αtim-3 b8.2c12 antibody  (Bio X Cell)

Journal: eLife

Article Title: Engineering PEG10assembled endogenous virus-like particles with genetically encoded neoantigen peptides for cancer vaccination

doi: 10.7554/eLife.98579

Figure Lengend Snippet: ( A ) Schematic diagram of vaccination-associated therapy in orthotopic HCC model (n=6 mice). The mice were sacrificed and sampled for analysis on the day of 28 after initiating treatment. ( B ) Tumor burden monitoring of PBS, αTIM-3 alone, ePAC alone, and ePAC plus αTIM-3 treated mice by bioluminescence imaging. ( C ) The photographs and H&E staining of tumor-bearing livers collected from mice after different treatments as indicated. ‘T’ represents tumor tissues, ‘L’ represents liver tissues, and there is the clear boundary with non-tumor sites. ( D–I ) Flow cytometry analysis of effector memory T cells in splenic CD8 + T cells ( D and G ), CD8 + T cell infiltration in the tumors ( E and H ), and Ptpn2 376-384 : H-2K b specific CD8 + T cells in tumor infiltrating CD8 + T cells ( F and I ). Data are presented as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Figure 6—source data 1. Excel file with raw data used to generate .

Article Snippet: For the orthotopic HCC cancer model, the mice were randomly divided into four groups (n=6): PBS, αTIM-3 (100 μg/mouse, Leinco Technologies, USA), ePAC (200 μg/mouse), ePAC (200 μg/mouse)+αTIM-3 (100 μg/mouse) after 10 days of tumor inoculation (recorded as day 0) and then the treatment was initiated.

Techniques: Imaging, Staining, Flow Cytometry

Journal: bioRxiv

Article Title: PD-1 blockade enhances HIV-1 vaccine-induced CD8⁺ T-cell responses in PWH early ART-treated

doi: 10.1101/2024.07.05.601669

Figure Lengend Snippet: Representative flow cytometry plots gated on CD8 + T-cells for a Chro individual showing A. Cellular proliferation (CFSElow), B. IFNγ production, and C . HLA-DR⁺CD38⁺ expression. Pair-wise comparison on the percentage of CFSE low , IFNγ and HLA-DR⁺CD38⁺ expression in response to the corresponding HIV peptides (HIV-1-Gag peptide pool for Et and Chro groups and HIVConsv peptide pool for Etvac) in the presence of αPD-1, αTIM-3, αPD-1+αTIM-3 (Combo) antibodies and isotype control antibodies in D. Etvac, E. Et and F. Chro study groups. The dots represent a sample tested. The Wilcoxon matched-pairs signed ranked test was used to calculate statistical differences. P-values: ns > 0.05, *<0.05, **<0.005 and ***<0.0005.

Article Snippet: Finally, we included αTIM-3+ αPD-1 and used anti-human RSV-IgG4 as an isotype control antibody (5 μg/mL, 60AGK S228P, Merck & Co., Inc., Rahway, NJ, USA).

Techniques: Flow Cytometry, Expressing, Comparison, Control

Journal: bioRxiv

Article Title: PD-1 blockade enhances HIV-1 vaccine-induced CD8⁺ T-cell responses in PWH early ART-treated

doi: 10.1101/2024.07.05.601669

Figure Lengend Snippet: Correlograms indicate correlations between parameters across study groups: Etvac (n=11), Et (n=13) and Chro (n=9) for each experimental condition (HIV-1 for peptide stimulation, HIV+αTIM-3, HIV+αPD-1 and HIV+Combo). The top of the diagonal shows Spearman’s rank correlation coefficient with p-value<0.05. The colour and size of the circles indicate the strength and direction of the correlation. The bottom of the diagonal represents the linear regression. p-values: ns>0.05, *<0.05, **<0.005 and ***<0.0005.

Article Snippet: Finally, we included αTIM-3+ αPD-1 and used anti-human RSV-IgG4 as an isotype control antibody (5 μg/mL, 60AGK S228P, Merck & Co., Inc., Rahway, NJ, USA).

Techniques:

Journal: bioRxiv

Article Title: Engineering PEG10 assembled endogenous virus-like particles with genetically encoded neoantigen peptides for cancer vaccination

doi: 10.1101/2024.04.25.591213

Figure Lengend Snippet: ( A ) Schematic diagram of vaccination-associated therapy in subcutaneous HCC model (n=6 mice). C57BL/6 mice were inoculated with Hepa1-6 cells on day −10 and immunized with the indicated vaccine formulations on day 0, 4 and 8 for a total of 3 treatments. (B) Growth curves of the average tumor volumes in the indicated groups (n=6 mice; two-way ANOVA). ( C ) Representative images of tumors harvested from tumor-bearing mice on day 20. ( D ) and ( E ) The percentage and statistical analysis of effector memory T cells in splenic CD8 + T cells detected by flow cytometry (n=6 mice; one-way ANOVA). ( F ) and ( G ) Immunohistochemical staining of CD4 + and CD8 + cells (brown) in the tumors collected at the day 20 and the quantitation of CD4 + and CD8 + cells in each field (n=3, two-way ANOVA). Scale bar: 100 μm in the lower panels. ( H ) and ( I ) The percentage and statistical analysis of activated T cells expressing 4-1BB from tumors detected by flow cytometry (n=6 mice; one-way ANOVA). ( J ) and ( K ) Representative images and the expression analysis of CD8, PD-1, TIM-3, and CTLA-4 in tumors by immunofluorescence (n=3; two-way ANOVA). Scale bar: 50 μm. Data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance.

Article Snippet: For the orthotopic HCC cancer model, the mice were randomly divided into 4 groups (n=6): PBS, αTIM-3 (100 µg/mouse, Leinco Technologies, USA), ePAC (200 μg/mouse), ePAC (200 µg/mouse) + αTIM-3 (100 µg/mouse) after 10 days of tumor inoculation (recorded as day 0) and then the treatment was initiated.

Techniques: Flow Cytometry, Immunohistochemical staining, Staining, Quantitation Assay, Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: Engineering PEG10 assembled endogenous virus-like particles with genetically encoded neoantigen peptides for cancer vaccination

doi: 10.1101/2024.04.25.591213

Figure Lengend Snippet: ( A ) Schematic diagram of vaccination-associated therapy in orthotopic HCC model (n=6 mice). ( B ) Tumor burden monitoring of PBS, αTIM-3 alone, ePAC alone, and ePAC plus αTIM-3 treated mice by bioluminescence imaging. (C) The photographs and H&E staining of tumor-bearing livers collected from mice after different treatments as indicated. “T” represents tumor tissues, “L” represents liver tissues, and there is the clear boundary with non-tumor sites. ( D-I ) Flow cytometry analysis of effector memory T cells in splenic CD8 + T cells (D and G), CD8 + T cell infiltration in the tumors (E and H), and Ptpn2 376-384 : H-2K b specific CD8 + T cells in tumor infiltrating CD8 + T cells (F and I). n=6 mice; one-way ANOVA. Data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: For the orthotopic HCC cancer model, the mice were randomly divided into 4 groups (n=6): PBS, αTIM-3 (100 µg/mouse, Leinco Technologies, USA), ePAC (200 μg/mouse), ePAC (200 µg/mouse) + αTIM-3 (100 µg/mouse) after 10 days of tumor inoculation (recorded as day 0) and then the treatment was initiated.

Techniques: Imaging, Staining, Flow Cytometry

Journal: PLOS Pathogens

Article Title: Influenza virus infection enhances tumour-specific CD8 + T-cell immunity, facilitating tumour control

doi: 10.1371/journal.ppat.1011982

Figure Lengend Snippet: B16F1 (B16) tumour-bearing mice were infected with Influenza A/PR/8/34 (IAV) (150 PFU/mL) as described. 8 days post infection mice were sacrificed for flow cytometry analysis. ( a ) Frequencies of PD-1 expressing gp100-specific CD8 + T-cells in the tumour. ( b ) Representative dot plots of CD8 + T-cells expressing PD-1 and TIM-3. ( c ) Frequencies of PD-1 high TIM-3 + or PD-1 int TIM-3 - of gp100-specific CD8 + T-cells in the tumour. ( d ) Frequencies of Granzyme B (GzmB) expressing cells of the populations described in c. ( e ) Frequencies of Thymocyte selection-associated high mobility group box protein (TOX) expressing gp100-specific CD8 + T-cells in the tumour. Data from 2 experiments are shown. Error bars represent SEM. Statistical tests between two groups were performed as Student’s t-tests. * = p<0.05, ** = p<0.01, *** = p<0.001.

Article Snippet: The following monoclonal antibodies were used: αCD4 (clone RM4-5), αCD8 (clone 53–6.7), αThy1.1 (clone OX-7) were obtained from BD Biosciences Pharmingen. αPD-1 (clone 29F.1A12), αIFNγ (clone XMG1.2) and αCD43, directed against its activation-associated glycosylated isoform (clone 1B11) were purchased from BioLegend. αGzmB (clone GB12) antibody was purchased from Invitrogen. αKi67 (clone SolA15), αCXCR3 (clone CXCR3-173) and αFoxP3 (clone FJK-16s) were purchased from eBioscience. αTIM-3 (clone 215008) was purchased from R&D systems.

Techniques: Infection, Flow Cytometry, Expressing, Selection

Journal: Journal for Immunotherapy of Cancer

Article Title: Combining toll-like receptor agonists with immune checkpoint blockade affects antitumor vaccine efficacy

doi: 10.1136/jitc-2024-008799

Figure Lengend Snippet: Combination of TLR agonists and αLAG-3 with peptide-activated CD8+T cells elicited prolonged suppression of E.G7-OVA-PDL1 high tumor growth. Ovalbumin-expressing E.G7 cells (eg,7-OVA-PD-L1 high ) were implanted in C57BL/6 mice and permitted to grow until tumors were palpable (7 days). As in , OT-I splenocytes were then adoptively transferred and mice were immunized subcutaneously the following day with SIINFEKL (OVA) peptide, with or without TLR agonists. Mice were then treated with immune checkpoint blockade, using αTIM-3, αTIGIT, αVISTA, αLAG-3, αCTLA-4, or IgG control, the day following immunization. (A) Shown are the tumor growth curves (mean+SE, n=7 animals per group). (B) Survival plots using the time to death or when tumors reached 2 cm 3 in size, whichever occurred first. (C) In a parallel study, animals were treated as in A but tumors were collected at day 16 and evaluated for the frequency of infiltrating CD3+CD8+ tetramer+ T cells per gram of tumor and the number of Tregs (CD3+CD4+ CD25+ FoxP3+). (D) Tumor-infiltrating CD3+CD8+ tetramer+ T cells were further evaluated for 4-1BB expression by flow cytometry. Asterisks indicate p<0.05 assessed by the mixed-effects model with Geisser-Greenhouse correction and Tukey’s multiple comparisons test with individual variances (A), by log-rank test (B), or by one-way ANOVA with Tukey’s multiple comparisons test (C, D). Error bars represent SEM. Results are from one experiment and are representative of data from three independent experiments (shown in ). ANOVA, analysis of variance; MFI, median fluorescence intensity; TLR, toll-like receptor.

Article Snippet: An αPD-1 hybridoma (clone G4, a gift by Dr. Lieping Chen) was produced by Envigo (Madison, WI). αCTLA-4 (BE0164), αLAG-3 (BE0174), αTIGIT (BE0274), αTIM-3 (BE0115), αVISTA (BE0310), and Armenian Hamster IgG (BE0091) were purchased from BioXCell.

Techniques: Expressing, Flow Cytometry, Fluorescence