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note
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5 thymocytes from the indicated mice in 96-well microcultures. After 24 hours, supernatants were harvested, and IL-2 production was quantitated by HT-2 cell proliferation. Proliferation was converted to IL-2 units with the use of a standard curve generated with recombinant IL-2 (Genzyme).
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1842355688
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note
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-/loW thymocytes were obtained by treatment of total thymocytes with antibody to HSA (clone J11d2) plus rabbit complement (Cederlane, Hornby, Ontario, Canada) for 45 min at 37°C, followed by isolation of viable cells on lympholyte M gradients (Cederlane). After Fc receptors were blocked by treatment with mAb 24G2, cells were stained with phycoerythrin-conjugated mAb specific for VβS.8.1/8.2 and fluorescein-conjugated mAb specific for NK1.1 or CD44, and then analyzed by flow cytometry. All mAbs were purchased from Pharmingen.
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-/low thymocytes in 1 ml of media. After 48 hours, supernatants were harvested and assayed for cytokine production (2). Recombinant IL-4 and IFN-γ (Genzyme) were used to generate standard curves.
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2M-deficient BALB/c mice, T. Laufer for help with intravenous injections, and K. Sahmel for technical assistance. Supported by a grant from NIH to M.J.G. (RO1 AI40171) and a gift from the Mathers Foundation.
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