Journal: PLoS ONE

Article Title: Characterizing responsive and refractory orthotopic mouse models of hepatocellular carcinoma in cancer immunotherapy

doi: 10.1371/journal.pone.0219517

Figure Lengend Snippet: Hep Cell tumors (day 21), Hep Frag tumors (day 28) and iAST tumors (day 56) were stained for: A, morphology (H&E); B, tumor stroma (FAP); C, proliferation (Ki67); D, immune cells (CD45); E, macrophages (F4/80); F, Kupffer cells (Clec4f); G, CD4 + T cells (CD4); H, CD8 + T cells (CD8) and I, tumor vasculature (CD31). E,F, Dashed lines in the images of F4/80 and Clec4F stainings indicate the border between vital liver (VL) and tumor (T). I, CD31 signal quantification (n = 5) indicates significantly higher tumor vessel density in iAST compared to Hep tumors. In the scatter plot, each dot represents the average of 5 images per tumor. Differences between groups were tested for significance using the one-way ANOVA followed by Tukey multiple comparison test (***p<0.001).

Article Snippet: The following anti-murine primary antibodies were used: Ki67 (rabbit monoclonal, clone 30–9, Ventana Medical Systems), F4/80 (rat monoclonal, clone BM8, Acris Antibodies GmbH), Clec4f (goat polyclonal, R&D Systems), CD4 (rabbit monoclonal, clone #1, Sino Biological Inc), CD8α (rat monoclonal, clone GHH8, Dianova), PD-L1 (rabbit monoclonal, clone E1L3N, Cell Signaling).

Techniques: Staining

Journal: PLoS ONE

Article Title: Characterizing responsive and refractory orthotopic mouse models of hepatocellular carcinoma in cancer immunotherapy

doi: 10.1371/journal.pone.0219517

Figure Lengend Snippet: A, Percentages of total immune cell infiltrate (CD45 + ) and subpopulations (CD4 + , CD8 + , NK cells, B cells, macrophages, MDSCs, DCs, other) in Hep Cell tumors (day 21), Hep Frag tumors (day 25) and iAST tumors (day 63). B, Exome sequencing of excised HCC tumors revealed considerably fewer lower number of missense genes in iAST (day 63) than in Hep Cell (day 21) and Hep Frag tumors (day 25). C, Venn diagrams show the mutated genes in each tumor with corresponding vital liver control (Liver Co). D, Venn diagram compares the mutated genes in the three tumors (Hep Cell , Hep Frag , iAST). Missense genes that were mutated in vital liver controls were subtracted from each tumor sample.

Article Snippet: The following anti-murine primary antibodies were used: Ki67 (rabbit monoclonal, clone 30–9, Ventana Medical Systems), F4/80 (rat monoclonal, clone BM8, Acris Antibodies GmbH), Clec4f (goat polyclonal, R&D Systems), CD4 (rabbit monoclonal, clone #1, Sino Biological Inc), CD8α (rat monoclonal, clone GHH8, Dianova), PD-L1 (rabbit monoclonal, clone E1L3N, Cell Signaling).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Characterizing responsive and refractory orthotopic mouse models of hepatocellular carcinoma in cancer immunotherapy

doi: 10.1371/journal.pone.0219517

Figure Lengend Snippet: A, Treatment schedule: anti-PD-1 (10 mg/kg, i.p.) and anti-CTLA-4 (5 mg/kg, i.p.) therapy started at day 25 after fragment implantation and was applied in 3 doses at days 0, 3 and 7. Necropsy of control (Co), anti-PD-1, anti-CTLA-4, and the anti-PD-1/anti-CTLA-4 combination groups (n = 5) was performed at day 10 after treatment initiation to assess tumor weight and immune cell infiltrates. B, Mice treated with combination of anti-PD-1 and anti-CTLA-4 show reduced tumor load compared to mice from the control and monotherapy groups. C, Representative images of explanted livers depicting the orthotopic tumor load of all groups. D, The total immune infiltrate in the tumors increased upon treatment with anti-CTLA-4 and the combination treatment with anti-PD-1. E, Similar increases were found for the total number of CD4 + T cells, CD8 + T cells and DCs upon combined treatment of anti-PD-1 and anti-CTLA-4. F, DC1 cells decrease upon treatment with anti-PD-1, anti-CTLA-4 and combination thereof, whereas an increase of DC2 cells was observed. G, The percentage of TRegs within CD4 + T cells decreases upon treatment with anti-CTLA-4 alone and in combination with anti-PD-1, whereas the content of CD4 + effector T cells and proliferating (Ki67 + ) CD4 + T cells did not significantly change upon treatment. H, Flow cytometry shows a significant increase in effector and proliferating (Ki67 + ) CD8 + T cells after anti-PD-1, anti-CTLA-4 and combined treatment. D-H, Differences among groups were tested for significance using the one-way ANOVA followed by Tukey multiple comparison test (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The following anti-murine primary antibodies were used: Ki67 (rabbit monoclonal, clone 30–9, Ventana Medical Systems), F4/80 (rat monoclonal, clone BM8, Acris Antibodies GmbH), Clec4f (goat polyclonal, R&D Systems), CD4 (rabbit monoclonal, clone #1, Sino Biological Inc), CD8α (rat monoclonal, clone GHH8, Dianova), PD-L1 (rabbit monoclonal, clone E1L3N, Cell Signaling).

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Characterizing responsive and refractory orthotopic mouse models of hepatocellular carcinoma in cancer immunotherapy

doi: 10.1371/journal.pone.0219517

Figure Lengend Snippet: A, Treatment schedule: anti-PD-1 (10 mg/kg, i.p.) and anti-CTLA-4 (5 mg/kg, i.p.) therapy started at day 53 after virus injection and was applied in three doses at days 0, 3, and 7. Necropsy of vehicle (Co), anti-PD-1, anti-CTLA-4 and combination group (n = 5) was performed at day 10 after treatment initiation. B, Treatment of mice with vehicle, anti-PD-1, anti-CTLA-4 or combination thereof did not alter tumor load. C, Representative images of explanted livers illustrate multinodular tumors after treatment with vehicle, anti-PD-1, anti-CTLA-4 or combination thereof. D, Analysis of the total immune infiltrate and subsets in the tumor nodules did not show any significant differences among the groups. E, The total number of CD4 + T cells, CD8 + T cells and DCs did not change after treatment with anti-PD-1, anti-CTLA-4 or combination thereof (n = 5). F, Percentage of TRegs within the CD4 + T cell population did not change upon treatment. G, The distribution of DC subsets was not affected by treatment. D-G, Differences among groups were tested for significance using the one-way ANOVA followed by Tukey multiple comparison test.

Article Snippet: The following anti-murine primary antibodies were used: Ki67 (rabbit monoclonal, clone 30–9, Ventana Medical Systems), F4/80 (rat monoclonal, clone BM8, Acris Antibodies GmbH), Clec4f (goat polyclonal, R&D Systems), CD4 (rabbit monoclonal, clone #1, Sino Biological Inc), CD8α (rat monoclonal, clone GHH8, Dianova), PD-L1 (rabbit monoclonal, clone E1L3N, Cell Signaling).

Techniques: Injection

Journal: Nature Communications

Article Title: Loss of HIV candidate vaccine efficacy in male macaques by mucosal nanoparticle immunization rescued by V2-specific response

doi: 10.1038/s41467-024-53359-2

Figure Lengend Snippet: a , b Comparison of mucosal B cells in rectal mucosa producing a IgG (V2-TTB NP, n = 12; TTB NP, n = 9; empty NP, n = 9) and b IgA (V2-TTB NP, n = 12; TTB NP, n = 9; empty NP, n = 9) antibodies that bind to ∆V1 gp120. c Comparison of plasma and mucosal IgG binding titer (AUC) against different SIV immunogens in all groups of animals (V2-TTB NP, n = 12; TTB NP, n = 9; empty NP, n = 9). d , e Comparison of d ADCC titers (the endpoint reciprocal dilution at which the ADCC killing was greater than control killing + 3 SD) and e ADCC activity among different groups of animals (V2-TTB NP, n = 12; TTB NP, n = 9; empty NP, n = 9; systemic vaccine, n = 14). f–h Correlation of f ADCC titer in the V2-TTB NP group ( n = 12, p = 0.02), g ADCC activity in the V2-TTB NP group ( n = 12, p = 0.02), and h ADCC titer in all animal groups combined ( n = 44, p < 0.0001) with number of intrarectal challenges. In h , red dots represent V2-TTB NP, black dots represent systemic vaccine, and blue squares represent TTB NP. Enlarged shapes represent multiple animals, as indicated. i , j Comparison of i V2 (NCI05)-specific ADCC activity and j frequency of V2 (NCI05)-specific ADCC activity in total ADCC among different groups of animals (V2-TTB NP, n = 12; TTB NP, n = 9; empty NP, n = 9; systemic vaccine, n = 14). k , l Correlation of V2 (NCI05)-specific ADCC activity in k the V2-TTB NP group ( n = 12, p = 0.003) and l in all animal groups combined ( n = 44, p = 0.003) with number of intrarectal challenges. m Correlation of V2 (NCI05)-specific ADCC activity at week 17 with VL at 15 wpi in the systemic vaccine group ( n = 9, p = 0.03). n Comparison of plasma antibody avidity score against ∆V1gp120 protein in different groups of animals (V2-TTB NP, n = 10; TTB NP, n = 7; empty NP, n = 8; systemic vaccine, n = 14). o Correlation of antibody response units against ∆V1gp120 (higher RU values indicated stronger binding of antibody to antigen) with number of intrarectal challenges in the V2-TTB NP+vaccine group ( n = 12, p = 0.02). Data shown in a – c , e , i , j , n were analyzed with the two-sided Mann-Whitney test. Data shown in d were analyzed with two-sided Cochran-Armitage tests for the comparisons between V2-TTB NP and TTB NP and Empty NP, separately. The two-sided Mann-Whitney test was used for the comparison between V2-TTB NP and systemic vaccine. Data shown in f – h , k – m , o were analyzed with the two-sided Spearman correlation test. Horizontal and vertical bars denote mean and SD. Source data are provided as a Source Data file. Here, the black, red, blue, and purple symbols represent male macaques immunized with a systemic vaccine, vaccine combined with V2-TTB NP, vaccine combined with TTB NP, and vaccine combined with empty NP, respectively.

Article Snippet: The primary antibodies were mouse (IgG2a) monoclonal anti-CD4 (clone OTI5D9, Novus Biologicals) and rabbit anti-Ki67 (Abcam).

Techniques: Comparison, Clinical Proteomics, Binding Assay, Control, Activity Assay, MANN-WHITNEY