Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Regulatory T cells play an important role in the prevention of murine melanocytic nevi and melanomas

doi: 10.1158/1940-6207.CAPR-20-0360

Figure Lengend Snippet: (A & B) Mice were sensitized on the abdomen with increasing doses of DMBA (100μl of 0.1, 0.5 and 1%) and challenged 5 days later on ears with 20μl of 0.1% DMBA. Ear draining LNs were isolated 3 days after DMBA challenge and stained with anti-CD4, CD8, CD25 and Foxp3 antibodies and analyzed by flow cytometry. One million cells from each group were stimulated with PMA/Ionomycin for 48h and supernatants were analyzed for IL-10 and IFNγ by ELISA. Histograms are 4 LNs pooled from two mice and experiments were done twice. (C) Mice were sensitized and challenged with DMBA as above. Three days after sensitization, ears were digested with collagenase D/DNase as described in the Materials and Methods. Cells were gated on CD45.2 as shown. Cells were stained for markers as shown in the figure. The ears from 3 mice were pooled and digested to release the cells. (D) WT and CD4KO mice were sensitized with increasing DMBA doses and challenged as described in A & B. (E) Mice were sensitized with increasing DMBA doses, 5 days later CD4 T cells were collected from each group and injected into WT mice. Twenty-four hours after the adoptive transfer, mice were sensitized with DMBA on the abdomen and were challenged on the ears 5 days later. Each group had 4–5 mice per group. *p<0.05, **p<0.01,***p<0.001, ****p<0.0001

Article Snippet: FOXP3-v450 (AB_10611728), IFNγ-PE-Cy7 (AB_396766), CD8-Alexa-647 (AB_396792), and CD8-PE (AB_394571) were obtained from BD-Pharmingen.

Techniques: Isolation, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Injection, Adoptive Transfer Assay

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Regulatory T cells play an important role in the prevention of murine melanocytic nevi and melanomas

doi: 10.1158/1940-6207.CAPR-20-0360

Figure Lengend Snippet: DMBA (100μg/ mouse) was applied on the shaved and naired backs of mice and one week later TPA (12.5 μg/mouse) was applied twice weekly for 25 weeks. Nevi were counted and the area was measured weekly. (A) WT and CD8 KO had nearly equal lesion numbers and these numbers were significantly greater than CD4KO mice. (B) Reduced lesion area per mouse in CD4KO but not CD8KO or WT mice. Like the lesion numbers, the area was significantly reduced in CD4KO mice as compared to WT and CD8KO mice. The lesion area per mouse between CD8KO and WT groups was equal. (C) Area of each individual lesion at Week 25 for each group was measured. The mean lesion area is greatly reduced in CD4KO mice as compared to both WT and CD8KO. CD8KO mice have lesion whose size is slightly larger than WT mice. (D) Stacked bar graph for lesion area at week 25 from each group, shows that in CD8KO mice there is a higher percentage of lesions greater than 3 mm2 or 6 mm2 as compared to WT and CD4KO have larger percentage in the range below 3mm2. (E) Day 3 allergic contact hypersensitivity responses (CHS) in CD4KO is increased significantly compared to WT and CD8KO mice. Each group had 8–10 mice per group. *p<0.05, **p<0.01,***p<0.001

Article Snippet: FOXP3-v450 (AB_10611728), IFNγ-PE-Cy7 (AB_396766), CD8-Alexa-647 (AB_396792), and CD8-PE (AB_394571) were obtained from BD-Pharmingen.

Techniques:

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Regulatory T cells play an important role in the prevention of murine melanocytic nevi and melanomas

doi: 10.1158/1940-6207.CAPR-20-0360

Figure Lengend Snippet: (A) Mice were sensitized on the abdomen with DMBA (100μl of 0.1%) and challenged 5 days later on ears with 20μl of 0.1% DMBA. Ear draining LNs were isolated 3 days after DMBA challenge and stained with anti-CD4, CD8, CD25 and Foxp3 antibodies and analyzed by flow cytometry. (B) One million cells from each group were stimulated with PMA/ionomycin for 6h and 48h for intracellular staining and ELISA, respectively, to detect IL-10 and IFNγ. Histograms are from four LNs pooled from two mice. The experiment was done twice. (C) Mice were sensitized and challenged as above. Three days after sensitization, ears were digested with collagenase D/DNAse as described in Materials and Methods. Cells were gated on CD45.2 as shown. Cells were stained for the markers as shown in the figure. The ears from 3 mice were pooled and digested to release the cells. **p<0.01

Article Snippet: FOXP3-v450 (AB_10611728), IFNγ-PE-Cy7 (AB_396766), CD8-Alexa-647 (AB_396792), and CD8-PE (AB_394571) were obtained from BD-Pharmingen.

Techniques: Isolation, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Biomedicines

Article Title: Therapeutic Potential of Chick Early Amniotic Fluid in Mitigating Ionizing-Radiation-Induced Damage

doi: 10.3390/biomedicines13051253

Figure Lengend Snippet: ceAF improves spleen immune function in IR-treated mice. ( A ) Schematic diagram of experiments. ( B , C ) ceAF effectively reduced CD4 + T cells, increased CD8 + T cells, and normalized the ratio of CD4 + /CD8 + T cell of spleen in IR-treated mice. n = 6 per group. Data represent the mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001. One-way ANOVA.

Article Snippet: To assess the proportion of CD4 + and CD8 + T cells, 1 × 10 6 cells were stained with CD3-APC antibody (1:100, Elabscience, Wuhan, China), CD4 FITC antibody (1:100, Elabscience, Wuhan, China), and CD8 PE antibody (1:100, Elabscience, Wuhan, China) for 30 min in the dark at room temperature.

Techniques:

Journal: PLoS ONE

Article Title: Effect of age on chronic inflammation and responsiveness to bacterial and viral challenges

doi: 10.1371/journal.pone.0188881

Figure Lengend Snippet: (A) Differences in the proportions of monocytes, T cells and NK cells with age. (B) Differences with age in the % of T cells that are CD4+ and CD8+, in the CD4/CD8 ratio with age, in the proportion of T cells that are Tregs, and in the proportion of CD4 and CD8 cells that are CD28+ and -. (C) Anti-CD3 + anti-CD28 stimulated proliferation (left panel) and IFNγ production (right panel). Results are expressed as the mean ± SEM. * indicates a statistically significant difference (P <0.05) between the young and old cohorts.

Article Snippet: The antibodies used were: CD14-PE (clone MØp9), CD8-PE (clone SK1) and CD3-FITC (clone SK7) from StemCell Technologies, Vancouver, BC; CD45-FITC (Hle1), CD28-APC (clone CD28.2), CD4-PE-Cy7 (clone SK3), CD25-BB515 (clone 2A3), CD127-AF647 (clone HIL-7R-M21) and FoxP3-PE (clone 236A/E7) from BD Biosciences, Mississauga, ON; CD56-APC (clone CMSSB) from eBioscience, San Diego, CA.

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