Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell proliferation through the IL-15 signaling pathway. ( A ) Flow cytometric analysis of CD8⁺ T cell infiltration in clinical OSCC tissues ( n = 6 per group). ( B ) Expression distribution of ALDH1L1 across cell subtypes in OSCC from the TISCH2 database ( GSE172577 dataset). ( C ) Immunohistochemical staining of ALDH1L1 and CD8 in OSCC sections (ALDH1L1 low , n = 23; ALDH1L1 high , n = 29). ( D ) Immunohistochemical staining of ALDH1L1 and CD8 in xenograft tumor sections ( n = 6 per group). ( E ) Quantitative analysis of CD8⁺ T cell proportions in xenograft tissues by flow cytometry ( n = 6). ( F - G ) Correlation analysis between ALDH1L1 and chemokine mRNA expression levels in clinical OSCC samples (qRT-PCR, n = 12). H. qRT-PCR analysis of IL-15 mRNA expression in NC and ALDH1L1-KD groups of CAL 27 and SCC-25 cells ( n = 3). I. Extracellular IL-15 concentration in OSCC cell cultures with different ALDH1L1 expression levels ( n = 3). ( J ) Flow cytometric analysis of IL-15-mediated CD8⁺ T-cell proliferation in vitro, ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry, Quantitative RT-PCR, Concentration Assay, In Vitro, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell function through metabolic reprogramming. ( A ) Quantification of PD-1⁺ CD8⁺ T cells in clinical OSCC specimens by flow cytometry ( n = 6 per group). ( B ) Quantification of PD-1⁺ CD8⁺ T cells in mouse xenografts by flow cytometry ( n = 6 per group). ( C ) Top 20 enriched KEGG pathways identified from transcriptomic analysis. ( D ) Chord diagram illustrating 29 key metabolites identified by metabolomic profiling. ( E - H ) Flow cytometric evaluation of CD8⁺ T cell marker expression after stimulation with metabolites derived from ALDH1L1-KD or control CAL 27 cells and SCC-25 cells ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: Cell Function Assay, Flow Cytometry, Metabolomic, Marker, Expressing, Derivative Assay, Control, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: L-glutamate accumulation mediates CD8⁺ T cell dysfunction in ALDH1L1-downregulated microenvironment. ( A - C ) Integrated multi-omics analysis reveals ALDH1L1-regulated metabolic signatures: Red boxes highlight co-enriched metabolic pathways from transcriptomic ( A ) and metabolomic ( B ) analysis, and key metabolites ( C ). ( D ) Extracellular L-glutamate concentration in conditioned media from CAL 27 and SCC-25 cell culture supernatants ( n = 3). ( E - F ) Flow cytometric analysis of L-glutamate effects on CD8⁺ T cell functional markers: ( E ) Proportion of IFN-γ⁺ CD8⁺ T cells; ( F ) Proportion of PD-1⁺ CD8⁺ T cells ( n = 3). ( G ) Cytotoxic activity of CD8⁺ T cells pretreated with conditioned medium from ALDH1L1-KD or NC OSCC cells medium with exogenous L-glutamate ( n = 3). (NC, negative control; KD, knockdown; Ctrl, control; Glu, L-glutamate; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: Biomarker Discovery, Metabolomic, Concentration Assay, Cell Culture, Functional Assay, Activity Assay, Negative Control, Knockdown, Control

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 transcriptionally regulates GLUL and interacts with GLUL to sustain L-glutamate metabolism. ( A ) Integrated multi-omics analysis of ALDH1L1-regulated metabolic network. Green circles: downregulated metabolites; Green squares: downregulated metabolic enzymes; Red squares: upregulated metabolic enzymes. ( B - C ) GLUL mRNA ( B ) and protein ( C ) expression in CAL 27 and SCC-25 cells assessed by qRT-PCR and western blot, respectively. ( D ) Protein-protein interaction (PPI) network between ALDH1L1 and glutamate-metabolizing enzymes predicted by the STRING database. ( E ) Three-dimensional structural model of ALDH1L1 (green)-GLUL (blue) interaction. Key residues are shown in stick representation; hydrogen bonds indicated by yellow dashed lines. ( F ) Co-immunoprecipitation analysis of endogenous interactions among ALDH1L1 and GLUL in CAL 27 cells. ( G ) ALDH1L1 enhances GLUL enzymatic activity in a dose-dependent manner in vitro ( n = 3). ( H ) Western blot validation of GLUL overexpression in plasmid-transfected CAL 27 cells. ( I ) Extracellular L-glutamate concentration following GLUL upregulation in ALDH1L1-KD CAL 27 cells ( n = 3). (NC: negative control; KD: ALDH1L1 knockdown; OE: GLUL overexpression; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Immunoprecipitation, Activity Assay, In Vitro, Over Expression, Plasmid Preparation, Transfection, Concentration Assay, Negative Control, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Stevioside enhances anti-PD-1 immunotherapy by targeting ALDH1L1. ( A ) Virtual screening workflow for identifying potential ALDH1L1-targeting compounds. ( B ) Two-dimensional chemical structures of the top 12 candidate compounds ranked by binding affinity to ALDH1L1 protein. ( C ) Western blot analysis demonstrating the effect of stevioside treatment on ALDH1L1 protein expression levels in CAL 27 cells (Ctrl, control). ( D ) Dose-response curve of stevioside (IC₅₀ = 195.3 µM). ( E ) Molecular docking analysis reveals the binding site of stevioside on ALDH1L1 protein. (Blue ribbon structure represents ALDH1L1 protein; green stick model represents stevioside molecule; stick structures indicate key interacting amino acid residues; yellow dashed lines denote hydrogen bond interactions.) ( F ) In vivo experiments confirm that stevioside combined with anti-PD-1 antibody treatment significantly enhances antitumor efficacy ( n = 5 per group). ( G - H ) Immunohistochemical staining of Ki-67 ( G ) and CD8 ( H ) in xenograft tumor sections ( n = 5 per group). (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: Binding Assay, Western Blot, Expressing, Control, In Vivo, Immunohistochemical staining, Staining

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Schematic model illustrating the mechanism by which ALDH1L1 reverses CD8⁺ T cell exhaustion in OSCC

Article Snippet: Recombinant Human ALDH1L1 Protein (Cat# bs-100230P, Biosschina) and Recombinant Human GLUL Protein (HY- P70294 , MedChemExpress) were obtained commercially.

Techniques: