Dear Anne, we got experience in using the secretion assay on human PBMC and T cells for the cytokines IL-4, IFN-y and IL-10. The power of this procedure is detecting and isolating very low percentages of living cytokine positive cells from heterogeneous populations. Even when the initial staining of the cells seems to be negative it is possible to isolate a substantial number of secreting cells by using the Miltenyi magnetic particle protocol (in contrast to the not stimulated population). We also used the protocol the other way around. Knowing IL-10 was produced upon stimulating PBMC with an 'antigen', we would like to know which cell type was responsible. By staining the three mayor lineage positive cells (CD3, CD14 and CD19) and using the IL-10 secretion assay. It turned out to be the monocytes instead of the 'expected' T cells that produced the IL-10. Kind regards, Joost ______________________________________________________________ J.H.N. Schuitemaker Senior Research Technician Dept. Cell Biology & Histology Cellular Immunology Group AMC, University of Amsterdam Meibergdreef 9, L3-351 PObox 22700 1105 AZ Amsterdam 1100 DE Amsterdam The Netherlands Interested? VISIT: (intranet) http://m3.amc.uva.nl/celbiologie (internet) http://www.amc.uva.nl/ and (internet) http://home.wanadoo.nl/flowcytometry/ Telephone: +31 (0)20 5664960 Fax: +31 (0)20 6974156 E-mail (privé): J.H.N.Schuitemaker@consunet.nl -----Original Message----- From: Husebekk Anne [mailto:anne.husebekk@rito.no] Sent: zaterdag 24 maart 2001 14:53 To: cyto-inbox Subject: cytokins Ikke sensitiv Dear flowers! I wonder if anybody has experience with the cytokine secretion assay system from Milteny -detection of cell-secreted IFN-gamma,IL-4, IL-10, IL-2? Anne Husebekk MD, PhD Dept. of Immunology and Transfusion Medicine University Hospital of Tromsoe, Norway Phone:+47 776 26299 Fax:+47 776 26304 anne.husebekk@rito.no
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:12 EST