Odds ratios and 95% confidence intervals comparing the chance of having the disease listed on the death certificate over time of Non-HIV related cancers (A), external causes (B), cardiovascular diseases (C), Diabetes Mellitus (D) and Tuberculosis (E); year as a categorical variable and 1999 as the baseline reference.

<p>Odds ratios and 95% confidence intervals comparing the chance of having the disease listed on the death certificate over time of Non-HIV related cancers (A), external causes (B), cardiovascular diseases (C), Diabetes Mellitus (D) and Tuberculosis (E); year as a categorical variable and 1999 as the baseline reference.</p

Relative number of cells during cell cycle events in MLNs of mice chronically-infected with <i>Schistosoma mansoni.</i>

<p>Data are reported as means ± SEM, They are representative of three independent experiments, Statistical analysis: Tukey's multiple comparison test (*, <i>P</i><0.05).</p

Immunohistochemistry to localize B lymphocyte and plasma cell niches in MLNs.

<p>(A) Immunoreactivity for B cells using anti-B220 antibody preferentially within of lymphoid follicles (LF) in chronically-infected WT wild type mice. (B) In galectin-3−/− mice, B220+ cells were randomly dispersed by the parenchyma forming numerous lymphoid follicles. In infected WT mice, CD138+ plasma cells and Blimp-1+ antibody-secreting cells were found in cellular cords in extrafollicular regions (C and E, respectively). In infected galectin-3−/− mice, CD138+ and Blimp-1+ plasma cells were randomly scattered throughout the parenchyma (D and F, respectively). A–F: Original magnification, 200x. Boxed images: original magnification, 400x. Data are reported as means + SEM and are representative of three independent experiments.</p

Quantification of phagocytic and non-phagocytic cells of MLNs of WT and galectin-3^−/− infected mice.

<p>Non-adherent lymph nodal cells were collected, induced to apoptosis by high temperature and co-cultured with adherent cells to be engulffed. Measurement of phagocytic and non-phagocytic cells after 24 h (A) and 72 h (B) of co-culture procedures. The solid bars indicate the wild-type mice and the open bars represent the gal-3^−/− mice. Phagocytic cells had a translucent vacuole and phagosomes (C, arrow), while non-phagocytic cells were identified by the absence of phagossomes and clear citoplasm (D, arrow). Data are reported as means ± SEM and are representative of three independent experiments using cells derived from chronically-infected mice. Statistical analysis: Tukey's multiple comparison test (*, <i>P</i><0.05).</p

Histological analysis of lymphoid follicles of MLNs of infected mice.

<p>(A) In wild type (WT) mice, section of lymphoid follicles showed scarce apoptotic bodies (arrow). In infected gal-3−/− mice (B), there was high number of cellular debris dispersed throughout the follicles (arrows). Immunofluorescence to MOMA-2+ macrophages. (C) Immunoreactivity for MOMA-2 Alexa 488 (green cells) in MLNs of WT and (D) in galectin-3−/− mice. (E) Detailed MOMA-2+ cell clusters in WT mice and (F) rare MOMA-2+ cells in the absence of galectin-3. The nuclei were stained with DAPI. Data are representative of three independent experiments, each carried out in three mice with chronic infection.</p

Histological analysis of MLNs of WTwild-type and galectin-3−/− mice.

<p>Midsagittal section of MLN showing lymphoid follicles preferentially within the cortex and scarcely in paracortex in uninfected and infected WT mice (A and C, respectively). Histological section from MLNs of uninfected and infected galectin-3−/− mice exhibiting lymphoid follicles randomly scattered throughout the cortex, paracortex and medulla (B and D, respectively). The samples were stained with hematoxilin and eosin. Lymphoid follicles were quantified by microscopic field in uninfected (E) and infected mice (F), with magnification of 25x. The solid bars indicate the WT mice and the open bars represent galectin-3−/− mice. Data are reported as means + SEM and are representative of three independent experiments. Statistical analysis: Tukey's multiple comparison test (*, P<0.05). A–D, original magnification: 200x.</p

Absolute number of the cell subsets in the mesenteric lymph nodes of WT and galectin-3−/− mice infected with <i>S.mansoni.</i>

<p>Data are reported as means ± SEM, They are representative of three independent experiments, Statistical analysis: Tukey's multiple comparison test (*, <i>P</i><0.05).</p

Cell cycle analysis and apoptosis index in MLNs of WT and galectin-3^−/− infected mice.

<p>Histograms represent the stages of cell cycle in WT (A) and Gal-3^−/− mice (B) infected with <i>S.mansoni</i>. In both graphs, the phases were described as bellow: M1 - sub G1/G0; M2 – G1/G0; M3 – S phase; M4 – G2/M and M5 – hyperploid cells. (C–D) Quantification of Annexin-V⁺/Propidium iodide (PI) ⁻ apoptotic cells (gated in R2 region) and Annexin-V⁺/PI⁺ dead cells (gated in R3 region), in WT (C) and Gal-3^−/− mice (D). (E) Quantification of apoptotic cells induced by high temperature. Solid bars represent WT mice and open bars indicate Gal-3^−/− mice. Data are reported as means ± SEM and are representative of three independent experiments. Statistical analysis: Tukey's multiple comparison test (*, <i>P</i><0.05). A–B, original magnification, 400x.</p

Phenotypic analysis of B lymphocytes in the MLNs.

<p>B220+ CD19+ cells were selected and quantified in uninfected wild typeWT and galectin-3−/− mice (A and B, respectively), and in chronically-infected wild typeWT and galectin-3−/− mice (C and D, respectively). (D) The arrow pointed to distinct B220low subpopulation found in the absence of galectin-3. (E) Histograms reflect the surface expression of CD138, a plasma cell marker. Full histogram: WT mice. Empty histogram: galectin-3−/− mice. (F) Absolute number of plasma cells in MLNs of infected WT (solid bars) and infected galectin-3−/− mice (open bars). Data are reported as means + SEM and are representative of three independent experiments, each carried out in five mice with chronic infection. Statistical analysis: Tukey's multiple comparison test (*, P<0.05).</p