anti mouse tim 3  (Bio X Cell)

Journal: Parasites & Vectors

Article Title: TGF-β regulated Tim-3 sustains macrophage phagocytic function and confers protection in Plasmodium yoelii NSM-infected mice

doi: 10.1186/s13071-026-07287-3

Figure Lengend Snippet: Tim-3 downregulation on splenic macrophages in P. yoelii NSM-infected mice. A C57BL/6 mice were infected with 1 × 10 6 P. yoelii NSM parasites by intraperitoneal injection, and 12 days after the infection, spleen lymphocytes were collected for single-cell sequencing. B UMAP visualization of immune cell subsets in the spleen. C Violin plot depicting immune-checkpoint expression dynamics in splenic macrophage populations. D Violin plot demonstrates Havcr2 expression in splenic macrophages at the single-cell level. E Tim-3 expression levels of splenic macrophages in naïve and infected groups. F mRNA expression level of Havcr2 in macrophages. G Tim-3 expression in RAW264.7 cells co-cultured with PBS, uRBCs, and iRBCs. H mRNA expression level of Havcr2 in RAW264.7. I Western blot analysis of Tim-3 protein expression. Data are mean ± SEM from three independent experiments ( n = 3–6 per group). ns not significant, P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001; D – F two-tailed unpaired t test; G , H one-way ANOVA

Article Snippet: For checkpoint blockade, mice received 200 μg of anti-mouse TIM-3 blocking antibody (BioXCell, Catalog No. BE0115) diluted in sterile PBS.

Techniques: Infection, Injection, Single Cell, Sequencing, Expressing, Cell Culture, Western Blot, Two Tailed Test

Journal: Parasites & Vectors

Article Title: TGF-β regulated Tim-3 sustains macrophage phagocytic function and confers protection in Plasmodium yoelii NSM-infected mice

doi: 10.1186/s13071-026-07287-3

Figure Lengend Snippet: Proinflammatory polarization of splenic Tim-3 + macrophages during P. yoelii NSM infection. A Heatmap of differentially expressed genes (DEGs) between naïve and infected Tim-3 + macrophages. B , C Enriched gene ontology (GO) terms and KEGG pathways in Tim-3 + macrophages post-infection. D CD86 expression on splenic Tim-3 + macrophages. E ROS accumulation in splenic Tim-3 + macrophages. F Flow cytometry gating strategy for cytokine (IFN-γ, IL-6, IL-10, TGF-β) expression in splenic macrophages. G – J Secretion levels of cytokine (IFN-γ, IL-6, IL-10, TGF-β) in splenic macrophages. Data are presented as mean ± SEM from three independent experiments ( n = 4–5 per group). ns not significant, P > 0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001; D , E , G – J two-tailed unpaired t test

Article Snippet: For checkpoint blockade, mice received 200 μg of anti-mouse TIM-3 blocking antibody (BioXCell, Catalog No. BE0115) diluted in sterile PBS.

Techniques: Infection, Expressing, Flow Cytometry, Two Tailed Test

Journal: Parasites & Vectors

Article Title: TGF-β regulated Tim-3 sustains macrophage phagocytic function and confers protection in Plasmodium yoelii NSM-infected mice

doi: 10.1186/s13071-026-07287-3

Figure Lengend Snippet: Inhibition of Tim-3 expression in splenic macrophages exacerbated P. yoelii NSM infection. A Mice received intraperitoneal injections of anti-Tim-3 blocking antibody (BE0115, 200 μg/mouse) on days 4 and 9 post-infection. B Spleen morphology of infected mice after injection. C The line graph illustrated the protozoan rate after Tim-3 blockade in C57BL/6 mice. D The line graph illustrated the rate of body weight change after Tim-3 blockade in C57BL/6 mice. E Histopathological analysis of spleen sections from infected mice. F Expression of Tim-3 on splenic macrophages in infected and anti-Tim-3 groups. G Expression of MHC-II on Tim-3 + macrophages. H Secretion levels of IFN-γ in CD4 + T cells. I Expression of CD69 in CD4 + T cells. Data are presented as mean ± SEM from three independent experiments ( n = 5–25 per group). ns not significant, P > 0.05, * P < 0.05, ** P < 0.01; B – D , F – J two-tailed unpaired t test

Article Snippet: For checkpoint blockade, mice received 200 μg of anti-mouse TIM-3 blocking antibody (BioXCell, Catalog No. BE0115) diluted in sterile PBS.

Techniques: Inhibition, Expressing, Infection, Blocking Assay, Injection, Two Tailed Test

Journal: Parasites & Vectors

Article Title: TGF-β regulated Tim-3 sustains macrophage phagocytic function and confers protection in Plasmodium yoelii NSM-infected mice

doi: 10.1186/s13071-026-07287-3

Figure Lengend Snippet: TGF-β modulates Tim-3 expression on splenic macrophages in mice of P. yoelii NSM infection. A The percentage of TGF-β in macrophages of naïve and infected groups. B Expression levels of TGF-β in macrophages. C AUCell score of the TGF-β pathway activity in macrophages. D Heatmap shows expression of TGF-β pathway genes in Tim-3 + and Tim-3 − macrophages. E Average expression (FPKM) of Havcr2 on Raw264.7 after TGF-β co-culture. F Expression levels of Tim-3 after co-culture with TGF-β and iRBCs. G Expression levels of Tim-3 after co-culture with galunisertib and iRBCs. H The expression of Tim-3 in the infected and the infection-injected galunisertib groups. Data are mean ± SEM from three independent experiments ( n = 4–5 per group). ns not significant, P > 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; B , C , H two-tailed unpaired t test; F , G one-way ANOVA

Article Snippet: For checkpoint blockade, mice received 200 μg of anti-mouse TIM-3 blocking antibody (BioXCell, Catalog No. BE0115) diluted in sterile PBS.

Techniques: Expressing, Infection, Activity Assay, Co-Culture Assay, Injection, Two Tailed Test

Journal: Parasites & Vectors

Article Title: TGF-β regulated Tim-3 sustains macrophage phagocytic function and confers protection in Plasmodium yoelii NSM-infected mice

doi: 10.1186/s13071-026-07287-3

Figure Lengend Snippet: Tim-3 + macrophages exhibit enhanced phagocytic activity during P. yoelii NSM infection. A The box plot of the AUCell score was used to illustrate the phagocytic function of Tim-3 + and Tim-3 − spleen macrophages. B The phagocytic function of Tim-3 + and Tim-3 − macrophages after co-culture with CFSE-labeled iRBCs. C The volcano plot shows the expression of genes related to phagocytosis in RAW264.7 cells after co-culture with TGF-β and iRBCs. D The enriched bubble chart presents the top ten differential enrichment pathways. E The proportion of phagocytosis of CFSE-iRBCs after adding galunisertib and TGF-β in the infected condition. F The level of apoptosis induced by TGF-β in the infected condition. Data are mean ± SEM from three independent experiments ( n = 4 per group). ns not significant, P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001; B , F Two-tailed unpaired t test; E one-way ANOVA

Article Snippet: For checkpoint blockade, mice received 200 μg of anti-mouse TIM-3 blocking antibody (BioXCell, Catalog No. BE0115) diluted in sterile PBS.

Techniques: Activity Assay, Infection, Co-Culture Assay, Labeling, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: IL-35 produced by dendritic cells via TIM-3-STAT3 signaling contributes to the development of visceral leishmaniasis

doi: 10.64898/2026.02.23.707416

Figure Lengend Snippet: ( A ) Top: putative STAT sites in the EBI3 and IL12A promoters. Bottom: ChIP-qPCR analysis of STAT3 recruitment to the indicated regions of EBI3 and IL12A promoters in BMDCs at 0.5 h after LDPm infection ( n = 6 replicates). Results are presented as fold enrichment relative to uninfected BMDCs. ( B ) Left: Details of EBI3 and IL12A promoter-specific oligonucleotides containing wild-type or mutated STAT sites (mutated bases in italics) used for the DNA pull-down assay. Right: DNA pull-down analysis using streptavidin (SA)-conjugated Dynabeads, followed by immunoblotting to assess the binding of STAT3 [present in the nuclear lysates of LDPm-infected (0.5 h) BMDCs] to the biotin (Btn)-labeled oligonucleotides shown in the left panel (representative of n = 3 experiments). ( C ) Immunoblot analysis confirming STAT3 silencing by siRNA; β-actin serves as a loading control (representative of n = 3 experiments). Ctrl siRNA, control siRNA. ( D ) IL-35 expression in uninfected and LDPm-infected BMDCs (48 h infection) transfected with the indicated siRNAs, analyzed by flow cytometry [representative data (left) and compiled data (right) from n = 3 experiments]. ( E ) Effect of TIM-3 blockade using an anti-TIM-3 antibody on IL-35 production by BMDCs infected with LDPm for 48 h, analyzed by flow cytometry [representative (left) and compiled (right) data from n = 3 experiments]. Uninfected BMDCs without any antibody treatment (no Ab) serve as controls. Each symbol represents data from one replicate (A) or one experiment (right panels of D and E). Horizontal bars (right panels of D and E) denote means; error bars (A) indicate SD. *** P < 0.001; ns, not significant.

Article Snippet: The TIM-3 blocking antibody (clone RMT3-23, BE0115) ( Chiba et al ., 2012 ) and the isotype control rat IgG2a,κ (BE0089) were obtained from Bio X Cell.

Techniques: ChIP-qPCR, Infection, Pull Down Assay, Western Blot, Binding Assay, Labeling, Control, Expressing, Transfection, Flow Cytometry

Journal: bioRxiv

Article Title: IL-35 produced by dendritic cells via TIM-3-STAT3 signaling contributes to the development of visceral leishmaniasis

doi: 10.64898/2026.02.23.707416

Figure Lengend Snippet: LD activates STAT3 in DCs via the TIM-3 receptor and its downstream signaling mediator Btk (as shown in our previous report ( Mishra et al ., 2023 )). Activated STAT3 then directly promotes IL-35 production in DCs by binding to the IL12A and EBI3 promoters ( IL12A and EBI3 encode the IL-35 subunits IL-12p35 and EBI3, respectively). DC-derived IL-35, in turn, inhibits the activation and maturation of bystander DCs by suppressing the NF-κB signaling pathway (inner green shaded box), promotes IL-35 expression in T cells, reduces T cell proliferation, and drives pathogenic type-2 T cell responses. Collectively, these events (summarized in the blue-outlined box) impair anti-leishmanial immunity and exacerbate disease pathogenesis. Notably, pharmacological blockade of STAT3 activation by WP1066 reduces IL-35 production by DCs, suppresses disease-promoting type-2 T cell responses, enhances host-protective type-1 T cell responses, and ultimately lowers parasite burden in vivo .

Article Snippet: The TIM-3 blocking antibody (clone RMT3-23, BE0115) ( Chiba et al ., 2012 ) and the isotype control rat IgG2a,κ (BE0089) were obtained from Bio X Cell.

Techniques: Binding Assay, Derivative Assay, Activation Assay, Expressing, In Vivo

Journal: The Journal of Biological Chemistry

Article Title: TIM-3 inhibition enhances breast tumor progression and metastasis: A paradoxical immune checkpoint response

doi: 10.1016/j.jbc.2025.111096

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of TIM-3 into 4T1 allograft breast tumors and detection of liver metastases following anti-TIM-3 monoclonal antibody treatment. A and B , TIM-3 immunohistochemical detection mostly observed into tumor infiltrating immune cells (indicated by red arrow ) in primary breast tumor tissue into both control tumor mice and anti-TIM3-treated mice, respectively, at × 400 magnification with a scale bar 20 μm. C , TIM-3 expression scoring into tumor (Control) and anti-TIM-3 treated group was determined semi-quantitatively by counting the percentage of TIM-3-positive cells and staining intensity of five × 400 magnification fields selected randomly and represented in the bar graph as mean ± SD, from three mice per group, ∗∗∗ p < 0.001 (Unpaired two-tailed t-test). D-G , primary breast tumor tissue section stained with H&E, ( D and E ) breast tumor tissue histology of control tumor mice and anti-TIM3 treated mice (at × 100 magnification respectively, with scale bar 100 μm), and ( F and G ) breast tumor tissue histology of control tumor mice and anti-TIM3 treated mice respectively (at × 400 magnification respectively, with scale bar 100 μm) resected after day 40. H – L , histological analysis of liver metastasis of tumor-bearing mice and treated group showed ( H ) tumor foci (indicated with a black circle ) of control tumor mice, and ( I ) anti-TIM3 mAb-treated tumor mice showed several tumor foci (indicated with several black circles ). J , the number of tumor metastatic foci into liver tissue section was determined by counting fifty high power field under × 400 magnification of each mouse of an experimental group (n = 3). Data represent mean ± SD. ∗ p < 0.05 (Unpaired two-tailed t -test). Infiltration of inflammatory cells into liver was observed and demarcated by ( K ) blue arrow indicated the moderate (not too severe) infiltration into control tumor-bearing mice, while ( L ) the green arrow indicated the severe inflammatory cell infiltration into the liver of anti- TIM-3 treated group. All data represent mean ± SD; n = 3.

Article Snippet: When tumor volume reached 200 to 250 mm 3 , mice were selected to receive the treatment of anti-TIM-3 monoclonal antibody (mAb) (Clone: RMT3-23, BioXcell, cat. # BE0115, RRID: AB_10949464 ).

Techniques: Immunohistochemical staining, Control, Expressing, Staining, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: TIM-3 inhibition enhances breast tumor progression and metastasis: A paradoxical immune checkpoint response

doi: 10.1016/j.jbc.2025.111096

Figure Lengend Snippet: Cytokine induction after Blockade of TIM3. Mouse cytokine array was performed to analyze the levels of cytokine production of IL-1β, IL-6, IL-10, IL-17A, IFN-γ and TNF-α, (n = 3/group) ( A – C ). A , serum samples were collected on day 40, at the end day of experiment, from normal healthy mice, control tumor and anti-TIM-3 treatment group of mice ( upper panel ). B , cytokine induction was determined from spleen tissue lysate ( middle panel ). Data represent mean ± SD, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, One-way ANOVA was performed. C , levels of cytokine induction was analyzed from tumor tissue lysate of tumor (Control) and anti-TIM3 treatment group ( lower panel ). Data represent mean ± SD, ∗ p < 0.05 (Unpaired t test).

Article Snippet: When tumor volume reached 200 to 250 mm 3 , mice were selected to receive the treatment of anti-TIM-3 monoclonal antibody (mAb) (Clone: RMT3-23, BioXcell, cat. # BE0115, RRID: AB_10949464 ).

Techniques: Control

Journal: The Journal of Biological Chemistry

Article Title: TIM-3 inhibition enhances breast tumor progression and metastasis: A paradoxical immune checkpoint response

doi: 10.1016/j.jbc.2025.111096

Figure Lengend Snippet: Anti-TIM3 treatment drives widespread changes in protein expression of tumor revealed by label-free quantification. Proteomics analysis of mice breast tumor tissue of both control tumor and anti-TIM-3-treated mice (n = 3/group) is shown. A , Volcano plot displayed the distribution of all differentially expressed significant up and down regulated proteins with fold change (±1) at log2 ratio were plotted against its significance level (negative log 10 p value) showing significantly ( p < 0.05) increased ( red colour ) and decreased ( blue colour ) proteins. B , heat map or cluster analysis of top 100 genes, where, top 90 up regulated and 10 down regulated genes were plotted. C , principal component analysis (PCA) of significantly expressed proteins in the treated and control tumor group. D , upregulation of Bat3 (Bag6) and Csk gene expression upon the blockade of TIM3 and inhibited the TIM-3 function. E and F , GSEA Plot evaluating major hallmark gene enrichment of PI3k-Akt and MTORC1 signaling pathway. G , relative increased expression of oncogene STAB1 and Bin1 (c-MYC) and ( H ) GSEA plot evaluating enrichment of hallmark genes related to MYC targeted pathways activation due to the impairment of FOXP3 in the anti-TIM3-treated group of mice.

Article Snippet: When tumor volume reached 200 to 250 mm 3 , mice were selected to receive the treatment of anti-TIM-3 monoclonal antibody (mAb) (Clone: RMT3-23, BioXcell, cat. # BE0115, RRID: AB_10949464 ).

Techniques: Expressing, Quantitative Proteomics, Control, Gene Expression, Activation Assay