Journal: PLoS ONE

Article Title: High-Anxious Individuals Show Increased Chronic Stress Burden, Decreased Protective Immunity, and Increased Cancer Progression in a Mouse Model of Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0033069

Figure Lengend Snippet: Percentages (%) and numbers (#) of lymphocytes in sentinel lymph nodes of low-anxious versus high-anxious mice.

Article Snippet: Directly conjugated rat anti-mouse CD3, CD4, CD8, and B220 (BD-Pharmingen, San Diego, CA) antibodies were used to label cells.

Techniques:

Journal: Frontiers in Immunology

Article Title: LigA formulated in AS04 or Montanide ISA720VG induced superior immune response compared to alum, which correlated to protective efficacy in a hamster model of leptospirosis

doi: 10.3389/fimmu.2022.985802

Figure Lengend Snippet: Evaluation of the long-term memory response induced by various vaccines. Mice were either bled and killed 6 months (day180) post-immunization or boosted and killed after 1 week (day 187) to analyze B-cell (antibodies) and T-cell memory response. (A) B-cell memory response. The antibodies including isotypes were analyzed in serum (obtained at day 180 and 187) by ELISA as described in Materials and Methods . (B) T-cell memory response. The splenocytes recovered from mice euthanized on days 180 and 187 were subjected to in vitro stimulation with recall antigen (LAV) and proliferation was assessed by counting the cells after 72 h. (C) Analysis of memory phenotype. Lymphocytes were stained with antibodies specific for CD3, CD4, CD8, CD62L, and CD44 and analyzed by flow cytometry. Representative flow cytometry plots show the gating strategy to analyze both central (CD44 high and CD62L high ) and effector (CD44 high , CD62L low ) population of CD4 and CD8 T cells. The gated population obtained were plotted as histograms to show each phenotype. (D) Analysis of germinal centers. Draining lymph nodes (DLNs) taken from mice at 28 days post-immunization were processed as frozen tissue sections. Consecutive sections were stained with fluorochrome-conjugated anti-mouse CD3/CD45R/B220/GL-7 antibodies, mounted, and then examined under a fluorescent microscope using the tile scanning and stitched using the ZEN software as described in Materials and Methods . Data are representative of three different experiments. Significant differences were calculated using the one-way ANOVA (****,***, **, *, and ns indicates P < 0.0001, P < 0.001, <0.01, P < 0.05 and non-significant respectively).

Article Snippet: After rehydration in Tris-buffered saline and blocking in Tris-buffered saline with 5% BSA and 0.05% Tween 20, the sections were stained with FITC-conjugated rat anti-mouse CD3 (555274, BD Biosciences, 1:300), Per-CP-conjugated CD45R (B220), Anti-mouse (130-102-815, Miltenyi Biotec, 1:100), and PE-conjugated T- and B-Cell Activation Antigen (GL-7) Rat Anti-Mouse (561530, BD Biosciences, 1:200).

Techniques: Enzyme-linked Immunosorbent Assay, In Vitro, Staining, Flow Cytometry, Microscopy, Software

Journal: PLoS ONE

Article Title: Mucosal immunity of mannose-modified chitosan microspheres loaded with the nontyepable Haemophilus influenzae outer membrane protein P6 in BALB/c mice

doi: 10.1371/journal.pone.0269153

Figure Lengend Snippet: A “Three-Color, Dual Anchor” gating strategy to identify the lymphocyte subsets (CD3 + , CD4 + and CD8 + ); the proportions are shown. Values are expressed as a percentage. Cells were stained with APC-Cy ™ 7 Rat Anti-Mouse CD3, FITC Rat Anti-Mouse CD4 and PE Rat Anti-Mouse CD8a. B Comparison of CD3 + T cell proportions in spleen lymphocytes. C Comparison of CD3 + CD4 + T cell proportions in spleen lymphocytes. D Comparison of CD3 + CD8 + T cell proportions in spleen lymphocytes. Mean ± SD, n = 3. * P <0.05, ** P <0.01.

