B cells and T cells are critical for the preservation of bone homeostasis and attainment of peak bone mass in vivo.
Journal: 2007/June - Blood
ISSN: 0006-4971
Abstract:
Bone homeostasis is regulated by a delicate balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclastogenesis is controlled by the ratio of receptor activator of NF-kappaB ligand (RANKL) relative to its decoy receptor, osteoprotegerin (OPG). The source of OPG has historically been attributed to osteoblasts (OBs). While activated lymphocytes play established roles in pathological bone destruction, no role for lymphocytes in basal bone homeostasis in vivo has been described. Using immunomagnetic isolation of bone marrow (BM) B cells and B-cell precursor populations and quantitation of their OPG production by enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), cells of the B lineage were found to be responsible for 64% of total BM OPG production, with 45% derived from mature B cells. Consistently B-cell knockout (KO) mice were found to be osteoporotic and deficient in BM OPG, phenomena rescued by B-cell reconstitution. Furthermore, T cells, through CD40 ligand (CD40L) to CD40 costimulation, promote OPG production by B cells in vivo. Consequently, T-cell-deficient nude mice, CD40 KO mice, and CD40L KO mice display osteoporosis and diminished BM OPG production. Our data suggest that lymphocytes are essential stabilizers of basal bone turnover and critical regulators of peak bone mass in vivo.
Relations:
Content
Citations
(134)
References
(55)
Diseases
(2)
Chemicals
(5)
Genes
(4)
Organisms
(4)
Processes
(3)
Anatomy
(4)
Affiliates
(2)
Similar articles
Articles by the same authors
Discussion board
Blood 109(9): 3839-3848

B cells and T cells are critical for the preservation of bone homeostasis and attainment of peak bone mass in vivo

Division of Endocrinology & Metabolism & Lipids, Emory University School of Medicine, Atlanta, GA;
Department of Internal Medicine, Section of Internal Medicine and Endocrine and Metabolic Sciences, University of Perugia, Italy
Corresponding author.
Received 2006 Jul 27; Accepted 2006 Dec 19.

Abstract

Bone homeostasis is regulated by a delicate balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclastogenesis is controlled by the ratio of receptor activator of NF-κB ligand (RANKL) relative to its decoy receptor, osteoprotegerin (OPG). The source of OPG has historically been attributed to osteoblasts (OBs). While activated lymphocytes play established roles in pathological bone destruction, no role for lymphocytes in basal bone homeostasis in vivo has been described. Using immunomagnetic isolation of bone marrow (BM) B cells and B-cell precursor populations and quantitation of their OPG production by enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcriptase–polymerase chain reaction (RT-PCR), cells of the B lineage were found to be responsible for 64% of total BM OPG production, with 45% derived from mature B cells. Consistently B-cell knockout (KO) mice were found to be osteoporotic and deficient in BM OPG, phenomena rescued by B-cell reconstitution. Furthermore, T cells, through CD40 ligand (CD40L) to CD40 costimulation, promote OPG production by B cells in vivo. Consequently, T-cell–deficient nude mice, CD40 KO mice, and CD40L KO mice display osteoporosis and diminished BM OPG production. Our data suggest that lymphocytes are essential stabilizers of basal bone turnover and critical regulators of peak bone mass in vivo.

Abstract

Trabecular indices, including TV, BV, BV/TV, Tb. Th., Tb. Sp., Tb. N., SMI, and Conn. D., and the cortical indices Co. Th. and Co. Vol. were computed from μCT scans. The data are presented as the mean ± SD of 8 mice per group. Percentage change between WT and KO mice is indicated as “% change.” Asterisks indicate significantly different results (P ≤ .05).

Trabecular indices, including TV, BV, BV/TV, Tb. Th., Tb. Sp., Tb. N., SMI, and Conn. D., and the cortical indices Co. Th. and Co. Vol. were computed from μCT scans. The data are presented as the mean ± SD of 8 mice per group. Percentage change between WT and nude mice is indicated as “% change.” Asterisks indicate significantly different results (P ≤ .05).

Trabecular indices, including TV, BV, BV/TV, Tb. Th., Tb. Sp., Tb. N., SMI, and Conn. D., and the cortical indices Co. Th. and Co. Vol. were computed from μCT scans of CD40 KO mice and their respective WT controls. The data are presented as the mean ± SD. Percentage change between WT and KO mice is indicated as “% change.” Asterisks indicate significantly different results (P ≤ .05); n = 12 WT and 13 CD40 KO mice.

Trabecular indices, including TV, BV, BV/TV, Tb. Th., Tb. Sp., Tb. N., SMI, and Conn. D., and the cortical indices Co. Th. and Co. Vol. were computed from μCT scans of CD40 and CD40L KO mice and their respective WT controls. The data are presented as the mean ± SD. Percentage change between WT and KO mice is indicated as “% change.” Asterisks indicate significantly different results (P ≤ .05); n = 11 mice per group for CD40L KO and WT mice.

Acknowledgments

This work was supported by a grant from the National Osteoporosis Foundation (M.N.W.), National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) ({"type":"entrez-nucleotide","attrs":{"text":"DK067389","term_id":"187695169","term_text":"DK067389"}}DK067389) (M.N.W.), National Cancer Institute (NCI) (U54CA119338) (M.N.W.), University Research Committee of Emory University (M.N.W.), a T32 training award from the NIDDK ({"type":"entrez-nucleotide","attrs":{"text":"DK007298","term_id":"187708774","term_text":"DK007298"}}DK007298) (Y.L.), and partial financial support from Dr Dwight A. Towler (Washington University) (G.T.).

The authors thank Dr Roberto Pacifici for critical reading of the manuscript and Dr Michaela Robbie-Ryan for helpful discussions. We thank Dr Dwight A. Towler for generous access to the μCT instrument of the Division of Bone and Mineral Diseases at Washington University.

Acknowledgments

Footnotes

The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked “advertisement” in accordance with 18 USC section 1734.

Footnotes

References

  • 1. Khosla SMinireview: the OPG/RANKL/RANK system. Endocrinology. 2001;142:5050–5055.[PubMed][Google Scholar]
  • 2. Teitelbaum SLBone resorption by osteoclasts. Science. 2000;289:1504–1508.[PubMed][Google Scholar]
  • 3. Simonet WS, Lacey DL, Dunstan CR, et al Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell. 1997;89:309–319.[PubMed][Google Scholar]
  • 4. Bucay N, Sarosi I, Dunstan CR, et al Osteoprotegerin-deficient mice develop early onset osteoporosis and arterial calcification. Genes Dev. 1998;12:1260–1268.[Google Scholar]
  • 5. Mizuno A, Amizuka N, Irie K, et al Severe osteoporosis in mice lacking osteoclastogenesis inhibitory factor/osteoprotegerin. Biochem Biophys Res Commun. 1998;247:610–615.[PubMed][Google Scholar]
  • 6. Suda T, Takahashi N, Udagawa N, Jimi E, Gillespie MT, Martin TJModulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev. 1999;20:345–357.[PubMed][Google Scholar]
  • 7. Takai H, Kanematsu M, Yano K, et al Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastogenesis inhibitory factor by bone marrow stromal cells. J Biol Chem. 1998;273:27091–27096.[PubMed][Google Scholar]
  • 8. Rifas L, Arackal S, Weitzmann MNInflammatory T cells rapidly induce differentiation of human bone marrow stromal cells into mature osteoblasts. J Cell Biochem. 2003;88:650–659.[PubMed][Google Scholar]
  • 9. Hofbauer LC, Khosla S, Dunstan CR, Lacey DL, Spelsberg TC, Riggs BLEstrogen stimulates gene expression and protein production of osteoprotegerin in human osteoblastic cells. Endocrinology. 1999;140:4367–4370.[PubMed][Google Scholar]
  • 10. Hofbauer LC, Khosla S, Dunstan CR, Lacey DL, Boyle WJ, Riggs BLThe roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption. J Bone Miner Res. 2000;15:2–12.[PubMed][Google Scholar]
  • 11. Cenci S, Toraldo G, Weitzmann MN, et al Estrogen deficiency induces bone loss by increasing T cell proliferation and lifespan through IFN-gamma-induced class II transactivator. Proc Natl Acad Sci U S A. 2003;100:10405–10410.[Google Scholar]
  • 12. Cenci S, Weitzmann MN, Roggia C, et al Estrogen deficiency induces bone loss by enhancing T-cell production of TNF-alpha. J Clin Invest. 2000;106:1229–1237.[Google Scholar]
  • 13. Roggia C, Gao Y, Cenci S, et al Up-regulation of TNF-producing T cells in the bone marrow: a key mechanism by which estrogen deficiency induces bone loss in vivo. Proc Natl Acad Sci U S A. 2001;98:13960–13965.[Google Scholar]
  • 14. Kong YY, Feige U, Sarosi I, et al Activated T cells regulate bone loss and joint destruction in adjuvant arthritis through osteoprotegerin ligand. Nature. 1999;402:304–309.[PubMed][Google Scholar]
  • 15. Kawai T, Matsuyama T, Hosokawa Y, et al B and T lymphocytes are the primary sources of RANKL in the bone resorptive lesion of periodontal disease. Am J Pathol. 2006;169:987–998.[Google Scholar]
  • 16. Grcevic D, Lee SK, Marusic A, Lorenzo JADepletion of CD4 and CD8 T lymphocytes in mice in vivo enhances 1,25- dihydroxyvitamin D3-stimulated osteoclast-like cell formation in vitro by a mechanism that is dependent on prostaglandin synthesis. J Immunol. 2000;165:4231–4238.[PubMed][Google Scholar]
  • 17. John V, Hock JM, Short LL, Glasebrook AL, Galvin RJA role for CD8+ T lymphocytes in osteoclast differentiation in vitro. Endocrinology. 1996;137:2457–2463.[PubMed][Google Scholar]
  • 18. Choi Y, Woo KM, Ko SH, et al Osteoclastogenesis is enhanced by activated B cells but suppressed by activated CD8(+) T cells. Eur J Immunol. 2001;31:2179–2188.[PubMed][Google Scholar]
