tim-3 blocking antibody  (Galectin Therapeutics)

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: International Journal of Clinical and Experimental Pathology

Article Title: IFNγ and TNFα synergistically promote galectin 9 secretion by human osteosarcoma cells MG-63 to prevent T cell killing

doi:

Figure Lengend Snippet: Galectin 9/Tim-3 pathway influenced CD4 and CD8 T cell apoptosis in the coculture of MG-63 cells and T cells. 10 ug/ml anti-human Tim-3 was applied to block Galectin 9/Tim-3 pathway. 1 ug/ml Galectin 9 was applied to activate Galectin 9/Tim-3 pathway. Apoptosis of CD4+ and CD8+ T cells were analyzed by flow cytometry. Annexin V+ 7-AAD+ populations refer to late apoptotic cells. *: P < 0.05, **: P < 0.01, ***: P < 0.001.

Article Snippet: Tim-3/Galectin 9 pathway is not the only immune checkpoint in tumor immune microenvironment, thus we applied Tim-3 blocking antibody and Galectin 9 to investigate the role of Tim-3/Galectin 9 pathway in the interaction of IFNγ + TNFα stimulated MG-63 cells and T cells.

Techniques: Blocking Assay, Flow Cytometry

Journal: International Journal of Clinical and Experimental Pathology

Article Title: IFNγ and TNFα synergistically promote galectin 9 secretion by human osteosarcoma cells MG-63 to prevent T cell killing

doi:

Figure Lengend Snippet: Serum concentrations of Galectin 9 and the proportion of Tim-3+ peripheral T cells presented in osteosarcoma patients. A. Serum concentration of Galectin 9 was detected by ELISA assay. B. Percentage of Tim-3+ cells on CD4+ and CD8+ T cells in patients and controls. Data were calculated from 17 patients and 14 healthy controls. *: P < 0.05, **: P < 0.01.

Article Snippet: Tim-3/Galectin 9 pathway is not the only immune checkpoint in tumor immune microenvironment, thus we applied Tim-3 blocking antibody and Galectin 9 to investigate the role of Tim-3/Galectin 9 pathway in the interaction of IFNγ + TNFα stimulated MG-63 cells and T cells.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay