mouse cd4 t cell enrichment column  (R&D Systems)

Journal: PLoS ONE

Article Title: Amiselimod (MT-1303), a novel sphingosine 1-phosphate receptor-1 functional antagonist, inhibits progress of chronic colitis induced by transfer of CD4 + CD45RB high T cells

doi: 10.1371/journal.pone.0226154

Figure Lengend Snippet: MT-1303 (0.3 mg/kg) or vehicle was orally administered to C57BL/6 mice for 3 days and S1P 1 receptor expression on CD4 + T cells from mesenteric lymph nodes was determined using flow cytometry. (A) Representative histograms of S1P 1 receptor expression in each group. Black, isotype control (rat IgG); red, vehicle-treated mice (S1P 1 ); blue, MT-1303-treated mice (S1P 1 ). (B) Mean fluorescence intensity (MFI) of the S1P 1 receptor on CD4 + T cells. Data are expressed as the mean ± S.E.M. (n = 3), and statistical differences were calculated using Student’s t -test after logarithmic transformation. ** p < 0.01, compared with the vehicle-treated group. (C) The number of CD4 + T cells in peripheral blood was measured using flow cytometry. Data are expressed as the mean ± S.E.M.; n = 3. Statistical differences were calculated using Student’s t -test. ** p < 0.01, compared with the vehicle-treated group.

Article Snippet: Rat anti-mouse S1P 1 antibody (713412) and mouse CD4 + T cell enrichment column were obtained from R&D Systems, and the Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec.

Techniques: Expressing, Flow Cytometry, Control, Fluorescence, Transformation Assay

Journal: PLoS ONE

Article Title: Amiselimod (MT-1303), a novel sphingosine 1-phosphate receptor-1 functional antagonist, inhibits progress of chronic colitis induced by transfer of CD4 + CD45RB high T cells

doi: 10.1371/journal.pone.0226154

Figure Lengend Snippet: SCID mice were injected with CD4 + CD45RB high T cells to induce colitis. MT-1303 or vehicle was orally administered to SCID mice every day from the day of CD4 + CD45RB high T cell transfer for 28 days. (A) Change in body weight over time. Body weight on each day is expressed as a percentage of the original weight. Each symbol represents the mean ± S.E.M. of body weight (%) in 14–15 mice (n = 14: normal group). Statistical differences were determined using Student’s t -test by comparing to the normal group (# p < 0.05, ## p < 0.01) or using Dunnett’s test by comparing with the vehicle-treated group (* p < 0.05, ** p < 0.01). (B) Clinical scores were determined the day after the final administration. Each column represents the mean ± S.E.M. of clinical scores in 14–15 mice. Statistical differences were determined using the Wilcoxon test by comparing with the normal group (## p < 0.01) or using Steel’s test by comparing with the vehicle-treated group (** p < 0.01). (C, D) Colon sections from vehicle- (C) or MT-1303 0.3 mg/kg (D)-treated mice were stained with hematoxylin-eosin.

Article Snippet: Rat anti-mouse S1P 1 antibody (713412) and mouse CD4 + T cell enrichment column were obtained from R&D Systems, and the Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec.

Techniques: Injection, Staining

Journal: PLoS ONE

Article Title: Amiselimod (MT-1303), a novel sphingosine 1-phosphate receptor-1 functional antagonist, inhibits progress of chronic colitis induced by transfer of CD4 + CD45RB high T cells

doi: 10.1371/journal.pone.0226154

Figure Lengend Snippet: SCID mice were injected with CD4 + CD45RB high T cells to induce colitis. MT-1303 was orally administered to SCID mice every day from day 7 after CD4 + CD45RB high T cell transfer for 21 days, and anti-mTNF-α mAb was intraperitoneally administered to SCID mice on day 7 and day 21 after cell transfer. (A, B) Change in body weight over time. Body weight on each day is expressed as a percentage of the original weight. Each symbol represents the mean ± S.E.M. of body weight (%) in 18 mice. Statistical differences were determined using Student’s t -test by comparing with the normal group (# p < 0.05, ## p < 0.01), using Dunnett’s test by comparing with the vehicle-treated group (* p < 0.05) or using Student’s t -test by comparing with the vehicle-treated group († p < 0.05, †† p < 0.01). (C) Clinical scores were determined on day 28 after cell transfer. Each column represents the mean ± S.E.M. of clinical scores in 18 mice. Statistical differences were determined using the Wilcoxon test by comparing with the normal group (## p < 0.01), using Steel’s test by comparing with the vehicle-treated group (** p < 0.01) or using the Wilcoxon test by comparing with the vehicle-treated group (†† p < 0.01). There were no significant differences between the anti-mTNF-α mAb-treated group and MT-1303-treated groups using Steel’s test.

Article Snippet: Rat anti-mouse S1P 1 antibody (713412) and mouse CD4 + T cell enrichment column were obtained from R&D Systems, and the Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec.

Techniques: Injection

Journal: PLoS ONE

Article Title: Amiselimod (MT-1303), a novel sphingosine 1-phosphate receptor-1 functional antagonist, inhibits progress of chronic colitis induced by transfer of CD4 + CD45RB high T cells

doi: 10.1371/journal.pone.0226154

Figure Lengend Snippet: MT-1303 or vehicle was orally administered to SCID mice every day from week 1 after CD4 + CD45RB high T cell transfer for 3–4 weeks. (A, B) The number of lymphocytes (A) and CD4 + T cells (B) in the lamina propria of the colon was measured using flow cytometry the day after the final administration. (C–E) LPLs were stimulated with PMA and ionomycin in the presence of brefeldin A for 6 h before intracellular cytokine staining was performed using anti-IFN-γ and anti-IL-17 mAbs (C). Th1 (D) and Th17 (E) cell counts in the lamina propria of the colon were determined. (F, G) After stimulation with PMA and ionomycin, the amount of IFN-γ (F) and IL-17 (G) in the culture supernatant was determined using a BD TM Cytometric Beads Array. Each bar represents the mean ± S.E.M. (n = 11: vehicle-treated group, n = 12: MT-1303-treated group). Statistical differences were determined using Student’s t -test by comparing with the normal group (* p < 0.05, ** p < 0.01).

Article Snippet: Rat anti-mouse S1P 1 antibody (713412) and mouse CD4 + T cell enrichment column were obtained from R&D Systems, and the Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec.

Techniques: Flow Cytometry, Staining

Journal:

Article Title: Primary Role for CD4 + T Lymphocytes in Recovery from Oropharyngeal Candidiasis

doi:

Figure Lengend Snippet: Time course of C. albicans clearance in BALB/c nu/nu mice reconstituted with 2.5 × 106 naive CD4+ or CD8+ T cells. The positive control group received 3 × 107 naive lymphocytes, while the negative control group received no cells. Bars represent scores (mean ± SEM) for a minimum of 10 mice/group. Each experiment was repeated twice. Comparison was made between animals that received either CD4+ or CD8+ T cells and negative controls. ∗, significantly different at P < 0.001 for CD4 group; •, significantly different at P < 0.05 for CD8 group. Positive control group was significantly different from negative control group at P < 0.001 from days 21 to 70.

Article Snippet: T-cell enrichment was carried out using commercially available CD4 + and CD8 + mouse T-cell enrichment columns (R&D, Minneapolis, Minn.), according to the manufacturer's instructions.

Techniques: Positive Control, Negative Control

Journal:

Article Title: Primary Role for CD4 + T Lymphocytes in Recovery from Oropharyngeal Candidiasis

doi:

Figure Lengend Snippet: Representative agarose gel stained with ethidium bromide showing CD4 and CD8 gene products in BALB/c nu/nu mice following lymphocyte reconstitution. RT-PCR was performed on BALB/c tongue and oral mucosa. Lane 1 shows size markers, while lane 2 shows the cDNA negative control for each gel. CD4 is not present in control nude mice (lane 3), whereas CD8 is present at all times (lanes 4, 6, and 8). CD4 is present in oral mucosa on day 4 (lane 5) and day 8 (lane 7) following lymphocyte transfer, but is not seen in the tongue until day 21 (data not shown).

Article Snippet: T-cell enrichment was carried out using commercially available CD4 + and CD8 + mouse T-cell enrichment columns (R&D, Minneapolis, Minn.), according to the manufacturer's instructions.

Techniques: Agarose Gel Electrophoresis, Staining, Reverse Transcription Polymerase Chain Reaction, Negative Control