Journal: Cells

Article Title: Oral Delivery of mRNA Vaccine by Plant-Derived Extracellular Vesicle Carriers.

doi: 10.3390/cells12141826

Figure Lengend Snippet: Figure 5. Rat vaccination with oral capsules. (A) Schematic representation of the experimental workflow: cells were isolated from rat spleen and stimulated or not with S1 antigen ex vivo be- fore cytofluorimetric analysis. (B) Representative plots of splenocytes isolated from rats treated with oEV or oEV-S1 and analyzed for CD3+ proliferation using CFSE assay. The boxes highlight proliferating cells. (C) Quantitative analysis of proliferating CD3+ lymphocytes in samples un- treated (NT) or treated with S1 antigen (S protein) expressed as percentage of events detected using CFSE assay. Samples were compared using TWO-way ANOVA with Tukey’s multiple com- parison test. (D) Cytofluorimetric quantification of total CD3+ lymphocytes in samples untreated (NT) or treated with S1 antigen (S protein) expressed as percentage of events. (E–L) Analysis of splenocyte immune populations expressed as the percentage of events detected for CD3+CD25+ (E), CD3+CD8+CD25+ (F), CD3+CD4+CD25+ (G), CD3+CD4+ (H), CD3+CD4+CD44+ (I), CD138+ (J), and IL-10+ (K), IL-4+ (L). Samples: rats treated with capsules containing unloaded oEVs (oEV) or S1 mRNA-loaded oEVs (oEV-S1). Data are presented as mean ± SD and were compared using t-test statistical analysis. ns, not statistically significant. * p < 0.05, ** p < 0.01, **** p < 0.001.

Article Snippet: Cells were incubated for 5 days and stained with CD3ε APC-Vio® 770 antibody (1:50, 130-122-015, Miltenyi, Bologna, Italy) and the associated isotype.

Techniques: Capsules, Isolation, Ex Vivo, CFSE Assay

Journal: Annals of Medicine and Surgery

Article Title: Da-yuan-yin decoction ameliorates inflammatory injury in severe pancreatitis by protecting intestinal mucosal barrier and immune function and preventing intestinal dysbiosis

doi: 10.1097/MS9.0000000000004873

Figure Lengend Snippet: Serum interleukin (IL)-1β and IL-10 expression levels (pg/ml) and cluster of differentiation (CD)3 + , CD4 + , and CD8 + cells from five ileal Peyer’s patches in the sham operation (SO), severe acute pancreatitis (SAP), and da-yuan-yin (DYY) + SAP groups, respectively. Serum IL-1β and IL-10 levels in the SAP and DYY + SAP groups were significantly higher than those in the SO group at 24, 48, and 72 h after surgery (all P < 0.05). Serum IL-1β and IL-10 levels in the DYY + SAP group were significantly lower than those in the SAP group 72 h postoperatively ( t = 4.37, P = 0.003; t = 2.41, P = 0.04, respectively). Levels of CD3 + , CD4 + , and CD8 + cells from ileal Peyer’s patches in the SAP and DYY + SAP groups decreased with time post-surgery. At 72 h postoperatively, the DYY + SAP group had significantly higher levels of CD3 + , CD4 + , and CD8 + cells than the SAP group ( t = 2.58, P = 0.036; t = 2.78, P = 0.027; t = 2.77, P = 0.028, respectively). There were no significant differences in early or late apoptosis in ileal Peyer’s patches 24, 48, and 72 h postoperatively in the SO, SAP, and DYY + SAP groups.

Article Snippet: Single-cell suspension is stained with the following antibodies: Live/Dead Fixable Violet Dead Cell (Cat# 2549272, Invitrogen), anti-Rat CD3 (Cat# G4.18, MultiSciences Biotech Co., Ltd), and anti-Rat CD4 (Cat# OX35, MultiSciences).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Desmoglein-4 Deficiency Exacerbates Psoriasiform Dermatitis in Rats While Psoriasis Patients Displayed a Decreased Gene Expression of DSG4

