Figure 3.
Figure 3. l-kynurenine affects NK-cell function. Freshly isolated NK cells were cultured for 2 days in the presence of IL-2 either alone or in combination with l-kynurenine at the final concentration of 0.3 mM and then assessed either for their cytolytic activity or for their ability to produce cytokines. (A) NK-cell cytotoxicity was assessed in a redirected killing assay against the FcγR+ P815 target cell line either in the absence of mAbs or in the presence of the mAbs to the indicated receptors. (Left) Gray bars indicate untreated NK cells; black bars, l-kynurenine–treated NK cells. (E/T ratio: 2:1.) (Right) Different E/T ratios are shown (indicated in the x-axis). Symbols indicate the specificity of the mAbs added in the assay. Diamonds indicate anti-CD16 mAb; squares, anti-NKp30 mAb; triangles, anti-NKp46 mAb; circles, anti-NKG2D mAb; X symbols, spontaneous lysis (ie, no mAbs added); white symbols, untreated NK cells; and black symbols, l-kynurenine–treated NK cells. (NK cells analyzed in the right panels are not the same as those shown in the left panel.) (B, upper panels) Treated (black diamonds) and untreated (white diamonds) NK cells were evaluated for their ability to kill the indicated target cell types. The E/T ratios are indicated in the horizontal axis. (Bottom panels) To determine which triggering receptors are involved in the NK-mediated recognition and killing of the various target cell types, the same (untreated) effector cells were assessed for their ability to kill the indicated targets either in the absence of mAbs or in the presence of mAbs that specifically mask the indicated receptors. (C) Treated (black bars) or untreated (gray bars) NK cells were stimulated overnight by plastic-bound goat antimouse antiserum plus one or another of the mAbs to the indicated receptors (anti-CD56 specific mAb was used as negative control). NKG2D was not analyzed since mAb-mediated triggering of this receptor does not result in cytokine release.37 Supernatants were harvested and assessed for cytokine content by specific ELISA.

l-kynurenine affects NK-cell function. Freshly isolated NK cells were cultured for 2 days in the presence of IL-2 either alone or in combination with l-kynurenine at the final concentration of 0.3 mM and then assessed either for their cytolytic activity or for their ability to produce cytokines. (A) NK-cell cytotoxicity was assessed in a redirected killing assay against the FcγR+ P815 target cell line either in the absence of mAbs or in the presence of the mAbs to the indicated receptors. (Left) Gray bars indicate untreated NK cells; black bars, l-kynurenine–treated NK cells. (E/T ratio: 2:1.) (Right) Different E/T ratios are shown (indicated in the x-axis). Symbols indicate the specificity of the mAbs added in the assay. Diamonds indicate anti-CD16 mAb; squares, anti-NKp30 mAb; triangles, anti-NKp46 mAb; circles, anti-NKG2D mAb; X symbols, spontaneous lysis (ie, no mAbs added); white symbols, untreated NK cells; and black symbols, l-kynurenine–treated NK cells. (NK cells analyzed in the right panels are not the same as those shown in the left panel.) (B, upper panels) Treated (black diamonds) and untreated (white diamonds) NK cells were evaluated for their ability to kill the indicated target cell types. The E/T ratios are indicated in the horizontal axis. (Bottom panels) To determine which triggering receptors are involved in the NK-mediated recognition and killing of the various target cell types, the same (untreated) effector cells were assessed for their ability to kill the indicated targets either in the absence of mAbs or in the presence of mAbs that specifically mask the indicated receptors. (C) Treated (black bars) or untreated (gray bars) NK cells were stimulated overnight by plastic-bound goat antimouse antiserum plus one or another of the mAbs to the indicated receptors (anti-CD56 specific mAb was used as negative control). NKG2D was not analyzed since mAb-mediated triggering of this receptor does not result in cytokine release.37  Supernatants were harvested and assessed for cytokine content by specific ELISA.

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