anti cd28  (Bio X Cell)

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A ) Circos plots showing the percentage of cells from HC (Healthy control, n = 6), ICI ( n = 2), RAC (RA control, n = 5), or irAE ( n = 5). ( B ) Expression of CD45RA and CCR7 on CD8 + T cells. Right: Summaries of the percentage of cells from HC ( n = 53), irAE ( n = 29), RAC ( n = 41), and ICI ( n = 26). ( C ) Expression of CXCR3 and CCR6 on CD8 + T cells. Right: Summaries of T cell subsets. HC ( n = 45), irAE ( n = 27), RAC ( n = 31), and ICI ( n = 26). ( D ) The cytotoxic score was evaluated using the gene list identified previously . ( E ) Specific genes were evaluated on CD8 + T cells. ( F ) Pathways that were significantly enriched in the CD8 + T cells between irAE and ICI. NF-κB, nuclear factor κB; STAT5, signal transducer and activator of transcription 5; DN, down. Gene set enrichment analysis (GSEA) plots of the allograft rejection ( G ), oxidative phosphorylation ( H ), IFN-α response ( I ), and IFN-γ response ( J ) between irAE and ICI. NES, normalized enrichment score ( K to M ) PBMCs were stimulated with plate-coated anti-CD3 and anti-CD28 (10 μg/ml) for 5 days. Mean fluorescence intensities (MFIs) of MitoTracker Green (MTG) (K), MitoTracker deep red (MTDR) (L) [HC ( n = 31), irAE ( n = 18), RAC ( n = 32), and ICI ( n = 16)] or Cy5-linked-1-amino-glucose (GluCy5) (M) [HC ( n = 35), irAE ( n = 20), RAC ( n = 36), and ICI ( n = 18)] in CD8 + T cells were presented. Expression was normalized to the HC in each experiment. ( N ) UMAP shows the presence or absence of T cell receptor (TCR) in the major immune cells across all the samples. ( O ) Pie charts showing the distribution of the top 100 TCR clones across different T cell subsets. Data in graphs represent mean ± SEM. Significance was tested by one-way analysis of variance (ANOVA). [(A) to (E) and (G) to (O)] ICI, ICI control.

Article Snippet: A total of 0.5 million isolated CD4 + T cells or CD8 + T cells was stimulated with plate-coated anti-CD3 (10 μg/ml; BioXCell, catalog no. BE0001-2) and anti-CD28 (BioXCell, catalog no. BE0291), rhIL-6 (100 ng/ml), rhIFN-α 2 (100 ng/ml), rhIL-12 (100 ng/ml), or the combination of rhIL-6, rhIFN-α 2 , and rhIL-12 for 5 days.

Techniques: Control, Expressing, Phospho-proteomics, Fluorescence, Clone Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: A total of 0.5 million isolated CD4 + T cells or CD8 + T cells was stimulated with plate-coated anti-CD3 (10 μg/ml; BioXCell, catalog no. BE0001-2) and anti-CD28 (BioXCell, catalog no. BE0291), rhIL-6 (100 ng/ml), rhIFN-α 2 (100 ng/ml), rhIL-12 (100 ng/ml), or the combination of rhIL-6, rhIFN-α 2 , and rhIL-12 for 5 days.

Techniques: Cell Culture, Control, Expressing

Journal: Oncoimmunology

Article Title: The LXRβ/NF-κB axis reprograms CAR-T cells to resist exhaustion in the tumor microenvironment

doi: 10.1080/2162402X.2025.2611615

Figure Lengend Snippet: mRNA expression and RNA-seq analysis in pMIG and LXRβ OE CAR-T cells. (a) Comparison of LXRβ mRNA expression during CAR-T cell generation. LXRβ mRNA levels were analyzed in naïve T cells, T cells stimulated with anti-CD3/CD28, and untransduced or CAR-transduced T cells to assess the transcriptional changes associated with CAR transduction. Data were presented as the mean ± S.E.M. of n = 3 technical replicates. Statistical significance was calculated with one-way ANOVA followed by Tukey's multiple comparisons. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001; and ns, not significant (not shown). (b–e) RNA-seq analysis comparing transcriptomic differences between pMIG and LXRβ OE CAR-T cells. (b) Principal component analysis (PCA) plot illustrating global transcriptomic differences between pMIG and LXRβ OE CAR-T cells. (c) Volcano plot illustrating differentially expressed mRNAs between pMIG and LXRβ OE CAR-T cells. A total of 169 genes were identified based on the criteria of |fold change| ≥ 2 and raw p -value < 0.05. The x-axis represents log₂ fold change, and the y-axis shows –log₁₀( p -value). Red and blue dots indicate significantly upregulated and downregulated genes in LXRβ OE CAR-T, respectively, while gray dots represent non-significant genes. (d) Heatmap of normalized RNA-seq reads (as z scores) showing the expression of genes involved in the inflammatory response, comparing pMIG and LXRβ OE CAR-T cells. (e) Heatmap of normalized RNA-seq reads (as z scores) illustrating the expression of genes associated with TNFα signaling via NF-κB, highlighting the transcriptional changes induced by LXRβ overexpression.

Article Snippet: Purified CD8+ T cells were activated for 24 h with 5 μg/mL plate-bound anti-CD3 and anti-CD28 antibodies (Bio X Cell) before CAR-T cell generation.

Techniques: Expressing, RNA Sequencing, Comparison, Transduction, Over Expression

Journal: bioRxiv

Article Title: SLAMF1-peptide mediated epigenetic priming reprograms innate immune responses in sepsis

doi: 10.64898/2025.12.29.696918

Figure Lengend Snippet: Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with anti-CD28) for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).

Article Snippet: T cell responses were assessed after 72 h, under non-stimulated conditions (RPMI alone) and RPMI with a synthetic superantigen (CytoStim, 2 μl/ml) from Miltenyi Biotec (cross-binding TCR and MCH) in combination with co-stimulating anti-CD28 mAb (InVivoMAb, BioXCell, 300 ng/ml).

Techniques: Ex Vivo, Clinical Proteomics