Journal: International immunopharmacology

Article Title: Modulation of T lymphocyte and eosinophil functions in vitro by natural tetranortriterpenoids isolated from Carapa guianensis Aublet.

doi: 10.1016/j.intimp.2010.09.010

Figure Lengend Snippet: Fig. 4. Effect of TNTPs on T lymphocyte activation. Pretreated splenocytes (105 cells/well) with pooled TNTPs 50 μg/ml for 1 h before stimulation were cultured in α-CD3 mAb-coated wells (10 μg/ml in 5% CO2 at 37 °C) for 24 or 72 h. (A) Splenocyte proliferation assessed 72 h after α-CD3 mAb stimulation. Percentages (B, E) and mean fluorescence intensity (MFI) (C, F) of the expression of CD25 and CD69, respectively, by CD3+ T lymphocytes 24 h after stimulation is shown. Data represents the mean±SEM from quadruplicate wells per group, as statistically analyzed by means of analysis of variance (ANOVA), followed by Newman–Keuls–Student test or Student t test. * indicates statistically significant differences (p≤0.05) between non-stimulated and stimulated animals, whereas + indicates significant differences between the stimulated and treated groups. Representative histograms of proliferation (A), CD25 (D) and CD69 (G) expression on T lymphocytes are shown.

Article Snippet: Antimurine eotaxin and anti-IL-2 mAbs, biotinylated anti-CCL11 and antiIL2 polyclonal antibodies (pAb), recombinant CCL11 and IL-2, FITCconjugated hamster IgG anti-murine CD3, PE-conjugated hamster IgG anti-murine CD25, PE-conjugated rat IgG anti-murine CD69, PEconjugated IgG and FITC-conjugated IgG were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Activation Assay, Cell Culture, Expressing

Journal:

Article Title: 111 Indium-labelled human gut-derived T cells from healthy subjects with strong in vitro adhesion to MAdCAM-1 show no detectable homing to the gut in vivo

doi: 10.1111/j.1365-2249.2004.02578.x

Figure Lengend Snippet: Phenotype of infused cultured gut-derived T cells (unseparated cultures)

Article Snippet: Antibodies The following antibodies were obtained from Becton Dickinson: CD25 (mouse IgG 1 , 2A3), CD69 (mouse IgG 1 , L78), CD4 (mouse IgG 1, SK3), CD3 (mouse IgG 1 , SK7), CD62L (mouse IgG 2A , SK11), Integrin β 1 (mouse IgG 1 , MAR4), Integrin β 7 (rat IgG 2A , FIB504); from Immunotech: Integrin α 4 (mouse IgG 1 , HP2/1), CD45RO (mouse IgG 2A , UCHL1), CD11a (mouse IgG 1 , SK7); and ACT-1 (mouse IgG) against heterodimer α4β7 [ 12 ], donated by LeukoSite (Cambridge, MA, USA), was labelled with secondary antibody Alexa(488)Fluor®-goat-anti-mouse (Molecular Probes, Leiden, the Netherlands).

Techniques: Cell Culture

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: Numbers and activation status of Foxp3+ regulatory T cells were determined on various days post-infection with PyL or PyNL in (A–D) WT mice or (E–F) Foxp3-GFP transgenic mice. (A) Representative dot plot of intracellular staining in CD4+ T cells. On selected days post-infection with PyL or PYNL, splenic CD4 + lymphocytes were analysed for (B) numbers of Foxp3 + cells,(C) intensity (MFI) of Foxp3 staining and (D) ratio of Foxp3 + to Foxp3 − cells. (E) Representative dot plots showing GFP (Foxp3) expression in CD4+ splenocytes from 7 day PyL and PyNL-infected animals and uninfected controls. (F) Numbers of splenic GFP + (Foxp3 + ) CD4 + regulatory T cells on day 5 post infection. 3–5 mice per group. Results are representative of 3 independent experiments. Symbols represent significant differences (p<0.05) between groups: # PyL vs PyNL; * PyL vs uninfected; ∼ PyNL vs uninfected.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Activation Assay, Infection, Transgenic Assay, Staining, Expressing

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: (A–C) Mice were given either a single dose of 0.75 mg 7D4 (IgM clone) or 0.25 mg 7D4 combined with 0.75 mg PC61 (IgG1 clone) on 3 days prior to infection with 10 4 PyL pRBC. The effect of anti-CD25 treatment on the course of PyL infection was examined in (A–C) C57BL/6 mice by following (A) parasitaemia, (B) anaemia and (C) weight loss. Groups consisted of 3–5 mice and data are representative of 2 independent experiments. Symbols represent significant differences (p<0.05) between groups: (A–D) # 7D4 vs 7D4/PC61; * 7D4 vs PBS; ∼ 7D4/PC61 vs PBS; (G–H) # 7D4 vs 7D4/PC61; * 7D4 vs 7D4×3; ∼ 7D4 vs PBS; + 7D4/PC61 vs 7D4×3; Φ 7D4/PC61 vs PBS; Δ 7D4×3 vs PBS. (D–F) Naive CD4 + CD25 − (non-Treg) and CD4 + CD25 hi (Treg) cells were purified by flow cytometric cell sorting and adoptively transferred alone or at a 10∶1 ratio (non Treg∶Treg) into RAG-1 −/− mice prior to infection with PyL parasites. (D) shows the purity of sorted CD4 + CD25 − and CD4 + CD25 hi populations prior to adoptive transfer and the relative expression of Foxp3 within the purified populations. The affect of adoptive transfer on the course of infection was determined by monitoring (E) parasitaemia and (F) survival for the duration of the experiment. Groups consisted of 5 mice and the results are representative of 2 independent experiments. Symbols represent significant differences (p<0.05) between groups: # CD25 − vs CD25 + ; * CD25 − vs CD25 − / CD25 + ; ∼ CD25 − vs control; + CD25 + vs CD25 − / CD25 + ; Φ CD25 + vs control; Δ CD25 − / CD25 + vs control.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Infection, Purification, FACS, Adoptive Transfer Assay, Expressing

