Journal:

Article Title: Infection with Toxoplasma gondii Increases Atherosclerotic Lesion in ApoE-Deficient Mice

doi: 10.1128/IAI.72.6.3571-3576.2004

Figure Lengend Snippet: (A) Total serum cholesterol of noninfected (n = 14, black bars) or T. gondii-infected ApoE KO mice (n = 17, gray bars) after 0, 4, and 5 weeks postinfection. (B) Lipoprotein profile of noninfected (n = 5, fine line) and T. gondii-infected (n = 5, bold line) ApoE KO mice after 5 weeks of experiment (VLDL, fractions 5 to 10; IDL + LDL, fractions 11 to 19; HDL, fractions 21 to 26). The averages ± standard errors of the VLDL, IDL + LDL, and HDL fractions were 355 ± 51, 99 ± 15, and 1.2 ± 15 for the control group and 175 ± 14, 109 ± 7, and 5.2 ± 2.1 for the T. gondii group, respectively. (C) IFN-γ concentration in serum of noninfected (n = 6, black bars) or T. gondii-infected ApoE KO mice (n = 6, gray bars) at the beginning of and during the experiment. (D) Serum nitrite/nitrate concentration (micromoles/liter) of noninfected (n = 9, black bar) and T. gondii-infected (n = 12, gray bar) ApoE KO mice after 5 weeks of experiment. *P < 0.05.

Article Snippet: Serum IFN-γ was assayed in a two-site enzyme-linked immunosorbent assay with the rat anti-IFN-γ monoclonal antibody R46A2 (American Type Culture Collection, Rockville, Md.) and a polyclonal rabbit serum specific for the cytokine, as previously described ( 4 ).

Techniques: Infection, Concentration Assay

Journal: PLoS ONE

Article Title: Plasmacytoid Dendritic Cells Are Inefficient in Activation of Human Regulatory T Cells

doi: 10.1371/journal.pone.0044056

Figure Lengend Snippet: A: Production of IFN-α by activated pDC. Supernatants of activated pDC (CpG/IL-3) or cDC (cytokine cocktail) cultures were harvested. Production of IFN-α was investigated by ELISA. Each circle resembles one individual experiment, bar represents mean; n.d.: not detectable, n = 4. B: Neutralization of cytokines in cocultures of Teff, Treg and pDC. Teff plus Treg (1∶1) were stimulated with allogeneic pDC at a ratio of 1∶5 in absence (Ø) or presence of neutralizing antibodies against IFN-α, IL-1β, IL-6 or TNF-α (10 µg/ml each). Proliferation in cocultures (mean ± SD of triplicates) is normalized to untreated cocultures (set to 100%). One representative experiment out of three is shown. C: Addition of supernatants from standard cultures. CD4 + Teff and Treg (1∶1) were stimulated with anti-CD3 mAb (0.5 µg/ml) plus irradiated TC-depleted PBMC. Supernatants from these cocultures were collected on day 3 and titrated (1∶2) to pDC-stimulated CD4 + Teff, Treg or cocultures. Proliferation was assessed on day 3 by 3 H-Tdr-incorporation. Diagrams show mean values (± SD) of three independent experiments. Proliferation of untreated and treated cocultures was normalized to untreated or treated Teff-proliferation (set to 100%).

Article Snippet: For determination of IFN-α production, activated 10 6 /ml pDC (stimulated with CPG plus IL-3) and cDC (stimulated with the cytokine cocktail) were cultured for 24 h. Supernatants were harvested and IFN-α was detected by using a commercial available IFN-α ELISA (Diaclone, Besancon, France, detection limit 3.16 pg/ml) as indicated by the manufacturer.

Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, Irradiation

Journal: PLoS ONE

Article Title: CAF01 Potentiates Immune Responses and Efficacy of an Inactivated Influenza Vaccine in Ferrets

doi: 10.1371/journal.pone.0022891

Figure Lengend Snippet: 6 to 10 months-old ferrets were immunized twice at two week-intervals with CAF01 adjuvanted (▴) and non-adjuvanted (▪) influenza vaccine Sanofi-Pasteur's Vaxigrip season 2005/2006 (n = 8 per group). A third group was mock-vaccinated (•). A. Serum IgG antibodies against the vaccine antigens measured by IgG specific ELISA at various timepoints. The titer was defined as the reciprocal value of the highest positive dilution. B. Hemagglutination inhibition assay using influenza A (New Caledonia 1999 (H1N1). HI titers were determined by the reciprocal dilution of the last well which contained non-agglutinated red blood cells. All ELISA values are expressed as geometric mean titers (GMT). C . Percentage of IFN-γ-positive lymphocytes: peripheral blood leucocytes were isolated and stimulated overnight with a recombinant H1 hemagglutinin from A/New Caledonia/20/99 (H1N1). Values marked with an asterisk are significantly different (*, p <0.05; **, p <0.01; ***, p <0.001), assessed by ANOVA.

Article Snippet: After culture, the PBLs were fixed in 4% paraformaldehyde and permeabilized with 0.1% saponin (Sigma, St. Louis, USA) and stained with 15 μl of PE-conjugated ferret cross-reactive mouse monoclonal antibody to bovine IFN-γ (clone CC302, AbD-Serotec, Denmark) .

Techniques: Enzyme-linked Immunosorbent Assay, HI Assay, Isolation, Recombinant

Journal: PLoS ONE

Article Title: CAF01 Potentiates Immune Responses and Efficacy of an Inactivated Influenza Vaccine in Ferrets

doi: 10.1371/journal.pone.0022891

Figure Lengend Snippet: 6 to 10 months-old ferrets were immunized twice at two week- intervals with CAF01 adjuvanted (▴) and non-adjuvanted (▪) influenza vaccine Sanofi-Pasteur's Vaxigrip season 2005/2006 (n = 8 per group). A third group was mock-vaccinated (•). All ferrets were challenged with 10 7 TCID 50 of A/New Caledonia/20/99 (H1N1) four weeks after the second immunization (n = 8 per group). A. Relative amounts of viral RNA found in nasal washes of infected animals during the first six days of challenge, measured by quantitative RT-PCR. B. Vaccine-specific IgG titers measured in serum by ELISA C. Vaccine-specific IgA titers measured in nasal washes by ELISA. D. Hemagglutination inhibition assay titers using influenza A New Caledonia 1999 (H1N1). All ELISA values are expressed as geometric mean titers (GMT). E. Percentage of IFN-γ-positive lymphocytes after 4 hours stimulation with PMA and ionomycin. Values marked with an asterisk are significantly different (*, p <0.05; **, p <0.01; ***, p <0.001), assessed by ANOVA, except for B assessed by log-rank test.

Article Snippet: After culture, the PBLs were fixed in 4% paraformaldehyde and permeabilized with 0.1% saponin (Sigma, St. Louis, USA) and stained with 15 μl of PE-conjugated ferret cross-reactive mouse monoclonal antibody to bovine IFN-γ (clone CC302, AbD-Serotec, Denmark) .

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, HI Assay