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Tumor necrosis factor receptor-associated factors (TRAFs) are a recently established group of proteins involved in the intracellular signal transduction of several members of the tumor necrosis factor receptor (TNFR) superfamily. Recently, specific members of the TRAF family have been implicated in promoting cell survival as well as activation of the transcription factor NF-kappaB. We investigated the constitutive expression of TRAF1 and TRAF2 in Hodgkin and Reed-Sternberg (HRS) cells from archived paraffin-embedded tissues obtained from 21 patients diagnosed with classical Hodgkin's disease (HD). In a selective portion of cases, examination of HRS cells for Epstein-Barr virus (EBV)-encoded RNA was performed by in situ hybridization, and the results were compared with the magnitude of TRAF1 and TRAF2 staining. We also determined the TRAF profile in the classical HD cell lines L428, KMH2, and HS445 by Western blotting using a series of antibodies that specifically recognize the six individual TRAF family proteins (TRAF1-TRAF6). Moderate to high constitutive expression of TRAF1 and TRAF2 were found in 19 of 21 and 20 of 21 cases of classical HD, respectively. Of the remaining cases, one case showed weak expression of TRAF1, and another case showed weak expression of both proteins. No relationship was found between the staining intensity of the TRAF proteins and EBV expression in HRS cells. Strong constitutive expression of TRAF1 was also identified in the HD cell line L428, compared with the relatively weak expression observed in KMH2 and HS445. All three HD cell lines showed strong expression of TRAF2 protein and moderate, comparatively equal expression of TRAF4 and TRAF6. In contrast, TRAF3 was not expressed in the HD cell lines. Although KMH2 showed weak expression, the remaining HD cell lines also lacked TRAF5 protein. These data demonstrate that constitutive expression of TRAF1 and TRAF2 is a characteristic feature of HRS cells from both patient and cell line specimens. Furthermore, with the exception of TRAF1 expression, HRS cells from the three HD cell lines showed similar TRAF protein expression patterns. Overall, these findings demonstrate the expression of several TRAF proteins in HD. Significantly, the altered regulation of selective TRAF proteins may reflect HRS cell response to stimulation from the microenvironment and potentially contribute both to apoptosis resistance and cell maintenance of HRS cells.

Expression of the Tumor Necrosis Factor Receptor-Associated Factors (TRAFs) 1 and 2 is a Characteristic Feature of Hodgkin and Reed-Sternberg Cells