Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The APC function of B cells is closely associated with their FA metabolism in metastatic OvCa. A GSVA pathway enrichment analysis of APC function low and APC function high B cells in patients with metastatic OvCa ( GSE235951 , GSE147082 and GSE154600 dataset in the GEO database, n = 13). B Representative images of the IHC staining of TLS structure (composed of CD3 + T cells, CD19 + B cells, and CD21 + FDC), CD80, and CD86, respectively, in the area adjacent to or away from tumor or adipose tissues in clinical HGSOC specimens (n = 20 for TLS and n = 5 for CD80 or CD86). Red dashed line area: TLS structure. Magnification × 200. C Representative image of lymphoid aggregates (white dashed line area) by immunofluorescent staining in ascites of OvCa mice with 3 w and 6 w in tumor-bearing mice. D Mean fluorescence intensity of CD80, CD86, CD83, and MHC class II molecules in ascitic CD19 + B cells of OvCa mice were detected by flow cytometry. E Comparison of mRNA levels of FA metabolic genes in ascitic B cells in OvCa mice. The relative expression of each gene was calculated using β -actin as the internal reference. F Protein expression of β-actin and FA metabolic proteins in ascitic B cells in OvCa mice was assessed by WB. β -Actin was used as the internal control to calculate the relative expression level of FA metabolic proteins. G Mean fluorescence intensity of Bodipy C16 in ascitic B cells of OvCa mice detected by flow cytometry. H Expression of A-CoA in ascitic B cells of OvCa mice detected by ELISA. I Expression of ATP in ascitic B cells of OvCa mice detected by ELISA. J Mean fluorescence intensity of intracellular oxidized lipid in ascitic B cells of OvCa mice detected by flow cytometry. I Comparison of mRNA levels of FA metabolic genes in ascitic B cells in OvCa mice. The relative expression of each gene was calculated using β -actin as the internal reference. Data are presented as the mean ± SD of three independent experiments. FA, fatty acid; TLS, tertiary lymphoid structure. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: Immunohistochemistry, Staining, Fluorescence, Flow Cytometry, Comparison, Expressing, Control, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The APC function and resulting anticancer immunity of B cells can be enhanced by oleic acid (OA) via reprogramming FA metabolism in vitro. A Mean fluorescence intensity of CD80 and MHC Class II molecules in CD19 + B cells from peripheral blood of healthy volunteers (n = 3) treated with OA and PA (both 150 μM), respectively. B Mean fluorescence intensity of CD80, CD86, CD83, MHC Class II molecules, and Ki67 in splenic CD19 + B cells of WT mice treated with 150 μM OA. C Mean fluorescence intensity of CD80, CD86, CD83, and MHC Class II molecules in ascitic CD19 + B cells from 3 w OvCa-bearing mice when treated with 150 μM OA. D Analysis of FA metabolism-related signaling pathways based on RNA-seq results. GSEA was used to analysised the FA metabolic pathways. E Comparison of mRNA levels of main FA metabolic genes in ascitic B cells from 3 w tumor-bearing mice when treated with 150 μM OA. The relative expression of each gene was calculated using β -actin as the internal reference. F Experimental scheme to detect the influence of inhibiting OA uptake on ascitic B cells. G Comparison of protein expressions of main FA metabolic molecules in 3 w OvCa-bearing mouse ascitic B cells pretreated with BMS and treated with OA. β -Actin was used as the internal control to calculate the relative expression level of the main FA metabolic molecules. H Mean fluorescence intensity of Bodipy C16 in 3 w OvCa-bearing mouse ascitic B cells pretreated with BMS and treated with OA was detected by flow cytometry. I The expression of A-CoA, ATP and the FAO activity in 3 w OvCa-bearing mouse ascitic B cells pretreated with BMS treated with OA was detected by ELISA. J Mean fluorescence intensity of intracellular oxidized lipid in 3 w OvCa-bearing mouse ascitic B cells pretreated with BMS and treated with OA was detected by flow cytometry. K Mean fluorescence intensity of CD80, CD86, and CD83 in 3 w OvCa-bearing mouse ascitic CD19 + B cells pretreated with BMS and treated with OA was detected by flow cytometry. L Mean fluorescence intensity of Bodipy C16, CD80, CD86 and CD83 in FABP4-knockdown CD19 + B cells, which are from the ascites of 3 w OvCa-bearing mice, treated with OA, was detected by flow cytometry. PBMC, peripheral blood mononuclear cell; SP, spleen; AS, Ascites; OA, oleic acid; PA, palmitic acid; BMS, BMS309403. Data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: In Vitro, Fluorescence, Protein-Protein interactions, RNA Sequencing, Comparison, Expressing, Control, Flow Cytometry, Activity Assay, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: Reprogramming of the FA metabolism of B cells by OA can improve anticancer immunity in vitro. A Experimental scheme to detect the in vitro effects of ascitic B cells treated with OA on anticancer immunity. B Mean fluorescence intensity of CD80, CD86, and CD83 in 3 w OvCa-bearing mouse ascitic CD19 + B cells after pulsed with ID8-Luc-cell-prepared antigenic peptides and pretreated with BMS and treated with OA, T cells and ID8-Luc. C Levels of IFN- γ , GZMB, and TNF-α in the supernatant of the coculture system constructed by ID8-Luc cells and 3 w OvCa-bearing mouse ascitic B and splenic T cells were detected by ELISA. D Cytotoxicity of T cells in the coculture system mentioned above was detected by luciferase assay. AS, Ascites; Ag, Antigen; OA, oleic acid; BMS, BMS309403. Data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: In Vitro, Fluorescence, Construct, Enzyme-linked Immunosorbent Assay, Luciferase

