Journal: bioRxiv

Article Title: Using peptide-exchange systems to interrogate peptide-specific KIR binding to HLA Class I

doi: 10.64898/2026.03.03.708729

Figure Lengend Snippet: (A) Display of ULBP1 and CD155 on CombiCells detected by mAbs and NKG2D-Fc, and DNAM-1-Fc, respectively. (B&C) KIR2DL1+ and KIR2DL1-NK-cell degranulation (CD107a upregulation) in response to ULBP1, CD155, and HLA-C*05:01 displayed on CombiCells. (C) Three types of HLA-C*05:01 were tested, all containing P2 (IIDKSGSTV); wild-type, open and dipeptide exchanged. Data from three independent experiments with NK cells from separate donors are shown.

Article Snippet: Plasmids encoding the ectodomains of ULBP1 (pD649-HAsp-ULBP1-Fc(DAPA)-AviTag-6xHis) and CD155 (pMP71-hCD155) were obtained from Addgene. pMP71-hCD155 was a gift from Sébastien Walchli (Addgene plasmid # 118630; http://n2t.net/addgene:118630 ; RRID:Addgene_118630) [ ]. pD649-HAsp-ULBP1-Fc(DAPA)-AviTag-6xHis was a gift from Chris Garcia (Addgene plasmid # 156597; http://n2t.net/addgene:156597 ; RRID:Addgene_156597) [ ].

Techniques:

Journal: Cell Death & Disease

Article Title: Rewiring melanoma cell fate: TRPM8 modulators trigger apoptosis and boost NK cell cytotoxicity

doi: 10.1038/s41419-026-08469-8

Figure Lengend Snippet: A Representative images of crystal violet-stained colonies, derived from WM266-4 cells, after 21-day treatment with TRPM8 modulators. B Western blot analysis showing ULBP1 expression in WM266-4 cells after treatment with TRPM8 modulators. The α-tubulin was used as a loading control. C WM266-4 derived spheroids treated for 21 days as indicated, in absence (upper panel; -NK cells) or presence (lower panel; + NK cells) of NK cells. Images are representative of three different experiments. Bar, 100 µm. D The graph represents the dead cells/total cells. Values of dead (red stained cells) and total cells (green stained cells) were analyzed using NIH Image J. They derive from red fluorescence mean/green fluorescence mean intensity and are expressed as mean ± SD of 3 different experiments ( n = 3); ** p < 0.01; *** p < 0.001. E NK cell cytotoxicity assay. WM266-4 cells pre-treated with TRPM8 modulators were co-cultured with primary NK cells at the indicated effector:target (E:T) ratios. Where indicated, neutralizing antibodies against ULBP1 or NKG2D were added 1 h before co-culture to melanoma cells or NK, respectively. Data are presented as percentage of lysis. * p < 0.05; ** p < 0.01. F WM266-4 cells were transfected with control siRNA (siRNA ctrl) or TP53-targeting siRNA (siRNA p53) at two different concentrations (300 pmol and 400 pmol, respectively). After 4 days, cells were collected, lysed, and Western blot analysis was performed on cell lysates using the indicated antibodies. α-Tubulin was used as a loading control. G WM2664 cells transfected with control siRNA (siRNA ctrl) or TP53-targeting siRNA (siRNA p53; 300 pmol) were unstimulated or stimulated with compounds 4 and 9 (at 1 μM) for 4 days and then collected and lysed. Western blot analysis was performed using the indicated antibodies. α-Tubulin was used as a loading control. H Phosphorylated AKT (Ser473) levels in melanoma cells treated with the PI3K agonist, 740 Y-P, in absence or presence of TRPM8 modulators (used at 1 μM) for 4 days, analyzed by Western blot. GAPDH was used as a loading control.

Article Snippet: They were co-cultured with melanoma cells pre-treated with TRPM8 modulators for 21 days, at effector-to-target (E:T) ratios of 25:1 and 50:1, for 8 h. Where indicated, blocking antibodies against NKG2D (sc-53501) or ULBP1 (#A21161; Abclonal) were added 60 min prior to co-culture.

