anti lag3 antibody  (Bio X Cell)

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: Single-cell atlas reveals cellular heterogeneity and LAG-3 expression dynamics in cervical cancer. (A) UMAP projection of single-cell transcriptomes after Harmony-based batch correction and PCA dimensionality reduction identified 33 transcriptionally distinct clusters, reflecting pronounced intratumoral heterogeneity. (B) Cell type annotation using canonical lineage markers categorized these clusters into 10 major cellular compartments within the cervical tumor microenvironment. (C) Distribution of cells by sample origin showed distinct spatial segregation across normal cervix (N), HSIL (H), primary tumor (T), and metastatic lymph node (L) in the integrated UMAP landscape. (D) Within the immune compartment, NK/T cells were delineated and T cell subsets were further resolved, enabling refined dissection of T cell heterogeneity. (E) Isolation of CD8 + T cells provided the basis for focused analyses of cytotoxic lymphocyte states across disease stages. (F) Cross-sample comparison revealed that LAG-3 expression was markedly elevated in CD8 + T cells from primary tumors (T) and metastatic lymph nodes (L) relative to normal cervical tissues (N). (G) LAG-3 exhibited a subset- and context-specific expression pattern, being preferentially enriched in exhausted T cells (Tex) and displaying distinct distribution across clinical sample types.

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Single Cell, Expressing, Dissection, Isolation, Comparison

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: Functional landscape of CD8 + T cells and correlation with LAG-3 expression. (A) Distribution of CD8 + T-cell functional subsets across different pathological samples. The relative proportions of naïve, central memory, effector, stress-response, and exhausted CD8 + T-cell subsets are shown for normal cervix (N), high-grade squamous intraepithelial lesions (HSIL, H), and metastatic lymph nodes (L). (B) Correlation between LAG-3 expression and overall CD8 + T-cell functional scores. Pearson correlation analysis showing the relationship between LAG-3 transcript levels and scores for exhaustion, cytotoxicity, and effector function across the total CD8 + T-cell population. (C) Subset-specific correlation of LAG-3 expression with exhaustion scores. LAG-3 expression exhibits a strong positive correlation with exhaustion in CD8 + exhausted (Tex), effector, and stress-response T-cell subsets. (D) UMAP visualization of LAG-3 expression in CD8 + T cells. Each point represents a single cell, with color intensity reflecting relative LAG-3 transcript abundance. (E) UMAP projection of exhaustion scores within the CD8 + T-cell compartment. Color intensity indicates the functional exhaustion score for each cell, highlighting spatial enrichment of exhausted phenotypes. (F) UMAP visualization of cytotoxicity scores in CD8 + T cells. Cells are colored according to their cytotoxicity module score, revealing spatial patterns of effector potential. (G) UMAP projection of stemness scores in CD8 + T cells. Relative stemness scores are mapped onto the CD8 + T-cell population, showing enrichment of stem-like features in naïve subsets and their spatial separation from LAG-3-high exhausted cells.

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Functional Assay, Expressing, Single Cell

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: LAG-3 is upregulated in cervical cancer tissues and predominantly colocalizes with CD8 + T cells (A) Representative immunohistochemical staining of LAG-3 in cervical cancer tissues, high-grade squamous intraepithelial lesions (HSIL), and normal cervical tissues (upper panels, 100×; lower panels, 400×). (B) Quantitative analysis of LAG-3 expression across the three tissue groups, based on the percentage of positively stained area (n = 60 per group). (C) Western blot analysis of LAG-3 protein levels in cervical cancer versus normal cervical tissues, Semiquantitative assessment of LAG-3 protein expression based on optical density (OD) measurements. (D) Multiplex immunofluorescence staining (cervical cancer tissue sections) was performed to analyze the mean fluorescence intensity of CD4+ T cells and CD8+ T cells in the same field of view. (n=9) (E) Representative images of immunofluorescence co-staining of tissue sections from cervical cancer patients (n=9), showing 4’,6-diamidino-2-phenylindole (blue), CD8 or CD4 (red), LAG-3 (green), and merged images. The boxed area is the histological image at 200× magnification, and the arrows indicate CD8+LAG-3+ T cells or CD4+LAG-3+ T cells. All data are presented as mean ± SEM. *P < 0.05; ****P < 0.0001; n.s., not significant (P > 0.05). DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Immunohistochemical staining, Staining, Expressing, Western Blot, Multiplex Assay, Immunofluorescence, Fluorescence

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: Statistical analyses of LAG-3 expression across key clinicopathological features of cervical cancer. (A) Comparison of LAG-3-positive expression among patient groups stratified by FIGO stage. (B) Comparison of LAG-3 expression across groups stratified by histological differentiation. (C) Comparison of LAG-3 expression between patients with and without lymph node metastasis. (D) Comparison of LAG-3 expression between patients with and without lymphovascular or perineural invasion. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant.

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: Functional changes in CD8 + T cells following co-culture of lag3 + and lag-3 - CD8 + T cells with U14 cervical cancer cells. (A) Representative flow cytometry plots and quantification of CD69 expression on CD8 + T cells following co-culture.TEM. (B) Representative plots and statistical analysis of Ki67 and T-bet expression in CD8 + T cells. (C) Representative flow cytometry plots and quantification of naive (TN), central memory (TCM), and effector memory CD8 + T-cell subsets. (D) Representative plots and quantification of TNF-α, IFN-γ, IL-2, and IL-10 secretion by CD44 + CD8 + T cells. Data are presented as mean ± SEM. WT, lag-3 + CD8 + T cells; KO, lag-3 - CD8 + T cells; Statistical significance: ***P < 0.001, **** P < 0.0001; n.s., not significant (P > 0.05); n = 5 per group.

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Functional Assay, Co-Culture Assay, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: LAG-3–associated CD8 + T-cell dysfunction in the cervical cancer tumor microenvironment

doi: 10.3389/fimmu.2026.1750726

Figure Lengend Snippet: Functional analysis of CD8 + T cells in subcutaneous tumors of C57BL/6 wild-type (WT) and lag-3 knockout (KO) mice. (A) Representative photographs of subcutaneous tumors from WT and KO mice. (B) Tumor growth curves and body weight changes over 15 days for both groups. (C) Representative flow cytometry plots and quantitative analysis of CD8 + and CD4 + T cell proportions within the tumor-infiltrating lymphocyte populations. (D) Flow cytometric analysis of CD69 expression in CD8 + T cells within the tumor microenvironment, including representative plots and quantification. (E) Flow cytometric analysis of Ki67 and T-bet expression in CD8 + T cells, showing representative plots and statistical analysis. (F) Representative plots and quantification of effector memory (TEM) and central memory (TCM) CD8 + T cell subsets in the tumor microenvironment. (G) Representative flow cytometry plots and corresponding quantitative analysis of IFN-γ, IL-2, TNF-α, and IL-10 production by CD44 + CD8 + T cells within the subcutaneous tumor microenvironment of the two mouse cohorts. Each group included five mice aged 8~10 weeks. Data are presented as mean ± standard error of the mean (SEM). WT, C57BL/6 wild-type mice; KO, lag-3-deficient mice. Statistical significance is indicated as: **P < 0.01, **** P < 0.0001; n.s., not significant (P > 0.05).

Article Snippet: For separation of lag-3 + and lag-3 - CD8 + T cells, the enriched CD8 + T-cell fraction was subsequently incubated with a biotin-conjugated anti-mouse lag-3 antibody (clone C9B7W, Miltenyi Biotec) for 15 min at 4°C, followed by labeling with anti-biotin microbeads for an additional 15 min.

Techniques: Functional Assay, Knock-Out, Flow Cytometry, Expressing

Journal: bioRxiv

Article Title: Immune cells adapt to distinct stem cell niches to govern tissue homeostasis

