Journal: Frontiers in Immunology

Article Title: Evaluating mesothelin as an immunotherapeutic target for endogenous T cells

doi: 10.3389/fimmu.2026.1853409

Figure Lengend Snippet: Effector functions of MSLN-specific T cells. (A) representative donor data (left panel) and summary results (median ± IQR, n=21; right panel) showing CD8 + /IFNγ + and CD4 + /IFNγ + T cells, as assessed by ICS. (B) representative donor data (left panel) and summary results (median ± IQR; n=21; right panel) showing dual (IFNγ/TNFα)-producing CD4 + and CD8 + MSLN-specific T cells, as assessed by ICS. Box plots show the median and 25th and 75th data quartiles. (C) polyfunctionality of MSLN-specific T cells as measured by FluoroSpot (IFNγ/Granzyme B/TNFα) (n=6). Top panel shows the total frequency (SFC/2x105 cells) of specific T cells for each of the individual effector molecules, while the bottom panel shows the proportion of cells that are single, dual or triple analyte-producing. All data represent biological replicates (i.e. independent donors). TNFα, tumor necrosis factor alpha; ICS, intracellular cytokine staining; GrB, Granzyme B.

Article Snippet: For the quantitation of polyfunctional cells simultaneously secreting IFNγ, GrB and/or TNFα a commercial FluoroSpot assay was used (Human IFNγ/Granzyme B/TNFα FluoroSpot Plus, Mabtech, Inc.).

Techniques: Staining

Journal: bioRxiv

Article Title: Multifaceted immune resistance landscapes in human oligodendrocytes protect against cytotoxic T cells and are dysregulated in MS brain cell subsets

doi: 10.64898/2026.04.21.719872

Figure Lengend Snippet: (A) FACS analysis of antigen specificity of FluTC generated from HLA-A2 + healthy donors by repetitive expansions. All samples were gated on lymphocytes, single cells, and live cells. CD8 and Flu-Pentamer stainings were performed on days 7 and 14 of antigen-specific expansion (ASE) and day 14 of rapid expansion protocol (REP). During ASE, some CD8 + T cells were expanded in the presence of unpulsed feeder cells (w/o flu-peptide) as a negative control. (B) ELISA to determine cytokine secretion by FluTC. MO3.13-A2-Luc cells were pulsed with diluting concentrations of flu peptide and co-cultured with FluTC. After 20 h of co-culture, the supernatant of co-culture was analyzed by ELISA to detect IFNγ (left) and Granzyme B (right) secretion by FluTC.

Article Snippet: Supernatants of FluTC-MO3.13-A2-Luc co-cultures and anti-CD3/CD28 activated FluTC were further analyzed for the detection of IFNγ (Human IFN-γ ELISA Set, BD OptEIA, #555142), TNFα (Human TNF ELISA Set, BD OptEIA, #555212), Granzyme B (Human Granzyme B ELISA development kit, Mabtech, #3485-1H-20) and TRAIL (human TRAIL/TNFSF10 DuoSet ELISA, R&D Systems, #DY375-05) ( , ).

Techniques: Generated, Negative Control, Enzyme-linked Immunosorbent Assay, Cell Culture, Co-Culture Assay