Techniques: Staining, Derivative Assay, Western Blot, Expressing, Control, Fluorescence, Cytotoxicity Assay, Cell Culture, Co-Culture Assay, Lysis, Transfection

Journal: Frontiers in Immunology

Article Title: Erastin-induced ferroptosis enhances natural killer cell anti-tumor activity and offers therapeutic potential in neuroblastoma

doi: 10.3389/fimmu.2026.1739503

Figure Lengend Snippet: Transcriptomic profiling reveals upregulation of ULBP1 and ferroptosis-related genes following Erastin treatment. (A) Volcano plots showing differentially expressed genes in KELLY and SK-N-AS neuroblastoma cell lines treated with 1 µM and 2 µM Erastin for 24 hours, respectively, compared to DMSO control. Non-significant genes are shown in gray; significantly upregulated and downregulated genes are shown in red and blue, respectively. (B) KEGG pathway enrichment analysis of DEGs in KELLY and SK-N-AS cells, showing significantly enriched pathways including ferroptosis and amino acid metabolism. (C) Gene Ontology (GO) enrichment network highlighting clusters associated with oxidative stress response, amino acid metabolic processes, and signaling receptor binding, with key genes indicated.

Article Snippet: Separately, NB cells (KELLY, SK-N-AS, SK-N-BE (2), and SK-N-FI) were seeded and washed with PBS, then stained for 30 minutes at 4°C with anti-ULBP1 primary antibody (1:100, clone 170818, R&D systems, #MAB-1380-500) prior to washing and 30 min incubation with secondary antibody (1:500, Invitrogen, # A55747 ).

Techniques: Control, Binding Assay

Journal: Frontiers in Immunology

Article Title: Erastin-induced ferroptosis enhances natural killer cell anti-tumor activity and offers therapeutic potential in neuroblastoma

doi: 10.3389/fimmu.2026.1739503

Figure Lengend Snippet: Low ULBP1 expression correlates with poor prognosis in neuroblastoma patients from the SEQC cohort. Kaplan–Meier overall survival analysis was performed using the SEQC neuroblastoma dataset (n = 498) via the R2 Genomics Analysis and Visualization Platform (A) Complete cohort; (B) MYCN -amplified subset; (C) Patients with progressive disease; (D) INSS stage 4 subset.

Article Snippet: Separately, NB cells (KELLY, SK-N-AS, SK-N-BE (2), and SK-N-FI) were seeded and washed with PBS, then stained for 30 minutes at 4°C with anti-ULBP1 primary antibody (1:100, clone 170818, R&D systems, #MAB-1380-500) prior to washing and 30 min incubation with secondary antibody (1:500, Invitrogen, # A55747 ).

Techniques: Expressing, Amplification

Journal: Frontiers in Immunology

Article Title: Erastin-induced ferroptosis enhances natural killer cell anti-tumor activity and offers therapeutic potential in neuroblastoma

doi: 10.3389/fimmu.2026.1739503

Figure Lengend Snippet: Erastin treatment increases ULBP1 surface expression and enhances NK cell-mediated cytotoxicity in neuroblastoma cells. (A) Surface expression of ULBP1 was measured by flow cytometry in KELLY, SK-N-AS, SK-N-BE (2), and SK-N-FI cells treated with DMSO (control) or Erastin. (B) NK cell-mediated cytotoxicity assay using NK-92 cells co-cultured with neuroblastoma cells (E:T ratio was 8:1) pre-treated with either DMSO (Control), Erastin, or Erastin in combination with Liproxstatin-1 (Lip-1). Cell lysis percentages were quantified in KELLY, SK-N-AS, and SK-N-FI cells. n = 3 biologically independent experiments. *P<0.05, **P<0.01, ***P<0.001; Paired t test.

Article Snippet: Separately, NB cells (KELLY, SK-N-AS, SK-N-BE (2), and SK-N-FI) were seeded and washed with PBS, then stained for 30 minutes at 4°C with anti-ULBP1 primary antibody (1:100, clone 170818, R&D systems, #MAB-1380-500) prior to washing and 30 min incubation with secondary antibody (1:500, Invitrogen, # A55747 ).

Techniques: Expressing, Flow Cytometry, Control, Cytotoxicity Assay, Cell Culture, Lysis