  • 19. Toraldo G, Roggia C, Qian WP, Pacifici R, Weitzmann MNIL-7 induces bone loss in vivo by induction of receptor activator of nuclear factor kappa B ligand and tumor necrosis factor alpha from T cells. Proc Natl Acad Sci U S A. 2003;100:125–130.[Google Scholar]
  • 20. Manabe N, Kawaguchi H, Chikuda H, et al Connection between B lymphocyte and osteoclast differentiation pathways. J Immunol. 2001;167:2625–2631.[PubMed][Google Scholar]
  • 21. Heider U, Langelotz C, Jakob C, et al Expression of receptor activator of nuclear factor kappaB ligand on bone marrow plasma cells correlates with osteolytic bone disease in patients with multiple myeloma. Clin Cancer Res. 2003;9:1436–1440.[PubMed][Google Scholar]
  • 22. Giuliani N, Colla S, Sala R, et al Human myeloma cells stimulate the receptor activator of nuclear factor-kappa B ligand (RANKL) in T lymphocytes: a potential role in multiple myeloma bone disease. Blood. 2002;100:4615–4621.[PubMed][Google Scholar]
  • 23. Giuliani N, Colla S, Morandi F, et al Myeloma cells block RUNX2/CBFA1 activity in human bone marrow osteoblast progenitors and inhibit osteoblast formation and differentiation. Blood. 2005;106:2472–2483.[PubMed][Google Scholar]
  • 24. Miyaura C, Onoe Y, Inada M, et al Increased B-lymphopoiesis by interleukin 7 induces bone loss in mice with intact ovarian function: similarity to estrogen deficiency. Proc Natl Acad Sci U S A. 1997;94:9360–9365.[Google Scholar]
  • 25. Masuzawa T, Miyaura C, Onoe Y, et al Estrogen deficiency stimulates B lymphopoiesis in mouse bone marrow. J Clin Invest. 1994;94:1090–1097.[Google Scholar]
  • 26. Erlandsson MC, Jonsson CA, Islander U, Ohlsson C, Carlsten HOestrogen receptor specificity in oestradiol-mediated effects on B lymphopoiesis and immunoglobulin production in male mice. Immunology. 2003;108:346–351.[Google Scholar]
  • 27. Sato T, Shibata T, Ikeda K, Watanabe KGeneration of bone-resorbing osteoclasts from B220+ cells: its role in accelerated osteoclastogenesis due to estrogen deficiency. J Bone Miner Res. 2001;16:2215–2221.[PubMed][Google Scholar]
  • 28. Horowitz MC, Xi Y, Pflugh DL, et al Pax5-deficient mice exhibit early onset osteopenia with increased osteoclast progenitors. J Immunol. 2004;173:6583–6591.[PubMed][Google Scholar]
  • 29. Weitzmann MN, Cenci S, Haug J, Brown C, DiPersio J, Pacifici RB lymphocytes inhibit human osteoclastogenesis by secretion of TGFbeta. J Cell Biochem. 2000;78:318–324.[PubMed][Google Scholar]
  • 30. Chenu C, Pfeilschifter J, Mundy GR, Roodman GDTransforming growth factor beta inhibits formation of osteoclast-like cells in long-term human marrow cultures. Proc Natl Acad Sci U S A. 1988;85:5683–5687.[Google Scholar]
  • 31. Hughes DE, Dai A, Tiffee JC, Li HH, Mundy GR, Boyce BFEstrogen promotes apoptosis of murine osteoclasts mediated by TGF-beta. Nat Med. 1996;2:1132–1136.[PubMed][Google Scholar]
  • 32. Thirunavukkarasu K, Miles RR, Halladay DL, et al. Stimulation of osteoprotegerin (OPG) gene expression by transforming growth factor-beta (TGF-beta). Mapping of the OPG promoter region that mediates TGF-beta effects. J Biol Chem. 2001;276:36241–36250.[PubMed]
  • 33. Klausen B, Hougen HP, Fiehn NEIncreased periodontal bone loss in temporarily B lymphocyte-deficient rats. J Periodontal Res. 1989;24:384–390.[PubMed][Google Scholar]
  • 34. Choi Y, Kim JJB cells activated in the presence of Th1 cytokines inhibit osteoclastogenesis. Exp Mol Med. 2003;35:385–392.[PubMed][Google Scholar]
  • 35. Yun TJ, Chaudhary PM, Shu GL, et al OPG/FDCR-1, a TNF receptor family member, is expressed in lymphoid cells and is up-regulated by ligating CD40. J Immunol. 1998;161:6113–6121.[PubMed][Google Scholar]
  • 36. Grewal IS, Flavell RACD40 and CD154 in cell-mediated immunity. Annu Rev Immunol. 1998;16:111–135.[PubMed][Google Scholar]
  • 37. Wuthrich M, Fisette PL, Filutowicz HI, Klein BSDifferential requirements of T cell subsets for CD40 costimulation in immunity to blastomyces dermatitidis. J Immunol. 2006;176:5538–5547.[PubMed][Google Scholar]
  • 38. Gao Y, Qian WP, Dark K, et al Estrogen prevents bone loss through transforming growth factor beta signaling in T cells. Proc Natl Acad Sci U S A. 2004;101:16618–16623.[Google Scholar]
  • 39. Livak KJ, Schmittgen TDAnalysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods. 2001;25:402–408.[PubMed][Google Scholar]
  • 40. Beamer WG, Donahue LR, Rosen CJ, Baylink DJGenetic variability in adult bone density among inbred strains of mice. Bone. 1996;18:397–403.[PubMed][Google Scholar]
  • 41. Kitamura D, Roes J, Kuhn R, Rajewsky KA B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin mu chain gene. Nature. 1991;350:423–426.[PubMed][Google Scholar]
  • 42. Uchida J, Lee Y, Hasegawa M, et al Mouse CD20 expression and function. Int Immunol. 2004;16:119–129.[PubMed][Google Scholar]
  • 43. Colovai AI, Giatzikis C, Ho EK, et al Flow cytometric analysis of normal and reactive spleen. Mod Pathol. 2004;17:918–927.[PubMed][Google Scholar]
  • 44. Fluckiger AC, Li Z, Kato RM, et al Btk/Tec kinases regulate sustained increases in intracellular Ca2+ following B-cell receptor activation. EMBO J. 1998;17:1973–1985.[Google Scholar]
  • 45. Yamada Y, Ando F, Niino N, Shimokata HAssociation of polymorphisms of the osteoprotegerin gene with bone mineral density in Japanese women but not men. Mol Genet Metab. 2003;80:344–349.[PubMed][Google Scholar]
  • 46. Langdahl BL, Carstens M, Stenkjaer L, Eriksen EFPolymorphisms in the osteoprotegerin gene are associated with osteoporotic fractures. J Bone Miner Res. 2002;17:1245–1255.[PubMed][Google Scholar]
  • 47. Rammensee HG, Falk K, Rotzschke OPeptides naturally presented by MHC class I molecules. Annu Rev Immunol. 1993;11:213–244.[PubMed][Google Scholar]
  • 48. Grossman Z, Paul WESelf-tolerance: context dependent tuning of T cell antigen recognition. Semin Immunol. 2000;12:197–203.[PubMed][Google Scholar]
  • 49. Sakaguchi S, Sakaguchi NRegulatory T cells in immunologic self-tolerance and autoimmune disease. Int Rev Immunol. 2005;24:211–226.[PubMed][Google Scholar]
  • 50. Eghbali-Fatourechi G, Khosla S, Sanyal A, Boyle WJ, Lacey DL, Riggs BLRole of RANK ligand in mediating increased bone resorption in early postmenopausal women. J Clin Invest. 2003;111:1221–1230.[Google Scholar]
  • 51. Sezer O, Heider U, Jakob C, Eucker J, Possinger KHuman bone marrow myeloma cells express RANKL. J Clin Oncol. 2002;20:353–354.[PubMed][Google Scholar]
  • 52. Wortis HH, Teutsch M, Higer M, Zheng J, Parker DCB-cell activation by crosslinking of surface IgM or ligation of CD40 involves alternative signal pathways and results in different B-cell phenotypes. Proc Natl Acad Sci U S A. 1995;92:3348–3352.[Google Scholar]
  • 53. Phipps RP, Koumas L, Leung E, Reddy SY, Blieden T, Kaufman JThe CD40-CD40 ligand system: a potential therapeutic target in atherosclerosis. Curr Opin Investig Drugs. 2001;2:773–777.[PubMed][Google Scholar]
  • 54. Ahdjoudj S, Lasmoles F, Holy X, Zerath E, Marie PJTransforming growth factor beta2 inhibits adipocyte differentiation induced by skeletal unloading in rat bone marrow stroma. J Bone Miner Res. 2002;17:668–677.[PubMed][Google Scholar]
  • 55. Pearson TC, Trambley J, Odom K, et al Anti-CD40 therapy extends renal allograft survival in rhesus macaques. Transplantation. 2002;74:933–940.[PubMed][Google Scholar]
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.