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: (A) On selected days post-infection, CD4 + T cells were purified from PyL and PyNL infected mice by MACS sorting and levels of IL-10 mRNA were determined by real time PCR (Taqman) relative to the house keeping gene GAPDH. The results are shown as the fold change in expression relative to uninfected naïve CD4+ T cells. In separate experiments the levels of IL-10 in the plasma of WT and RAG-1 −/− mice were determined by ELISA on selected days of (B) PyL and (C) PyNL infection. Groups consisted of 3–5 mice and the results are representative of 2 separate experiments. For Taqman analysis, purified cells from several mice in each group were pooled; no significant differences between groups were identified. * indicates significant differences (p<0.05) between WT and RAG-1−/− infected mice.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Infection, Purification, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: Transgenic, IL-10-GFP knockin tiger mice were infected with PyL or PyNL. (A) Splenic lymphocytes from infected or uninfected control mice were analysed for expression of CD4+ and GFP. (B,C) 7 days post infection, splenic CD4 + T cells were analysed for (B) expression of CD25 and Foxp3, or (C) GFP (IL-10) and CD25, CD69, CD62L or CD127. (D) The frequency and number of GFP+ (IL-10+) CD4 + T cells was calculated 7 days post infection in infected or uninfected mice. Groups consisted of 3–5 mice and the results are representative of 2 independent experiments.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Transgenic Assay, Knock-In, Infection, Expressing

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: Splenic CD4 + T cells were isolated from transgenic IL-10-GFP mice on day 7 of PyL or PyNL infection and were purified by flow cytometric cell sorting into GFP + (IL-10 + ) and GFP − (IL-10 − ) populations. (A) shows the purity of the purified GFP + and GFP − populations. (B) Expression of IL-10, Foxp3, IFN-γ, IL-4, IL-13 and IL-17A mRNA was determined by real time PCR (Taqman) relative to the house keeping gene, GAPDH. The results are shown as the fold change in expression relative to uninfected naïve CD4+ T cells. For Taqman analysis, purified cells from several mice in each group were pooled.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Isolation, Transgenic Assay, Infection, Purification, FACS, Expressing, Real-time Polymerase Chain Reaction

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: Prior to infection with (A–D) PyNL and (E–L) PyL with (A–H) 10 4 or (I–L) 10 3 pRBC's, RAG-1 −/− mice received naïve CD4 + T cells that had been purified from either WT or IL-10 −/− mice by magnetic bead sorting. The course of infection was followed by monitoring (A, E, I) parasitaemia (B, F, J) anaemia and (C, G, K) weight loss and (D, H, L) survival. Groups consisted of 4 mice and the results are representative of 2 independent experiments. * indicates significant differences (p<0.05) between mice receiving WT and IL-10−/− CD4 T cells.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Infection, Purification

Journal: PLoS Pathogens

Article Title: IL-10 from CD4 + CD25 − Foxp3 − CD127 − Adaptive Regulatory T Cells Modulates Parasite Clearance and Pathology during Malaria Infection

doi: 10.1371/journal.ppat.1000004

Figure Lengend Snippet: Liver pathology was examined (A) in WT and IL-10 −/− mice that were either uninfected or had been infected with P. yoelii 7 days (PyL, PyNL) or 14 days (PyNL) previously, (B) on day 25 post-infection with PyNL in RAG-1 −/− mice reconstituted with either WT or IL-10 −/− CD4+ T cells prior to infection. Groups consisted of 4–5 mice and the slides shown are representative of mice from 2 independent experiments. Arrows highlight areas of interest: c = central vein periportal infiltration, pv = vessels packed with inflammatory cells, p = pigmented kupffer cells, n = necrosis, i = inflammation in parenchymia, pt = necrosis and gross infiltration in portal triad.

Article Snippet: For flow cytometric analysis, cells were surface stained with anti-mouse CD4 (RM4-5; Rat IgG 2a ; BD Biosciences), anti-mouse CD25 (PC61; Rat IgG 1 ; eBioscience), anti-mouse CD69 (H1.2F3; Armenian Hamster IgG; eBioscience), anti-mouse CD62L (MEL-14; Rat IgG2a; eBioscience) or anti-mouse CD127 (A7R34; Rat IgG2a; eBioscience).

Techniques: Infection