Journal: Journal for Immunotherapy of Cancer

Article Title: Oligoclonal tumor-specific CD8 T-cell revival and IRE1α/XBP1-GDF15-mediated immunosuppressive niches determine neoadjuvant chemoimmunotherapy efficacy in cervical cancer

doi: 10.1136/jitc-2025-012630

Figure Lengend Snippet: ER stress activation drives chemoresistance and immune evasion in cervical cancer. SiHa and ME180 cells were treated with low-dose cisplatin (1 µM), high-dose cisplatin (2 µM), low-dose oxaliplatin (2.5 µM) or high-dose oxaliplatin (5 µM) for 24 hours in 6-well plates, and alterations in ER stress-related genes or EP8 signature genes at the mRNA ( A ) or protein ( B ) level were further detected. ( A ) Heatmap visualization of miRNA levels in SiHa and ME180 cervical cancer cells treated with chemotherapy for 24 hours. The values represent log₂-transformed fold changes relative to untreated controls. The data were normalized to those of ACTB (mean±SEM, n=3). ( B ) Western blot showing dose-dependent upregulation of ER stress and EP8-associated secretory proteins in cells treated as described in ( A ). β-actin served as a loading control. ( C ) CCK-8 viability assays after 24 hours of drug exposure demonstrating synergistic chemosensitivity in ME180 cells treated with cisplatin or oxaliplatin combined with the IRE1α inhibitor 4μ8C (10 µM). ( D ) Schematic of the in vivo experimental design. ( E ) Endpoint volumes in TC-1 syngeneic mice (n=5) treated with vehicle, chemotherapy only (cisplatin 1.5 mg/kg+paclitaxel 15 mg/kg, weekly), combination therapy (chemotherapy+anti-PD-1 200 µg biweekly), or triple therapy (combination therapy+4μ8C 10 mg/kg biweekly). All regimens were injected intraperitoneally. ( F ) Tumor growth curves (mean±SEM) of the tumors in the mice described in ( e ). Endpoint tumor volumes were analyzed by two-way ANOVA with Tukey’s post hoc test (n=5). P values are indicated in the figure. ( G ) Endpoint tumor weights (means±SEMs) of the tumors in the mice described in ( e ). Statistical significance was analyzed by two-way ANOVA with Tukey’s post hoc test (n=5). P values are indicated in the figure. ( H ) Western blot showing dose-dependent upregulation of ER stress and EP8-associated secretory proteins in the tumors of the mice treated as described in ( D ). β-actin served as a loading control. ( I–J ) Boxplots showing the GSVA scores of the CD8, EP8 subset signatures ( I ) and myeloid subsets ( J ) in the TC-1 syngeneic mice (n=3). The boxplots display the median, upper quartile, and lower quartile. Statistical significance was analyzed via the Kruskal-Wallis test. ( L ) Heatmap displaying the expression profiles of ER stress signaling and EP8 signature genes. The samples are grouped in (n=3). Gene expression values were Z score normalized. ( K ) IGV snapshot illustrating the chromatin occupancy of XBP1 at the GDF15 locus. ANOVA, analysis of variance; ATF6, activating transcription factor 6; CDDP, cisplatin; EP8, epithelial subcluster 8; ER, endoplasmic reticulum; GSVA, gene set variation analysis; IGV, Integrative Genomics Viewer; i.p., intraperitoneal; IRE1α, inositol‑requiring enzyme 1 alpha; ME180, human cervical cancer cell line; PD-1, programmed cell death protein 1; PERK, protein kinase R-like ER kinase; SiHa, human cervical cancer cell line; TC-1, mouse lung epithelial cell line transfected with HPV‑16 E6/E7 oncogenes; .

Article Snippet: Cisplatin (1.5 mg/kg, MCE, cat No. HY-17394), anti-PD-1 antibody (200 μg/mouse, MCE, cat No. HY- P99144 ), IgG isotype control (200 μg/mouse, MCE, cat No. HY-P990679) and the IRE1α inhibitor 4μ8C (10 mg/kg, MCE, cat No. HY-19707) were formulated in sterile saline, while paclitaxel (15 mg/kg, MCE, cat No. HY-B0015) was prepared in a vehicle containing 1% DMSO, 4% PEG300, 0.5% Tween-80, and 94.5% saline.

Techniques: Activation Assay, Transformation Assay, Western Blot, Control, CCK-8 Assay, In Vivo, Injection, Expressing, Gene Expression, Transfection

Journal: Journal for Immunotherapy of Cancer

Article Title: Oligoclonal tumor-specific CD8 T-cell revival and IRE1α/XBP1-GDF15-mediated immunosuppressive niches determine neoadjuvant chemoimmunotherapy efficacy in cervical cancer

doi: 10.1136/jitc-2025-012630

Figure Lengend Snippet: ER stress activation drives chemoresistance and immune evasion in cervical cancer. SiHa and ME180 cells were treated with low-dose cisplatin (1 µM), high-dose cisplatin (2 µM), low-dose oxaliplatin (2.5 µM) or high-dose oxaliplatin (5 µM) for 24 hours in 6-well plates, and alterations in ER stress-related genes or EP8 signature genes at the mRNA ( A ) or protein ( B ) level were further detected. ( A ) Heatmap visualization of miRNA levels in SiHa and ME180 cervical cancer cells treated with chemotherapy for 24 hours. The values represent log₂-transformed fold changes relative to untreated controls. The data were normalized to those of ACTB (mean±SEM, n=3). ( B ) Western blot showing dose-dependent upregulation of ER stress and EP8-associated secretory proteins in cells treated as described in ( A ). β-actin served as a loading control. ( C ) CCK-8 viability assays after 24 hours of drug exposure demonstrating synergistic chemosensitivity in ME180 cells treated with cisplatin or oxaliplatin combined with the IRE1α inhibitor 4μ8C (10 µM). ( D ) Schematic of the in vivo experimental design. ( E ) Endpoint volumes in TC-1 syngeneic mice (n=5) treated with vehicle, chemotherapy only (cisplatin 1.5 mg/kg+paclitaxel 15 mg/kg, weekly), combination therapy (chemotherapy+anti-PD-1 200 µg biweekly), or triple therapy (combination therapy+4μ8C 10 mg/kg biweekly). All regimens were injected intraperitoneally. ( F ) Tumor growth curves (mean±SEM) of the tumors in the mice described in ( e ). Endpoint tumor volumes were analyzed by two-way ANOVA with Tukey’s post hoc test (n=5). P values are indicated in the figure. ( G ) Endpoint tumor weights (means±SEMs) of the tumors in the mice described in ( e ). Statistical significance was analyzed by two-way ANOVA with Tukey’s post hoc test (n=5). P values are indicated in the figure. ( H ) Western blot showing dose-dependent upregulation of ER stress and EP8-associated secretory proteins in the tumors of the mice treated as described in ( D ). β-actin served as a loading control. ( I–J ) Boxplots showing the GSVA scores of the CD8, EP8 subset signatures ( I ) and myeloid subsets ( J ) in the TC-1 syngeneic mice (n=3). The boxplots display the median, upper quartile, and lower quartile. Statistical significance was analyzed via the Kruskal-Wallis test. ( L ) Heatmap displaying the expression profiles of ER stress signaling and EP8 signature genes. The samples are grouped in (n=3). Gene expression values were Z score normalized. ( K ) IGV snapshot illustrating the chromatin occupancy of XBP1 at the GDF15 locus. ANOVA, analysis of variance; ATF6, activating transcription factor 6; CDDP, cisplatin; EP8, epithelial subcluster 8; ER, endoplasmic reticulum; GSVA, gene set variation analysis; IGV, Integrative Genomics Viewer; i.p., intraperitoneal; IRE1α, inositol‑requiring enzyme 1 alpha; ME180, human cervical cancer cell line; PD-1, programmed cell death protein 1; PERK, protein kinase R-like ER kinase; SiHa, human cervical cancer cell line; TC-1, mouse lung epithelial cell line transfected with HPV‑16 E6/E7 oncogenes; .

Article Snippet: Cisplatin (1.5 mg/kg, MCE, cat No. HY-17394), anti-PD-1 antibody (200 μg/mouse, MCE, cat No. HY- P99144 ), IgG isotype control (200 μg/mouse, MCE, cat No. HY-P990679) and the IRE1α inhibitor 4μ8C (10 mg/kg, MCE, cat No. HY-19707) were formulated in sterile saline, while paclitaxel (15 mg/kg, MCE, cat No. HY-B0015) was prepared in a vehicle containing 1% DMSO, 4% PEG300, 0.5% Tween-80, and 94.5% saline.

Techniques: Activation Assay, Transformation Assay, Western Blot, Control, CCK-8 Assay, In Vivo, Injection, Expressing, Gene Expression, Transfection

Journal: Journal of Translational Medicine

Article Title: IGFBP2 promotes immunosuppression by regulating macrophage PD-L1 expression in NEUROD1-high small cell lung cancer

doi: 10.1186/s12967-026-08112-2

Figure Lengend Snippet: IGFBP2 is selectively up-regulated in SCLC-N subtype and associated with poor prognosis. ( A ) Single cell sequencing data show IGFBP2 is up-regulated in SCLC-N. ( B ) A scatter plot showing the correlation between the transcriptional levels of IGFBP2 and NEUROD1 in the GSE60052 dataset. ( C ) Comparison of IGFBP2 levels between the NEUROD1 high-expression group and the NEUROD1 low-expression group in the GSE60052 dataset. ( D - E ) The expression levels of IGFBP2 and NEUROD1 in tissue specimens of SCLC patients were detected by mIHC ( D ) and RT-qPCR ( E ), respectively. ( F - G ) The mRNA and protein expression levels of IGFBP2 in different subtypes of SCLC cell lines. “a” denotes comparison with H446, “b” with H82, and “c” with H524. ( H ) High IGFBP2 expression is associated with poor prognosis. ( I ) Kaplan-Meier analysis of patient survival based on IGFBP2 and NEUROD1 expression levels in four groups

Article Snippet: Experimental mice received recombinant mouse IGFBP2 (rmIGFBP2) protein (MCE, USA; Cat#HY- P74846 ) (50 μg/kg) via tail-vein injection; controls received an equal volume of sterile saline.

