Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Concomitant immunity against superimposed homologous Echinostoma caproni infections in mice is mediated by interleuquin-25

doi: 10.1590/0074-02760250004

Figure Lengend Snippet: primary and superimposed infection induce different patterns of macrophage activation. (A) Indirect immunofluorescence with recombinant anti-CD16-2/FCGR4/Fc for M1 and antibody MR/CD206/Mannose Receptor for M2 (red) on intestinal tissue of Institute of Cancer Research (ICR) naïve and mice after Echinostoma caproni primary, superimposed infection in mice and superimposed infections in anti-interleuquin-25 (α-IL-25)-treated mice (scale bar: 100 mm). (B) Quantitation of the percentage of area covered by anti-CD16-2/FCGR4/Fc for M1 and antibody MR/CD206/Mannose Receptor staining. Vertical bars show standard deviation. a: significant differences with respect to negative controls; b: significant differences with respect to primary infections; c: significant differences with respect to superimposed infection- in α-IL-25-treated mice (p < 0.05).

Article Snippet: A rabbit anti-CD16-2/FCGR4/Fc antibody (Sino Biological, clone no. 012), and a goat antibody MR/CD206/Mannose Receptor (Novus Invitro Technologies Ltd, Colorado, USA) were used for labelling M1 and M2, respectively.

Techniques: Infection, Activation Assay, Immunofluorescence, Recombinant, Quantitation Assay, Staining, Standard Deviation

Journal: Journal for Immunotherapy of Cancer

Article Title: Agonist anti-GITR monoclonal antibody and stereotactic radiation induce immune-mediated survival advantage in murine intracranial glioma

doi: 10.1186/s40425-016-0132-2

Figure Lengend Snippet: Survival benefit conferred by anti-GITR (1)/SRS treatment of murine glioma requires CD4+ T cells. C57/BL6 mice were inoculated with intracranial GL261-luc tumor, randomized to ≥7 mice per group and administered anti-GITR (1) and SRS as in Fig. . Mice were injected i.p. with 200 μl anti-CD4 (10 mg/kg) a and anti-CD8 (10 mg/kg) b depleting antibodies on days 5–7, 14, and 28 and followed for survival. Curve-adjacent asterisks compare indicated curve to control. The same control, anti-GITR (1), and anti-GITR (1)/SRS groups were used in ( a ) and ( b ) as the experiments were performed concurrently. The difference in survival between control mice and mice depleted of CD8+ T cells was statistically significant, but was not significant in the absence of CD4+ cells. FoxP3 DTR mice were inoculated intracranially with 1.3 × 10 5 GL261-luc cells, randomized, injected i.p. with 200 μl of diphtheria toxin (50 ng/g) on day 8, 9 and every 2 days thereafter in order to achieve FoxP3+ cell depletion, administered anti-GITR (1) or SRS as in Fig. ., and followed for tumor growth by bioluminescent imaging c . Tumor size was quantified by bioluminescence d . * P < .05; ** P < .01. NS, non-significant

Article Snippet: The following antibodies were used: LIVE/DEAD Aqua (Life Technologies, L34957), CD3 APC/Cy7 (BioLegend, 300317), CD3e PerCP/Cy5.5 (BD, 561108), CD4 PB (Invitrogen, MHCD0428), CD8 BV605 (BD, 564115), CD25 PE-Cy7 (eBioscience, 25025942), Foxp3 PE (eBioscience, 12477182), IFN-γ PE-Cy7 (eBioscience, 25731141), TNF-α FITC (BD, 554418), IL-2 APC (BD, 562041), F4/80 PE-Cy7 (BioLegend, 123113), CD16/CD32 APC (eBioscience, 17016181), CD16-2/FCGR4 PE (SinoBiological, 50036R012P10), CD11b FITC (eBiosciences, 11011281), CD45 AF700 (BioLegend, 103127).

Techniques: Injection, Imaging

Journal: Journal for Immunotherapy of Cancer

Article Title: Agonist anti-GITR monoclonal antibody and stereotactic radiation induce immune-mediated survival advantage in murine intracranial glioma

doi: 10.1186/s40425-016-0132-2

Figure Lengend Snippet: Tumor infiltrating lymphocytes (TIL) in the anti-GITR (1)/SRS group have a Th1 immunophenotype and elevated effector to Treg ratio. C57BL/6 mice were inoculated with GL261-luc tumor, randomized to groups of ≥5, and dosed with anti-GITR (1) and SRS as in Fig. . Mice were sacrificed on day 21, tumor infiltrating lymphocytes were isolated, cells were stimulated with PMA/Ionomycin, fixed, and permeabilized for staining of intracellular markers. a CD8+ and b CD4+ cell populations were analyzed by flow cytometry for expression of IFNγ, TNFα, and IL-2. Intratumoral effector CD8+ and CD4+ to Treg ratios in the anti-GITR (1)/SRS group were calculated c . Symbol and horizontal bar denote single mouse and average value, respectively. * P < .05

Article Snippet: The following antibodies were used: LIVE/DEAD Aqua (Life Technologies, L34957), CD3 APC/Cy7 (BioLegend, 300317), CD3e PerCP/Cy5.5 (BD, 561108), CD4 PB (Invitrogen, MHCD0428), CD8 BV605 (BD, 564115), CD25 PE-Cy7 (eBioscience, 25025942), Foxp3 PE (eBioscience, 12477182), IFN-γ PE-Cy7 (eBioscience, 25731141), TNF-α FITC (BD, 554418), IL-2 APC (BD, 562041), F4/80 PE-Cy7 (BioLegend, 123113), CD16/CD32 APC (eBioscience, 17016181), CD16-2/FCGR4 PE (SinoBiological, 50036R012P10), CD11b FITC (eBiosciences, 11011281), CD45 AF700 (BioLegend, 103127).

Techniques: Isolation, Staining, Flow Cytometry, Expressing