Opposite effects of the CD30 ligand are not due to CD30 mutations: Results from cDNA cloning and sequence comparison of the CD30 antigen from different sources

doi:10.1016/0161-5890(94)90051-5

Molecular Immunology

Volume 31, Issue 17, December 1994, Pages 1329-1334

https://doi.org/10.1016/0161-5890(94)90051-5 Get rights and content

Abstract

CD30L, the ligand for the activation antigen CD30, is a member of the tumor necrosis factor family of cytokines. Binding of CD30L to CD30, which is a member of the nerve growth factor/tumor necrosis factor receptor family, induces proliferation in peripheral blood lymphocytes and Hodgkin's derived cell lines with a T-cell phenotype such as HDLM-2 and L540, while cell lines derived from anaplastic large cell lymphomas, such as Karpas 299, undergo cell death. In order to investigate whether mutations of the CD30 antigen are responsible for these opposite effects, we cloned the open reading frame of CD30 cDNAs from the cell lines L540 and Karpas 299 and from peripheral blood lymphocytes by reverse transcriptase polymerase chain reaction. Sequencing of independent plasmid clones revealed that these cells have a silent transition (A→G) at position 771 of the open reading frame compared to the published sequence derived from the HTLV-1⁺ cell line HUT-102. As published data have shown that crosslinking of CD30 induces an elevation of cytosolic free calcium ([Ca²⁺]_i) in TCR positive Jurkat cells, we have analysed the effect of crosslinking of CD30 on L540 and Karpas 299 cells. No elevations of [Ca²⁺]_i have been observed in these cell lines after crosslinking of CD30 with HRS-4. We conclude (i) that the different functional effects of CD30 in PBL, L540 and Karpas 299 are not due to differences in the primary structure of the receptor; and (ii) that the different responses observed upon engagement with CD30L for the cell lines L540 and Karpas 299 do not correlate with differences in mobilization of [Ca²⁺]_i after crosslinking of CD30.

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Cited by (11)

