IL-5 responses to larval <i>Strongyloides stercoralis</i> antigens are decreased in patients with strongyloidiasis and HTLV-1 co-infection (n = 13) compared to HTLV-1 negative subjects with strongyloidiasis (n = 27).

<p>Peripheral blood mononuclear cells from newly diagnosed strongyloidiasis patients were cultured with or without <i>Strongyloides stercoralis</i> infective-stage larval antigen. Culture supernatants were analyzed for IL-5 production. Patients with HTLV-1 co-infection had significantly decreased IL-5 responses (* p = 0.0004, Mann Whitney test).</p

Patients with strongyloidiasis and HTLV-1 co-infection (n = 12) had more <i>Strongyloides stercoralis</i> larvae found in stool when compared to strongyloidiasis-only patients (n = 25).

<p>A semi-quantitative measure of <i>Strongyloides stercoralis</i> parasite load was determined at the time of diagnosis according to the number of larvae observed in stool sediment after Baermann concentration (range from1+ to 4+). Parasite load determination was made before the HTLV-1 status was known. HTLV-1 carriers had higher parasite loads (3+ and 4+) when compared to the non HTLV-1 group (10 out of 12 vs. 4 out of 25 respectively; p<0.001, Chi square test, Lima, Peru).</p

Increased proportions of regulatory T cells in strongyloidiasis/HTLV-1 co-infected patients.

<p>Comparison of regulatory T cell proportions (CD25+FoxP3+ among CD4+ T-cells) between healthy controls (blue, n = 17), patients with strongyloidiasis without HTLV-1 infection (red, n = 27), HTLV-1 asymptomatic carriers (orange, n = 9) and strongyloidiasis/HTLV-1 co-infected patients (green, n = 13). Co-infected patients had a significant increase of regulatory T cells compared to non-HTLV-1 patients. (* = P<0.05, One-way ANOVA). Lima, Peru.</p

Blood analysis, flow cytometry and interleukin-5 responses to <i>Strongyloides stercoralis</i> larval antigens A peripheral blood sample was obtained at enrollment from all strongyloidiasis patients.

<p>Samples were analyzed before HTLV-1 status was known. Numbers represent median of all parameters analyzed. Complete peripheral blood cell count showed lower hemoglobin and hematocrit levels in the HTLV-1 co-infected group. White blood cell, neutrophil, eosinophil, and lymphocyte proportions were not significantly different among groups, although a tendency to lower absolute eosinophil count was observed in the HTLV-1 infected group. Regulatory T cells were significantly increased in patients with strongyloidiasis and HTLV-1 co-infection (* = p<0.05, One way ANOVA, Lima, Peru).</p

Increased numbers of regulatory T cells correlates with reduced IL-5 responses to <i>Strongyloides stercoralis</i> larval antigens.

<p>Patients with higher regulatory T cell numbers showed lower IL-5 responses to <i>Strongyloides stercoralis</i> infective-stage crude antigen. An inverse correlation was determined by Sperman non-parametric correlation analysis (Spearman r = −0.38, p = 0.03).</p

Demographics and frequency of clinical signs and symptoms of strongyloidiasis patients with and without HTLV-1 co-infection.

<p>Clinical data was obtained from newly diagnosed patients with strongyloidiasis by stool modified Baermann sedimentation method and tested for HTLV-1 co-infection. Age and gender were similar among strongyloidiasis patients with or without HTLV-1 co-infection. Gastrointestinal signs and symptoms were more frequently reported among HTLV-1 co-infected patients. Table shows proportion of patients reporting symptoms. Frequencies were compared between <i>S. stercoralis</i> groups by Chi square test, * = p<0.05 (Lima, Peru).</p

Flow cytometry analysis showing increased proportions of CD4+CD25hi T cells and CD4+CD25+FoxP3+ regulatory T cells in peripheral blood of patients with strongyloidiasis and HTLV-1 co-infection.

<p>Representative flow cytometry analysis of peripheral blood mononuclear cells comparing a patient with strongyloidiasis without HTLV-1 co-infection (left) to strongylioidiasis/HTLV-1 co-infected patient (right). Upper figures show CD4 and CD25 staining of cells within the lymphocyte gate. Note population of CD4+CD25hi cells in the HTLV-1 co-infected patient suggesting regulatory T cell phenotype. Lower figures show CD4+ lymphocytes stained with CD25 and intracellular staining with FoxP3 confirming that CD4+CD25hi cells are indeed FoxP3+ (regulatory T cells).</p

Synergistic effects of GDC0941-AZD6244 combination therapy on cell viability and proliferation.

<p>BxPC-3 (A) and MIA PaCa-2 cells (B) were treated with GDC0941 alone, AZD6244 alone or GDC0941-AZD6244 in combination. Results were analyzed according to the Chou-Talalay method [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077243#B18" target="_blank">18</a>]. The combination index (CI) values were calculated by using CalcuSyn software. PANC-1 (C) and Capan-2 cell line (D) were treated with GDC0941 at 400 nM, AZD6244 at 200 nM or combination. * p value <0.05 compared with control or single agent alone.</p

Effects of GDC0941-AZD6244 combination therapy on PI3K/AKT and MEK/ERK pathways.

<p>All four pancreatic cancer cell lines were treated with the GDC0941-AZD6244 combination for 24 hours, Cell lysates were then harvested to detect p-AKT (S473)/AKT, p-ERK (T202/Y204)/ERK, p-S6 (S240/244)/S6 and p-4E-BP1 (S65)/4E-BP1.</p

Effects of GDC0941 and AZD6244 on PI3K/AKT and MEK/ERK pathways of MIA PaCa-2 and PANC-1 cell lines.

<p>Both MIA PaCa-2 and PANC-1 cells were treated with 400 nm GDC0941, 200 nM AZD6244 or a combination at these doses for 24 hours. Cell proteins were then harvested to detect p-AKT(S473)/AKT, p-ERK(T202/Y204)/ERK, p-S6(S240/244)/S6 and p-4E-BP1(S65)/4E-BP-1.</p