Journal: British Journal of Cancer

Article Title: T cells in testicular germ cell tumors: new evidence of fundamental contributions by rare subsets

doi: 10.1038/s41416-024-02669-9

Figure Lengend Snippet: a Representative analysis of immune cells in tissue samples from an individual patient with embryonal carcinoma, obtained from different localizations of both, tumor-bearing testis (tumor central [Tumor]; tumor-adjacent [Tumor-Adj]; tumor-distant [Tumor-Dis]) and contralateral testis ([Contralateral 1; Contralateral 2]). Percentage of positive cells with individual immune cell markers in total live cells is indicated. b–d Overview of individual values measured for different immune cell markers in tumor-bearing and contralateral testes from n = 24 patients [seminoma (red): Tumor n = 12; Tumor-Adj n = 10; Tumor-Dis n = 9; Contralateral 1 n = 5; Contralateral 2 n = 6; embryonal carcinoma (blue): Tumor n = 6; Tumor-Adj n = 5; Tumor-Dis n = 4; Contralateral 1 n = 4; Contralateral 2 n = 4; mixed tumors (green): Tumor n = 6; Tumor-Adj n = 6; Tumor-Dis n = 6; Contralateral 1 n = 4; Contralateral 2 n = 4]. e , f Respective percentages of CD25 + FOXP3+ Treg and CXCR5 + BCL6+ Tfh cells in total live cells [seminoma (red): Tumor n = 8; Tumor-Adj n = 7; Tumor-Dis n = 5; Contralateral 1 n = 5; Contralateral 2 n = 6; embryonal carcinoma (blue): Tumor n = 4; Tumor-Adj n = 4; Tumor-Dis n = 3; Contralateral 1 n = 4; Contralateral 2 n = 4; mixed tumors (green): Tumor n = 5; Tumor-Adj n = 6; Tumor-Dis n = 6; Contralateral 1 n = 4; Contralateral 2 n = 4]. Columns indicate median values for each subgroup; significance tested by ordinary one-way ANOVA including Tukey’s Honest Significant Difference Test; * p < 0.05, ** p < 0.01, *** p < 0.001 (see Supplementary Table ).

Article Snippet: After washing in MACs Quant buffer, cells were incubated with FOXP3 staining buffer and then washed with permeabilization buffer (both from Miltenyi Biotec).

Techniques:

Journal: Journal of Nanobiotechnology

Article Title: Optogenetic engineered umbilical cord MSC-derived exosomes for remodeling of the immune microenvironment in diabetic wounds and the promotion of tissue repair

doi: 10.1186/s12951-023-01886-3

Figure Lengend Snippet: Remodeling of the immune microenvironment of the chronic wounds in diabetic mice by UCMSCs-exo/eNOS. I: Treatment with PBS. II: Treatment with UCMSCs-exo. III: Treatment with UCMSCs-exo/eNOS. a Immunohistochemistry of CD3 and CD11c in the skin wounds of diabetic mice treated with PBS, UCMSCs-exo, and UCMSCs-exo/eNOS on postoperative day 14. Scale bar: 100 μm. b , c Quantification of immunohistochemical staining of CD3 and CD11c (n = 6 in each group, two-tailed Student’s t-test). d M1 macrophages in the wound tissue. immunofluorescence images of F4/80 and CD86. Scale bar: 100 μm. e M2 macrophages in the wound tissue: immunofluorescence images of F4/80 and CD163. Scale bar: 100 μm. f , g Quantitative immunofluorescence analysis of M1 and M2 macrophages(n = 6 in each group, two-tailed Student’s t-test). h Representative flow cytometry plots of CD25 + Foxp3 + Treg cells in wound tissue. i Representative flow cytometry image of CD69 + CD103 + T RM cells in wound tissues. j Quantitative analysis of the proportion of CD25 + Foxp3 + Treg cells in wound tissues (n = 4 in I group, n = 4 in II group and n = 3 in III group, Mann–Whitney U test). k Quantitative analysis of the proportion of CD69 + CD103 + T RM cells in wound tissues (n = 4 in each group, two-tailed Student’s t-test). Data represent means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. PBS group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. UCMSCs-exo group

Article Snippet: Staining of intracellular markers, such as the Treg marker FoxP3 (MultiSciences, China), were performed on ice.

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining, Two Tailed Test, Immunofluorescence, Flow Cytometry, MANN-WHITNEY

Journal: Virology Journal

Article Title: Japanese Encephalitis Virus wild strain infection suppresses dendritic cells maturation and function, and causes the expansion of regulatory T cells

doi: 10.1186/1743-422X-8-39

Figure Lengend Snippet: Effects of P3 infection on DCs-induced differentiation of regulatory T cells . 1 × 10 5 mock-, P3-, UV-P3- or LPS-treated bmDCs were incubated with 1 × 10 6 allogeneic naïve T cells for 5 days. T cells were purified and doubly labeled for CD4 and Foxp3, and assessed by FACS. The in vivo Treg in splenocytes were purified and examined by FACS from mice inoculated with 1 × 10 5 PFU P3 or identical UV-P3 i.p. for 5 days. Representative result was shown from three independent experiments (A). The percentage represented the ratio of CD4+ Foxp3+ cells in CD4+ T cells. P3-infected bmDCs elicited the Treg differentiation in vitro (B). After P3 infection or UV-P3 stimulation of mice i.p., Treg differentiation in vivo was analyzed immediately (C). Results were expressed as the means ± SD of triplicates. *, P < 0.05.

Article Snippet: After 5 days of co-culture, in vitro Treg cells (CD4+ and Foxp3+) were isolated (StemCell, Vancouver, BC, Canada) and stained with Mouse Regulatory T Cell Staining Kit (eBioscience Inc., San Diego, CA) in accordance with the manufacturer's instructions and analyzed by FACS.

Techniques: Infection, Incubation, Purification, Labeling, In Vivo, In Vitro