Osteoclasts expressing the measles virus nucleocapsid gene display a pagetic phenotype
Noriyoshi Kurihara, … , Dirk Anderson, G. David Roodman
Noriyoshi Kurihara, … , Dirk Anderson, G. David Roodman
Published March 1, 2000
Citation Information: J Clin Invest. 2000;105(5):607-614. https://doi.org/10.1172/JCI8489.
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Article

Osteoclasts expressing the measles virus nucleocapsid gene display a pagetic phenotype

Abstract

Osteoclasts (OCLs) in Paget’s disease are markedly increased in number and size, have increased numbers of nuclei per multinucleated cell, and demonstrate increased resorption capacity and increased sensitivity to 1,25-(OH)2D3, the active form of vitamin D. These cells also contain nuclear inclusions, reminiscent of those seen in paramyxovirus-infected cells, which cross-react with antibodies to measles virus nucleocapsid (MVNP) antigen. To elucidate the role of MV in the abnormal OCL phenotype of Paget’s disease, we transduced normal OCL precursors with retroviral vectors expressing MVNP and the MV matrix (MVM) genes. The transduced cells were then cultured with 1,25-(OH)2D3 for14 or 21 days to induce formation of OCL-like multinucleated cells. The MVNP-transduced cells formed increased numbers of multinucleated cells, which contained many more nuclei and had increased resorption capacity compared with multinucleated cells derived from empty vector–transduced (EV-transduced) and MVM-transduced or normal bone marrow cells. Furthermore, MVNP-transduced cells showed increased sensitivity to 1,25-(OH)2D3, and formed OCLs at concentrations of 1,25-(OH)2D3 that were 1 log lower than that required for normal, EV-transduced, or MVM-transduced cells. These results demonstrate that expression of the MVNP gene in normal OCL precursors stimulates OCL formation and induces OCLs that express a phenotype similar to that of pagetic OCLs. These results support a potential pathophysiologic role for MV infection in the abnormal OCL activity and morphology that are characteristic of pagetic OCLs.

Authors

Noriyoshi Kurihara, Sakamuri V. Reddy, Cheikh Menaa, Dirk Anderson, G. David Roodman

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Figure 1

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Transduction of OCL precursors. (a) Immunostaining of human bone marrow ...
Transduction of OCL precursors. (a) Immunostaining of human bone marrow cells for MV expression. Two days after transduction of the MVNP and MVM genes into human bone marrow cells, gene expression was evaluated by immunostaining using the anti-MVNP and anti-MVM monoclonal antibodies. The dark staining denotes positive reactivity. A similar pattern of immunostaining was detected in 3 separate experiments. (b) CFU-GM colony formation from transduced normal human bone marrow cells. The cells were cultured for CFU-GM, the earliest identifiable OCL precursor, with or without G418 (500 μg/mL). Transduction efficiency was assayed by counting developed CFU-GM on day 7. Results represent those from a typical experiment. *P < 0.05 compared with nontransduced cells; **P < 0.01 compared with nontransduced cells cultured with 500 μg/mL of G418. A similar pattern of results was seen in 5 separate experiments. (c) Western blot analysis for MVNP and MVM gene expression in marrow mononuclear cells transduced with retroviral constructs containing the MVNP or MVM gene. Marrow mononuclear cells were transduced with the MVNP or MVM gene; after 3 days, the cells were lysed and the lysates were subjected to Western blot analysis. MVNP-transduced cells expressed only the MVNP protein, and MVM-transduced cells expressed only the MVM protein.