TY - JOUR
T1 - Tumor necrosis factor production and receptor expression by a human malignant glioma cell line, D54-MG
AU - Bethea, John R.
AU - Yancey Gillespie, G.
AU - ^Yup Chung, I.
AU - Benveniste, Etty N.
N1 - Funding Information:
Society( E.N.B), grant BNS-8708233fr om the National Science Foundation (E.N.B.), grant AI 27290 from the National Institutes of Health (E.N.B.), grantsC A 30479a nd NS 20023f rom the NationalI nstituteso f Health (G.Y.G.) and a grant from the Preuss Foundationfo r Brain Tumor Research (G.Y.G.). J.R.B. is affiliated with the GraduateT rainingP rogramin Neuroscience.
Funding Information:
This work was supportedin part by grant RG 1954-A2 from the National Multiple Sclerosis
PY - 1990/11
Y1 - 1990/11
N2 - Human malignant gliomas possess some of the same immune-related functions as astrocytes do. For instance, they are capable of secreting various immunoregulatory molecules and expressing HLA-DR antigens on their surface. The human malignant glioma cell line, D54-MG, was used to investigate the proliferation effects of tumor necrosis factor-α (TNF-α) and the expression of specific surface receptors for TNF-α. Additionally, we were interested in examining whether D54-MG cells are capable of synthesizing and secreting biologically active TNF-α. D54-MG cells responded in a mitogenic fashion upon incubation with TNF-α for 48 h under serum-free conditions. 125I-labeled TNF-α was used in this study to investigate the expression of receptors specific for TNF-α on D54-MG cells. Scatchard analysis of our receptor binding data produced curvilinear plots indicating there are two distinct receptor sites for TNF-α. From these data, we calculated that there are approximately 3500 high affinity and 24,666 low affinity binding sites per cell. Pretreating these cells with interferon-γ (IFN-γ) resulted in a 2-fold increase in the number of high affinity binding sites and a moderate increase in the number of low affinity binding sites, with no appreciable change in binding affinity (Kd) of either site. D54-MG cells were unable to constitutively secrete TNF-α; however, upon stimulation, these cells synthesize and secrete biologically active TNF-α. Polyclonal antisera reactive with human macrophage-derived TNF-α neutralized the cytotoxicity of D54-MG-derived TNF-α, demonstrating that the cytotoxic activity was in fact due to TNF-α. Our observations indicate that TNF-α could act in an autocrine fashion to induce the proliferation of this malignant glioma cell line and that TNF-α exerts its effect by binding to specific TNF-α receptors whose expression was enhanced by IFN-γ.
AB - Human malignant gliomas possess some of the same immune-related functions as astrocytes do. For instance, they are capable of secreting various immunoregulatory molecules and expressing HLA-DR antigens on their surface. The human malignant glioma cell line, D54-MG, was used to investigate the proliferation effects of tumor necrosis factor-α (TNF-α) and the expression of specific surface receptors for TNF-α. Additionally, we were interested in examining whether D54-MG cells are capable of synthesizing and secreting biologically active TNF-α. D54-MG cells responded in a mitogenic fashion upon incubation with TNF-α for 48 h under serum-free conditions. 125I-labeled TNF-α was used in this study to investigate the expression of receptors specific for TNF-α on D54-MG cells. Scatchard analysis of our receptor binding data produced curvilinear plots indicating there are two distinct receptor sites for TNF-α. From these data, we calculated that there are approximately 3500 high affinity and 24,666 low affinity binding sites per cell. Pretreating these cells with interferon-γ (IFN-γ) resulted in a 2-fold increase in the number of high affinity binding sites and a moderate increase in the number of low affinity binding sites, with no appreciable change in binding affinity (Kd) of either site. D54-MG cells were unable to constitutively secrete TNF-α; however, upon stimulation, these cells synthesize and secrete biologically active TNF-α. Polyclonal antisera reactive with human macrophage-derived TNF-α neutralized the cytotoxicity of D54-MG-derived TNF-α, demonstrating that the cytotoxic activity was in fact due to TNF-α. Our observations indicate that TNF-α could act in an autocrine fashion to induce the proliferation of this malignant glioma cell line and that TNF-α exerts its effect by binding to specific TNF-α receptors whose expression was enhanced by IFN-γ.
KW - Glioma
KW - Interferon-γ
KW - Proliferation
KW - Tumor necrosis factor-α
KW - Tumor necrosis factor-α receptor
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U2 - 10.1016/0165-5728(90)90047-Q
DO - 10.1016/0165-5728(90)90047-Q
M3 - Article
C2 - 2172302
AN - SCOPUS:0025103845
VL - 30
SP - 1
EP - 13
JO - Journal of Neuroimmunology
JF - Journal of Neuroimmunology
SN - 0165-5728
IS - 1
ER -