Table 1.
Surface marker antibodies for flow cytometry.
Table 2.
Primers for qPCR.
Fig 1.
Male B10.RIII mice develop arthritis after hydrodynamic injection of IL-23 EEV.
(A) Experimental layout for IL-23 EEV-induced arthritis experiments. (B) 8–12 week-old male B10.RIII mice received 500 ng IL-23 EEV on day 0 via hydrodynamic tail vein injection; control mice received no injection (n = 12–15 mice/group). Clinical score (mean ± SD). Paw swelling (mean ± SD). Body weight (mean ± SD). (C) Serum IL-23 concentrations on day 14 were determined by ELISA (n = 9–11 mice/group). (D) Representative H&E stained sections of day 14 wrists from control and IL-23 EEV injected mice, scale bar 250 μm. (E) Gene expression analysis in the wrists on day 14 (n = 5–6 mice/group). qPCR data for individual samples were first normalized by Hprt expression and then divided by the mean of the uninjected control group. P values on day 14 were determined by unpaired t-test.
Fig 2.
Male B10.RIII mice injected with IL-23 EEV develop colitis and psoriasis-like skin disease but not spondylitis or uveitis.
(A) Colon weight of the animals in the experiment shown in Fig 1. (B) Representative H&E stained colon sections from control and IL-23 EEV injected mice, scale bar 50 μm. (C) Expression of Tnf, Il1b, Il17a, and Il22 in the colon was analyzed by qPCR (n = 5–6 mice/group). (D) Representative H&E stained sections of day 14 ears from control and IL-23 EEV injected mice, scale bar 50 μm. (E) Expression for Tnf, Il1b, Il17a, and Il22 in skin from the forepaws was analyzed by qPCR. P values were determined by unpaired t-test. (F) Representative H&E stained sections of day 14 lumbar spine from control and IL-23 EEV injected mice, scale bar 100 μm. (G) Representative H&E stained sections of day 14 eyes from control and IL-23 EEV injected mice, scale bar 125 μm.
Fig 3.
Systemic overexpression of IL-23 results in expansion of myeloid cells in the spleen.
(A) Spleen weight of the mice from the experiment in Fig 1. (B) For a subset of animals, single cell suspensions were prepared from the spleen and analyzed by flow cytometry, n = 5 mice/group. P values were determined by unpaired t-test.
Fig 4.
Male B6 do not develop arthritis after hydrodynamic injection of IL-23 EEV.
8–12 week-old male B10.RIII and C57BL/6 mice received 500 ng IL-23 EEV on day 0 via hydrodynamic tail vein injection; control mice received no injection (n = 12–15 mice/group). (A) Clinical score (mean ± SD). Paw swelling (mean ± SD). Body weight (mean ± SD). (B) Serum IL-23 and IL-17A concentrations on day 14 were determined by ELISA. (C) Representative H&E stained sections of day 14 wrists from IL-23 EEV injected B10.RIII and B6 mice, scale bar 250 μm. (D) Gene expression analysis in the wrists on day 14 (n = 5–6 mice/group). qPCR data for individual samples were first normalized by Hprt expression and then divided by the mean of all (B10.RIII and B6) uninjected control mice. P values were determined by unpaired t-test.
Fig 5.
Male B6 mice develop colitis and mild psoriasis-like skin disease after hydrodynamic injection of IL-23 EEV.
Data are from the animals in the experiment shown in Fig 4. (A) Colon weight on day 14. (B) Representative H&E stained sections of day 14 colon from IL-23 EEV injected B10.RIII and B6 mice, scale bar 50 μm. (C) Gene expression for Tnf, Il1b, Il17a, and Il22 in the colon was analyzed by qPCR. (D) Representative H&E stained sections of day 14 ears from IL-23 EEV injected B10.RIII and B6 mice, scale bar 50 μm. (E) Gene expression for Tnf, Il1b, Il17a, and Il22 in forepaw skin was analyzed by qPCR.
Fig 6.
Male B6 mice accumulate significantly fewer myeloid cells than B10.RIII mice in the spleen after hydrodynamic injection of IL-23 EEV.
(A) Spleen weight of the mice from the experiment in Fig 4. (B) For a subset of animals, single cell suspensions were prepared from the spleens and analyzed by flow cytometry, n = 5 mice/group. P values were determined by unpaired t-test.
Fig 7.
Both B10.RIII and B6 mice lose trabecular and cortical bone after IL-23 EEV injection.
(A) μCT trabecular and cortical bone parameters from femurs of control and IL-23 EEV injected B10.RIII and B6 14 days after IL-23 EEV injection (n = 4–7 mice/group). Trabecular bone volume/total volume (Tb. BV/TV), trabecular bone total mineral density (Tb. TMD), cortical thickness (Ct. Th), and cortical total mineral density (Ct. TMD). P values were determined by unpaired t-test. (B) Representative μCT images of trabecular bone and (C) representative μCT images of cortical bone from control and IL-23 EEV injected B10.RIII and B6 mice.
Fig 8.
Female B10.RIII mice develop more severe arthritis than males but do not lose weight after hydrodynamic injection of IL-23 EEV.
8–12 week-old male and female B10.RIII mice received 500 ng IL-23 EEV on day 0 via hydrodynamic tail vein injection; control mice received no injection (n = 14 mice in IL-23 EEV injected groups, n = 5 mice in control groups). (A) Clinical score (mean ± SD). Paw swelling (mean ± SD). Body weight (mean ± SD). (B) Serum IL-23 on day 14 was determined by ELISA. (C) Representative H&E stained sections of day 14 wrists, scale bar 250 μm. (D) Gene expression analysis in the wrists on day 14 (n = 9–11 mice/group). qPCR data for individual samples were first normalized by Hprt expression and then divided by the mean of the IL-23 EEV injected males. P values were determined by unpaired t-test. (E) Colon weight on day 14. (F) Representative H&E stained colon sections from male and female B10.RIII IL-23 EEV injected mice, scale bar 50 μm.
Fig 9.
Female B10.RIII mice exhibit greater expansion of myeloid cells in the spleen after hydrodynamic injection of IL-23 EEV.
(A) Spleen weight of the mice from the experiment in Fig 8. (B) For a subset of animals, single cell suspensions were prepared from the spleens and analyzed by flow cytometry, n = 4–5 mice/group. P values were determined by unpaired t-test.