Techniques: Single Cell, Sequencing, Comparison, Expressing, Quantitative RT-PCR

Journal: Journal of Translational Medicine

Article Title: IGFBP2 promotes immunosuppression by regulating macrophage PD-L1 expression in NEUROD1-high small cell lung cancer

doi: 10.1186/s12967-026-08112-2

Figure Lengend Snippet: SCLC-N-driven macrophage polarization is IGFBP2-dependent. ( A ) Bar plot showing the correlation between IGFBP2 expression and immune cell infiltration in SCLC, based on CIBERSORT analysis of the GSE60052 dataset. ( B - C ) Protein and transcript levels of CD163 and IGFBP2 in tumor tissues were examined by mIHC ( B ) and RT-qPCR ( C ), respectively. ( D - F ) M0-THP-1 cells were treated with rhIGFBP2, and the mRNA and protein levels of M2-TAM markers (CD163 and HLA) were assessed by RT-qPCR and Western blotting ( D - E ), while cytokine levels (IL-10 and IL-12) in the culture supernatants were measured by ELISA ( F ). ( G - L ) M0-THP-1 cells were co-cultured with the indicated SCLC cells with IGFBP2 knockdown (sgIGFBP2). The mRNA and protein levels of M2-TAM markers (CD163 and HLA) were assessed by RT-qPCR and Western blot ( G - J ), while cytokine levels (IL-10 and IL-12) in the culture supernatants were measured by ELISA ( K - L )

Article Snippet: Experimental mice received recombinant mouse IGFBP2 (rmIGFBP2) protein (MCE, USA; Cat#HY- P74846 ) (50 μg/kg) via tail-vein injection; controls received an equal volume of sterile saline.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, Knockdown

Journal: Journal of Translational Medicine

Article Title: IGFBP2 promotes immunosuppression by regulating macrophage PD-L1 expression in NEUROD1-high small cell lung cancer

doi: 10.1186/s12967-026-08112-2

Figure Lengend Snippet: IGFBP2 promotes the expression of PD-L1 in macrophages. ( A ) Correlation analysis showing the association between PD-L1 and FOXP3 transcriptional levels in SCLC tissues. ( B - E ) RT-qPCR and Western blotting were used to assess the impact of IGFBP2 on PD-L1 expression in M0-THP-1 and M0-Mc macrophages (Mc, macrophages derived from human peripheral blood monocytes). ( F - H ) RT-qPCR, Western blotting and IF assay were used to assess IGFBP protein accumulation in M0-PM (PM, mouse peritoneal macrophages). ( I - J ) RT-qPCR and Western blotting showed that the SCLC-N subtype elicited a markedly greater up-regulation of PD-L1 in macrophages than any other subtype

Article Snippet: Experimental mice received recombinant mouse IGFBP2 (rmIGFBP2) protein (MCE, USA; Cat#HY- P74846 ) (50 μg/kg) via tail-vein injection; controls received an equal volume of sterile saline.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Derivative Assay

Journal: Journal of Translational Medicine

Article Title: IGFBP2 promotes immunosuppression by regulating macrophage PD-L1 expression in NEUROD1-high small cell lung cancer

doi: 10.1186/s12967-026-08112-2

Figure Lengend Snippet: IGFBP2 modulates CD8 + T-cell function by promoting PD-L1 expression on macrophages. ( A ) Schematic of the co-culture system. ( B - F ) Flow cytometry confirmed that PD-L1 blockade can reverse the suppression of CD8 + T-cell infiltration caused by high IGFBP2 expression in SCLC cells. ( G ) Tumor size of mice in different treatment groups. ( H ) Growth curves of tumors from mice with different treatments

Article Snippet: Experimental mice received recombinant mouse IGFBP2 (rmIGFBP2) protein (MCE, USA; Cat#HY- P74846 ) (50 μg/kg) via tail-vein injection; controls received an equal volume of sterile saline.

Techniques: Cell Function Assay, Expressing, Co-Culture Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: IGFBP2 promotes immunosuppression by regulating macrophage PD-L1 expression in NEUROD1-high small cell lung cancer

doi: 10.1186/s12967-026-08112-2

Figure Lengend Snippet: The mechanism of IGFBP2 regulating PD-L1 expression in macrophages. ( A ) Western blotting showed that IGFBP2 modulates PD-L1 in macrophages independently of the IGF axis and PI3K-AKT signaling pathway. ( B - C ) NLS structure domain and cell localization diagram. ( D - E ) RT-qPCR analysis of IGFBP2 and PD-L1 expression in cells after transfection. ( F ) Western blot showing IGFBP2 and PD-L1 protein levels following transfection. ( G ) IF showing IGFBP2 and PD-L1 expression in M0-THP-1 cells after transfection

Article Snippet: Experimental mice received recombinant mouse IGFBP2 (rmIGFBP2) protein (MCE, USA; Cat#HY- P74846 ) (50 μg/kg) via tail-vein injection; controls received an equal volume of sterile saline.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection