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Arch Histol Cytol. 2003 Dec;66(5):397-406.

Localization of lymphocyte apoptosis in murine lymphoid tissues after stimulation of natural killer T cells with alpha-galactosylceramide.

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Departments of Anatomy, Osaka City University Medical School, Osaka, Japan.


Alpha-galactosylceramide (alpha-GalCer) has been reported to induce activation-induced cell death (AICD) in natural killer T (NKT) cells. We undertook this study to demonstrate the distribution of AICD of NKT cells in the lymphoid tissues and differences in frequency between the central and peripheral lymphoid tissues, histologically and quantitatively analyzing the apoptotic figure in the murine spleen, lymph node, and thymus after alpha-GalCer treatment. Lymphocyte apoptosis was identified as a cluster of nuclei with chromatin condensation in hematoxylin-eosin-stained sections, and was confirmed by TUNEL staining, double staining for TUNEL and CD4, and electron microscopy. In the spleen, it appeared at 12 h after alpha-GalCer administration, increased in number, reaching a peak at 24 h, and then falling to a normal level at 72 h. It was preferentially found in the white pulp, especially in the periarterial lymphoid sheath, but was sparse in the red pulp. Alpha-GalCer-induced lymphocyte apoptosis was seen in tumor-necrosis factor (TNF)-deficient mice as well, but was not in lpr/lpr (Fas-deficient) or gld/gld (Fas ligand-deficient) mice. As for the tissue distribution, lymphocyte apoptosis was frequently seen in the paracortex of the lymph node, whereas it was rare in the thymus. These data indicate that alpha-GalCer-induced AICD of NKT cells takes place in the T-cell area of peripheral lymphoid tissues (i.e., the spleen and lymph node) through the Fas/Fas ligand, but not the TNF pathway.

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