Hospice resources and utilization from 2000–2006.

<p>Hospice resources and utilization from 2000–2006.</p

Urban-rural disparities of cancer patients with first-time hospice care from 2000–2006.

<p>Urban-rural disparities of cancer patients with first-time hospice care from 2000–2006.</p

Features of cancer patients with hospice care during 2000–2006, stratified by location of residence.

<p>Features of cancer patients with hospice care during 2000–2006, stratified by location of residence.</p

PbT-I cells proliferate in response to irradiated sporozoites.

<p>B6 mice were adoptively transferred with 2×10⁶ CFSE-labeled PbT-I cells. The next day, mice were injected i.v. with 5×10⁴–10⁵ radiation attenuated sporozoites (RAS) or equivalent salivary extract. Four days later spleens were harvested and the proliferation of PbT-I cells was analyzed. (A) Representative histograms showing the proliferation of PbT-I cells in response to irradiated sporozoites. (B) Pooled data showing the number of divided PbT-I cells from four experiments. The lines represent the mean and each data point represents a mouse. Mice similarly treated but left until day 7 post-challenge showed no breakthrough in blood-stage infection indicating full attenuation of sporozoites. Data were compared by one-way ANOVA and Tukey's multiple comparison test (*, p&lt;0.05).</p

PbT-I cells respond in the spleen to i.v. blood-stage PbA.

<p>B6 mice were adoptively transferred with 2×10⁶ Ly5.1⁺ PbT-I cells and the next day infected i.v. with 10⁶ blood-stage PbA. Spleens were harvested three or five days later and the proliferation of PbT-I cells was analyzed. The gating strategy to identify PbT-I cells was similar to that shown in <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004135#ppat.1004135.s013" target="_blank">Figure S13</a></b>. (A) Representative histograms showing the proliferation of PbT-I cells on day three or five post-infection. (B) Number of proliferating PbT-I cells in the spleen and blood of mice infected with PbA for three or five days. Data are pooled from three experiments. Each data point represents a mouse and the lines represent the mean. Data were compared using student t test (**, p&lt;0.01; ***, p&lt;0.001). (C) B6 mice were adoptively transferred with 10⁶ CFSE-labeled Ly5.1⁺ PbT-I cells. The next day, mice were injected i.v. with 10⁶ blood-stage PbA. Various tissues (spleen, blood, celiac lymph node (ceLN), portal LN (pLN), inguinal LN (iLN), mesenteric LN (mLN)) were harvested after 2 days and PbT-I cells examined for CD69 and CFSE expression. Profiles are gated on PbT-I cells. This experiment was performed three times (two-three mice per group) with similar results. Typical profiles are shown. (D) The mean percentage of CD69⁺ PbT-I cells for the analysis shown in (C). Histograms represent values from infected animals minus mean values from uninfected animals. Error bars represent standard error of the mean.</p

PbT-I cells infiltrate the brain and accelerate ECM.

<p>B6 mice were adoptively transferred with 2×10⁶ or 2×10⁴ Ly5.1⁺ PbT-I cells or 2×10⁶ herpes simplex virus-specific gBT-I cells or left uninjected. The next day mice were infected i.v. with 10⁶ blood-stage PbA. (A) Mice were monitored for the development of ECM. Data are pooled from three independent experiments. The differences between 2×10⁶ PbT-I and the group that did not receive any transgenic cells or the group that received gBT-I cells are statistically significant (p&lt;0.0001) as determined by a Log-rank test. (B) Mice were adoptively transferred with PbT-I cells (filled circle) or gBT-I cells (filled square) or no cells (open circle) and were sacrificed on days 4, 5 or 6 post-infection. Their brains were then analysed for the infiltration of PbT-I cells (left) or gBT-I cells (right). Data are pooled from 2–4 experiments. Data were compared using student t test (*, p&lt;0.05).</p

Characterization of T cells from the spleen and lymph node of PbT-I mice.

<p>Cells were harvested from the spleen and the lymph nodes of PbT-I transgenic or littermate control mice (WT). FACS analysis was performed to characterize the expression of CD8, CD4 and the transgenic TCR alpha (Vα8.3) and beta (Vβ10) chains. (A) Representative dot-plots showing the proportions of CD8 versus CD4 cells in the spleen and lymph node of PbT-I and WT mice. (B) Representative histograms showing the expression of the transgenic TCR Vα8.3 and Vβ10 chains on the CD8 or CD4 single-positive cells from the spleen. This experiment was repeated three times with two mice per experiment.</p

PbT-I cells are primed mainly in the spleen when irradiated sporozoites are delivered by the i.v. route.

<p>B6 mice were adoptively transferred with 10⁶ CFSE-labeled PbT-I cells. The next day, mice were injected i.v. with 10⁵ irradiated PbA sporozoites. Various organs (spleen, blood, liver, different lymph nodes) were harvested on days 1–4 post-vaccination and the activation and proliferation of PbT-I cells was analyzed by flow cytometry. For gating strategy to identify PbT-I cells see <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004135#ppat.1004135.s013" target="_blank">Figure S13</a></b>. (A) Representative histograms showing the proliferation of PbT-I cells versus the upregulation of CD69 in the various organs. (B) Number of divided PbT-I cells in the different organs. The insert shows a more sensitive scale to identify the few divided PbT-I cells in the liver and lymph nodes on days three and four post-infection. (C) Percentage of CD69⁺ PbT-I cells in each organ on days 1-2. Error bars represent standard error of the mean. Data are pooled from three independent experiments.</p

CD8α⁺ DC are required for presentation of irradiated sporozoites delivered by the i.v. route.

<p>B6 or Batf3^-/- mice were adoptively transferred with 10⁶ CellTracker Violet-labeled PbT-I cells then infected with irradiated PbA sporozoites. (A) Representative histograms show the proliferation of PbT-I cells in B6 and Batf3^-/- mice on day 5 after injection with irradiated sporozoites. (B) Pooled data showing the proliferation of PbT-I cells from three experiments. The lines represent the mean and each data point represents a mouse. Data were compared by one-way ANOVA and Tukey's multiple comparison test (***, p&lt;0.001).</p

PbT-I cells respond to <i>P. chabaudi</i> AS.

<p>B6 mice were adoptively transferred i.v. with 2×10⁶ CFSE-labeled PbT-I cells. The next day, mice were injected i.v. with 10⁵<i>P. chabaudi</i> AS. Six or seven days later, spleens were harvested and the proliferation of PbT-I was analyzed. (A) Representative histograms of CFSE-labeled PbT-I cells in the spleen of uninfected or <i>P. chabaudi</i> infected mice. (B) Number of PbT-I cells in the spleens of mice on days 6–7. The lines represent the mean and each data point represents a mouse. Data are pooled from three experiments. Data were compared using student t test (***, p&lt;0.001).</p