monoclonal antibody 215015  (R&D Systems)

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