CD30-induced signaling is absent in Hodgkin's cells but present in anaplastic large cell lymphoma cells
2008, American Journal of Pathology
Citation Excerpt :
Because CD30 ligand and Ki-1 mAb induced identical effects (data not shown), we performed CD30 stimulation with Ki-1 mAb, and control treatment with IgG3, mAb binding was verified by flow cytometric analysis. All cell lines express wild-type CD30 (B. Hirsch and H. Dürkop, unpublished data).20 The pan-caspase inhibitor Z-VAD-FMK (Calbiochem, Schwalbach, Germany) was used at 50 μmol/L.
High CD30 expression in classical Hodgkin's lymphoma and anaplastic large cell lymphoma (ALCL) suggests an important pathogenic role of this cytokine receptor. To test this hypothesis, we investigated CD30 signaling in Hodgkin's and ALCL cell lines by different approaches: 1) CD30 stimulation, 2) CD30 down-regulation, and 3) a combination of both. The effects were determined at the RNA (microarray and real-time quantitative RT-PCR), protein (electrophoretic mobility shift analysis, immunoblot, and flow cytometry), and cellular/functional (proliferation and apoptosis) levels. We demonstrate that Hodgkin's cells are virtually CD30 unresponsive. Neither CD30 stimulation nor CD30 silencing of Hodgkin's cells had any significant effect. In contrast, CD30 stimulation of ALCL cells activated nuclear transcription factor-κB (NF-κB), induced major transcriptional changes, and decreased proliferation. These effects could be abrogated by down-regulation of CD30. Stimulation of CD30 in ALCL cells, stably transfected with a dominant-negative NF-κB inhibitor, induced pronounced caspase activation and massive apoptosis. Our data indicate that 1) CD30 signaling is not effective in Hodgkin's cell lines but is effective in ALCL cell lines, 2) CD30 is probably not significantly involved in the pathogenesis of classical Hodgkin's lymphoma, and 3) CD30 stimulation triggers two competing effects in ALCL cells, namely activation of caspases and NF-κB-mediated survival. These data suggest that CD30-targeted therapy in ALCL should be combined with NF-κB inhibitors to induce effective cell killing.
Systemic and primary cutaneous anaplastic large cell lymphomas
2003, Seminars in Hematology
Citation Excerpt :
CD30 activation leads to growth arrest in cell lines from ALK-positive ALCL, but causes proliferation in some Hodgkin’s cell lines and some cutaneous ALCL cell lines.38,81,84,95,100 CD30 mutations are not responsible for the different cellular responses to CD30 activation, since a Hodgkin’s line (L540) that proliferates in response to CD30 and an ACLC line (Karpas) that undergoes growth arrest in response to CD30, both have wild-type CD30 genes.51 Little is known about CD30 signaling in ALCL, and some of the data appear to be contradictory.
Anaplastic large cell lymphoma (ALCL) is a neoplasm of activated lymphocytes, commonly expressing T-cell antigens and cytotoxic proteins. Histopathology reveals distinctive infiltration of sinuses and paracortical T-cell-rich regions of lymph nodes by tumor cells which have abundant cytoplasm and large irregular/convoluted nuclei, and which are frequently multinucleated with prominent nucleoli. ALCL often presents in advanced clinical stages with B symptoms; extranodal disease occurs in 40% of patients. The pathogenesis of systemic ALCL is linked to phosphorylation of a tyrosine kinase (ALK) resulting in unregulated growth of affected lymphoid cells. ALK is activated through chromosomal translocations/inversions with any of several partner genes, most commonly nucleophosmin (NPM). Downstream signal transduction pathway(s) are not fully defined but appear to involve phospholipase Cγ, phosphatidylinositol (PI)3K/Akt, and STAT-3 and STAT-5 proteins. Primary cutaneous ALCL appears to have a different pathogenesis and better prognosis than does systemic ALCL, presenting as one or more skin tumors, usually localized. Excision or local irradiation is usually effective treatment. A clinically benign variant of primary cutaneous ALCL is lymphomatoid papulosis (LyP), characterized by recurrent crops of papules/nodules up to 2 cm in diameter which undergo spontaneous regression. LyP is managed by observation, ultraviolet light therapy, or low-dose methotrexate. LyP patients have a predisposition to develop malignant lymphomas, including Hodgkin’s lymphoma, mycosis fungoides, and non-Hodgkin’s lymphoma, by as yet unknown mechanisms. The prognosis for patients with LyP is otherwise excellent.
High-level expression of BCL3 differentiates t(2;5)(p23;q35)-positive anaplastic large cell lymphoma from Hodgkin disease
2003, Blood
Citation Excerpt :
The previous studies and our present work suggest that t(2;5)+ALCL cells are characterized by the presence of NPM/ALK fusion protein predominantly in the cytoplasm, expression of the CD30 molecules on the cell surface, and high-level expression of Bcl-3 in the nucleus. Because there is no link between the signaling cascades activated by NPM/ALK and those by CD30 stimulation, t(2;5) leading to the generation of NPM/ALK may occur at a stage of T-cell differentiation that expresses CD30, and therefore CD30 expression itself may not be involved in the development of ALCL9; indeed, ALCL as well as HD cells carry the CD30 gene with a germ-line configuration.50 In contrast, theBCL3gene of ALCL cells showed genetic and epigenetic modifications, both of which are likely associated with oncogenesis.
Anaplastic large cell lymphoma (ALCL) with t(2;5)(p23;q35) and Hodgkin disease (HD) share many cellular features, including expression of CD30. We compared gene expression profiles of 4 ALCL (Karpas 299, SU-DHL-1, DEL, SR-786) and 3 HD cell lines and found thatBCL3,which encodes a nuclear protein belonging to the IκB family of inhibitors of nuclear factor–κB (NF-κB) transcriptional factors, was expressed at higher levels in ALCL than HD. Northern and Western blotting analyses confirmed the high-level expression ofBCL3in ALCL at both mRNA and protein levels. We established a real-time reverse transcriptase–mediated polymerase chain reaction assay to measure theBCL3mRNA level and found a predominant level ofBCL3expression in t(2;5)⁺ALCL; the levels of cell lines and clinical materials were comparable to or higher than that of a B-cell chronic lymphocytic leukemia carrying t(14;19)(q32;q13). Southern blotting and fluorescence in situ hybridization disclosed that theBCL3gene copies were amplified in SU-DHL-1, whereas Karpas 299 carried 4BCL3gene loci. TheBCL3gene contains 2 cytosine-guanine dinucleotide (CpG) islands, and the intragenic 3′ CpG was entirely demethylated in SU-DHL-1 and DEL. In contrast to HD, in which NF-κB was constitutively activated, ALCL cells consistently showed (p50)₂homodimer binding activity on electrophoretic mobility shift assay. It is suggested that the high-level nuclear Bcl-3 sequestrates the (p50)₂homodimer to the nucleus, which may account for the contradictory effect of CD30 stimulation on ALCL and HD. We propose thatBCL3is overexpressed by genetic and epigenetic modifications, potentially contributing to the development of t(2;5)⁺ALCL.
The tyrosine kinase NPM-ALK, associated with anaplastic large cell lymphoma, binds the intracellular domain of the surface receptor CD30 but is not activated by CD30 stimulation
1999, Experimental Hematology
The heterogenous group of anaplastic large cell lymphomas (ALCLs) is characterized by expression of the Ki-1/CD30 antigen, a member of the tumor necrosis factor receptor superfamily. About 40 to 50% of cases diagnosed as ALCL contain a specific chromosomal rearrangement, t(2;5)(p23;q35), resulting in expression of the chimeric tyrosine kinase NPM-ALK. As NPM-ALK–positive lymphomas define a distinct subtype within the group of ALCL, the chimeric protein might be responsible for certain pathogenetic and clinicopathologic characteristics. To better elucidate the function of NPM-ALK, we investigated a possible mechanism for regulation of its activity. We demonstrate that NPM-ALK specifically binds to the intracellular domain of the cytokine receptor CD30. In vitro binding assays revealed that the ALK portion of NPM-ALK mediates interaction of the two proteins. Stimulation of the CD30 receptor by cross-linking with immobilized anti-CD30 antibody results in complete growth inhibition of Karpas 299, an NPM-ALK–positive ALCL cell line, but does not alter proliferation of HDLM-2, a Hodgkin's lymphoma-derived cell line lacking t(2;5). Western blot analysis of coimmunoprecipitated CD30 and NPM-ALK proteins from stimulated Karpas 299 cells showed that the interaction of the proteins is not modified by stimulation. Activation of CD30 neither enhanced NPM-ALK activity measured by autophosphorylation of the chimeric tyrosine kinase nor phosphorylation of phospholipase C-γ, an NPM-ALK substrate. We conclude that NPM-ALK is not stimulated by CD30 activation, but exists as a constitutively hyperactivated protein. Interaction with CD30 may extend the subcellular localization of NPM-ALK to the microenvironment of membrane-associated proteins.
A review of CD30 expression in cutaneous neoplasms
2021, Journal of Cutaneous Pathology
Galectin-1-mediated cell death is increased by CD30-induced signaling in anaplastic large cell lymphoma cells but not in Hodgkin lymphoma cells
2012, Laboratory Investigation

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Opposite effects of the CD30 ligand are not due to CD30 mutations: Results from cDNA cloning and sequence comparison of the CD30 antigen from different sources

Abstract

Mutation Res.

Analyt. Biochem.

Cell

Blood

Blood

J. biol. Chem.

Cell

Cell

Blood

Human macrophages can express the Hodgkin's cellassociated Ki-1 (CD30)

Am. J. Pathol.