Journal: PLoS ONE

Article Title: CXCL10/IP-10 Neutralization Can Ameliorate Lipopolysaccharide-Induced Acute Respiratory Distress Syndrome in Rats

doi: 10.1371/journal.pone.0169100

Figure Lengend Snippet: Monoclonal antibodies used for the analysis of inflammatory cells.

Article Snippet: CD3 , PerCP-Vio700 , Miltenyi Biotec , REA223 , 130-103-128 , 1 in 100.

Techniques: Bioprocessing, Control

Journal: Annals of Medicine and Surgery

Article Title: Da-yuan-yin decoction ameliorates inflammatory injury in severe pancreatitis by protecting intestinal mucosal barrier and immune function and preventing intestinal dysbiosis

doi: 10.1097/MS9.0000000000004873

Figure Lengend Snippet: Serum interleukin (IL)-1β and IL-10 expression levels (pg/ml) and cluster of differentiation (CD)3 + , CD4 + , and CD8 + cells from five ileal Peyer’s patches in the sham operation (SO), severe acute pancreatitis (SAP), and da-yuan-yin (DYY) + SAP groups, respectively. Serum IL-1β and IL-10 levels in the SAP and DYY + SAP groups were significantly higher than those in the SO group at 24, 48, and 72 h after surgery (all P < 0.05). Serum IL-1β and IL-10 levels in the DYY + SAP group were significantly lower than those in the SAP group 72 h postoperatively ( t = 4.37, P = 0.003; t = 2.41, P = 0.04, respectively). Levels of CD3 + , CD4 + , and CD8 + cells from ileal Peyer’s patches in the SAP and DYY + SAP groups decreased with time post-surgery. At 72 h postoperatively, the DYY + SAP group had significantly higher levels of CD3 + , CD4 + , and CD8 + cells than the SAP group ( t = 2.58, P = 0.036; t = 2.78, P = 0.027; t = 2.77, P = 0.028, respectively). There were no significant differences in early or late apoptosis in ileal Peyer’s patches 24, 48, and 72 h postoperatively in the SO, SAP, and DYY + SAP groups.

Article Snippet: Single-cell suspension is stained with the following antibodies: Live/Dead Fixable Violet Dead Cell (Cat# 2549272, Invitrogen), anti-Rat CD3 (Cat# G4.18, MultiSciences Biotech Co., Ltd), and anti-Rat CD4 (Cat# OX35, MultiSciences).

Techniques: Expressing

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:

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: Frontiers in Immunology

Article Title: Desmoglein-4 Deficiency Exacerbates Psoriasiform Dermatitis in Rats While Psoriasis Patients Displayed a Decreased Gene Expression of DSG4

doi: 10.3389/fimmu.2021.625617

Figure Lengend Snippet: Dsg-4 deficiency induces high mRNA expression of inflammatory genes in response to IMQ. RNA from skin was extracted, and subsequently real-time PCR and cDNA synthesis were performed to evaluate mRNA abundance of cytokines: IL-1β (A) , IL-8 (B) , IL-17 (C) , IL-10 (D) , and TGF-β (E) performing Kruskal–Wallis tests analysis. Additionally, TGF-β/IL-17 mRNA abundance means ratio was calculated (F) . Similarly, chemokine receptors mRNA levels were measured: CCR1 (G) , CCR2 (H) , CCR3 (I) , CCR5 (J) , and CXCR5 (K) . Kruskal–Wallis and ANOVA tests were used; data represent the median with values range; * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets. (L) Pseudocolor dot plots represent counts of infiltrating CD45 + cells and CD3 + cells from total skin cells analyzed by flow cytometry derived from SD and OFA rats. (M,N) Graphs represent the percentage and absolute cell counts of CD45 + CD3 + cells from total skin cells. ( n rats /group = 6; two independent experiments). (O,P) Spleen and inguinal lymph node weights. FACS and lymphatic tissue weight data are represented as the mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001 with ANOVA and Bonferroni post hoc test was used. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets.

Article Snippet: The cells were then resuspended in PBS/2% FBS and stained with PE-Cy5-conjugated anti-rat CD45 (clone OX-1; isotype mouse IgG1 k, BD Biosciences, San Jose, USA) and FITC-conjugated anti-rat CD3 (clone IF4; isotype mouse IgM, Cedarlane Laboratories, US).

Techniques: Expressing, Real-time Polymerase Chain Reaction, cDNA Synthesis, Flow Cytometry, Derivative Assay