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 promotes tumor immune evasion by upregulating PD-L1 expression within TME (A–C) scRNA-seq analysis of two paired tumor and paratumor samples from LUAD patients showing cell clustering (A), WFDC2 /HE4-expressing cell populations (B), and quantification of WFDC2 /HE4 expression in epithelial/malignant clusters (C). (D) Online dataset analysis of WFDC2 mRNA expression in tumor versus normal LUAD tissues. (E and F) HE4 overexpression (E) or knockout (F) respectively promoted or suppressed the growth of subcutaneous LLC tumors in mice. (G) Survival of mice intraperitoneally inoculated with HE4-overexpressing or control ID8 cells. (H) HE4 knockout attenuated ID8 peritoneal tumor progression, shown by representative abdominal images and ascites volume. (I) SDS-PAGE with Coomassie blue staining showing the purity of Fc and mouse HE4-Fc (mHE4-Fc) recombinant proteins. (J) Administration of mHE4-Fc promoted MC38 subcutaneous tumor growth. (K) mHE4-Fc administration reversed the growth suppression of HE4-KO LLC subcutaneous tumors. (L) mHE4-Fc failed to reverse tumor growth suppression in HE4-KO LLC tumors implanted in Rag1 -deficient mice. (M) Extracellular HE4 did not promote LLC cell proliferation in vitro , as assessed by CCK8 assay. (N) mHE4-Fc administration suppressed CD8 + T cell activation in the TME of HE4-KO LLC tumors, assessed by IFN-γ + and granzyme B + CD8 + T cells. (O) HE4 upregulated PD-L1 expression on macrophages in the microenvironment of LLC, MC38, and HE4-KO LLC tumors. (P–R) mHE4-Fc administration failed to reverse tumor suppression of HE4-KO LLC subcutaneous tumors in Cd274 -deficient mice, with treatment scheme (P), tumor growth (Q), and tumor weight (R). Statistical analyses were performed using Wilcoxon rank-sum test (D), two-way ANOVA (E, F, J–L, and Q), log rank (Mantel-Cox) test (G), and two-tailed paired (C) or unpaired t tests (H, M, N, O, and R). Data represent two independent experiments (E and M).

Article Snippet: Mouse Granzyme B ELISA kit , Elabscience , Cat# E-EL-M0594.

Techniques: Expressing, Over Expression, Knock-Out, Control, SDS Page, Staining, Recombinant, In Vitro, CCK-8 Assay, Activation Assay, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 competes with IFN-γ for IFN-γR binding and modulates downstream gene expression (A and B) Raw264.7 cells were stimulated with HE4-Fc (20 μg/mL) or IFN-γ (100 ng/mL) for 3 h, followed by RNA-seq; volcano plots of HE4- (A) or IFN-γ-regulated genes (B) are shown. (C) Genes commonly upregulated by HE4 and IFN-γ. (D) AlphaFold-3-predicted interfaces of HE4-IFNGR1/2 and IFN-γ-IFNGR1/2 complexes, with shared receptor-contact residues highlighted. (E) Competitive binding assay: His-tagged IFNGR1/2 was incubated with Flag-HE4 in the presence or absence of IFN-γ, followed by Ni-TED pull-down and immunoblotting. (F and G) SPR sensorgrams showing binding of mHE4-Fc (F) or mIFN-γ-Fc (G) to mIFNGR1-His. (H and I) ELISA quantification of HE4 and/or IFN-γ levels in ascites from ID8-tumor-bearing mice (H) and LLC-tumor-conditioned media (I). (J) High concentrations of HE4 reduced IFN-γ binding to IFNGR1/2 in competitive pull-down assays. (K) PCA of RNA-seq profiles from Raw264.7 cells treated with HE4-Fc, HE4-Fc plus IFN-γ, or IFN-γ for 12 h. (L) Expression (TPM) of representative STAT1- or STAT3-associated genes following the indicated treatments. Statistical analyses were performed using two-tailed paired Student’s t tests (H and I). Data in (E) and SPR sensorgrams (F and G) are representative of three independent experiments.

Article Snippet: Mouse Granzyme B ELISA kit , Elabscience , Cat# E-EL-M0594.

