Journal: Scientific Reports

Article Title: Enhancing cancer immunotherapy using cordycepin and C ordyceps militaris extract to sensitize cancer cells and modulate immune responses

doi: 10.1038/s41598-024-72833-x

Figure Lengend Snippet: Changes in cytokines, cytotoxic mediators, and subpopulations of effector immune cells following treatment with cordycepin (CD) and Cm-EE. Non-adherent cells were exposed to 100 µM of cordycepin (CD) or 100 µg/mL Cm-EE for 24 h. Culture supernatants were collected for determination of cytokine and cytotoxic mediator production by using cytokine bead array. The fold changes in cytokine ( a ) and cytotoxic mediator ( b ) production from the cordycepin or Cm-EE treatments were compared to the appropriate control. Subpopulations of treated immune cells ( c ), including CD4 (CD3 + and CD4+), CD8 (CD3 + and CD8+), NK cells (CD3- and CD56+), and B cells (CD3- and CD19+) were characterized using specific fluorescence dye-conjugated monoclonal antibodies. Data are presented as the mean ± SEM. Statistically significant values (* p < 0.05) were determined by Student’s t-test ( N = 3).

Article Snippet: The non-adherent cells were seeded at a density of 1 × 10 6 cells/well in 12-well plate and treated with either 100 µM cordycepin or 100 µg/mL Cm-EE for 24 h. Following treatment, cells were harvested and stained with monoclonal fluorescent-conjugated antibodies, including anti-CD3 FITC (Clone: UCHT-1), anti-CD4 APC (Clone: OKT-4), anti-CD8 APC (Clone: UCHT-4), anti-CD16 APC (Clone: LNK16), anti-CD19 APC (Clone: LT19) (all sourced from Immunotools, Friesoythe, Germany), and anti-CD56 PE (Clone: HCD56; BioLegend).

Techniques: Control, Fluorescence

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: Archives of Iranian Medicine

Article Title: Clinicopathologic Effects of Xenogeneic GvHD Induced by Adoptively Transferred Human-Derived T Cells in Severely Immunodeficient Mice

doi: 10.34172/aim.28597

Figure Lengend Snippet: Plotting Intervention Categories and Outcomes against Histopathological and Clinical xGvHD Scoring (n = 50)

Article Snippet: For this purpose, T cells were re-suspended in 100 μL PBS and incubated with the recommended amount of anti-human CD3-PerCP (Miltenyi Biotec, Germany), anti-human CD4-FITC/CD8-PE (Cytognos SL, Salamanca, Spain) (Figure S1, ).

Techniques:

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: