Journal: bioRxiv

Article Title: TcrDesign: De novo design of epitope specific full-length T cell receptors

doi: 10.64898/2026.01.15.699824

Figure Lengend Snippet: ( A ) UMAP dimensionality reduction was performed on generated TCRs for two distinct epitopes, utilizing TCR embedding extracted by TcrDesign-B. ( B ) Binding assay of HLA-A*02:01-ELAGIGILTV tetramer to the nine generated TCRs expressing Jurkat cells was analyzed by flow cytometry. Three candidate TCRs ELA-TCR3, ELA-TCR4 and ELA-TCR6 exhibited specific binding to the tetramer. ( C ) Activation markers NFAT and CD69 levels of reporter Jurkat NFAT-ZsGreen cells expressing candidate TCRs ELA-TCR3, ELA-TCR4, ELA-TCR6 and ELApositive TCR after incubation with ELAGIGILTV-pulsed T2 cell for 24 hours, each sample was tested in triplicate. ( D ) The NFAT-ZsGreen signal and CD69 level of ELApositive-TCR and ELA-TCR3 were determined by fluorescence imaging and flow cytometry analysis. ( E ) Engineered Jurkat T cells were co-cultured with T2 cells and serially diluted peptides for 24 h. Co-cultured supernatants were analyzed by ELISA for secreted IL-2. ( F ) TCR-T cells at various E:T ratios were co-cultured with luciferase-transduced T2 cells. The % specific lysis of T2-luciferase (black) and 1ug/mL ELAGIGILTV pulsed T2-luciferase cells (blue) obtained by bioluminescence assay is plotted against multiple E:T ratios. Dots in the figure represents three replicates.

Article Snippet: For 1 x 10 6 T-lymphocytes, 25 μL of Dynabeads coated with anti-CD3 and anti-CD28 antibodies was added to the RPMI 1640 media supplemented with 200U human recombinant IL-2 (#11848-HNAH1-E, Sino Biological).

Techniques: Generated, Binding Assay, Expressing, Flow Cytometry, Activation Assay, Incubation, Fluorescence, Imaging, Cell Culture, Enzyme-linked Immunosorbent Assay, Luciferase, Lysis, ATP Bioluminescent Assay

Journal: Frontiers in Immunology

Article Title: Single-cell atlas of human skin implicates APOE pro-inflammatory signaling in diabetic foot ulcers

doi: 10.3389/fimmu.2025.1591944

Figure Lengend Snippet: APOE+ fibroblasts promote fibrosis and inflammation in DFU. (A) Representative images showing histological differences between human healthy donor (HD) and diabetic foot ulcer (DFU) skin tissues assessed by HE staining (left) and APOE expression visualized by IHC (right). Insets display magnifications of different regions in the skin. Scale bar in the original images: 500 μm, 2000 μm. Scale bar in the magnified images: 50 μm. Upper: Cellular infiltration was quantified in HE-stained sections by measuring the percentage of infiltrated area. Lower: APOE expression was quantified through positive area percentage analysis of IHC staining. (B) Western blot analysis and quantitative comparison of APOE protein in human HD and DFU skin tissues. (C) RT-qPCR analysis of mRNA levels of α-SMA, COL1A1 and COL3A1 in human HD and DFU skin tissues. (D) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human HD and DFU skin tissues. (E) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human fibroblasts treated with APOE3 at concentrations of 10 ng/mL or 25 ng/mL. (F) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human fibroblasts treated with APOE3 at a concentration of 10 ng/mL for 6 h, 12 h or 24 h. Data are presented as mean ± SD from three independent biological replicates (n = 3). * p < 0.05, *** p < 0.001.

Article Snippet: Cells were treated with human recombinant APOE3 protein (MedChemExpress, China) at 10 ng/mL or 25 ng/mL for 24 hours, or at 10 ng/mL for 6, 12, or 24 hours.

Techniques: Staining, Expressing, Immunohistochemistry, Western Blot, Comparison, Quantitative RT-PCR, Protein-Protein interactions, Concentration Assay