/1J genetic background (Bomholtgaard, Ry, Denmark) for six generations. Animals heterozygous for the respective null mutation were intercrossed to generate homozygous C3-deficient (C3−/−), FB-deficient (FB−/−), and control lines. Mice were maintained at the transgenic core facility at Göteborg University under specific pathogen-free conditions. Routine screening of the mice for murine pathogens according to the recommendations of the Federation of European Laboratory Animal Science Associations was negative.

Bovine type II collagen (BCII) was prepared from nasal cartilage by pepsin digestion and subsequent purification as described previously (19). BCII was solubilized to a concentration of 2 mg/ml in 0.01 M acetic acid at 4°C with constant mixing overnight. For induction of CIA, BCII was emulsified with an equal volume (1:1) of CFA (Difco, Detroit, MI).

Male C3−/−, FB−/−, and control mice at the age of 8–12 wk were injected intradermally at the base of the tail with 50 μl of the BCII/CFA emulsion under light isofluorane anesthesia. In the booster group, the injection was repeated after 21 days.

Arthritis development was assessed by inspection three times a week by a blinded examiner. Clinical severity of arthritis was quantified according to a graded scale from 0 to 3 as follows: 0, normal; 1, detectable swelling in one joint; 2, swelling in more than one but not in all joints; and 3, severe swelling of the entire paw and/or ankylosis. Each paw was graded, and each mouse could achieve a maximum score of 12. A mean arthritic score value was calculated.

Mice were bled from the tails at different time points after immunization, and individual sera were analyzed for CII-specific IgG, IgG1, and IgG2a Abs by ELISA. Microtiter plates (Maxisorp; Nunc, Roskilde, Denmark) were coated overnight at 4°C with 10 μg of native BCII in PBS. Plates were washed with PBS containing 0.05% Tween 20 (PBS/Tween), and serum samples were added in serial dilution with PBS/Tween and incubated for 2 h at room temperature. The plates were then washed and incubated for 2 h at room temperature with 50 μl of sheep anti-mouse IgG conjugated to HRP peroxidase (DAKO, Glostrup, Denmark) diluted 1/1000, anti-mouse IgG1, or anti-mouse IgG2a (Southern Biotechnology Associates, Birmingham, AL) diluted 1/2000 in PBS/Tween. After additional washings, the enzyme reaction was started by the addition of chromogen (20 mg of 1,2-phenylenediamine dihydrochloride (Sigma-Aldrich, St. Louis, MO)) and 10 μl of H2O2 in 75 ml of 0.1 M citrate/phosphate buffer, pH 5). The reaction was stopped by addition of 1 M H2SO4 after 4 min. The staining was quantified spectrophotometrically at 490 nm. A polyclonal anti-BCII standard with known Ab concentration was included on every microtiter plate to allow calculation of the Ab content using Softmax software (Molecular Devices, Menlo Park, CA). The standard was purified by affinity chromatography from pooled sera obtained from BCII-hyperimmunized mice.

All mice were killed on day 72, hind paws were fixed in 4% phosphate-buffered formaldehyde solution, and decalcified in Parengy’s decalcification solution overnight. The tissue was then processed and embedded in paraffin. Tissue sections (4 μm) were stained with H&E using standard methodology. The joints were studied by two blinded examiners (M.A.H. and I.-M.J.). Synovial hypertrophy was defined as a synovial membrane thickness of more than two cell layers (20). Pannus formation as well as cartilage and bone erosion were registered in the knee, ankle, talar, and toe joints. Each parameter was given a score of 0–3.

DNA samples were collected from tail biopsies and purified by phenol-chisam extraction. The primers for H2-specific amplification were derived from Eb gene sequences (21): the forward primer 5′-CGACTGTAGAACCTTAGCCTG-3′ and the reverse primer 5′-GTGGACACAATTCCTGTTTT-3′. The PCR amplification was conducted using 50–100 ng of purified DNA in the presence of 1.9 mM MgCl2 0.1% Tween 20, 10 mM of each dNTP, 12.5 pmol/μl each primer, 2 U Taq DNA polymerase including 10× PCR Buffer (Lifetech/InVitrogen, Carlsbad, CA), in a final volume of 40 μl. The PCR scheme was as follows: initial denaturation step at 94°C for 3 min, 35 cycles including a denaturation step at 93°C for 1 min, annealing at 58°C for 2 min, extension at 72°C for 2 min followed by a final extension at 72°C for 4 min in a thermal cycler (PerkinElmer, Norwalk, CT).

Following amplification, the DNA was precipitated with 99.5% ethanol and resuspended in 25 μl of water. Twenty microliters was treated with 2 U of Fnu4HI (New England Biolabs, Beverly, MA) overnight at 37°C in the buffer supplied by the manufacturer. The DNA were electrophoresed through a 2% agarose gel. The H2b allele gave fragments of 85 and 208 bp, respectively, while the H2q allele yielded two fragments of 186 and 115 bp, respectively.

Data were analyzed using the SPSS 10.0 and Statview 5.0 software. Repeated measure ANOVA was used for statistical analysis of the clinical arthritic score and anti-BCII Ab response. For histological scores, the Mann-Whitney U two-sample rank test was used. Values of p < 0.05 were considered to be significant.

Single immunization of C3−/− mice with BCII did not trigger development of arthritis. Indeed, their arthritic score reached a maximum of 0.25 ± 0.13 (mean ± SEM) on day 39 and declined thereafter. In contrast, control mice developed progressive arthritis with the arthritic score of 10.0 ± 0.71 (mean ± SEM) by the end of the experiment. (Fig. 1,a). The C3−/− mice reached 25% incidence of arthritis on day 42 and 12.5% at the termination of the experiment (Fig. 2 a). Both the control and FB−/− animals developed CIA with 100% incidence within 28 days after the BCII injection. However, at termination of the experiment, the FB−/− mice did not show any clinical signs of arthritis.

FIGURE 1.

Clinical severity of arthritis. Clinical arthritic score after single dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice, (mean ± SEM; n, number of mice). By repeated measure ANOVA, there was a significant effect of genotype, the p values refer to the difference between the control group vs C3−/− and FB−/− group, respectively. Difference between C3−/− and FB−/− groups in b, p < 0.01.

FIGURE 1.

Clinical severity of arthritis. Clinical arthritic score after single dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice, (mean ± SEM; n, number of mice). By repeated measure ANOVA, there was a significant effect of genotype, the p values refer to the difference between the control group vs C3−/− and FB−/− group, respectively. Difference between C3−/− and FB−/− groups in b, p < 0.01.

Close modal
FIGURE 2.

Incidence of arthritis. Incidence of CIA after single-dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice.

FIGURE 2.

Incidence of arthritis. Incidence of CIA after single-dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice.

Close modal

Booster immunization with BCII elicited rapid development of arthritis both in the control and in the FB−/− mice. However, despite the booster immunization, the C3−/− animals displayed only mild arthritis with a delayed onset (Fig. 1,b). At the end of the experiment, the arthritic score was 11.1 ± 0.40 in the control group vs 7.17 ± 0.98 in the FB−/− group and 1.63 ± 1.49 in the C3−/− group (mean ± SEM). Only one animal of the nine mice in the C3−/− group developed arthritis of the same severity as the control mice (arthritic score of 12 at the end of the experiment). The incidence for both the control and FB−/− mice was 100% already 23 days after immunization and remained unchanged for the rest of the experiment. The C3−/− mice showed a peak incidence of 88% at day 46 but by the end of the experiment the incidence was reduced to 25% (Fig. 2 b).

The control mice injected with a single dose of BCII exhibited substantial pannus formation and thickening of the synovium. Booster immunization led to severe erosion of both cartilage and bone and massive infiltration of the joints with inflammatory cells. The FB−/− mice showed no pathological signs in the joint after single-dose immunization, whereas the booster immunization of FB−/− mice triggered a slight thickening of the synovium. Irrespective of the immunization protocol, the C3−/− mice did not show any signs of inflammation or destruction of the joints (Figs. 3 and 4 and Table I). The affected joints of the single C3−/− mouse that displayed severe arthritis when evaluated by clinical scoring exhibited morphological changes that were similar to those of the control mice.

