A simple assay for tumor necrosis factor using HEp-2 target cells.
We developed a sensitive bioassay system for the determination of tumor necrosis factor-alpha (TNF) using HEp-2 adherent human epipharynx carcinoma cells as targets. TNF from separated human monocytes was triggered by lipopolysaccharide (LPS). In a 24 hr 3H-thymidine incorporation assay, TNF-like activity was seen to reproducibly destroy radiolabeled target cells, i.e., inhibits thymidine incorporation and causes detachment of adherent HEp-2 cells. HEp-2 cells were insensitive to human interleukin-1 (IL-1) and interleukin-2 (IL-2). In contrast, human interferon-alpha and gamma were also cytotoxic for target cells. Monocyte supernatants stimulated by LPS, however, failed to contain detectable amounts of interferons.
Müzes
G
G
2nd Department of Medicine, Semmelweis University, Budapest, Hungary.
Vien
C V
CV
Gonzalez-Cabello
R
R
Gergely
P
P
Feher
J
J
D001681:Biological Assay / Q000379:methods*; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D006801:Humans; D007372:Interferons / Q000032:analysis / Q000494:pharmacology; D007378:Interleukins / Q000494:pharmacology; D007822:Laryngeal Neoplasms / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation; D011994:Recombinant Proteins / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism
D007378:Interleukins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007372:Interferons