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Amelioration of inflammatory arthritis by targeting the pre-ligand assembly domain of tumor necrosis factor receptors


Tumor necrosis factor (TNF)-α has an important role in the pathogenesis of autoimmune and inflammatory diseases such as rheumatoid and septic arthritis. The biological effects of TNF-α are mediated by binding to TNF receptors TNFR1 (also known as P60) or TNFR2 (also known as P80). The pre-ligand assembly domain (PLAD) is a portion of the extracellular region of TNFRs that mediates receptor-chain association essential for signaling. We found that soluble versions of PLAD, especially those derived from P60, block the biochemical effects of TNF-α in vitro and potently inhibit arthritis in animal models. Thus, targeting the PLAD may have clinical value in the treatment of human arthritis and other disorders involving receptors of the TNFR superfamily.

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Figure 1: Expression of recombinant bacterial PLAD proteins.
Figure 2: Effects of PLAD proteins in TNF-α–induced cell death.
Figure 3: Effects of P60 and P80 PLAD proteins on arthritis induced by intra-articular injection of TNF-α in BALB/c mice and bacterial CpG DNA in C3H/HeJ mice.
Figure 4: Effects of P60 and P80 PLAD proteins on CIA in DBA/1J mice.
Figure 5: TNFR expression in arthritic joints and PLAD protein inhibition of TNF-α binding and NF-κB activation.
Figure 6: P60 PLAD protein inhibits osteoclastogenesis and RANK and RANKL expression.

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We thank H. Su, C. Trageser, N. Bidere and L. Yu for experimental assistance and R. Siegel, D. Bolton and K. Sakai for critically reading the manuscript. G.-M.D. is supported by a Pharmacology Research Associate Training fellowship of the National Institute of General Medical Science. We thank O. Schwartz and the microscopy group of the National Institute of Allergy and Infectious Disease, National Institutes of Health for confocal microscopy help and J. Qin for statistical help. This work was supported by the intramural program of the National Institute of Allergy and Infectious Disease, US National Institutes of Health.

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Correspondence to Michael Lenardo.

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Supplementary information

Supplementary Fig. 1

Amino acid sequences of human PLAD-GST fusion proteins in standard single-letter code. (PDF 141 kb)

Supplementary Fig. 2

Effects of P60 PLAD protein on TNF-α–induced cell death in L929 cells. (PDF 286 kb)

Supplementary Fig. 3

Effects of GST protein and P80 PLAD protein on inflammatory arthritis. (PDF 1414 kb)

Supplementary Fig. 4

Photomicrographs of immunohistochemistry. (PDF 5160 kb)

Supplementary Fig. 5

Immunogenicity and half-life of the P60 PLAD protein. (PDF 196 kb)

Supplementary Fig. 6

PLAD proteins inhibit TNF-α–induced IκBα degradation. (PDF 462 kb)

Supplementary Fig. 7

Gel electrophoresis of 50 ng of human TNF-α immunoprecipitated (IP) with different amounts of etanercept and P60 PLAD protein. (PDF 207 kb)

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Deng, GM., Zheng, L., Ka-Ming Chan, F. et al. Amelioration of inflammatory arthritis by targeting the pre-ligand assembly domain of tumor necrosis factor receptors. Nat Med 11, 1066–1072 (2005).

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