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. 2003 May;62(5):465–471. doi: 10.1136/ard.62.5.465

Skewed balance in basal expression and regulation of activating v inhibitory Fcγ receptors in macrophages of collagen induced arthritis sensitive mice

A Blom 1, P L E M van Lent 1, A Holthuysen 1, C Jacobs 1, W B van den Berg 1
PMCID: PMC1754518  PMID: 12695162

Abstract

Background: Recently, it has been found that collagen type II arthritis susceptible mouse strains are hyperreactive to immune complexes (ICs), locally deposited into their knee joints.

Objective: To investigate whether this strain specific knee joint hyperreactivity is related to a disturbed regulation of activatory and inhibitory FcγR on their macrophages before and after stimulation with ICs.

Methods: Macrophages from collagen induced arthritis susceptible strains (DBA/1 and B10.RIII) and non-susceptible strains (C57BL/6 and BALB/c) were compared. FcγR levels on macrophages were detected at protein level by flow cytometric analysis and at mRNA level by reverse transcriptase-polymerase chain reaction. Macrophages were stimulated with ICs, and production of cytokines and enzymes was measured at different times.

Results: On synovial and peritoneal macrophages of DBA/1 mice a higher basal FcγRII and III expression was found, which was skewed towards the activating FcγRIII. In B10.RIII macrophages, however, FcγRIII levels were much lower. Regulation of FcγR mRNA levels in macrophages was tested after stimulation with ICs for one and three days. DBA/1 and B10.RIII macrophages showed a prolonged up regulation of activating FcγRI and III, whereas the inhibiting FcγRII was significantly down regulated compared with non-susceptible strains. In line with this, DBA/1 and B10.RIII macrophages showed a higher interleukin 1 (IL1) and matrix metalloproteinase (MMP) production after IC exposure, whereas IL6 production was significantly reduced.

Conclusions: This study indicates that macrophages derived from collagen type II arthritis susceptible mice show a disregulated FcγR expression before, and even more clearly, after activation by ICs involved in inflammation and cartilage degradation, resulting in prolonged expression of activatory FcγRI and III, down regulation of inhibitory FcγRII and increased release of IL1 and MMP.

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Figure 1.

Figure 1

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Expression of FcγRs on macrophages of C57BL/6, BALB/c, DBA/1, and B10.RIII mice. The expression of FcγRII/III was significantly higher in DBA/1 and B10.RIII mice than in the other strains. However when FcγR expression was tested on peripheral blood mononuclear cells (PBMC) of these different strains, no difference in expression was found (A). When K9-361 was used to detect FcγRII, we found higher expression of this receptor in DBA/1 and B10.RIII mice (B). To detect FcγRIII, the binding of anti-FcγRII/III (2.4G2) to FcγRII was blocked using K9-361. DBA/1 mice express high levels of FcγRIII than the other strains (C).

Figure 2.

Figure 2

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Regulation of expression of FcγRI (A), II (B), and III (C) by macrophages of C57BL/6, BALB/c, DBA/1, and B10.RIII after stimulation with HAGG. The number of PCR cycles needed to detect these receptors in unstimulated cells was subtracted from the number needed in stimulated cells, after correction for GAPDH content. Note that CIA sensitive mice show a prolonged up regulation of FcγRI and III and a prolonged down regulation of FcγRII, whereas BALB/c and C57BL/6 mice show an up regulation of FcγRII at a later time.

Figure 3.

Figure 3

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IL1α (A) and IL6 (B) production by macrophages of C57BL/6, BALB/c, DBA/1, and B10.RIII after stimulation by HAGG. IL1α production was measured by ELISA and was higher in DBA/1 mice at day 1 and in DBA/1 and B10.RIII at day 2, indicating higher cytokine production after stimulation with HAGG by macrophages of CIA sensitive strains. This is in contrast with IL6 levels produced by peritoneal macrophages after HAGG stimulation. DBA/1 and B10.RIII mice produced significantly less IL6 than the other strains.

Figure 4.

Figure 4

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Collagenase/gelatinase production by macrophages of C57BL/6, BALB/c, DBA/1, and B10.RIII mice after stimulation with HAGG. Collagenase/gelatinase production was measured at day 2 after stimulation with HAGG, using a specific fluorescent substrate. The activity of the enzyme was measured by following fluorescence at different time points. Basal enzyme activity was subtracted from the levels of enzyme activity after stimulation with HAGG. Interestingly, enzyme activity of DBA/1 mice is largely increased at day 2 after stimulation with HAGG, compared with other strains.

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