S100A8/A9 increases the mobilization of pro-inflammatory Ly6C^high monocytes to the synovium during experimental...

Monocytes are dominant cells present within the inflamed synovium during osteoarthritis (OA). In mice, two functionally distinct monocyte subsets are described: pro-inflammatory Ly6C^high and patrolling Ly6C^low monocytes. Alarmins S100A8/A9 locally released by the synovium during inflammatory OA for prolonged periods may be dominant proteins involved in stimulating recruitment of Ly6C^high monocytes from the circulation to the joint. Our objective was to investigate the role of S100A8/A9 in the mobilization of Ly6C^high and Ly6C^low monocytic populations to the inflamed joint in collagenase-induced OA (CiOA).

S100A8 was injected intra-articularly to investigate monocyte influx. CiOA was induced by injection of collagenase into knee joints of wild-type C57BL/6 (WT), and S100a9^-/- mice. Mice were sacrificed together with age-matched saline-injected control mice (n = 6/group), and expression of monocyte markers, pro-inflammatory cytokines, and chemokines was determined in the synovium using ELISA and RT-qPCR. Cells were isolated from the bone marrow (BM), spleen, blood, and synovium and monocytes were identified using FACS.

S100A8/A9 was highly expressed during CiOA. Intra-articular injection of S100A8 leads to elevated expression of monocyte markers and the monocyte-attracting chemokines CCL2 and CX3CL1 in the synovium. At day 7 (d7) after CiOA induction in WT mice, numbers of Ly6C^high, but not Ly6C^low monocytes, were strongly increased (7.6-fold) in the synovium compared to saline-injected controls. This coincided with strong upregulation of CCL2, which preferentially attracts Ly6C^high monocytes. In contrast, S100a9^-/- mice showed a significant increase in Ly6C^low monocytes (twofold) within the synovium at CiOA d7, whereas the number of Ly6C^high monocytes remained unaffected. In agreement with this finding, the Ly6C^low mobilization marker CX3CL1 was significantly higher within the synovium of S100a9^-/- mice. Next, we studied the effect of S100A8/A9 on release of Ly6C^high monocytes from the BM into the circulation. A 14% decrease in myeloid cells was found in WT BM at CiOA d7. No decrease in myeloid cells in S100a9^-/- BM was found, suggesting that S100A8/A9 promotes the release of myeloid populations from the BM.

Induction of OA locally leads to strongly elevated S100A8/A9 expression and an elevated influx of Ly6C^high monocytes from the BM to the synovium.