Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: TriKE binds CD16 + NK cells and induces NK cell proliferation and activation (A–C) Schematic of the human VRC (A) and CD4 (B) TriKEs and the rhesusized CD4 (C) TriKE. (D) Whole PBMCs from PWOH (black, n = 3) and PWH (red, n = 3) were incubated with or without TriKE (10 nM), and NK cells (live CD56 + CD3 − ) were assessed for CD16 expression and bound TriKE (anti-His) by flow cytometry. (E–I) Whole PBMCs from PWOH (black, n = 5) and PWH (red, n = 5) were CellTrace labeled and cultured with listed treatments for 7 days, and (E) CD56 + CD3 − NK cell proliferation (CTB dilution) and activation by CD38 (F), HLA-DR (G), CD69 (H), and CD57 (I) was assessed by flow cytometry. NT, no treatment, IL-15 (10 nM), TriKEs (10 nM). Statistical assessment was performed with a matched repeated measures two-way ANOVA with a Geisser-Greenhouse correction. Multiple comparisons were controlled for by a Dunnett’s multiple comparisons test. Each dot represents a unique donor, and error bars represent mean ± SEM.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Activation Assay, Incubation, Expressing, Flow Cytometry, Labeling, Cell Culture

Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: HIV TriKEs mediate enhanced NK cell function and killing of HIV-infected cells Primary human PBMCs were co-cultured with CellTrace-labeled HIV-uninfected (A3, blue diamonds), productively infected (8E5, red circles), or latent HIV (ACH-2, green squares) or no targets (PB alone, black triangles) at an E:T of 2:1 with listed treatments. After 5 h, NK cells were evaluated for CD107a (A and B), IFN-γ (C and D), and TNF-α (E and F) expression and killing of target cells (G and H) by flow cytometry. NT, no treatment, IL-15 (10 nM), VRC01 bNab (10 μg/mL), TriKEs (10 nM). Statistical assessment was performed with a matched repeated measures two-way ANOVA with a Geisser-Greenhouse correction. Multiple comparisons were controlled for by a Dunnett’s multiple comparisons test. Data are represented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005. N = 5 for each PWOH and PWH.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Cell Function Assay, Infection, Cell Culture, Labeling, Expressing, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: CD38 + CD57 + NK cells exhibit the greatest polyfunctional response mediated by HIV-targeting TriKEs against 8E5 HIV-infected cells Polyfunctional NK cell responses were assessed by co-expression of CD38, CD57, CD107a, IFN-γ, and TNF-α and analyzed using SPICE (Simplified Presentation of Incredibly Complex Evaluations). Whole PBMCs from HIV− (A–E) and HIV+ (F–J) donors were co-cultured with 8E5 target cells at a ratio of 2:1 and either left untreated or treated with 10 nM IL-15 (A and F), 10 μg/mL VRC01 bNAb (B and G), IL-15+bNAb (C and H), 10 nM VRC TriKE (D and I), or 10 nM CD4 TriKE (E and J). A Wilcoxon rank-sum test was run to assess statistical differences between each condition compared to the IL-15 control. (K–N) Bar plots showing polyfunctional NK cell responses (CD107a+IFN-γ+TNF-α+) within NK cell subsets that are CD38 + CD57 + (K), CD38 + CD57 − (L), CD38 − CD57 + (M), and CD38 − CD57 − (N). Each dot represents a unique donor, and error bars represent mean ± SEM. N = 5 for each PWOH and PWH.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Infection, Expressing, Cell Culture, Control

Journal: Frontiers in Immunology

Article Title: Comparative profiling of MAIT cells across gastrointestinal cancers identifies conserved NK receptor dysregulation and HCC-specific defects in cytotoxicity and XBP1s signaling

