Trypanosomiasis associated elimination of parasite specific host antibody responses.

<p>Survival of mice was recorded after intra-peritoneal infection with of 5000 living parasites, using pleomorphic AnTat 1.1 (A), cloned monomorphic AnTat 1.1 (B), and unrelated cloned monomorphic MITat 1.4 (C) parasites (MS: Median survival). Re-challenge experiments were performed as presented in the insert box. Survival was recorded for the primary pleomorphic AnTat 1.1 infection (▪), and mice re-challenged with the cloned monomrophic AnTat 1.1 (□) or MiTat 1.4 (*) parasites on day 10, in WT mice (D), T-cell deficient nu/nu mice (E), and B-cell deficient µMT mice (F). A re-challenge was subsequently also performed on day 17 in WT mice (G). In each experiment 10 mice per experimental condition were used. One out of three representative experiments is shown for every experimental condition.</p

Alteration of Follicular and Plasma B cell numbers during <i>T. brucei</i> infections.

<p>The number of follicular B cells (A) as well as plasma B cells (B) per spleen was calculated on different days after <i>T. brucei</i> AnTat 1.1E infection. Calculations were performed on cells harvested from 3 individual spleens per time point. Values represent the mean±SD. One of four representative experiments is shown. Plasma spleen B cells were stained with a B220/CD138 combination (C).</p

Infection-induced apoptosis of MZB cells. CD21^HighCD23^Low cells were gated and analyzed for 7AAD and Annexin V staining.

<p>The percentage of positive cells was calculated in MZ B cells from naïve mice, as well as from <i>T. brucei</i> AnTat 1.1E infected mice on days 4, 7 and 10 (A). CD21^HighCD23^Low cells were FACS sorted on day 7, lyzed and analyzed in Western Blot with an anti-caspase 3 antibody, showing bands at 32 kD (pro-caspase 3), and cleaved forms 17 kD and 12 kD (B). RT-PCR for Casp3, BAFF-R, and Bcl-2 was performed on CD19⁺ spleen cells isolated from naïve mice as well as from <i>T. brucei-</i>infected mice on day 4 and day 7 (C). Results present the means of 3 mice per time point ±SD. One of three representative experiments is shown.</p

Destruction of spleen marginal zones.

<p>The MZ macrophage populations of non-infected (upper panel) and day 10 <i>T. brucei</i> AnTat 1.1 infected (lower panel) C57Bl/6 mice are visualized by section staining with ER-TR9 (anti-MZM) (A,C) and MOMA-1 (anti-MMM) antibodies (B,D) (400x magnification).</p

Trypanosomiasis associated elimination of non-related host antibody responses.

<p>Mice were vaccinated with the commercial DPTa vaccine and boosted after three weeks. 14 days after the vaccine boost, mice were infected with 5000 <i>T. brucei</i> AnTat 1.1E parasite by intra-peritoneal injection, followed 10 days later by an intranasal challenged with 5×10⁶ CFU of <i>B. pertussis/</i>mouse (▪). Control groups consisted of non-vaccinated <i>B. pertussis</i> challenged mice (×), and DPTa vaccinated mice that were challenged with <i>B. pertussis</i> 24 days after the second DTPa boost (□). Mice were sacrificed 3h and 3, 5 and 8, days after challenge and lung homogenates were prepared and plated on the Bordet-Gongou agar plates. CFU's were measured after 72 h incubation. Values are represented as the mean±SD of 3 individual mice per time point.</p

<i>T. brucei</i> induced abrogation of B cell proliferation.

<p>CD19⁺ MACS sorted cells derived form control mice or <i>T.brucei</i> AnTat 1.1E infected mice (day 10 post infection) were incubated for 24 h in the presence of different doses of anti-IgM Fab (A,B), or different doses of LPS (C,D). Proliferation was measure by thymidine incorporation. Results were obtained using spleen cell preparations of four individual mice, and represent the mean % of CPM increase ±SD, with the 100% showing the mean CPM level of non-stimulated cells.</p

Alterations of Marginal Zone (MZ) B cells during <i>T. brucei</i> infections.

<p>MZ B cells were detected using FACS as CD21^HighCD23^Low (R1), IgD^IntIgM^High (R2) or B220⁺CD1d⁺ (R3) on spleen cells derived from non- infected mice (A, upper FACS panel ) or day 10 <i>T. brucei</i> AnTat 1.1E-infected mice (A, lower FACS panel). The decrease in total number of MZ B-cells per spleen was calculated for different time points during infection (B), based on the total amount of cells harvested per spleen at each time point (C). Calculations were performed on cells harvested from 3 individual spleens per time point. Values represent the mean±SD. One of four representative experiments is shown.</p

Levels of lung IL-17, IL-1β and TNFα.

<p>C57/Bl6 and IL-17R^−/− mice were exposed to air or to ozone. Data shown as mean ± SEM for n = 5 in each group; *p<0.05 compared to air in the same species; ^#p<0.05 compared to C57/Bl6 mice with corresponding exposure.</p

Lung levels of phosphorylated ERK1/2, p38 MAPK and JNK 1/2/3.

<p>C57/Bl6 and IL-17R^−/− mice were exposed to air or to ozone. Data shown as mean ± SEM for n = 5 in each group; *p<0.05 compared to air in the same species; ^#p<0.05 compared to C57/Bl6 mice with corresponding exposure. RFU: Relative fluorescence unit.</p

Effect of dexamethasone (10⁻⁶ M) on acetylcholine (ACh)-induced bronchial contractile responses.

<p>Air-exposed C57/BL6 mice (n = 6; Panel A) and IL-17R^−/− mice (n = 6; Panel B) and ozone-exposed C57/BL6 mice (n = 9; Panel C) and IL-17R^−/− mice (n = 6; Panel D) were studied. Under each condition, the effect of dexamethasone has been compared to responses in the absence of this inhibitor. Data presented as mean±SEM. *p<0.05 compared with dexamethasone-treated tissues.</p