Techniques: Binding Assay, Gene Expression, RNA Sequencing, Competitive Binding Assay, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 blockade exerts therapeutic efficacy across multiple mouse tumor models (A–C) Mice bearing subcutaneous LLC tumors were treated intraperitoneally with isotype immunoglobulin G (IgG) or anti-mHE4 mAbs (clones #1 or #117). Treatment scheme (A), tumor growth curves (B), and representative tumors with weights (C) are shown. (D and E) Anti-mHE4 (clone #117) treatment suppressed urethane-induced lung tumorigenesis, shown by representative H&E staining and tumor quantification. Scale bar, 1,000 μm. (F and G) HE4 blockade attenuated ID8 intraperitoneal tumor progression, shown by representative abdominal images and ascites volume. (H and I) Therapeutic efficacy of anti-mHE4 (clone #117) in an orthotopic HGS-1 ovarian tumor model, shown by treatment scheme and representative tumors with weights. (J) Serum-free HE4 levels measured by ELISA in LLC-tumor-bearing mice one day after the final antibody treatment. (K and L) Anti-mHE4 (clone #117) reduced PD-L1 expression on tumor-associated macrophages in the LLC subcutaneous and ID8 intraperitoneal models. (M and N) Anti-mHE4 (clone #117) failed to suppress LLC tumor growth in Cd274 -deficient mice. (O) Combination therapy with anti-mHE4 and anti-mCTLA-4 enhanced tumor suppression in the LLC subcutaneous model. (P) Combination therapy with anti-mHE4 and paclitaxel (PTX) enhanced antitumor efficacy in the LLC subcutaneous model. (Q) Serum cytokines and clinical chemistry parameters following treatment with isotype IgG, anti-mHE4 mAb, or anti-mPD-1 mAb. (R and S) Safety assessment showing representative H&E images and incidence of inflammation in heart, liver, lung, and colon after anti-mHE4 or anti-mPD-1 treatment. Statistical analyses were performed using two-way ANOVA (B and N–P), one-way ANOVA (C, K, and N–Q), two-tailed unpaired t tests (E, G–J, and L), and chi-squared test (S). Data in (A–P) are pooled from two independent experiments. Scale bars, 100 μm.

Article Snippet: Mouse Granzyme B ELISA kit , Elabscience , Cat# E-EL-M0594.

Techniques: Drug discovery, Clone Assay, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 neutralization exerts therapeutic activity in human cancer models (A–C) PMA-differentiated THP-1 macrophages were stimulated with Fc or hHE4-Fc, and PD-L1 expression was assessed by flow cytometry (A), immunoblotting (B), and RT-qPCR (C); a commercial hHE4-Fc was included as an independent control. (D) Binding of hHE4 to PMA-differentiated THP-1 cells assessed by flow cytometry. (E and F) PMA-differentiated THP-1 cells were pretreated with ruxolitinib, fludarabine, or Stattic, followed by hHE4-Fc stimulation; PD-L1 was quantified by RT-qPCR (E) and flow cytometry (F). (G and H) Anti-hHE4 monoclonal antibodies inhibited hHE4-induced PD-L1 upregulation in PMA-differentiated THP-1 cells, assessed by RT-qPCR (G) and flow cytometry (H). (I) Anti-hHE4 mAb clone #10 blocked hHE4 binding to PMA-differentiated THP-1 cells. (J) Binding of wild-type or epitope-mutant hHE4-Fc to anti-hHE4 mAb clone #10 was quantified by ELISA. (K) Pharmacokinetic analysis of anti-hHE4 mAb clone #10 in C57BL/6 mice following intravenous administration. (L–O) Fresh human LUAD tumor cell suspensions were treated with anti-hHE4 mAb clone #10, followed by flow cytometric analysis of PD-L1 and ELISA measurement of IFN-γ and granzyme B. (P) Recombinant HE4 suppressed IFN-γ production in human LUAD tumor cell suspensions. (Q–T) HE4 blockade enhanced PBMC-mediated antitumor activity in humanized C-NKG mice bearing OVCAR3 or NCI-H358 tumors, shown by treatment scheme, representative tumors, and tumor weights. Schematics (L and Q) were created using BioRender. Statistical analyses were performed using one-way ANOVA (C, E, and G), paired t tests (M–P), or unpaired t tests (S and T). Data in (A–J) are representative of three independent experiments; data in (Q–T) are pooled from two independent experiments.

Article Snippet: Mouse Granzyme B ELISA kit , Elabscience , Cat# E-EL-M0594.

Techniques: Neutralization, Activity Assay, Expressing, Flow Cytometry, Western Blot, Quantitative RT-PCR, Control, Binding Assay, Bioprocessing, Mutagenesis, Enzyme-linked Immunosorbent Assay, Recombinant