FIGURE 3.

Histopathology of the knee joint. H&E staining of the knee joint of control mice (a and b), FB−/− mice (c and d), and C3−/− mice (e and f) after single or repeated immunization with BCII. Control mice show erosion of cartilage and proliferation of synovial cells (a), which is accentuated in the mice immunized twice, with severe erosion of cartilage and bone, massive infiltration of inflammatory cells, and severe synovial cell hyperplasia (b). Mild synovial hypertrophy is seen in FB−/− mice only after repeated immunization (d). Normal histopathology in the knee joint (c, e, and f). Original magnification, ×100. sh, Synovial hyperplasia; pf, pannus formation; ce, cartilage erosion; be, bone erosion; ic, inflammatory cells. The arrows in b depict a severe lesion on both cartilage and bone.

FIGURE 3.

Histopathology of the knee joint. H&E staining of the knee joint of control mice (a and b), FB−/− mice (c and d), and C3−/− mice (e and f) after single or repeated immunization with BCII. Control mice show erosion of cartilage and proliferation of synovial cells (a), which is accentuated in the mice immunized twice, with severe erosion of cartilage and bone, massive infiltration of inflammatory cells, and severe synovial cell hyperplasia (b). Mild synovial hypertrophy is seen in FB−/− mice only after repeated immunization (d). Normal histopathology in the knee joint (c, e, and f). Original magnification, ×100. sh, Synovial hyperplasia; pf, pannus formation; ce, cartilage erosion; be, bone erosion; ic, inflammatory cells. The arrows in b depict a severe lesion on both cartilage and bone.

Close modal
FIGURE 4.

Histopathological severity of arthritis. Histopathological arthritic score was assessed as synovial hypertrophy, cartilage destruction, bone destruction, and pannus formation; mean ± SEM. The statistical significance of the effect of genotype on the difference between the control group vs C3−/− and FB−/− groups is summarized in Table I.

FIGURE 4.

Histopathological severity of arthritis. Histopathological arthritic score was assessed as synovial hypertrophy, cartilage destruction, bone destruction, and pannus formation; mean ± SEM. The statistical significance of the effect of genotype on the difference between the control group vs C3−/− and FB−/− groups is summarized in Table I.

Close modal
Table I.

Summary of the statistical significance of the effect of genotype on the four components of histopathological arthritic scorea

Synovial Hypertrophy (p)Cartilage Destruction (p)Bone Destruction (p)Pannus (p)
Single immunization     
C3−/− 0.0002 0.0002 0.0045 0.0006 
FB−/− 0.0172 0.016 0.0603 0.0182 
Repeated immunization     
C3−/− 0.0005 0.0003 0.0044 0.0018 
FB−/− 0.0126 0.0097 0.0202 0.0198 
Synovial Hypertrophy (p)Cartilage Destruction (p)Bone Destruction (p)Pannus (p)
Single immunization     
C3−/− 0.0002 0.0002 0.0045 0.0006 
FB−/− 0.0172 0.016 0.0603 0.0182 
Repeated immunization     
C3−/− 0.0005 0.0003 0.0044 0.0018 
FB−/− 0.0126 0.0097 0.0202 0.0198 
a

Mann-Whitney U test analysis of control vs C3−/− and FB−/− mice, respectively.

Single immunization with 50 μg of BCII triggered low CII-specific IgG response in the C3−/− and FB−/−mice while the corresponding response in the control mice was strong (Fig. 5,a). The Ab levels reached the peak on day 56 after immunization in all groups: 12,028 ± 2,678 μg/ml in the control mice, 4,190 ± 937 μg/ml in the C3−/− mice, and 4,716 ± 622 μg/ml in the FB−/− mice (mean ± SEM). Repeated immunization of the C3−/− and FB−/− mice with BCII 3 wk later led to increased production of CII-specific IgG, fully comparable to that of control animals that received only single doses of BCII (9,946 ± 2,275 μg/ml in the C3−/− mice and 14,004 ± 1,899 μg/ml in the FB−/− mice on day 35 after the initial dose (mean ± SEM; Fig. 5 b). To assess the IgG subclass composition of the Ab response, we measured the levels of the CII-specific IgG1 and IgG2a isotypes in sera on day 35 after the first immunization. The ratio of the CII-specific IgG1 and IgG2a subclasses (IgG1:IgG2a) revealed that to single BCII immunization, the C3−/− mice responded predominantly by IgG1 production while the control and FB−/− mice responded predominantly by IgG2a production (IgG1:IgG2a ratio, 1.84 vs 0.57 and 0.14, respectively). There were no differences in the IgG isotype pattern between the C3−/− and control mice after repeated immunization (IgG1:IgG2a ratio 0.91 vs 1.00), while the response of FB−/− still contained a predominant IgG2a component (IgG1:IgG2a ratio 0.25).

FIGURE 5.

Specific anti-CII Ab response. Specific anti-CII Ig levels after single dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice (mean ± SEM; n, number of mice). The p values refer to the difference between the control group vs the C3−/− and FB−/− groups, respectively.

FIGURE 5.

Specific anti-CII Ab response. Specific anti-CII Ig levels after single dose immunization with BCII (a) and repeated immunization (b) of C3−/−, FB−/−, and control (+/+) mice (mean ± SEM; n, number of mice). The p values refer to the difference between the control group vs the C3−/− and FB−/− groups, respectively.

Close modal

Murine C3 and FB are both encoded by genes located on chromosome 17 at a distance of 13 and <1 cM, respectively (22) from H-2, a major CIA susceptibility locus (23). To rule out the possibility that the differences in the response to CII immunization between the control and C3−/− mice are due to differences in the H-2 alleles, the mice were genotyped at the H-2 locus. Although all the control and C3−/− mice were found to carry only the H-2q allele, the FB−/− mice still carried the H-2b allele from the original C57BL/6 × 129Ola strain (data not shown).

The complement system plays a crucial role in the humoral immune response with functions ranging from initiation of inflammation, regulation of Ab response, clearance of immune complexes, to elimination of pathogens and cell lysis. In this study, we investigated whether deficiency for C3 or FB affects the susceptibility to CIA. We found that mice deficient in either C3 or FB were highly resistant to CIA. The protection against arthritis seems to be only partly mediated by the impaired Ab response in the complement-deficient animals.

Both humoral and cell-mediated immunity are required for active induction of CIA (2). T as well as B cells have been implicated in the pathogenesis of arthritis with self-reactive T cells indirectly provoking the disease via B lymphocytes and Igs (reviewed in Ref. 24). The severity of murine arthritis is positively correlated with the IgG autoantibody response to CII (3) and the predominance of autoreactive IgG2a Abs (25). Complement has been shown to enhance the immunogenicity of both T cell-independent (26) and T cell-dependent Ags (27, 28, 29, 30). It was suggested that the requirement for complement activation products and receptors for the Ab response may be partly reversed by an increased dose of Ag (27). Here, we show that both pathways of complement activation are required for a normal Ab response to immunization with BCII since a single dose of BCII triggered deficient Ab response in both mice lacking C3 and FB. The singly immunized C3−/− mice were found to respond by an aberrant CII-specific IgG1:IgG2a isotype ratio which may point to a role of the C3 activation products in the Th1- and Th2-induced switch from IgM to IgG2a/b and IgG1, respectively, in activated B lymphocytes. We have also found that the impaired Ab response, including the IgG1:IgG2a isotype ratio, is reversed in complement-deficient mice by repeated immunization with BCII. Thus, complement plays a role in the induction phase of CIA by controlling both the quantity and the isotypic pattern of the specific Ab production but the requirement for complement activation in the production of CII-specific Abs can be overcome by repeated immunization.

The importance of anti-CII Abs and Fc receptors for IgG in the regulation of CIA was recently demonstrated by Kleinau et al. (31). The mild arthritic response of complement-deficient mice cannot be solely explained by the impaired B cell response leading to low levels of BCII Abs with an aberrant IgG isotype pattern, since the C3−/− mice exhibited very low mean arthritic scores also after repeated immunization, despite high titers of the CII-specific IgG and IgG1:IgG2a isotype ratio comparable to that of the control mice. Thus, complement activation seems to play a critical role in the inflammatory reaction triggered by the Abs reactive with CII. The fact that the FB−/− mice exhibited reduced arthritic scores compared with the control mice after both single and booster immunization indicates that the alternative pathway of complement activation plays a role in the pathogenesis of CIA. The classical pathway of complement activation is therefore most plausibly triggered by the anti-CII Abs and the activation and generation of inflammatory molecules is further amplified by the alternative pathway. Indeed, in a passive Ab transfer model of arthritis, Ji et al. (32) have recently shown that the alternative pathway is critical for the recruitment and/or activation of polymorphonuclear leukocytes and development of arthritis while components of the classical pathway are entirely dispensable for the effector phase of arthritis. Altogether our results indicate that complement deficiency affects both the induction and the effector phases of CIA.

It has earlier been shown that the H-2 MHC locus on chromosome 17 is important for the total IgG autoantibody response (25) and that it determines the susceptibility of mice to CIA (23) but not the isotypic pattern of the autoantibody response (25). Although the C3−/− mice in our study carried the H-2q allele, the FB−/− mice carried the H-2b allele of the C57BL/6 × 129Ola strain on which the null mutation was originally generated. Although C57BL/6 mice (H-2b) do not develop CIA even after repeated immunization (data not shown) and six generations of backcrossing to DBA/1J strain imply that ∼97% of loci in the FB−/− mice are homozygous for the DBA/1J allele, we cannot exclude that the differences at the H-2 and perhaps even additional loci could have contributed to the impaired specific autoantibody and inflammatory response in the singly immunized FB−/− mice but presumably not to the relative resistance of the FB−/− mice to CIA after repeated immunization. Although rather demanding, the reconstitution of the FB−/− mice with FB would clearly provide the most appropriate control group to allow the assessment of the alternative pathway contribution to both the induction and effector phases of CIA. Because passive anti-collagen Ab transfer induction of arthritis is independent of the H-2 haplotype (4), this approach or induction of arthritis by monoclonal anti-CII Ab transfer would resolve the question of the involvement of the alternative pathway in the effector phase and generation of tissue damage. Recently, C3−/− as well as FB−/− mice but not C4−/− mice were found to be highly resistant to arthritis induced by transfer of serum containing anti-GPI Abs (32). Although this model differs from CIA, the results from both studies implicate the alternative pathway as an important player in the pathogenesis of arthritis.

Although the mean arthritic score of the C3−/− mice was substantially lower compared with both the FB−/− and control mice, after booster immunization the incidence of arthritis in the C3−/− mice was comparable to that of the FB−/− and control mice. Thus, the C3 deficiency ameliorates CIA but does not fully protect against its development. Our observation that one of nine C3−/− mice (i.e., 11%) developed severe arthritis suggests that the inhibition of complement activation increases the threshold for the development of CIA. Our findings are in line with the recent observations of Wang et al. (14), reporting that C5-deficient mice are highly resistant to CIA. Interestingly, 15% of the C5−/− mice in that study developed arthritis clinically and pathologically indistinguishable from that seen in the control mice.

In summary, the C3−/− and FB−/− mice respond by a low specific Ab production to single doses of BCII but this impaired Ab response can be overcome by repeated immunization. Complement deficiency ameliorates CIA in mice. Complement inhibition at the level of C3 activation thus appears to be a rational therapeutic approach in patients with RA.

We acknowledge Dr. Christer Betsholtz for his support of the project and Dr. Milos Pekny for critical reading of this manuscript.

1

This work was supported by grants from the Swedish Medical Council (Project 13470), King Gustaf V’s 80-Year Foundation, Swedish Society of Medicine, Swedish Society for Medical Research, Göteborg Medical Society, and Sigurd and Elsa Golje’s Foundation.

3

Abbreviations used in this paper: RA, rheumatoid arthritis; CIA, collagen-induced arthritis; CII, type II collagen; FB, factor B; BCII, bovine type II collagen.

1
Courtenay, J. S., M. J. Dallman, A. D. Dayan, A. Martin, B. Mosedale.
1980
. Immunisation against heterologous type II collagen induces arthritis in mice.
Nature
283
:
666
2
Seki, N., Y. Sudo, T. Yoshioka, S. Sugihara, T. Fujitsu, S. Sakuma, T. Ogawa, T. Hamaoka, H. Senoh, H. Fujiwara.
1988
. Type II collagen-induced murine arthritis. I. Induction and perpetuation of arthritis require synergy between humoral and cell-mediated immunity.
J. Immunol.
140
:
1477
3
Williams, P. J., R. H. Jones, T. W. Rademacher.
1998
. Correlation between IgG anti-type II collagen levels and arthritic severity in murine arthritis.
Autoimmunity
27
:
201
4
Stuart, J. M., F. J. Dixon.
1983
. Serum transfer of collagen-induced arthritis in mice.
J. Exp. Med.
158
:
378
5
Wooley, P. H., H. S. Luthra, C. J. Krco, J. M. Stuart, C. S. David.
1984
. Type II collagen-induced arthritis in mice. II. Passive transfer and suppression by intravenous injection of anti-type II collagen antibody or free native type II collagen.
Arthritis Rheum.
27
:
1010
6
Holmdahl, R., L. Jansson, A. Larsson, R. Jonsson.
1990
. Arthritis in DBA/1 mice induced with passively transferred type II collagen immune serum: immunohistopathology and serum levels of anti-type II collagen auto-antibodies.
Scand. J. Immunol.
31
:
147
7
Svensson, L., J. Jirholt, R. Holmdahl, L. Jansson.
1998
. B cell-deficient mice do not develop type II collagen-induced arthritis (CIA).
Clin. Exp. Immunol.
111
:
521
8
Hedberg, H..
1964
. The depressed synovial complement activity in adult and juvenile rheumatoid arthritis.
Acta Rheum. Scand.
10
:
109
9
Pekin, T. J., N. J. Zvaifler.
1964
. Hemolytic complement in synovial fluid.
J. Clin. Invest.
43
:
1372
10
Gulati, P., D. Guc, C. Lemercier, D. Lappin, K. Whaley.
1994
. Expression of the components and regulatory proteins of the classical pathway of complement in normal and diseased synovium.
Rheumatol. Int.
14
:
13
11
Morgan, B. P., R. H. Daniels, B. D. Williams.
1988
. Measurement of terminal complement complexes in rheumatoid arthritis.
Clin. Exp. Immunol.
73
:
473
12
Lens, J. W., W. B. van den Berg, L. B. van de Putte, J. H. Berden, S. P. Lems.
1984
. Flare-up of antigen-induced arthritis in mice after challenge with intravenous antigen: effects of pre-treatment with cobra venom factor and anti-lymphocyte serum.
Clin. Exp. Immunol.
57
:
520
13
Wang, Y., S. A. Rollins, J. A. Madri, L. A. Matis.
1995
. Anti-C5 monoclonal antibody therapy prevents collagen-induced arthritis and ameliorates established disease.
Proc. Natl. Acad. Sci. USA
92
:
8955
14
Wang, Y., J. Kristan, L. Hao, C. S. Lenkoski, Y. Shen, L. A. Matis.
2000
. A role for complement in antibody-mediated inflammation: C5-deficient DBA/1 mice are resistant to collagen-induced arthritis.
J. Immunol.
164
:
4340
15
Goodfellow, R. M., A. S. Williams, J. L. Levin, B. D. Williams, B. P. Morgan.
2000
. Soluble complement receptor one (sCR1) inhibits the development and progression of rat collagen-induced arthritis.
Clin. Exp. Immunol.
119
:
210
16
Dreja, H., A. Annenkov, Y. Chernajovsky.
2000
. Soluble complement receptor 1 (CD35) delivered by retrovirally infected syngeneic cells or by naked DNA injection prevents the progression of collagen-induced arthritis.
Arthritis Rheum.
43
:
1698
17
Pekna, M., M. A. Hietala, T. Rosklint, C. Betsholtz, M. Pekny.
1998
. Targeted disruption of the murine gene coding for the third complement component (C3).
Scand. J. Immunol.
47
:
25
18
Pekna, M., M. A. Hietala, A. Landin, A. K. Nilsson, C. Lagerberg, C. Betsholtz, M. Pekny.
1998
. Mice deficient for the complement factor B develop and reproduce normally.
Scand. J. Immunol.
47
:
375
19
Miller, E. J..
1972
. Structural studies on cartilage collagen employing limited cleavage and solubilization with pepsin.
Biochemistry
11
:
4903
20
Bremell, T., A. Abdelnour, A. Tarkowski.
1992
. Histopathological and serological progression of experimental Staphylococcus aureus arthritis.
Infect. Immun.
60
:
2976
21
Saha, B. K..
1996
. Typing of murine major histocompatibility complex with a microsatellite in the class II Eb gene.
J. Immunol. Methods
194
:
77
22
Lyon, M. F., S. Rastan, S. D. M. Brown.
1996
. In
Genetic Variants and Strains of the Laboratory Mouse
Vol. 2
:
1807
Oxford Univ. Press, Oxford.
23
Brunsberg, U., K. Gustafsson, L. Jansson, E. Michaelsson, L. Ahrlund-Richter, S. Pettersson, R. Mattsson, R. Holmdahl.
1994
. Expression of a transgenic class II Ab gene confers susceptibility to collagen-induced arthritis.
Eur. J. Immunol.
24
:
1698
24
Benoist, C., D. Mathis.
2000
. A revival of the B cell paradigm for rheumatoid arthritis pathogenesis?.
Arthritis Res.
2
:
90
25
Watson, W. C., A. S. Townes.
1985
. Genetic susceptibility to murine collagen II autoimmune arthritis: proposed relationship to the IgG2 autoantibody subclass response, complement C5, major histocompatibility complex (MHC) and non-MHC loci.
J. Exp. Med.
162
:
1878
26
Ochsenbein, A. F., D. D. Pinschewer, B. Odermatt, M. C. Carroll, H. Hengartner, R. M. Zinkernagel.
1999
. Protective T cell-independent antiviral antibody responses are dependent on complement.
J. Exp. Med.
190
:
1165
27
Fischer, M. B., M. Ma, S. Goerg, X. Zhou, J. Xia, O. Finco, S. Han, G. Kelsoe, R. G. Howard, T. L. Rothstein, et al
1996
. Regulation of the B cell response to T-dependent antigens by classical pathway complement.
J. Immunol.
157
:
549
28
Molina, H., V. M. Holers, B. Li, Y. Fung, S. Mariathasan, J. Goellner, J. Strauss-Schoenberger, R. W. Karr, D. D. Chaplin.
1996
. Markedly impaired humoral immune response in mice deficient in complement receptors 1 and 2.
Proc. Natl. Acad. Sci. USA
93
:
3357
29
Croix, D. A., J. M. Ahearn, A. M. Rosengard, S. Han, G. Kelsoe, M. Ma, M. C. Carroll.
1996
. Antibody response to a T-dependent antigen requires B cell expression of complement receptors.
J. Exp. Med.
183
:
1857
30
Gustavsson, S., T. Kinoshita, B. Heyman.
1995
. Antibodies to murine complement receptor 1 and 2 can inhibit the antibody response in vivo without inhibiting T helper cell induction.
J. Immunol.
154
:
6524
31
Kleinau, S., P. Martinsson, B. Heyman.
2000
. Induction and suppression of collagen-induced arthritis is dependent on distinct Fcγ receptors.
J. Exp. Med.
191
:
1611
32
Ji, H., K. Ohmura, U. Mahmood, D. M. Lee, F. M. Hofhuis, S. A. Boackle, K. Takahashi, V. M. Holers, M. Walport, C. Gerard, et al
2002
. Arthritis critically dependent on innate immune system players.
Immunity
16
:
157