doi: 10.64898/2026.01.28.701831

Figure Lengend Snippet: a , UMAP plots displaying distribution of Ltb, Areg and Lag3 transcripts across different cell types in mouse skin. Each dot is one cell, color-coded based on transcript expression levels. b, Schematic of the Ltb-CreER x R26-tdT mouse line. c, Left: Representative whole mount immunofluorescence showing that double positive (tdT + and TCRγδ + ) LTB-expressing DETCs reside primarily within the uHF compartment as do other LTB-expressing lymphocytes (tdT + and TCRγδ neg ). Epithelial SCs are marked by K14. Scale bar 30μm. Right: quantifications of (left) the probability of finding LTB + DETCs (tdT + and TCRγδ + ) in uHF vs IFE and (right) probability of finding any LTB + cell (tdT + ) in the uHF region vs IFE. Each circle represents data from one mouse (3-8 fields of view). d-e , Representative FACS plots and quantifications showing that most DETCs expressing AREG (d) and LAG-3 (e) are uLIPSTIC neg (IFE-DETC) and not uLIPSTIC + (uHF-DETC). MFI, mean fluorescence intensity. f, Percentage of LTB-tdT + , AREG + or LAG-3 + DETC out of total DETC as quantified by FACS of the skins of mice at the indicated postnatal time points. Each point represents the mean with SEM of 2-7 mice. g, Percentage of AREG + DETC in the embryonic thymus of E17-18 pups and from the back skin of adult second telogen mice as quantified by FACS. h, Quantifications of total skin intraepithelial Ltb transcripts determined by qPCR, and AREG + or LAG-3 + DETC determined by FACS when mice are kept in specific pathogen free (SPF) or germ-free (GF) facility. i, Schematic summarizing the compartmentalization of the two DETC niche programs in steady state. Data are representative of two-four (panel h), two (panel e), three (panel c, d, g) or three-seven (panel f) independent experiments and, unless indicated, each circle represents one mouse. Data in c, d, g and h are analysed by unpaired two-tailed Student’s t- test. p- values are indicated in each figure and data are represented as mean with SEM. Further details on statistics and reproducibility in Methods. See for additional supporting experiments.

Article Snippet: monoclonal antibodies against mouse LAG-3 (clone C9B7W, BioXcell) were injected intraperitoneally every other day (for a total of three injections) in wild-type C57BL/6 animals.

Techniques: Expressing, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Immune cells adapt to distinct stem cell niches to govern tissue homeostasis

doi: 10.64898/2026.01.28.701831

Figure Lengend Snippet: a , Percentage of LAG-3 + cells gated on DETC, ILC or TCRαβ + T cells in wild type back skin as determined by FACS. b, Left: strategy to block LAG-3 in vivo . Right: quantifications reveal that DETCs but not TCRαβ + T cells increase in numbers in the skin when anti-LAG-3 antibodies are delivered intraperitoneally to mice. c, Left: representative max projections of whole mount immunofluorescence of IFE and uHF compartments showing that the increase in DETC (in yellow) resulting from anti-LAG-3 blocking antibodies occurs largely in the IFE-SC and not the uHF-SC niche. Scale bar 50μm. * denote autofluorescence. d, Normalized concentration of AREG determined by ELISA shows elevated AREG in the intraepithelial tissue, consistent with the elevation in DETCs caused by LAG-3 inhibition. e, Normalized percentage of EdU + SCs determined by FACS after a 3-hour pulse in mice injected with either PBS or anti-LAG-3 blocking antibodies. f, Top: strategy to deliver recombinant AREG (rAREG) systemically in vivo by osmotic pump implantation; bottom: concentration of AREG determined by ELISA in the intraepithelial tissue of mice implanted with pumps containing either rAREG or PBS as a control. g, Percentage of EdU + SCs determined by FACS after a 4-hour pulse in mice implanted with pumps containing either rAREG or PBS as a control. Data are representative of two (panel f), three (panel c, d, g), five (panel a, e) or seven (panel b) independent experiments and each circle represents one mouse. Data in b, c, d, e, f and g are analyzed by unpaired two-tailed Student’s t- test. Data in panel a are analyzed by ordinary one-way ANOVA with Tukey’s multiple comparison post-test. p- values are indicated in each figure and data are represented as mean with SEM. Further details on statistics and reproducibility in Methods. See and for additional supporting experiments.

Article Snippet: monoclonal antibodies against mouse LAG-3 (clone C9B7W, BioXcell) were injected intraperitoneally every other day (for a total of three injections) in wild-type C57BL/6 animals.

