Abstract
It is known that two major cytotoxins, lymphotoxin (LT) and tumour necrosis factor (TNF), are produced by T cells and macrophages, respectively. We report in this paper that a human T-cell hybridoma, AC5-8, produces TNF in addition to LT depending on the kind of stimuli used. When phorbol myristate acetate (PMA) and concanavalin A (Con A) or PMA alone were used to induce AC5-8 cells to secrete a cytotoxin, we found that the cytotoxin secreted was antigenically indistinguishable from LT. On the other hand, when AC5-8 cells were activated by PMA and A23187, they secreted another cytotoxin which was antigenically indistinguishable from TNF, in addition to LT. These two cytotoxins could be separated by affinity chromatography on a column of Con A-Sepharose. A cytotoxin, which did not bind to the Con A-Sepharose column and showed about 70-95% of the total cytotoxic activity secreted from AC5-8 cells stimulated with PMA and A23187, was found to be antigenically identical to TNF. The other cytotoxin, which bound to the Con A-Sepharose column and showed only 5-30% of the total activity induced by PMA and A23187, was found to be antigenically identical to LT.
MeSH Terms
Antibody Specificity
Calcimycin/pharmacology
Concanavalin A/pharmacology
Glycoproteins/biosynthesis
Growth Inhibitors/biosynthesis
Humans
Hybridomas/metabolism
Immune Sera/immunology
Lymphotoxin-alpha/biosynthesis
T-Lymphocytes/metabolism
Tetradecanoylphorbol Acetate/pharmacology
Tumor Necrosis Factor-alpha
Chemicals
Glycoproteins
Growth Inhibitors
Immune Sera
Lymphotoxin-alpha
Tumor Necrosis Factor-alpha
Concanavalin A
Calcimycin
Tetradecanoylphorbol Acetate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kobayashi Y
Asada M
Osawa T
References (10)
10 references, click to expand
-
Human T cell hybridomas producing lymphokines. I. Establishment and characterization of human T cell hybridomas producing lymphotoxin and migration inhibitory factor.
J Immunol. 1982 Jun;128(6):2714-8
PMID: 7042845
-
Human T-cell hybridomas producing lymphokines. II. Enhancement of lymphotoxin secretion from human T-cell hybridomas by phorbol myristate acetate.
Cell Immunol. 1983 Apr 1;77(1):150-60
PMID: 6340837
-
Human lymphotoxin. Production by a lymphoblastoid cell line, purification, and initial characterization.
J Biol Chem. 1984 Jan 10;259(1):686-91
PMID: 6608523
-
Cloning and expression of cDNA for human lymphotoxin, a lymphokine with tumour necrosis activity.
Nature. 1984 Dec 20-1985 Jan 2;312(5996):721-4
PMID: 6334807
-
Cloning and expression of human lymphotoxin mRNA derived from a human T cell hybridoma.
J Biochem. 1986 Sep;100(3):727-33
PMID: 3536896
-
Primary structure of human lymphotoxin derived from 1788 lymphoblastoid cell line.
J Biol Chem. 1985 Feb 25;260(4):2334-44
PMID: 3882692
-
Human tumor necrosis factor. Production, purification, and characterization.
J Biol Chem. 1985 Feb 25;260(4):2345-54
PMID: 3871770
-
Effect of interleukin 2, interferon-gamma, and mitogens on the production of tumor necrosis factors alpha and beta.
J Immunol. 1985 Oct;135(4):2492-7
PMID: 3928750
-
Activation of IL 1-dependent and IL 1-independent T cell lines by calcium ionophore and phorbol ester.
J Immunol. 1986 Feb 1;136(3):1033-7
PMID: 3079793
-
Human tumour necrosis factor: precursor structure, expression and homology to lymphotoxin.
Nature. 1984 Dec 20-1985 Jan 2;312(5996):724-9
PMID: 6392892