[2] Preparation and characterization of the different types of collagen

Objective. To compare the genetic regulation of collagen-induced arthritis (CIA) with that of pristaneinduced arthritis (PIA) in rats.Methods. A genome-wide linkage analysis of an (E3 ؋ DA)DA backcross of rats with CIA (n ‫؍‬ 364 male rats; the same strain combinations as previously used to determine the genetic control of PIA) was performed. The strongest loci in both CIA and PIA (i.e., Cia12/Pia4 and Cia13/Pia7) were isolated in congenic strains. Susceptibility in both congenic strains was tested in rats with CIA and in rats with PIA.Results. We found a striking, although not complete, similarity of the arthritis-controlling loci in CIA and in PIA, as well as the previously defined loci associated with cartilage destruction, antibody production, and the acute-phase response. All major PIA quantitative trait loci (QTLs) identified in early severe arthritis were also strong regulators of CIA. The 2 strongest QTLs, Cia12/Pia4 on chromosome 12 and Cia13/Pia7 on chromosome 4, were also analyzed in congenic strains with DA or E3 as the background genome. Consistent with the results of linkage analysis, the congenic strain experiments showed that the chromosome 4 locus was more penetrant in CIA than in PIA, while the chromosome 12 locus almost completely dominated the control of PIA severity.Conclusion. The underlying genetic control of CIA was found to have many, but not all, pathogenic mechanisms in common with PIA, despite the use of a cartilage-specific antigen (type II collagen) to induce CIA but not PIA.

Section: Methodsmentioning

confidence: 99%

A comparative genetic analysis between collagen‐induced arthritis and pristane‐induced arthritis

Olofsson¹,

Lu²,

Holmberg³

et al. 2003

“…Articular cartilage was obtained from the metacarpophalangeal joints of 1-2-year-old cows. Bovine type I1 collagen was prepared according to the method of Miller and Rhodes (37). Collagen was dissolved in 0.05M acetic acid (10 mg/ml) and stored at -70°C.…”

Section: Methodsmentioning

confidence: 99%

Role of interleukin‐4 and interleukin‐10 in murine collagen‐induced arthritis. Protective effect of interleukin‐4 and interleukin‐10 treatment on cartilage destruction

Joosten

Lubberts

Durez

et al. 1997

374

246

Objective. To examine the role of endogenous interleukin‐4 (IL‐4) and interleukin‐10 (IL‐10) and the therapeutic effect of the addition of IL‐4 and IL‐10 in early and established murine collagen‐induced arthritis (CIA). Methods. Murine recombinant IL‐4, IL‐10, or the combination was given intraperitoneally twice daily from the day of arthritis onset up to 7–10 days of CIA in DBA/1 mice. Anti‐IL‐4, anti–IL‐10, or both antibodies were given intraperitoneally before or after the onset of CIA. The effect of cytokine or anticytokine treatment was monitored visually by macroscopic scoring. Histology and reverse transcription‐polymerase chain reaction (RT‐PCR) analyses were performed at the end of the treatment period. Results. IL‐4 alone did not provoke any effect, IL‐10 slightly suppressed the arthritis, but a more pronounced amelioration was found with the combination. This cooperative effect was noted after early treatment but also occurred when the start of treatment was delayed until 1 week after onset. Apart from suppression of macroscopic signs of inflammation, combined treatment with IL‐4/IL‐10 also reduced cellular infiltrates in the synovial tissue and caused pronounced protection against cartilage destruction. Moreover, levels of mRNA for tumor necrosis factor α (TNFα) and IL‐1 were highly suppressed both in the synovial tissue and in the articular cartilage. In contrast, levels of IL‐1 receptor antagonist (IL‐1Ra) mRNA remained elevated, which suggests that the mechanism of protection may be related to suppressed production of TNFα and IL‐1, with concomitant up‐regulation of the IL‐1Ra/IL‐1 balance. However, accelerated onset of CIA and increased severity could be achieved with neutralizing anti–IL‐10 antibodies. This expression could be further optimized with a combination of anti–IL‐4 and anti–IL‐10 antibodies, although anti–IL‐4 alone was without effect. Conclusion. Our data are consistent with a dominant role of IL‐10 in the natural suppression of arthritis expression, whereas combined treatment with IL‐4 and IL‐10 appears of potential therapeutic value, not only at the onset, but also in established arthritis.

“…Preparation of type I1 collagen from the chicken xiphoid process was performed as described previously (13 acids comprising N-terminal fragment al(II)3/4 TCA was obtained by gel chromatography. Purity of al(II)3/4 TCA was confirmed by autoradiography of the radiolabeled purified fragment and by size-exclusion chromatography.…”

Section: Methodsmentioning

confidence: 99%

Characterization of the binding region for the yersinia enterocolitica adhesin yada on types i and ii collagen

Schulze‐Koops

Burkhardt

Heesemann

et al. 1995

Objective. The plasmid-encoded adhesin YadA confers pathogenic functions on Yersinia enterocolitica, a microorganism associated with reactive arthritis. While emerging evidence has indicated that the persistence of the bacteria in individuals with reactive arthritis is a prerequisite for the development of the disease, the tissue specificity of this immunologic disease sequela remains elusive. The present study was undertaken to investigate YadA-mediated binding of Y enterocolitica to the most abundant collagens in joints, types I and I1 collagen.Methods. Binding studies were performed with recombinant Y enterocoliticu strains and highly purified type I1 collagen and the d ( I ) chain of type I collagen, or fragments of these collagens generated by various enzymatic and nonenzymatic cleavage procedures. Interactions of bacteria with the proteins were determined in binding assays with radiolabeled proteins.Results. Binding regions for YadA were identified at the 181-amino acid fragment d(I)78CBN of type I collagen and the CBlO fragment of type I1 collagen. From binding and blocking experiments with al(1) fragments, cyanogen bromide-derived or mammalian collagenasderived type I1 collagen fragments, and synthetic peptides with collagen-like structures, it was ~~~