Techniques: Blocking Assay, In Vivo, Immunofluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Injection, Recombinant, Control, Two Tailed Test, Comparison

Journal: bioRxiv

Article Title: Immune cells adapt to distinct stem cell niches to govern tissue homeostasis

doi: 10.64898/2026.01.28.701831

Figure Lengend Snippet: a , UMAP plots showing expression pattern of H2-ab1 (MHC-II) and Lgals3 (Galectin-3) across different cell type from mouse back skin. b , Quantification of the percentage of MHC-II + and mean fluorescence intensity (MFI) of Galectin-3 in Bulge-SC, uHF-SC and IFE-SC from wild type mice determined by FACS. Each dot represents one mouse. c , Percentage of LAG-3 + cells gated on CD8 + CD44 + splenocytes from wild type mice injected with PBS or with anti-LAG3 blocking antibodies. Each dot represents one mouse from two independent experiments. d , qPCR analysis of mKi67 expression in the intraepithelial compartment of wild type mice injected with PBS or with anti-LAG3 blocking antibodies. Each dot represents one mouse from three independent experiments. Data in c and d are analysed by unpaired two-tailed Student’s t- test. Data in b are analysed by ordinary one-way ANOVA with Tukey’s multiple comparison post-test. p -values are indicated in each figure and data are represented as mean with SEM.

Article Snippet: monoclonal antibodies against mouse LAG-3 (clone C9B7W, BioXcell) were injected intraperitoneally every other day (for a total of three injections) in wild-type C57BL/6 animals.

Techniques: Expressing, Fluorescence, Injection, Blocking Assay, Two Tailed Test, Comparison

Journal: bioRxiv

Article Title: Immune cells adapt to distinct stem cell niches to govern tissue homeostasis

doi: 10.64898/2026.01.28.701831

Figure Lengend Snippet: Representative whole mount max projections of the IFE compartment confirms that immigrant T cells (CD3+) occupy the IFE in FVB-TAC (mutant for DETC-cognate ligand, Skint1 ). FVB-JAX mice are shown as WT controls. Scale bar 100μm. b, Bar plot showing relative proportions of Vg5+, Vg5neg, TCRab+ T cells and ILCs in the intraepithelial compartment of FVB JAX or TAC mice. c, Left: percentage of AREG+ Vg5neg TCRgd T cells detected by FACS in the intraepithelial compartment of FVB JAX or TAC mice. Middle: ELISA showing comparable AREG levels in the intraepithelial fractions from FVB JAX and FVB TAC mice. Right: Representative FACS plots and quantifications of percentage of LAG-3+ Vg5neg TCRgd T cells from the intraepithelial fraction of FVB JAX or TAC mice. d, Representative whole mount max projections of the IFE compartment from wild type and Rag2-/-mice showing intraepithelial CD45+ immune cells (red) and CD3+ T cells (green). Scale bar 100μm. * denote autofluorescence. e, Quantifications reveal that ILCs (CD45+CD90+ TCRgdneg TCRbneg) compensate for the loss of DETCs (CD45+CD90+ TCRgdhigh) when all T cells are ablated. f, Left: FACS analyses of the intraepithelial compartment reveal that ILCs become AREG+ when they occupy the IFE-SC niche vacated by DETC loss. Middle: ELISA reveals that the level of AREG in the intraepithelial fraction is largely independent of the immune resident in the IFE niche. Right: Representative FACS plots and quantifications of percentage of LAG-3+ ILCs from the intraepithelial fraction of wild type (WT) and Rag2-/- mice. g, Schematic showing immune adaptation within distinct SC niches and homeostatic mechanisms to keep niche size and activity in check. Data are representative of two (panel b, c, e) or three (f) independent experiments, and each circle represents one mouse. Data in c and f are analysed by unpaired two-tailed Student’s t- test. p- values are indicated in each figure and data are represented as mean with SEM. Further details on statistics and reproducibility in Methods. See for additional supporting experiments.

Article Snippet: monoclonal antibodies against mouse LAG-3 (clone C9B7W, BioXcell) were injected intraperitoneally every other day (for a total of three injections) in wild-type C57BL/6 animals.

Techniques: Mutagenesis, Enzyme-linked Immunosorbent Assay, Activity Assay, Two Tailed Test