51 research outputs found
Trypanosomosis drastically affects the titers of circulating anti-CII IgG Abs.
<p>DBA/1 mice were immunized primarily with CII in CFA on day 0 and secondarily with CII in IFA on day 21. One group of mice was infected with <i>T</i>. <i>brucei</i> (filled and open triangle) on day 14 i.p. and the other was left untreated (filled and open square). Serial serum dilution levels of anti-CII (<b>A</b>) IgG1, (<b>B</b>) IgG2a, (<b>C</b>) IgG2b and (<b>D</b>) IgM Abs were measured by ELISA on day 14 (before infection, filled square and triangle) and day 35 (open square and triangle). Graphs show the mean ± SD from at least n = 6 mice per group and the data are representative of two independent experiments.</p
Berenil treatment restores the appearance of the CIA-associated clinical symptoms.
<p>DBA/1 mice were immunized primarily with CII in CFA on day 0 and secondarily with CII in IFA on day 21. One group of mice out of two was infected with <i>T</i>. <i>brucei</i> (filled and open triangle) on day 14 either i.p. or using tsetse flies and the other was left untreated (filled and open square). Both groups were treated with the trypanocidal drug, Berenil, at (<b>A</b>,<b>B</b>) day 38 (i.p.) or (<b>C,D</b>) day 28 (tsetse). (<b>A</b>) The average clinical per mouse score was followed from day 0. (<b>B</b>) Serial serum dilution levels of anti-CII IgG2a and IgG2b Abs were measured by ELISA on day 38 (before Berenil, filled square and triangle) and day 56 (open square and triangle). (<b>C</b>) Clinical score was followed from day 0 and (<b>D</b>) serial serum dilution levels of anti-CII IgG2a and IgG2b Abs were measured by ELISA on day 28 (before Berenil, filled square and triangle) and day 42 (open square and triangle). Arrows represent the day when trypanocidal drug Berenil was administered. Graphs show the mean ± SD from at least n = 6 mice per group and the data are representative of two independent experiments.</p
Trypanosomosis substantially delays the onset of CIA in DBA/1 prone mice.
<p>DBA/1 mice were immunized primarily with CII in CFA on day 0 and secondarily with CII in IFA on day 21. One group of mice was infected with <i>T</i>. <i>brucei</i> (filled triangle) on day 14 i.p. and the other was left untreated (filled square). (<b>A</b>) Average clinical score and (<b>C</b>) the number of arthritic limb per mouse were followed starting at day 0. (<b>B</b>) Representative pictures of arthritic limbs at day 42. Graphs show the mean ± SD from at least n = 6 mice per group and the data are representative of three independent experiments. (D&E) DBA/1 mice were infected with <i>T</i>. <i>brucei</i> i.p. and (<b>D</b>) survival and (<b>E</b>) blood parasite counts were monitored. Graphs show the mean ± SD from at least n = 6 mice per group and the data are representative of two independent experiments.</p
MCMV Splenic Tropism
<div><p>All analysis was performed 3 d post virus infection.</p><p>(A) Splenic stromal cells from mock or MCMV (2 × 10<sup>5</sup> pfu, Smith strain) infected spleens were separated from hematopoietic cells by extrusion through a nylon mesh filter (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0020016#s4" target="_blank">Materials and Methods</a>). Total cell RNA was then isolated from either the stromal cells (S) or hematopoietic cells (H) fractions, and quantitative RT-PCR was performed to detect expression of CCL21-ser, CCL19, and CXCL13 as well as expression of the MCMV immediate early-1 (IE1) transcript.</p><p>(B) Spleens from MCMV-GFP infected mice (2 × 10<sup>5</sup> pfu) were examined by immunohistochemistry. Panels are color-coded with the text for the antigen examined, and field magnification is shown to the right of the panels. No background staining was seen in uninfected mouse spleen when directly detecting GFP expression or when using an anti-GFP antibody (αGFP) for signal amplification.</p><p>WP, white pulp.</p></div
Reduced CCL21-ser Expression after MCMV Infection
<div><p>Spleens from B6 mice mock-infected or infected with 3.2 × 10<sup>5</sup> (A,B,D) or 1.5 × 10<sup>5</sup> (C) pfu of MCMV were harvested at day 3 post-infection. Gene expression was measured by quantitative PCR and was normalized to 18S RNA and presented as relative mRNA levels compared to uninfected mice (A and D) or as absolute values (C). When presented as relative values, mRNA levels from three uninfected mice (mock) were determined and averaged for comparison for each experiment.</p><p>Open symbols in (A) represent mice infected with UV-inactivated MCMV (equivalent to 3.2 × 10<sup>5</sup> pfu).</p><p>(B) Immunohistochemical analysis of CCL21 expression in the spleen of a mock or MCMV-infected B6 mouse 3 d post-infection; the location of the periarteriolar lymphoid sheath “PALS” and B-cell follicles “B” is noted, and the interrupted white line indicates the outer edge of the MZ. This section is representative of three experiments (<i>n</i> = 3 mice per group).</p><p>In (C), primers used for analysis of CCL21 levels amplified either the serine (CCL21-ser), leucine (CCL21-leu), or both (CCL21u) isoforms.</p><p>The data in (D) is an average of three infected mice ± SEM (<i>p</i> < .001).</p></div
MCMV-Induced Suppression of CCL21 Abolishes T Cell Localization to the White Pulp
<div><p>Mock- or MCMV-infected B6 mice (1.5 Ă— 10<sup>5</sup> pfu) were injected with CFSE labeled naive T cells, and spleens were removed for analysis 2 h later.</p><p>(A) Flow cytometry was used to assess the total number of CFSE-labeled cells in the spleen after adoptive transfer into mice 3 d post-infection.</p><p>(B) Immunohistochemical analysis of T cell localization in the spleen at 3 d post-infection using the MOMA-1 marker to identify MMM (shown in blue, magnification 10Ă—). WP, white pulp; RP, red pulp.</p></div
MCMV Modulates Splenic Architecture
<div><p>(A) Spleens from PBS (mock) or MCMV-infected B6 mice (3.2 Ă— 10<sup>5</sup> pfu) were harvested at day 3 post-infection for immunohistochemical analysis. Spleen sections were incubated with markers specific to MMM (MOMA-1, MMM, blue), MZM (ER-TR9, MZM, red), granulocytes (Gr-1/Ly-6G, green), stromal cell populations (BP-3, blue, to identify B-cell-zone stromal cells and gp38, red, to identify T-cell-zone stromal cells, in uninfected mice), and B (B220, blue), and T lymphocytes (Thy1.2, red) (magnification 20Ă—). These sections are representative of four independent experiments with <i>n</i> = 3 mice per group with two or more sections taken from each spleen. White pulp (WP) and red pulp (RP) and the location of the central arteriole (ca) are indicated.</p><p>(B) Similar immunohistochemical analysis performed on Balb/c mice, 3 d post-MCMV-infection (1 Ă— 10<sup>5</sup> pfu).</p></div
Pharmacologic Activation of LTβR Signaling Partially Restores CCL21 Expression and T Cell Localization in the Spleen of MCMV-Infected Mice
<div><p>(A) Quantitative PCR was used to determine the relative levels of CCL21 gene expression 3 d post-MCMV infection in spleens from control- or virus- (V) infected mice ± agonistic anti-LTβR antibody (Ab) treatment.</p><p>Left panel, 3.2 × 10<sup>5</sup> pfu of MCMV was used for infection, and only total CCL21 mRNA levels were analyzed using CCL21u primers (see below).</p><p>Right Panel, mice were infected with 1.5 × 10<sup>5</sup> pfu MCMV and expression of the various CCL21 isoforms was determined. CCL21u, universal CCL21 primers that amplify both the leucine (leu) and serine (ser) isoforms; <i>p</i> = 0.02</p><p>(B) Shown are representative spleen sections from the same mice that were analyzed in the right panel of (A). Sections were incubated with an antibody to detect MMM (MOMA-1, MMM, blue), and adoptively transferred, CFSE-labeled T cells are green. The number of CFSE labeled T cells located per WP area after transfer was analyzed (<i>n</i> = number of WP regions analyzed per spleen), and differences between groups of infected mice ± antibody is statistically significant to a <i>p-</i>value < 0.0001.</p></div
<i>T. brucei</i> induced abrogation of B cell proliferation.
<p>CD19<sup>+</sup> 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<sup>High</sup>CD23<sup>Low</sup> (R1), IgD<sup>Int</sup>IgM<sup>High</sup> (R2) or B220<sup>+</sup>CD1d<sup>+</sup> (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
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