Article Snippet: Then, the lymphocytes were adjusted to a concentration of 1×10 7 cells/ml, and 100 μl of cells were stained with APC-Cy ™ 7 Rat Anti-Mouse CD3, FITC Rat Anti-Mouse CD4 and PE Rat Anti-Mouse CD8a (BD Biosciences, San Diego, US) at room temperature for 30 min.

Techniques: Staining

Journal: Journal of Immunology Research

Article Title: Amiselimod (MT-1303), a Novel Sphingosine 1-Phosphate Receptor-1 Modulator, Potently Inhibits the Progression of Lupus Nephritis in Two Murine SLE Models

doi: 10.1155/2019/5821589

Figure Lengend Snippet: Prophylactic effect of MT-1303 on the development of proteinuria in MRL/ lpr mice. MT-1303 was administered to MRL/ lpr mice for 18 weeks from 8 weeks of age. (a) Proteinuria was assessed once a week using Ames urinalysis strips and was scored on a scale of 0–4 based on urinary protein concentrations as described in Materials and Methods. Each symbol represents the mean ± S.E.M. score of 12 animals. Statistical significance was calculated using the Shirley-Williams test by comparison with the vehicle-treated control group ( ∗ p < 0.05, ∗∗ p < 0.01). (b, c) Kidney sections from vehicle- (b) or MT-1303 0.3 mg/kg- (c) treated mice were labeled with anti-mouse CD3 mAb. (d, e) The axillary lymph nodes (d) and spleen (e) were weighed on the day after final administration. Results are expressed as the mean ± S.E.M. of 8 mice. (f–h) The number of T cells (d), B cells (e), and abnormal T cells (CD3 + B220 + ) (f) was measured by flow cytometry. Results are expressed as the mean ± S.E.M. of 4 mice. (d–h) Statistical differences were calculated using the Williams test by comparison with the control ( ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: The sections were incubated with rat anti-mouse CD3 mAb (555273) followed by secondary mAb conjugated to amino acid polymer and peroxidase (Histofine®, Nichirei Biosciences, Tokyo, Japan).

Techniques: Comparison, Control, Labeling, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Amiselimod (MT-1303), a Novel Sphingosine 1-Phosphate Receptor-1 Modulator, Potently Inhibits the Progression of Lupus Nephritis in Two Murine SLE Models

doi: 10.1155/2019/5821589

Figure Lengend Snippet: Therapeutic effects of MT-1303 and FK506 on established proteinuria in aged MRL/ lpr mice. MT-1303 and FK506 were orally administered to MRL/ lpr mice with a proteinuria score of 2 from 20 to 26 weeks of age. Proteinuria was assessed once a week using Ames urinalysis strips. (a, b) Proteinuria scores following administration of MT-1303 (a) and FK506 (b) are expressed as the mean ± S.E.M. of 16 mice. Statistical differences were calculated using the Shirley-Williams test by comparison with the control ( ∗ p < 0.05, ∗∗ p < 0.01). (c, d) The incidence of proteinuria in MT-1303 groups (c) and FK506 groups (d) is expressed as the proportion of proteinuria-positive mice (proteinuria score ≥ 2) from a total of 16 mice. Statistical differences were calculated using Fisher's exact test with Hommel's multiple comparison test by comparison with the control ( ∗ p < 0.05, ∗∗ p < 0.01). (e–h) Kidney sections from vehicle- (e, g) or MT-1303 0.3 mg/kg- (f, h) treated mice were stained with anti-mouse CD3 mAb (e, f) or HE (g, h).

Article Snippet: The sections were incubated with rat anti-mouse CD3 mAb (555273) followed by secondary mAb conjugated to amino acid polymer and peroxidase (Histofine®, Nichirei Biosciences, Tokyo, Japan).

Techniques: Comparison, Control, Staining