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The enhanced APC function of B cells by OA in vitro is achieved through H3K27ac-mediated upregulation of PPAR γ expression. A Protein expression of β -actin and H3K27ac in 3 w OvCa-bearing mouse ascitic CD19 + B cells pretreated with BMS and treated with OA was assessed by WB. β -Actin was used as the internal control to calculate the relative expression level of H3K27ac. B The enrichment percentage of H3K27ac at the PPAR γ , CD80, CD86, and CD83 promoter regions was quantified using ChIP-seq analysis. C Correlation analysis of PPAR γ and APC function-related genes (CD80, CD86, CD83, MHC II) in B cells in OvCa patients in the TCGA database (n = 426). D Analysis of protein interaction among FABP4, PPAR γ , CD80, CD86, and CD83. E The enrichment percentage of PPAR γ at the CD80, CD86, and CD83 promoter regions was quantified using ChIP-seq analysis. F Mean fluorescence intensity of CD80, CD86, and CD83 in 3 w OvCa-bearing mouse ascitic CD19 + B cells pretreated with BMS, GW9662/Trog, and treated with OA was detected by flow cytometry. AS, Ascites; Ag, Antigen; OA, oleic acid; BMS, BMS309403. Data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: In Vitro, Expressing, Control, ChIP-sequencing, Fluorescence, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The combination of adoptive immunotherapy with APC function-enhanced B cells and LDC improved anticancer immunity in metastatic OvCa mice. A Experimental scheme to evaluate the effects of combining adoptive immunotherapy with APC function-enhanced B cells and LDC in metastatic OvCa mice. B Representative BLI images and comparison of OvCa progression in each group (n = 3). C Representative images ( a ) and the number of tumor nodules ( b ) in the abdominal wall of mice in each group (n = 3). Areas marked by yellow dashed lines: Representative tumor nodules in the abdominal wall. D (a) Representative images of lymphoid aggregates (composed of CD3 + T cells, CD19 + B cells, and CD21 + FDC) in the ascitic cells of mice in each group. (b) Comparison of the amount of CD3 + T, CD8 + T, Ki67 + CD3 + T, Ki67 + CD8 + T, CD19 + B, CD80 + B, CD86 + B, and lymphoid aggregates respectively in the ascitic cells of mice in each group (n = 3) in frozen sections detected by immunofluorescence. E Comparison of the proportion of CD3 + T, CD8 + T, CD19 + B, CD19 − CD138 + B cells, mean fluorescence intensity of Ki67 in CD3 + T and CD8 + T, and mean fluorescence intensity of CD80, CD86, CD83, MHC II in CD19 + B, respectively, in ascitic cells of each group of mice (n = 3) detected by flow cytometry. F Mean fluorescence intensity values of IL-2, IFN- γ , GZMB, and CTLA-4 in CD8 + T cells, respectively. G Kaplan Meier analysis of survival time of each group of mice (n = 7). LDC, low-dose chemotherapy; B OA , B cells that have been treated with OA for 24h; L-DDP, low-dose DDP (1 mg/kg); H-DDP, high-dose DDP (2 mg/kg); BLI, bioluminescence imaging. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: Comparison, Immunofluorescence, Fluorescence, Flow Cytometry, Imaging

Journal: PLoS Pathogens

Article Title: Inhibition of polo-like kinase 1 (PLK1) facilitates reactivation of gamma-herpesviruses and their elimination

doi: 10.1371/journal.ppat.1009764

Figure Lengend Snippet: (A) Total RNAs were extracted from the fixed diffuse large B-cell lymphoma (DLBCL) tissue samples of HIV-negative (HIV-, n = 3) and HIV-positive (HIV+, n = 5) individuals. mRNA level of PLK1 in these tissue samples was measured by RT-qPCR and normalized to GAPDH. (B) A PE-conjugated CD19 antibody was used for B cell enrichment from PBMCs of HIV-infected, aviremic patients (HIV+, n = 5) or healthy donors (HIV-, n = 5). Purity of isolated CD19 + B cells was confirmed by flow cytometry analysis. (C) mRNA level of PLK1 in above isolated B cells (B) was measured by RT-qPCR and normalized to GAPDH. (D) Protein level of intracellular PLK1 in above isolated B cells (B) was measured by immunofluorescence. Percentage of PLK1-expressing cells was determined by flow cytometry analysis. (F) HIV-1 IIIB infected Jurkat cells were co-cultured with BJAB cells that were pre-stained with CFSE. Protein level of intracellular PLK1 in CFSE-labeled BJAB cells was measured by immunofluorescence. Percentage of PLK1-expressing cells was determined by flow cytometry analysis. (G-I) Protein level of PLK1 in BJAB (G), Akata-BX1 (H), or TREx BCBL1-Rta (I) cells treated with recombinant Nef (rNef) protein at the increasing dose or DMSO was measured by immunoblotting. GAPDH was used as loading control. Results were calculated from n = 3 independent experiments and presented as mean ± SD (* p<0.05; two-tailed paired Student t-test).

Article Snippet: Portion of isolated B cells was incubated with CD19 antibody (1/200 of stock, Milteny) for 30 mins to determine the purity using BD Accuri C6 Plus with corresponding optical filters.

Techniques: Quantitative RT-PCR, Infection, Isolation, Flow Cytometry, Immunofluorescence, Expressing, Cell Culture, Staining, Labeling, Recombinant, Western Blot, Control, Two Tailed Test