Fig 1.
Time course of human lymphocyte reconstitution in peripheral blood of EBV-infected hu-NOG mice.
Following inoculation with EBV, the percentages of human CD45+, CD4+, CD8+, and CD19+ cells among the peripheral blood mononuclear cells were measured weekly. Upon EBV infection, the percentage of human CD8+ T cells increased rapidly, and the ratio of CD4+ cells to CD8+ cells decreased to below 1. Values represent mean ± standard error.
Fig 2.
3D-CT images of knee joints in EBV-infected and -uninfected hu-NOG mice.
(A) Joint from an EBV-infected mouse. Arrowheads indicate bone erosion at the distal portion of the femur and proximal portion of the tibia. (B) Joint of another EBV-infected mouse. (C) Joint of an EBV-uninfected mouse (control). Republished from [Nagasawa Y, Ikumi N, Nozaki T, Inomata H, Imadome K, et al. Human osteoclasts are mobilized in erosive arthritis of Epstein-Barr virus-infected humanized NOD/Shi-scid/IL2Rγnull mice. 2014 ACR/ARHP Annual Meeting. Abstract number:2340.] under a CC BY license, with permission from [American College of Rheumatology], original copyright [2014].
Table 1.
Bone erosion in hu-NOG mice.
Fig 3.
Histochemistry of the knee joint section of EBV-infected hu-NOG mice.
(A–D) Joint sections from an EBV-infected mouse (N 87–6). (A and B) Joint sections stained for TRAP. TRAP observed in multinucleated cells at the bone erosion site stained red violet. (C) Joint section immunostained for human cathepsin K. Multinucleated cells positive for human cathepsin K stained brown with the anti-human cathepsin K antibody. (D) Joint section immunostained for human MMP-9. Multinucleated cells positive for MMP-9 stained brown with the anti-human MMP-9 antibody (dilution, 1:25). (E–H) Joint sections from another EBV-infected mouse (N 70–13). (E and F) Joint sections stained for TRAP. TRAP-positive multinucleated cells were found in the knee joint tissue. Joint sections immunostained for human cathepsin K (G) and human MMP-9 (dilution, 1:50) (H). These multinucleated cells stained positively with the anti-human cathepsin K antibody and the anti-human MMP-9 antibody. Original magnification, A: 200×, others: 400×. Republished from [Nagasawa Y, Ikumi N, Nozaki T, Inomata H, Imadome K, et al. Human osteoclasts are mobilized in erosive arthritis of Epstein-Barr virus-infected humanized NOD/Shi-scid/IL2Rγnull mice. 2014 ACR/ARHP Annual Meeting. Abstract number:2340.] under a CC BY license, with permission from [American College of Rheumatology], original copyright [2014].
Fig 4.
Cytochemistry of cultured long bone marrow cells from EBV-infected and -uninfected hu-NOG mice in the presence of M-CSF and RANKL differentiation signals.
(A–D) Cultured cells from an EBV-infected mouse (NOG 12–6). (A) Cultured cells stained for TRAP. TRAP in the cultured multinucleated cell stained red violet. (B) Cultured cells immunostained for human cathepsin K. The positive multinucleated cell stained brown with the anti-human cathepsin K antibody (dilution, 1:1800). (C) Cultured cells immunostained for human mitochondrial protein. Multinucleated cell positively stained brown with anti-human mitochondria antibody (dilution, 1:40). (D) Cultured cells immunostained using isotype control rabbit immunoglobulin (dilution, 1:4500). (E–H) Cultured cells from another EBV-infected mouse (NOG 12–7). The multinucleated cells positively stained for TRAP (E), and with anti-human cathepsin K antibody (dilution, 1:600) (F) and anti-human mitochondria antibody (dilution, 1:10) (G). (H) Cultured cells immunostained using isotype control mouse immunoglobulin (dilution, 1:5). (I) Cultured cells from another EBV-infected mouse (NOG 18–1) on a pit formation assay plate. TRAP in the cultured multinucleated cells stained red violet. Calcium phosphate coating around the TRAP-positive multinucleated cell was eliminated (arrow). (J and K) Cultured cells from an EBV-uninfected mouse (NOG 24–1). (J) Cultured cells immunostained for human MMP-9. The multinucleated cell positive for human MMP-9 stained brown with anti-human MMP-9 antibody (dilution, 1:25). (K) Cultured cells immunostained for human mitochondrial protein. The multinucleated cell positive for human mitochondrial protein stained brown with anti-human mitochondria antibody (dilution, 1:10). (L and M) Cultured cells from another EBV-uninfected mouse (NOG 26–1). The multinucleated cells were considered positive when stained with anti-human MMP-9 antibody (dilution, 1:50) (L) and anti-human mitochondria antibody (dilution, 1:40) (M). (original magnification, 400×) Republished from [Nagasawa Y, Ikumi N, Nozaki T, Inomata H, Imadome K, et al. Human osteoclasts are mobilized in erosive arthritis of Epstein-Barr virus-infected humanized NOD/Shi-scid/IL2Rγnull mice. 2014 ACR/ARHP Annual Meeting. Abstract number:2340.] under a CC BY license, with permission from [American College of Rheumatology], original copyright [2014].
Fig 5.
Cytochemistry of cultured mouse osteoclast progenitor cells in the presence of M-CSF and RANKL differentiation signals.
(A–D) Cultured cells from mouse osteoclast progenitor cells. (A) Cultured cells immunostained with an antibody that reacts with mouse MMP-9. Multinucleated cells positive for MMP-9 stained brown with the anti-MMP-9 antibody (dilution, 1:100). (B) Cultured cells immunostained with an antibody specific to the human MMP-9. Multinucleated cells were not stained (antibody dilution, 1:12.5). (C) Cultured cells immunostained with an antibody that reacts with mouse mitochondrial protein. Multinucleated cells positive for the mouse mitochondrial protein stained brown with the anti-mitochondrial antibody (dilution, 1:360). (D) Cultured cells immunostained with an antibody specific to human mitochondrial protein. Multinucleated cells were not stained (antibody dilution, 1:10). Original magnification, 400×.
Fig 6.
Cytochemistry of cultured pelvic bone marrow cells from EBV-infected hu-NOG mice in the absence of differentiation signals.
(A–C) Cultured cells from an EBV-infected mouse (NOG 22–7) in glass-bottom dishes. (A and B) Cultured cells stained for TRAP. (A) Multinucleated cells were confirmed in the bone marrow (arrow). (B) The same cultured cell stained for TRAP. TRAP in the cultured multinucleated cell stained red violet. (C) The same cultured cell immunostained for human MMP-9. The TRAP-positive multinucleated cell stained red with the anti-human MMP-9 antibody. (D and E) Cultured cells from another mouse (NOG 25–3). (D) Cultured cell stained for TRAP. The multinucleated cell stained positive for TRAP. (E) The same cultured cell immunostained for human MMP-9. Multinucleated cell positive for anti-human MMP-9 antibody were stained. (F and G) Cultured cells from another mouse (NOG 25–4). (F) Cultured cell stained for TRAP. (G) The same cultured cell immunostained for human MMP-9. Original magnification, A: 100×, others: 400×.