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. 2009 Feb 6;106(8):2741–2746. doi: 10.1073/pnas.0813367106

Fig. 4.

Fig. 4.

Defective dendritic cell maturation underlies the diminished CD8 T-cell response in B7-H1−/− Mice. (A) Defective DC maturation in B7-H1−/− mice is restored by adoptive transfer of B7-H1+/+ T cells. WT and B7-H1−/− mice were instilled i.n. with CFSE. Some of the B7-H1−/− mice were also given i.v. 1 × 106 purified B7-H1+/+ F5 T cells (>98% CD8+) at the time of CFSE instillation. Six hours later mice were infected with WSN-SIY virus, and 24 hours postinfection MLN were pooled from four mice and digested with collagenase D/DNase I. The resulting single-cell suspensions were assayed for CD11c, CFSE plus CD80, CD86, or CD40. The histograms show CD80, CD86, and CD40 staining intensity of CD11c+ CFSE+ DCs. As control, CD11c+ CFSE− DCs from spleen of WT mice were used. (B) Restoration of DC maturation in RAG1−/− mice by transferred B7-H1+/+, but not B7-H1−/−, T cells. Purified B7-H1+/+ or B7-H1−/− 2C T cells (>98% CD8+) were transferred i.v. into RAG1−/− mice (1 × 106 per mouse). Twenty-four hours later, mice were infected i.p. with 1 × 106 pfu of WSN virus. As control, RAG1−/− mice without T-cell transfer were infected at the same time. Twenty-four hours postinfection, cells from the mesenteric lymph node were isolated and assayed for CD11c plus CD40, CD80, or CD86. Histograms show CD80, CD86, and CD40 staining intensity of CD11c+ DCs. (C and D) Restoration of CD8 T-cell response in B7-H1−/− mice by transferred B7-H1+/+, but not B7-H1−/−, DCs. The DCs from the spleen of either WT or B7-H1−/− mice were purified (> 95% CD11c+) and transferred into hind footpads of either WT mice or B7-H1−/− mice (5 × 105 per mouse). Twenty-four hours later, mice were infected with 100 pfu of WSN-SIY virus i.n. and 7 dpi cells from the lung were stained with anti-CD8 and SIY-Kb. (C) Representative SIY-Kb versus CD8 staining profiles of live cells are shown. Numbers indicate percentages (mean ± SEM, n = 3) of CD8+ SIY-Kb+ cells among all live cells. (D) Comparison of the number of SIY-specific CD8 T cells in different tissues. Values represent mean ± SEM (n = 3). *, P < 0.05. (E) Restoration of CD8 T-cell response in B7-H1−/− mice by conditioned DCs from RAG1−/− mice. Purified B7-H1+/+ or B7-H1−/− T cells (>98% CD8+) were transferred i.v. in RAG1−/− mice (1 × 106 per mouse). Three days later, DCs were purified from these RAG1−/− mice (>98% CD11c+) and injected into hind footpads of B7-H1−/− mice (5 × 105 per mouse). Twenty-four hours after DC transfer, mice were infected i.n. with 100 pfu of WSN-SIY virus. As control, B7-H1−/− mice without DC transfer were infected at the same time. Seven days postinfection, cells from the lung, MLN, and spleen were stained with anti-CD8 and SIY-Kb. Comparison of the numbers of SIY-specific CD8 T cells in different tissues of recipient mice. Values represent the mean ± SEM (n = 3). *, P < 0.05.