Alteration of the cellular response to interleukin-1 beta by SV40 large T antigen in rheumatoid synovial fibroblasts.
The large T antigen of SV40 (LT) has been widely used to immortalize primary cells for various studies. In this study, synovial fibroblasts of a patient from rheumatoid arthritis (RA) were transformed with LT gene to analyze the effect of SV40-mediated transformation on the production of cytokines, such as IL-6, IL-8, and GM-CSF, that are under the control of interleukin-1 beta (IL-1 beta), a physiological inducer of nuclear factor kappa B (NF-kappa B). It was noted that the basal levels of GM-CSF and IL-8 were upregulated, whereas that of IL-6 was downregulated. Moreover, the extents of induction of these cytokines in response to IL-1 beta were markedly downregulated in synovial fibroblasts transformed by LT as compared from parental cells. Although IL-1 beta could translocate NF-kappa B to the nucleus in all cells, some of the transformed cells exhibited nuclear translocation of NF-kappa B even before the stimulation with IL-1 beta, suggesting that transformation of LT resulted in the constitutive activation of NF-kappa B, either directly or indirectly. In order to examine whether LT downregulate the kappa B-dependent gene expression, we performed the transient luciferase gene expression assay. We found that cotransfection of LT did not downregulate the kappa B-dependent gene expression that was stimulated with L-1 beta. These observations suggest that the apparent inhibitory effect of LT on the IL-1-induced expression of cytokines may not be through its direct action on the NF-kappa B transactivation.
Asamitsu
K
K
Department of Molecular Genetics, Nagoya City University Medical School, Japan.
Sakurada
S
S
Mashiba
K
K
Nakagawa
K
K
Torikai
K
K
Onozaki
K
K
Okamoto
T
T
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000502:physiology*; D001172:Arthritis, Rheumatoid / Q000378:metabolism / Q000473:pathology; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D016207:Cytokines / Q000187:drug effects / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D016228:Endothelial Growth Factors / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008222:Lymphokines / Q000187:drug effects / Q000378:metabolism / Q000494:pharmacology; D008875:Middle Aged; D011994:Recombinant Proteins / Q000494:pharmacology; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D013583:Synovial Membrane / Q000166:cytology / Q000378:metabolism; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000952:Antigens, Polyomavirus Transforming; D016207:Cytokines; D004274:DNA, Recombinant; D016228:Endothelial Growth Factors; D007375:Interleukin-1; D015850:Interleukin-6; D016209:Interleukin-8; D008222:Lymphokines; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor