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Demonstration of IL-1 alpha, and IL-1 beta secretion by the monocytic leukemia cell line, THP-1.
Gaffney, E V; Stoner, C R; Lingenfelter, S E; Wagner, L A.
Afiliación
  • Gaffney EV; Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
J Immunol Methods ; 122(2): 211-8, 1989 Sep 01.
Article en En | MEDLINE | ID: mdl-2507640
This study examined the secretion of IL-1 alpha and IL-1 beta by THP-1 leukemia cells following activation with mezerein and promotion of synthesis by interferon (IFN-gamma). Interleukin-1 (IL-1) was not detected by co-mitogenic thymocyte assays of crude supernates. Isoelectrofocusing of concentrated medium showed that all biologically active IL-1 migrated at a pH of 6.8-7.2, indicating that the major secreted form was IL-1 beta. Double antibody ELISA confirmed the presence of IL-1 beta, but failed to detect IL-1 alpha in isofocused fractions. Although it appeared that THP-1 cells do not secrete IL-1 alpha; an inhibitor of thymocyte response to IL-1 was present in conditioned medium, migrated in an acidic pH range and masked the expression of biologically active rIL-1 alpha and rIL-1 beta. In contrast, IL-1 alpha was detected using a cell blotting assay. This technique permitted visualization of subpicogram levels of IL-1 when secreted by cells attached to an immunoblotting paper. Cell blotting showed that a greater proportion of attached cells incubated for 24 h in medium containing mezerein and IFN-gamma secreted IL-1 than cells in control medium. In conclusion, the amount of immunoreactive or biologically active IL-1 alpha secreted by stimulated THP-1 cells appeared to be much lower than that reported for human peripheral blood monocytes.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Leucemia Monocítica Aguda / Interleucina-1 / Diterpenos Límite: Humans Idioma: En Revista: J Immunol Methods Año: 1989 Tipo del documento: Article Pais de publicación: Países Bajos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Leucemia Monocítica Aguda / Interleucina-1 / Diterpenos Límite: Humans Idioma: En Revista: J Immunol Methods Año: 1989 Tipo del documento: Article Pais de publicación: Países Bajos