Journal: Cancer gene therapy

Article Title: Anti-tumor immunity elicited by direct intratumoral administration of a recombinant adenovirus expressing either IL-28A/IFN-λ2 or IL-29/IFN-λ1.

doi: 10.1038/cgt.2016.29

Figure Lengend Snippet: Figure 6. Treatment with either AdIL-28A or AdIL-29 was associated with increased interferon (IFN)-γ production in splenic CD8+ T cells and augmented tumor-specific cytotoxic T-cell activity. (a, b) Groups of three mice, inoculated with either (a) MCA205 or (b) B16-F10 cells, were treated with AdNull, AdIL-28A or AdIL-29 on days 6 and 9. On day 11, CD8+ T cells were separated from spleen cells and stimulated with anti- CD3 mAb for 48 h. IFN-γ levels in culture supernatants were measured using ELISA. Data are shown as mean ± s.d. (n = 3). The result is representative of two independent experiments. (a) AdNull versus AdIL-28A; Po0.0005, AdNull versus AdIL-29; Po0.004, (b) AdNull versus AdIL-28A; Po0.0004, AdNull versus AdIL-29; Po0.001. (c, d) MCA205 tumors were treated with intratumoral injection of AdIL-28A, AdIL-29, AdNull or phosphate-buffered saline (PBS) on days 6 and 9. On day 14, spleen cells were isolated and restimulated with mitomycin C-treated MCA205 cells for 5 days. The restimulated effector cells were assayed for cytolytic activity by using (c) MCA205 or (d) 3LL cells as targets in a standard 51Cr release assay. All results are shown as ± s.d. (n = 4 per data point). The result is representative of two independent experiments. (e, f) B16-F10 tumors were treated with AdIL-28A, AdIL-29, AdNull or PBS on days 6 and 9. On day 14, spleen cells were isolated and restimulated with mitomycin C-treated B16-F10 cells for 5 days. The restimulated effector cells were assayed for cytolytic activity by using (e) B16-F10 or (f) 3LL cells as targets in a standard 51Cr release assay. All results are shown as ± s.d. (n = 4 per data point). The result is representative of two independent experiments.

Article Snippet: CD8+ T cells (1 × 106 per well) were cultured for 48 h in a 48-well culture plate precoated with 1.0 μg of anti-mouse CD3 epsilon mAb (R&D Systems).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Injection, Saline, Isolation, Release Assay

Journal:

Article Title: Expression and regulation in the brain of the chemokine CCL27 gene locus

doi: 10.1016/j.jneuroim.2010.04.019

Figure Lengend Snippet: Digital images at 200 times magnification showing FITC immunofluorescent signals corresponding to CD3 positive cells (A) in the meningeal spaces of the olfactory bulbs of mice sensitized and challenged with albumin from chicken egg (OVA). Images obtained from Hoechst staining (B) were merged with those of CD3 to confirm co-localization of the CD3 signal with nuclear DNA (C).

Article Snippet: Sections were incubated with the same goat anti-mouse CCL27 antibody used in the fresh frozen protocol for 24 hours at 4° C (1:1,000 dilution) in combination with mouse monoclonal anti-NeuN (1:10,000 dilution) or rabbit polyclonal anti-glial fibrillary acidic protein (GFAP) (1:5000 dilution) or anti-CD3 antibody (Serotec; MCA 1477at 1:1000 dilution).

Techniques: Staining

Journal:

Article Title: Expression and regulation in the brain of the chemokine CCL27 gene locus

doi: 10.1016/j.jneuroim.2010.04.019

Figure Lengend Snippet: Digital images at 100 times magnification (A, B, C) or 400 times magnification (D, E, F) of FITC immunofluorescent signals corresponding to CD3 positive cells (A, D) in the olfactory bulbs of mice sensitized and challenged with albumin from chicken egg (OVA). Images obtained from Hoechst staining (B, E) were merged with those of CD3 to confirm co-localization of the CD3 signal with nuclear DNA (C, F). Arrows indicate some identified CD3 positive cells in the glomerular cell layer (Gl). EPl: external plexiform cell layer; Mi: mitral cell layer; IGr: internal granular cell layer.

Article Snippet: Sections were incubated with the same goat anti-mouse CCL27 antibody used in the fresh frozen protocol for 24 hours at 4° C (1:1,000 dilution) in combination with mouse monoclonal anti-NeuN (1:10,000 dilution) or rabbit polyclonal anti-glial fibrillary acidic protein (GFAP) (1:5000 dilution) or anti-CD3 antibody (Serotec; MCA 1477at 1:1000 dilution).

Techniques: Staining

Journal:

Article Title: Expression and regulation in the brain of the chemokine CCL27 gene locus

doi: 10.1016/j.jneuroim.2010.04.019

Figure Lengend Snippet: Digital image of double fluorescent immunohistochemistry of a CD3 positive cell (FITC, green) in close proximity of CCL27 positive (Cy3, red) cells in the glomerular cell layer of the olfactory bulbs of mice sensitized and challenged with albumin from chicken egg (OVA). Sections were counter stained with Hoechst to reveal nuclear DNA (blue). Scale bar 10 μm.

Article Snippet: Sections were incubated with the same goat anti-mouse CCL27 antibody used in the fresh frozen protocol for 24 hours at 4° C (1:1,000 dilution) in combination with mouse monoclonal anti-NeuN (1:10,000 dilution) or rabbit polyclonal anti-glial fibrillary acidic protein (GFAP) (1:5000 dilution) or anti-CD3 antibody (Serotec; MCA 1477at 1:1000 dilution).

Techniques: Immunohistochemistry, Staining

Journal: The Journal of Experimental Medicine

Article Title: Neuroprotective intervention by interferon-γ blockade prevents CD8 + T cell–mediated dendrite and synapse loss

doi: 10.1084/jem.20122143

Figure Lengend Snippet: Deafferentation in viral déjà vu disease requires CTL contact with infected neurons. (A–D) Carrier mice (C57BL/6 WT or YFP reporter mice) and noncarrier controls (without neonatal infection) were challenged with LCMVwt (10 4 PFU) i.v. in adulthood. (A) 10 d later, animals were sacrificed and brains were processed for histological analysis Left: representative section stained for synaptophysin + perisomatic boutons (arrowheads) in the DCN of carrier and noncarrier mice 10 d after challenge. Right: quantification of perisomatic bouton density. Symbols represent individual animals. (B) On day 8 after challenge, CNS sections of DCN were triple immunostained for T cells, synaptophysin, and LCMV-NP antigen ( n = 4 animals). Perisomatic bouton density was quantified in LCMV-NP–positive (+) neurons that were in juxtaposition to infiltrating T cells. (C) Dendritic projections into the spinal cord white matter in inflamed and noninflamed areas were quantified on MAP-2–immunostained sections ( n = 4). (D) T cell infiltrates (CD3 + , red) in the spinal cord of YFP-H mice (yellow subset of neurons). Proximity of T cells to somata of ventral horn neurons (left) and dendrites (middle) is shown. Dendrites without neighboring T cells (right) are also shown. Monochromatic YFP images (middle and right only) illustrate dendrite morphology. Nuclei were visualized with neurotrace (NT). Error bars in B and C represent the mean + SEM of 4 mice per group. Bars: (A) 20 µm; (C, top) 200 µm; (C, bottom) 50 µm; (D, left) 20 µm; (D, middle and right) 10 µm. One representative dataset out of two similar experiments is shown for A–D. **, P < 0.01.

Article Snippet: PFA-fixed sections were stained with primary antibodies: rat anti–human CD3 (cross-reactive with murine T cells; Serotec), mouse anti-neuronal nuclei NeuN (EMD Millipore), rabbit anti–Iba-1 (microglia/macrophages; Wako Chemicals USA), rat anti-LCMV nucleoprotein sera (generated by prime-boost immunization against purified LCMV-NP), rabbit anti–phospho-STAT1 (detecting only phosphorylated STAT1 at position Tyrosine 701; Cell Signaling Technology), mouse anti–rat synaptophysin (cross-reactive with mouse; clone SY38c; Dako), and mouse anti–human CD8 (clone C8/114B; Dako).

Techniques: Infection, Staining

Journal: Food Science & Nutrition

Article Title: Combining Network Pharmacology, Molecular Docking, and Integrative Studies to Explore the Mechanism of Helminthostachys zeylanica in Alleviating Ulcerative Colitis

doi: 10.1002/fsn3.71139

Figure Lengend Snippet: (A) TNF‐α concentration in the plasma of mice from all groups. * p < 0.05 versus DSS mice; ** p < 0.01 versus DSS mice. (B) IL‐6 concentration in the colon and intestinal tissues of mice from all groups. * p < 0.01 (colon tissue); # p < 0.05 (intestinal tissue). (C) TNF‐α concentration in the medium of IEC‐6 and T84 cells from all groups. (D) IL‐6 concentration in the medium of IEC‐6 and T84 cells from all groups. (E) Immunofluorescent staining with CD3 in colon tissues from mice. Red staining indicates CD3, while DAPI (blue) is a nuclear stain. All images are magnified ×400.

Article Snippet: Monoclonal antibodies to CD3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA; Cat# sc‐20,047).

Techniques: Concentration Assay, Clinical Proteomics, Staining

Journal: Food Science & Nutrition

Article Title: Combining Network Pharmacology, Molecular Docking, and Integrative Studies to Explore the Mechanism of Helminthostachys zeylanica in Alleviating Ulcerative Colitis

doi: 10.1002/fsn3.71139

Figure Lengend Snippet: The mechanism of HZ in UC involves the TLR4/NF‐κB pathway. The anti‐inflammatory properties of HZ on UC are mediated through the TLR4/NF‐κB signaling pathway, encompassing the inhibition of cytokine production and prevention of T cell activation and differentiation. HZ achieves a downregulation of TLR4 and NF‐κB, resulting in reduced cellular release of TNF and IL‐6. Subsequent molecular docking simulation analysis reveals that the pure compounds of HZ, namely UgoninM, O, R, and K, exert their inhibitory effects on the TLR4/NF‐κB signaling pathway by influencing proteins such as TAK1, IKKβ, and RELA. Consequently, this impedes the degradation of the NF‐κB trimer, preventing its entry into the cell nucleus and activating pro‐inflammatory effects. On another note, CD3+ T cells, when associated, play a role in enhancing the production of specific antibodies by immune cells through the secretion of cytokines such as IL‐6 and TNF, thereby amplifying the extent of humoral immune responses.

Article Snippet: Monoclonal antibodies to CD3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA; Cat# sc‐20,047).

Techniques: Inhibition, Activation Assay