Figure 1.
Joint inflammation is differentially affected by Clo-Lip and genetic deficiencies in MoPh. (A) Quantification of the number of indicated cells in 50 μl mouse blood 24 h after i.v. injection of Clo-Lip or PBS (n = 5 per group). Data are mean ± SD; two-tailed Student’s t test; ***, P = 0.0003. (B) Clinical evaluation of K/BxN STA in C57BL/6 mice treated with 200 μl Clo-Lip or PBS 24 h before arthritis induction using a semiquantitative clinical index and the corresponding area under the curve (AUC; n = 5 per group). Data are mean ± SEM; two-tailed Student’s t test of AUC; ***, P = 0.0027. The experiment was performed twice. (C) Flow cytometry of CD45+CD11b+CD115+ monocytes in the peripheral blood of NR4a1−/− and C57BL/6 wild-type mice demonstrating the absence of the CD45+CD11b+CD115+ Ly6Clow monocyte subset in NR4a1−/− mice. Numbers in the flow plot correspond to the percentage of the parent population. (D) Clinical development of STA in NR4a1−/− and C57BL/6 wild-type mice quantified by a semiquantitative clinical index and the corresponding AUC (n = 3 per group). (E) Treatment regimen for continuous monocyte and macrophage depletion of Cx3cr1cre;iDTR mice or iDTR control mice. Bottom: Representative flow cytometry plots demonstrating the depletion of blood monocytes 1 d after intraperitoneal injection of 500 ng DT. During STA, mice were treated daily starting 1 d before arthritis induction. (F) Quantification of neutrophil and monocyte numbers in 100 μl blood of Cx3cr1cre;iDTR mice (n = 5) and iDTR control mice (n = 3) 1 d after first DT injection. (G) Clinical course of STA in Cx3cr1cre;iDTR (n = 9) and iDTR control mice (n = 8) upon daily treatment with DT quantified by a semiquantitative index and the corresponding AUC. (H) Treatment regimen for predepletion of synovial macrophages of Cx3cr1cre;iDTR or iDTR control mice and representative flow cytometry plots demonstrating recovery of blood monocytes 5 d after two intraperitoneal injections of 500 ng DT. (I) Representative images of confocal laser scanning fluorescence microscopy of the synovial membrane of knee joints of Cx3cr1cre;iDTR mice or iDTR control mice 5 d after DT administration showing depletion of CD68+ (red) synovial macrophages in Cx3cr1cre;iDTR mice (scale bar, 10 µm). sc, synovial cavity; st, synovial tissue. (J) Clinical course of STA in predepleted Cx3cr1cre;iDTR (n = 8) and iDTR control mice (n = 9) quantified by the clinical index and the corresponding AUC. Data are mean ± SEM; two-tailed Student’s t test of AUC; *, P = 0.0142. Experiments (A–J) were performed at least two times. Numbers in flow plots correspond to percentages of parent population (C, E, and H).

Joint inflammation is differentially affected by Clo-Lip and genetic deficiencies in MoPh. (A) Quantification of the number of indicated cells in 50 μl mouse blood 24 h after i.v. injection of Clo-Lip or PBS (n = 5 per group). Data are mean ± SD; two-tailed Student’s t test; ***, P = 0.0003. (B) Clinical evaluation of K/BxN STA in C57BL/6 mice treated with 200 μl Clo-Lip or PBS 24 h before arthritis induction using a semiquantitative clinical index and the corresponding area under the curve (AUC; n = 5 per group). Data are mean ± SEM; two-tailed Student’s t test of AUC; ***, P = 0.0027. The experiment was performed twice. (C) Flow cytometry of CD45+CD11b+CD115+ monocytes in the peripheral blood of NR4a1−/− and C57BL/6 wild-type mice demonstrating the absence of the CD45+CD11b+CD115+ Ly6Clow monocyte subset in NR4a1−/− mice. Numbers in the flow plot correspond to the percentage of the parent population. (D) Clinical development of STA in NR4a1−/− and C57BL/6 wild-type mice quantified by a semiquantitative clinical index and the corresponding AUC (n = 3 per group). (E) Treatment regimen for continuous monocyte and macrophage depletion of Cx3cr1cre;iDTR mice or iDTR control mice. Bottom: Representative flow cytometry plots demonstrating the depletion of blood monocytes 1 d after intraperitoneal injection of 500 ng DT. During STA, mice were treated daily starting 1 d before arthritis induction. (F) Quantification of neutrophil and monocyte numbers in 100 μl blood of Cx3cr1cre;iDTR mice (n = 5) and iDTR control mice (n = 3) 1 d after first DT injection. (G) Clinical course of STA in Cx3cr1cre;iDTR (n = 9) and iDTR control mice (n = 8) upon daily treatment with DT quantified by a semiquantitative index and the corresponding AUC. (H) Treatment regimen for predepletion of synovial macrophages of Cx3cr1cre;iDTR or iDTR control mice and representative flow cytometry plots demonstrating recovery of blood monocytes 5 d after two intraperitoneal injections of 500 ng DT. (I) Representative images of confocal laser scanning fluorescence microscopy of the synovial membrane of knee joints of Cx3cr1cre;iDTR mice or iDTR control mice 5 d after DT administration showing depletion of CD68+ (red) synovial macrophages in Cx3cr1cre;iDTR mice (scale bar, 10 µm). sc, synovial cavity; st, synovial tissue. (J) Clinical course of STA in predepleted Cx3cr1cre;iDTR (n = 8) and iDTR control mice (n = 9) quantified by the clinical index and the corresponding AUC. Data are mean ± SEM; two-tailed Student’s t test of AUC; *, P = 0.0142. Experiments (A–J) were performed at least two times. Numbers in flow plots correspond to percentages of parent population (C, E, and H).

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