doi: 10.3389/fimmu.2026.1924000

Figure Lengend Snippet: Cytotoxic capacity of HCC-derived MAIT cells. (A, B) Representative flow cytometry plots illustrating the cytotoxic potential of MAIT and NK cells ex vivo and following 96 hours of IL-15 stimulation. (C, D) Differences in cytotoxicity of blood-derived MAIT and NK cells from healthy participants (HD) and HCC patients (HCC), analyzed ex vivo and after IL-15 stimulation at an effector-to-target ratio of 20:1, n =7-8/group. (E, F) Differences in cytotoxicity of peritumour and tumour-derived MAIT and NK cells from HCC patients. The cytotoxicity assay was conducted either ex vivo or over a 96-hour period with IL-15 at a ratio of 1:20, n =5-7/group. (G, H) Induction of NKG2D and Granzyme B on peripheral blood, peritumour and tumour-derived MAIT cells after 96 hours of IL-15 stimulation. (I) Inhibition of the cytotoxic capacity of peripheral MAIT and NK cells from HCC patients following 96-hour IL-15 stimulation, using a neutralizing antibody against NKG2D or the Granzyme B inhibitor concanamycin A (CMA), at an ratio of 1:20, n = 5-6/group. (J, K) Inhibition of MAIT and NK cell cytotoxicity from HCC peritumoral and tumour samples using CMA after 96-hour IL-15 stimulation at an ratio of 1:20 (n = 5–6 per group). HD: n = 6–9; HCC: n = 5–6. Non-significant (n.s.): p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001, as calculated using ordinary one-way ANOVA, Kruskal–Wallis test (C–F, I), a two-tailed paired t-test or a two-tailed Wilcoxon matched-pairs signed-rank test (G, H).

Article Snippet: Cells were then cultured in RPMI 1640 medium supplemented with 0.3 mg/mL glutamine, 100 U/mL penicillin, 0.1 mg/mL streptomycin, 10% human AB serum (Pan Biotech, REF: P30-2701), and recombinant human IL-15 (50 ng/mL; Miltenyi Biotec, REF: 130-093-955) or Phorbol-12-myristat-13-acetat (25 ng/ml; Sigma-Aldrich, REF: P8139)/Ionomycin calcium salt (1μg/ml; Sigma-Aldrich, REF: I0634) (PMA/Iono).

Techniques: Derivative Assay, Flow Cytometry, Ex Vivo, Cytotoxicity Assay, Inhibition, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Comparative profiling of MAIT cells across gastrointestinal cancers identifies conserved NK receptor dysregulation and HCC-specific defects in cytotoxicity and XBP1s signaling

doi: 10.3389/fimmu.2026.1924000

Figure Lengend Snippet: XBP1s activation in peripheral and tissue-derived MAIT cells. (A) Representative flow cytometry contour plots illustrating XBP1s expression in MAIT cells isolated from peripheral blood, peritumoral tissue, and tumour tissue of patients with HCC. (B) Frequencies of XBP1s + MAIT cells in peripheral blood from age-matched healthy donors (HD, n = 8) and in peripheral blood- and tissue-derived (peritumoral and tumour) MAIT cells from patients with HCC, n=6-10/tissue. (C) Frequencies of peripheral XBP1s + MAIT cells - isolated from healthy donors (HD, n = 8) and patients with HCC (n = 6–8), or from (D) peritumour and tumour HCC tissue - analyzed ex vivo or following functional stimulation to modulate cytotoxic capacity using the ER stress inducers thapsigargin (TG) and tunicamycin (TM), or IL-15 for 24 or 96 hours; n = 8–12/group. (E) Frequencies of peripheral MAIT cells expressing NKG2D or Granzyme B - isolated from healthy donors (HD, n = 8) and patients with HCC (n = 6–8), or from (F) peritumour and tumour HCC tissue - analyzed ex vivo or following functional stimulation to modulate cytotoxic capacity using the ER stress inducers TG and TM or IL-15 for 24 or 96 hours; n = 8–12/group. (G) Correlation analysis between XBP1s and Granzyme B in peripheral MAIT cells from age-matched healthy donors and HCC patients (left panel), and in peritumoral and tumour MAIT cells from HCC tissue (right panel) after IL-15 stimulation for 96 hours. (H) Cytotoxic capacity of peripheral MAIT cells isolated from healthy donors ex vivo or following 96-hour stimulation with IL-15 or the ER stress inducers TG and TM (n = 3–7 per group). Non-significant (n.s.): p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001, as calculated by a two-tailed paired t-test, two-tailed Wilcoxon matched-pairs signed-rank test (C–F), ordinary one-way ANOVA or Kruskal–Wallis test (B, H) and simple linear regression.

Article Snippet: Cells were then cultured in RPMI 1640 medium supplemented with 0.3 mg/mL glutamine, 100 U/mL penicillin, 0.1 mg/mL streptomycin, 10% human AB serum (Pan Biotech, REF: P30-2701), and recombinant human IL-15 (50 ng/mL; Miltenyi Biotec, REF: 130-093-955) or Phorbol-12-myristat-13-acetat (25 ng/ml; Sigma-Aldrich, REF: P8139)/Ionomycin calcium salt (1μg/ml; Sigma-Aldrich, REF: I0634) (PMA/Iono).

Techniques: Activation Assay, Tissue, Derivative Assay, Flow Cytometry, Expressing, Isolation, Ex Vivo, Functional Assay, Analysis, Two Tailed Test