doi: 10.3389/fimmu.2021.625617

Figure Lengend Snippet: Dsg-4 deficiency induces high mRNA expression of inflammatory genes in response to IMQ. RNA from skin was extracted, and subsequently real-time PCR and cDNA synthesis were performed to evaluate mRNA abundance of cytokines: IL-1β (A) , IL-8 (B) , IL-17 (C) , IL-10 (D) , and TGF-β (E) performing Kruskal–Wallis tests analysis. Additionally, TGF-β/IL-17 mRNA abundance means ratio was calculated (F) . Similarly, chemokine receptors mRNA levels were measured: CCR1 (G) , CCR2 (H) , CCR3 (I) , CCR5 (J) , and CXCR5 (K) . Kruskal–Wallis and ANOVA tests were used; data represent the median with values range; * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets. (L) Pseudocolor dot plots represent counts of infiltrating CD45 + cells and CD3 + cells from total skin cells analyzed by flow cytometry derived from SD and OFA rats. (M,N) Graphs represent the percentage and absolute cell counts of CD45 + CD3 + cells from total skin cells. ( n rats /group = 6; two independent experiments). (O,P) Spleen and inguinal lymph node weights. FACS and lymphatic tissue weight data are represented as the mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001 with ANOVA and Bonferroni post hoc test was used. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets.

Article Snippet: The cells were then resuspended in PBS/2% FBS and stained with PE-Cy5-conjugated anti-rat CD45 (clone OX-1; isotype mouse IgG1 k, BD Biosciences, San Jose, USA) and FITC-conjugated anti-rat CD3 (clone IF4; isotype mouse IgM, Cedarlane Laboratories, US).

Techniques: Expressing, Real-time Polymerase Chain Reaction, cDNA Synthesis, Flow Cytometry, Derivative Assay

Journal: bioRxiv

Article Title: Optimizing 3D-printed Scaffold Geometry Decreases Foreign Body Response and Enhances Allogeneic Islet Transplant Outcomes

doi: 10.64898/2026.02.02.701816

Figure Lengend Snippet: Digital spatial profiling of 150 µm and 300 µm 3D-printed scaffold prototypes explants after transplantation into the epididymal fat pad of C57BL/6 mice for 14 and 30 days. Schematic depicting the segmentation of cross-sectioned explants into major and minor pore ROIs, cutout depicts staining of cells with fluorescently tagged antibodies for visualization and UV-photocleavable oligo conjugated antibodies for immune cell quantification (A). Cross-sectional images of CD3 + T cells (pink), α-smooth muscle actin (green), and nuclei (blue) infiltration into the 150 µm and 300 µm scaffolds prototypes on explant day 14 and 30 (B; SS = silicone scaffold rung). Fold change in immune cell markers on day 14 vs. day 30 within the major pores (grey) vs. the minor pores (blue) in the 300 µm scaffold (C) or the 150 µm scaffold (D). Fold change in immune cell markers within the 300 vs. 150 µm scaffold on day 14 (grey) or day 30 (blue) within the major pores (E) or minor pores (F). Fold change in immune cell markers within major vs. minor pores on day 14 (grey) or day 30 (blue) within the 300 µm scaffold (G) or the 150 µm scaffold (H). (Significantly up- or down-regulated immune cell phenotypes are indicated by the outlined, labeled data points; N = 1 scaffold per explant day, n ≥ 9 minor or major pore ROIs; Linear mixed model with Benjamini-Hochberg multiple comparisons test for comparisons between scaffold prototypes and explant days, Multiple unpaired T test for comparisons between pore type; p < 0.05).

Article Snippet: Samples were stained with a multiplexed cocktail of primary immune-centric antibodies tagged with unique UV-photocleavable oligonucleotides (30 proteins in total, Cat. No. 121300106 & 121300118; Nanostring Technologies; see Supplemental Table S1 ), rat anti-human CD3 antibody conjugated with AlexaFluor 647 (Cat. No. MCA1477A647; Bio-Rad Antibodies), mouse anti-rat α-Smooth Muscle Actin antibody conjugated with AlexaFluor 488 (Cat. No. AB184675; Abcam), and Syto83 nucleic acid stain (Cat. No. S11364; Life Technologies).

Techniques: Transplantation Assay, Staining, Labeling