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.
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.
Jung
W
W
Med. Klinik und Poliklinik, Innere Medizin I Universität des Saarlandes, Homburg, Germany.
Krueger
S
S
Renner
C
C
Gause
A
A
Sahin
U
U
Trümper
L
L
Pfreundschuh
M
M
D001483:Base Sequence; D053285:CD30 Ligand; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D015698:Genomic Library; D006801:Humans; D017730:Ki-1 Antigen / Q000737:chemistry* / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D016366:Open Reading Frames; D010835:Phytohemagglutinins; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D053285:CD30 Ligand; D017730:Ki-1 Antigen; D008562:Membrane Glycoproteins; D010835:Phytohemagglutinins; C505774:TNFSF8 protein, human; D012313:RNA; D002118:Calcium