Figure 1.
CD103 deficiency attenuates intestinal GVHD mediated by donor CD8 T cells.
CD8 T cells primed to A/J alloantigens from either BALB/c-WT (WT) or BALB/c-CD103 KO donors were adoptively transferred into lethally irradiated A/J recipient mice in combination with WT BMC. A. Data shown are survival rates of irradiation control mice (diamonds), recipients of WT BMC plus purified CD8 T cells from either WT (WT CD8, squares, n = 12) or KO donors (KO CD8, triangles, n = 13), and recipients of BMC only (circles, n = 3). These data derive from two separate experiments with similar results. **p-value = 0.003, compared to the survival of hosts receiving WT CD8 T cells (log-rank test). B. H&E sections of the host intestine (upper panel) and spleen (lower panel) taken from recipients of WT or KO CD8 T cells. Arrows indicate apoptotic bodies. C. Mean number of apoptotic bodies per hundred intestinal villi ± SEM in recipients of CD8 T cells from WT donors (WT, closed bar, n = 3) or KO donors (KO, open bar, n = 3). **p<0.01.
Figure 2.
CD103 is not required for effective clearance of solid tumor by donor CD8 T cells.
A. Lethally irradiated A/J mice were inoculated with SaI/N fibrosarcoma (H-2a) plus BALB/c-WT (WT) either alone (BMC only, n = 6) or in combination with purified CD8 T cells from primed WT (n = 8) or BALB/c-CD103 KO donors (n = 8). Data shown are the mean volumes of solid tumor (cm3) measured at the indicated time points ± SEM. N.D. indicates that no tumor was detected. This is a combination of two separate experiments with similar results. Since some experimental mice died early of GVHD, data of 6, 5 or 2 recipient mice of each respective group are shown in last time point. B. Sal/N fibrosarcoma cells (1.0×106) were transferred into BALB.scid mice alone (closed circles; n = 4) or together with primed CD8 T cells (1.0×107) from either WT (triangles; n = 4) or KO donors (open circles; n = 4). Data show mortality of recipient mice. Surviving mice were sacrificed at day 70 after adoptive transfer.
Figure 3.
CD103 expression is not required for accumulation of donor CD8 T effectors at the site of tumor growth.
Equal numbers of CD8 T cells from BALB/c-WT (WT, Thy1.1+) and BALB/c-CD103 KO, (KO, Thy1.1+Thy1.2+) donors were adoptively transferred into a group of lethally irradiated A/J mice together with BALB/c-WT BMC plus SaI/N tumor cells. Recipient mice were sacrificed at days 7, 14, 21 and 28. Lymphocytes isolated from the host intestinal epithelium and the tumor were subjected to flow cytometric analyses. A. Representative dot plots of Thy1.2 expression by gated CD8 T cells in the two compartments at the indicated time points. B. Mean ratios ± SEM (n = 5) of WT versus KO CD8 T cells in the intestinal epithelim (IEL, solid circles) and tumor (TIL, open circles). *p<0.05 compared with TIL.
Figure 4.
CD103 is preferentially expressed by CD8 T cells in the intestinal epithelium as compared to the tumor.
Equal numbers of CD8 T cells from BALB/c-WT (WT, Thy1.1+) and BALB/c-CD103 KO (KO, Thy1.1+Thy1.2+) donors were adoptively transferred into a group of lethally irradiated A/J mice together with BALB/c-WT BMC plus SaI/N tumor cells. Recipient mice were sacrificed at days 7, 14, 21 and 28. Lymphocytes isolated from the indicated host compartments were then subjected to flow cytometric analyses. A. Mean percentage ± SEM (n = 5) of CD8 T cells in the intestinal epithelium (IEL closed circles) and the tumor (open circles) that expressed significant levels of CD103 at the indicated time points. B. Representative dotplot of CD103 expression by gated CD8+ cells in the intestinal epithelium (IEL) and tumor (TIL).
Figure 5.
CD103 is not required for accumulation of CD8 T cells in the lamina propria or mucosal lymphoid compartments during GVHD.
Equal numbers of CD8 T cells from BALB/c-WT (WT, Thy1.1+) and BALB/c-CD103 KO (KO, Thy1.1+Thy1.2+) donors were adoptively transferred into a group of lethally irradiated A/J mice together with BALB/c-WT BMC plus SaI/N tumor cells. Recipient mice were sacrificed at day 21. Lymphocytes isolated from the indicated host compartments were subjected to flow cytometric analyses. A. Representative dot plots of WT and KO CD8 T cells in the host intestinal epithelium (IEL), lamina propria (LPL), Peyer's patch (PPL), and mesenteric lymph node (MLN) at d21 post alloSCT. B. Mean ratios ± SEM (n = 5) of WT to KO CD8 T cells in the indicated compartments. *p<0.05 compared with IEL; **p-value<0.01 compared with IEL.
Figure 6.
CD103 expression is not required for cytokine production by gut infiltrating CD8 T cells.
Equal numbers of CD8 T cells from BALB/c-WT (WT, Thy1.1+Thy1.2-) and BALB/c-CD103 KO donors (KO, Thy1.1+Thy1.2+) were adoptively transferred into a group (n = 3) of lethally irradiated A/J mice together with WT BMC plus SaI/N tumor cells. Lymphocytes were isolated from the host intestinal epithelium at day 7 post-transplant and subjected to intracellular staining for INFγ, TNFα and IL-2. A. Representative histograms of cytokine staining by gated CD8+ lymphocytes of either WT or KO origin (thin line) vs. isotype control staining (heavy line). B. Mean production (±SEM) of the indicated cytokines by gated WT (n = 3, solid bars) or KO CD8 T cells (n = 3, open bars). Data represent two independent experiments with similar results.