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Measurement of mouse and human interleukin 9
Jean-Christophe Renauld1, Jacques Van Snick1
1Ludwig Institute for Cancer Research and Experimental Medicine Unit, Catholic University of Louvain, Brussels, Belgium.
This unit describes two proliferation assays to detect or quantitate human and murine interleukin 9 (IL-9). The first is based on the ability of IL-9 to stimulate the proliferation of the TS1h9RA3 cell line, a murine IL-9-dependent cell line transfected with the human IL-9 receptor. An alternate protocol is based on the ability of IL-9 to stimulate the proliferation of the human megakaryoblastic leukemia cell line, M-O7e. M-O7e cells depend on either human IL-3 or granulocyte/macrophage colony-stimulating factor (GM-CSF) for growth, although other cytokines including IL-2, -4, -6, and -9 and steel factor are also weakly mitogenic (relative to IL-3 and GM-CSF). Thus, although M-O7e cells can readily be used to quantitate levels of IL-9 in the absence of other cytokines, analysis in the presence of complex mixtures of cytokines (e.g., natural sources) requires the use of specific antibodies against IL-9 and the other cytokines, as described in this unit.
This unit describes two proliferation assays to detect or quantitate human and murine interleukin 9 (IL-9). The first is based on the ability of IL-9 to stimulate the proliferation of the TS1h9RA3 cell line, a murine IL-9-dependent cell line transfected with the human IL-9 receptor. An alternate protocol is based on the ability of IL-9 to stimulate the proliferation of the human megakaryoblastic leukemia cell line, M-O7e. M-O7e cells depend on either human IL-3 or granulocyte/macrophage colony-stimulating factor (GM-CSF) for growth, although other cytokines including IL-2, -4, -6, and -9 and steel factor are also weakly mitogenic (relative to IL-3 and GM-CSF). Thus, although M-O7e cells can readily be used to quantitate levels of IL-9 in the absence of other cytokines, analysis in the presence of complex mixtures of cytokines (e.g., natural sources) requires the use of specific antibodies against IL-9 and the other cytokines, as described in this unit.
