Journal: Frontiers in Immunology

Article Title: Reorganization and functional divergence of the CD4+ memory T cell compartment in hidradenitis suppurativa

doi: 10.3389/fimmu.2026.1831664

Figure Lengend Snippet: Expansion of central memory T cells (T CM ) in HS skin. Flow cytometric analysis of total CD4+ and CD8+ T cell subsets in (A) the skin and (B) peripheral blood. Frequency of naïve and memory subsets within the CD4+ and CD8+ compartments are shown for (C) skin and (D) peripheral blood. Each dot represents an individual donor (Healthy, diamonds; HS, circles); bars indicate the mean frequency of the population relative to the indicated gate ± standard deviation (SD). Healthy PBMCs, n = 8; Healthy skin, n = 5; HS skin and PBMCs matched samples ( n = 5). Detailed gating strategies for T cell memory subsets (Naïve, T CM , T EM and T RM ) are provided in Supplementary Figure 1 . * P < 0.05, ** P < 0.01 and *** P < 0.001 (Two-way ANOVA with Šídák’s multiple comparisons test).

Article Snippet: Peripheral blood mononuclear cells (PBMCs) from blood were isolated with StraightFrom ® Whole Blood PBMC Isolation Kit, human (130-126-359, Miltenyi Biotec), and resuspended in the same culture medium as skin cells.

Techniques: Standard Deviation

Journal: Frontiers in Immunology

Article Title: Reorganization and functional divergence of the CD4+ memory T cell compartment in hidradenitis suppurativa

doi: 10.3389/fimmu.2026.1831664

Figure Lengend Snippet: Heightened pro-inflammatory potential of skin vs peripheral blood CD4+ memory T cells. Frequencies of cytokine-expressing memory T cell subsets from matched skin and PBMC samples ( n = 4). Bars represent the mean percentage (± SD) of cytokine-expressing memory T cells vs parent, under unstimulated baseline conditions (“Unstim”, blue bars/circles) and following stimulation with anti-CD3/anti-CD28 coated beads (“aCD3/aCD28”, red bars/triangles). Data are shown for TNF-α-producing CD4+ T EM (A) and T CM (D) cells, IFN-γ-producing CD4+ T EM (B) and T CM (E) , IL-17A-producing CD4+ T EM (C) and T CM (F) . Statistical differences between skin and PBMCs and stimulation condition were evaluated by 2-way repeated measures ANOVA and Fisher’s LSD test (* P < 0.05 and ** P < 0.01).

Article Snippet: Peripheral blood mononuclear cells (PBMCs) from blood were isolated with StraightFrom ® Whole Blood PBMC Isolation Kit, human (130-126-359, Miltenyi Biotec), and resuspended in the same culture medium as skin cells.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Expression of 5T4 and PD-L1 and anti-tumor effects of 53L10 on a panel of tumor cells. (A) The expression levels of 5T4 and PD-L1 on breast MDA-MB-231, lung A-549, gastric NCI-N87, pancreatic PANC-1, and ovarian SKOV-3 cancer cells were tested by cell ELISAs by using a commercial anti-PD-L1 or anti-5T4 antibody followed by their respective HRP-conjugated secondary Abs. The breast MCF-7 cell line was used as negative control for PD-L1 expression. Binding values were reported as the mean of determinations obtained in three independent biological experiments. Error bars depicted means ± SD (Table A). The table reports the expression levels (number of cell surface 5T4 and PD-L1) obtained by quantitative flow cytometry by QIFIKIT (Dako) analyses on MDA-MB-231, A-549, PANC-1, and MCF-7 cells. (B, C) Tumor cells were co-cultured with hPBMCs (5:1 effector-to-target cell ratio) and treated with 53L10 (B) or 53P (C) used at increasing concentrations for 48 h. The effects on tumor cell lysis were reported as dose–response curves. As a negative control, MCF-7 cells were tested. (D) Images of NCI-N87, PANC-1, and SKOV-3 cells untreated or treated with 53L10 in the presence of hPBMCs were acquired and processed by using a digital microscopy from Zen 3.2 cell discover. Cells co-cultured with hPBMCs and treated with an unrelated human IgG1 were used as a negative control. Scale bar, 200 μm.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing, Negative Control, Binding Assay, Flow Cytometry, Cell Culture, Lysis, Microscopy

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Cytotoxicity and target expression levels on other breast, prostate, pancreatic, epidermoid carcinoma cell lines and CHO-transfected cells. (A) The cytotoxic activity of 53L10 was evaluated in co-cultures of various cancer cell lines and human PBMCs pre-activated with anti-CD3/anti-CD28 beads. Cells were mixed at an effector-to-target cell ratio (E/T ratio) of 5:1 or 10:1. 53L10 was added to achieve a final concentration of 10 −4 to 10 −10 M. After 3 days of culture, the amount of LDH in the culture supernatant was measured with LDH-Glo™ Cytotoxicity Assay (Promega) following the manufacturer’s instructions. Measures were performed in triplicates at each point. Cytotoxicity percentage was calculated according to the following formula: C y t o t o x i c i t y ( % ) = ( E f f e c t o r t a r g e t c e l l m i x − E f f e c t o r c e l l c o n t r o l − L o w c o n t r o l ) H i g h c o n t r o l − L o w c o n t r o l × 100 (B, C) Cytotoxic effects of 53L10 on CHO-K1 cells stably transfected with DNA encoding 5T4 or PD-L1 and expressing different levels of the targets. CHO-K1 cell lines expressing varying levels of 5T4 or PD-L1 were established and subjected to a cytotoxicity assay in the presence of 53L10 and human PBMCs pre-activated with anti-CD3/anti-CD28 beads. LDH release was evaluated after 3 days of culture. The cytotoxic activity of 53L10 against CHO-K1 cells expressing human 5T4 or human PD-L1 is shown in (B) and (C) , respectively. The number of cell surface-expressed human 5T4 or human PD-L1 was determined using the QIFIKIT (Dako) and is indicated above each graph.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing, Transfection, Activity Assay, Co-Cultures, Concentration Assay, Cytotoxicity Assay, Stable Transfection

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Cell surface expression of 5T4 and PD-L1 and EC50 values for 53L10 effect on tumor cells. The values were calculated by fitting the dose–response curves reported in Figure 2 after treatment of the cells with hPBMCs from three different donors and performed in triplicates.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Analysis of tumor-infiltrating T cells in a PBMC-humanized A-549 xenograft model. (A) A-549 cells were transplanted s.c. into NOD-scid mice. When the average tumor volume reached 100 mm³, mice were treated with anti-sialo GM1 pAb. The next day, human PBMCs were introduced intraperitoneally, and 200 µg of 53P or 53L10 was subsequently administered intravenously once every 2 days. Tumors were resected on days 6 and 10. (B) Average tumor size on day 10 (mean of five mice per group). Error bars indicate standard deviations. Statistical significance was assessed by one-way ANOVA followed by Tukey’s post-hoc test and established as * p < 0.01. (C, D) On days 6 and 10, tumors from three mice per group were collected and pooled, and infiltrating lymphocytes were extracted. Human CD45 + lymphocytes were analyzed for CD4 and CD8 expression by flow cytometry.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Analysis, Xenograft Assay, Expressing, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Comparison of the antibody constructs, tumor models, PBMC conditions, treatment schedules, endpoints, and principal findings of the previous and current studies.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Comparison, Construct, Tumor Models, Injection, Transplantation Assay