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Expression of Cytokine Genes, Cytokine Receptor Genes, and Transcription Factors in Cultured Hodgkin and Reed-Sternberg Cells¹

Department of Internal Medicine I, University of Freiburg Medical Center, Freiburg [H-J. G., M. A. B., R. B., R. H. M., F. H.], and German Collection of Microorganisms and Cell Cultures [H-G. D.], Braunschweig, Germany

In the present study, we show by Northern blot analysis and enzyme linked immunosorbent assay that the Hodgkin's disease (HD)-derived cell lines HDLM-2 and KM-H2 express a variety of cytokine genes either constitutively or upon induction with phorbol ester 12-O-tetrade-canoylphorbol-13-acetate. Cytokine genes expressed by HD-derived lines include granulocyte-macrophage colony-stimulating factor (CSF), macrophage-CSF, interleukin (IL)-1-, IL-3, IL-5, IL-6, IL-8, leukemia inhibitory factor, tumor necrosis factor-, tumor necrosis factor-ß, and transforming growth factor-ß, while transcripts and the corresponding proteins for granulocyte-CSF, IL-1-ß, IL-2, IL-4, IL-7, IL-10, and the JE/macrophage chemoattractant and activating factor gene were not detectable in cytoplasmic RNA and culture supernatants obtained from both lines. In addition, IL-2 receptor (R) p55 and macrophage-CSF R (c-fms) genes were expressed by both lines. HDLM-2, but not KM-H2 cells, exhibited the IL-6 R p80 and the IL-2 R p75 chain. Analysis of nuclear proteins that bind to oligonucleotides containing the consensus sequences of the transcription factors activation protein 1, nuclear factor (NF) B, and NFAT 1 revealed a pattern for HD lines resembling that of activated T-cells: HDLM-2 and KM-H2 cells constitutively expressed NF binding to the NF of activated T-cells (type 1), previously described to be T-cell specific. In addition, NF B-binding proteins obtained from both lines showed, in electrophoretic mobility shift assays, the same migration pattern as T-cell-derived proteins but differed from monocyte- and B-cell-derived proteins. UV cross-linking experiments confirmed that NF B-binding proteins of M_r 85,000, 75,000, and 50,000/55,000 were detectable in nuclear extracts obtained from T-cells and both HD lines, while monocytes and B-cells displayed the M_r 50,000/55,000 and 75,000 NF B complex only. Both HD lines also constitutively expressed transcripts for c-fos and c-jun, which are involved in heterodimeric formation of the transcription factor activation protein 1, as well as for the NF B/KBF1 gene.

¹ Supported by the Deutsche Krebshilfe.

² To whom requests for reprints should be addressed, at Department of Internal Medicine I, University of Freiburg Medical Center, Hugstetter Str. 55, D-7800 Freiburg i.Br., Germany.

Received 12/10/91. Accepted 4/ 6/92.

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