Journal: Frontiers in Immunology

Article Title: Glycoengineering CAR-T cells to overcome galectin-3-mediated immunosuppression

doi: 10.3389/fimmu.2026.1766555

Figure Lengend Snippet: ST6 OE CAR-T cells exhibited effective tumoricidal activity in the presence of Gal-3 and evaded Gal-3-induced expression of IL-5, which compromised anti-tumor efficacy. Day 10-expanded T cells, CAR-T cells, and ST6 OE CAR-T cells were incubated with Gal-3 low Raji-luc + cells or Gal-3 high SUDHL-6-luc + cells at effector:target (E:T) ratios of 20:1. Raji cell cytotoxicity was evaluated (a) without rhGal-3 or (b) with rhGal-3, and (c) Gal-3 high SUDHL-6 cell cytotoxicity was examined without exogenous rhGal-3. Data are presented from n = 4 independent donors and analyzed using a two-way repeated-measures ANOVA with Sidak’s multiple-comparisons test, with ** p < 0.01, * p < 0.05. (d) Human cytokine proteome profiling of supernatants from Day 10-expanded T cells, CAR-T cells, and ST6 OE CAR-T cells incubated with rhGal-3 for 16h revealed a marked upregulation of IL-5 (red hatched box) in CAR-T cells that was absent in ST6 OE CAR-T cells. (e) Arrays were repeated 4 times and graphed as Relative Optical Density (Mean ± SEM). Intracellular IL-5 expression was assessed by flow cytometry in control Day 10-expanded T cells, CAR-T cells and ST6 OE CAR-T cells after rhGal-3 incubation for 16h and mean ± SEM fold change from n = 4 independent donors was graphed (f) (* p < 0.05). Statistical significance was determined using a Mann–Whitney U test. CAR-T cells and ST6 OE CAR-T cells were incubated with (g) Gal-3 low Raji-luc + cells or (h) Gal-3 high SUDHL6-luc + at E:T ratios of 20:1 (** p <0.01, *** p <0.001). Statistical significance was determined using two-way repeated-measures ANOVA with Sidak’s multiple-comparisons test.

Article Snippet: Human B-lymphoma Raji cell line expressing luciferase (Raji-Luc + ) purchased from the ATCC (Manassas, VA).

Techniques: Activity Assay, Expressing, Incubation, Flow Cytometry, Control, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Glycoengineering CAR-T cells to overcome galectin-3-mediated immunosuppression

doi: 10.3389/fimmu.2026.1766555

Figure Lengend Snippet: ST6 OE CAR-T cells exhibited potent in vivo anti-tumor activity and persisted longer in vivo than conventional CAR-T cells. Using a DLBCL xenograft model (a) , NSG mice were inoculated with Raji-luc + cells and, after 7 days, treated with media (control), Day 10-expanded T cells, CAR-T cells, or ST6 OE CAR-T cells. Tumor growth in NSG mice was monitored weekly by in vivo BL imaging system (b) , and BLI measurements reflecting tumor burden from respective groups were graphed as shown (n = 9 mice per group) (** p < 0.01) (c) . Xenografted mouse survival data were graphed as shown (Gehan-Breslow-Wilcoxon test – ** p = 0.0106) (d) , and flow cytometry of CAR + T cells isolated from xenografted mouse spleens expressed as Mean ± SEM were graphed as shown (Mann–Whitney U test- * p <0.05) (e) .

Article Snippet: Human B-lymphoma Raji cell line expressing luciferase (Raji-Luc + ) purchased from the ATCC (Manassas, VA).

Techniques: In Vivo, Activity Assay, Control, Imaging, Flow Cytometry, Isolation, MANN-WHITNEY

Journal: medRxiv

Article Title: TRAF1 S146 is constitutively phosphorylated in primary CLL cells by PKN1/2

doi: 10.64898/2026.02.11.26346036

Figure Lengend Snippet: 5 or 10 µg of whole cell lysates from WT, bulk PKN1 knockout (PKN1 KO), bulk PKN2 (KO) or bulk PKN1 and PKN2 double knockout (PKN1/2 KO) RAJI cells were immunoblotted for pTRAF1 S146 (p-TRAF1), total TRAF1 (TRAF1), PKN1, PKN2 and GAPDH. Lysates from TRAF1 WT and S146A overexpressing 293 cells were loaded separately as controls for pTRAF1 assay. These data are representative of 3 similar experiments, in which 2 of 3 experiments showed similar role for PKN1 and PKN2 and one experiment showed greater role for PKN1 than PKN2 in TRAF1 S146 phosphorylation.

Article Snippet: RAJI cells (ATCC) were cultured in RPMI with 25mM Hepes (purchased from Wisent Inc.) supplemented with 10%FCS from Wisent and 1% of 100X Glutamine-Pyruvate Penicillin-Streptomycin (GPPS) (Sigma-Aldrich, Oakville, Canada).

Techniques: Knock-Out, Double Knockout, Phospho-proteomics