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Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1β using a Whole Blood Stimulation Assay
Published on: March 1, 2011
Production of interleukin-1 beta by periodic acid-oxidized human peripheral blood mononuclear cells
G I Malinin1, F J Hornicek, T I Malinin
1Physics Department, Georgetown University, Washington, DC 20057.
Interleukin-1 (IL-1) production by periodic acid (H5IO6)-oxidized human peripheral blood mononuclear (PBMN) cells was assessed by the thymocyte co-mitogenesis assay. Maximum IL-1 levels (approximately 1.2 U/ml) in the conditioned media of PBMN cells were registered within the first 24 hrs post-oxidation, whereas no IL-1 was detected in the media from 24 hrs control cultures. Thymocyte proliferation, driven by periodic acid-induced IL-1, was abolished by an antibody to IL-1 alpha and IL-1 beta. Quantitative analysis of IL-1-containing medium by radioimmunoassay (RIA) indicated that IL-1 beta comprised about 80% of total IL-1. Partial characterization of H5IO6-induced IL-1 beta indicated that it was identical to IL-1 produced by lipopolysaccharide-stimulated macrophages. It is concluded that oxidation of human PBMN cells by H5IO6 triggers synthesis and release of IL-1, most of which was in its IL-1 beta form.
Interleukin-1 (IL-1) production by periodic acid (H5IO6)-oxidized human peripheral blood mononuclear (PBMN) cells was assessed by the thymocyte co-mitogenesis assay. Maximum IL-1 levels (approximately 1.2 U/ml) in the conditioned media of PBMN cells were registered within the first 24 hrs post-oxidation, whereas no IL-1 was detected in the media from 24 hrs control cultures. Thymocyte proliferation, driven by periodic acid-induced IL-1, was abolished by an antibody to IL-1 alpha and IL-1 beta. Quantitative analysis of IL-1-containing medium by radioimmunoassay (RIA) indicated that IL-1 beta comprised about 80% of total IL-1. Partial characterization of H5IO6-induced IL-1 beta indicated that it was identical to IL-1 produced by lipopolysaccharide-stimulated macrophages. It is concluded that oxidation of human PBMN cells by H5IO6 triggers synthesis and release of IL-1, most of which was in its IL-1 beta form.
