Journal: Journal of Nanobiotechnology

Article Title: Silver nanoparticle induced immunogenic cell death can improve immunotherapy

doi: 10.1186/s12951-024-02951-1

Figure Lengend Snippet: Therapeutic effects of Ag-citrate-5 nm on Renca tumors and syngergy with anti-PD1 ICB. ( A ) Relative photon flux (radiance) of Renca-luc + tumors either injected ( a ) peritumorally with PBS (100 µl/animal; control), ( b ) peritumorally with Ag citrate-5 nm (20 µg/mouse), ( c ) intraperitoneally with an anti-PD-1 antibody (150 µg/animal). The mice in the combination groups were injected peritumorally with Ag citrate-5 nm (20 µg/mouse) and intraperitoneally with anti-PD-1 antibody (150 µg/animal). The combination groups and the IT group were in total injected intraperitoneally with anti-PD-1 three times (boost) with an interval of 4 days between injections. Further control groups include (1) intraperitoneal administration of anti-CD8 (150 µg/ml), (2) anti-PD1 + anti-CD8, (3) Ag citrate-5 nm + anti-CD8 or Ag citrate-5 nm + anti-PD1 + anti-CD8. Representative bioluminescence images of mice with Renca Luc + flank tumors in different treatment groups. These groups also include treatment with anti-CD8 antibody either as monotherapy, or in combination with NPs, anti-PD1 or both where anti-CD8 antibody (150 µg/animal) was administered intraperitoneally three times with an interval of 4 days between injections. ( B ) Tumor flux of Renca Luc + cells in photon per flux. Subcutaneous Renca tumors were treated with higher concentration of Ag-citrate-5 nm (50 µg/mouse) and Anti-PD1 (200 µg/mouse). Representative bioluminescence images of mice with Renca Luc + flank tumors in different treatment groups. Representative image of the tumors at the final timepoint. ( C ) Bar graphs displaying the level of caspase and elastase at the tumor as measured through non-invasive optical imaging. Representative fluorescence images of mice with Renca Luc + flank tumors in different treatment groups. ( D ) Bar graphs showing the cancer cell-selective calreticulin translocation to the cell surface as evaluated through ImageStream based flow cytometry upon treatment with the various conditions. The results are presented as the mean of the animals/group ± SEM. The level of significance was indicated when appropriate (*: p < 0.05; **: p < 0.01; ***: p < 0.001)

Article Snippet: Anti-CD8 was based on monoclonal antibody against CD8 alpha ( In vivo MAb anti-mouse CD8α, Biocell).

Techniques: Injection, Control, Concentration Assay, Optical Imaging, Fluorescence, Translocation Assay, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Silver nanoparticle induced immunogenic cell death can improve immunotherapy

doi: 10.1186/s12951-024-02951-1

Figure Lengend Snippet: Effect of Ag-citrate-5 nm, with or without IT, on immune cell activation in the spleen and tumor tissue. Representative fluorescence images of tumor tissue section obtained from Renca luc+ tumors treated with ( A ) PBS, ( B ) Ag-citrate-5 nm monotherapy, ( C ) anti-PD1 monotherapy or ( D ) combination therapy. The images reveal tissue sections stained for F4/80 (green, macrophages), CD8 (CD8 + T cells, red) and counterstained with DAPI (cell nuclei, blue). Scale bars of 100 μm are indicated in the bottom left corner. Bar graphs displaying the levels of ( E ) CD4+ T cells in the spleen, ( F ) CD4 + CD38+ active T cells in the spleen, ( G ) CD4 + CD69+ active T cells in the spleen or ( H ) CD8+ T cells in the spleen. The results are presented as the normalized mean + SEM in percentages related to the control group (PBS =100%). The level of significance was indicated when appropriate (*: p <0.05; **: p < 0.01; ***: p <0.001; ****: p <0.0001). I ) Representative H&E stained images of kidney (top row), lung (middle row) and liver (bottom row) tissue sections of tumor-bearing mice treated with the respective agents indicated at the top

Article Snippet: Anti-CD8 was based on monoclonal antibody against CD8 alpha ( In vivo MAb anti-mouse CD8α, Biocell).

Techniques: Activation Assay, Fluorescence, Staining, Control

Journal: Cancer immunology research

Article Title: The Tumor Microenvironment Regulates Sensitivity of Murine Lung Tumors to PD-1/PD-L1 Antibody Blockade

doi: 10.1158/2326-6066.CIR-16-0365

Figure Lengend Snippet: CMT167 subcutaneous tumors induce a T-cell response that is skewed toward CD8+ T cells and are less sensitive than CMT167 orthotopic tumors to anti-PD-1/PD-L1 therapy. (A) CMT167 subcutaneous tumor-bearing mice were injected with anti-PD-1, anti-PD-L1, or isotype control antibody (n = 10 for IgG2a and anti-PD-1 groups, n = 5 for IgG2b and anti-PD-L1 groups). CMT167 orthotopic tumor-bearing lungs (n = 9) and CMT167 subcutaneous tumors (n = 7) were analyzed by flow cytometry for (B) CD8+ T cells, (C) CD4+ T cells, (D) CD4:CD8 ratio, (E) PD-1 expression on CD8+ T cells, (F) PD-1 expression on CD4+ T cells, and (G) Foxp3 expression in CD4+ T cells. Statistically significant differences are indicated as determined by Student unpaired t test; ** = P < 0.01, *** = P < 0.001. (H) PD-L1 expression was analyzed by flow cytometry in CMT167 subcutaneous tumors. Similar results were obtained in five independent mice.

Article Snippet: Rat monoclonal anti-mouse CD8 (clone 53-6.72) and rat IgG2a (clone 2A3) were purchased from BioXCell (Lebanon, NH).

Techniques: Injection, Control, Flow Cytometry, Expressing

Journal: Cancer immunology research

Article Title: The Tumor Microenvironment Regulates Sensitivity of Murine Lung Tumors to PD-1/PD-L1 Antibody Blockade

doi: 10.1158/2326-6066.CIR-16-0365

Figure Lengend Snippet: Both CMT167 orthotopic lung tumors and LLC orthotopic lung tumors generate an adaptive immune response and induce PD-1 expression on T cells. Tissue sections from CMT167 (representative image shown in Fig. 5A) and LLC (representative image shown in Fig. 5B) tumor-bearing mice were stained for CD3 (FITC, green) and nuclei were stained with DAPI (blue). Magnification x20. (C) Quantification of tumor-infiltrating lymphocytes in CMT167 tumors (n = 9) and LLC tumors (n = 6). (D) Effects of CD8+ T cell immunodepletion on growth of CMT167 (n = 7 each group) and LLC orthotopic tumors (n = 6 each group). CMT167 (n = 9, same mice as in Fig. 3B–F) or LLC tumor-bearing lungs (n = 9) were analyzed by flow cytometry and compared with lungs from naive mice (n = 8) for (E) CD8+ T cells, (F) CD4+ T cells, (G) PD-1 expression on CD8+ T cells, and (H) PD-1 expression on CD4+ T cells. Statistically significant differences are indicated as determined by Student unpaired t test (C-D) or by one-way ANOVA (E-H); * = P < 0.05, ** = P < 0.01, *** = P < 0.001.

Article Snippet: Rat monoclonal anti-mouse CD8 (clone 53-6.72) and rat IgG2a (clone 2A3) were purchased from BioXCell (Lebanon, NH).

Techniques: Expressing, Staining, Immunodepletion, Flow Cytometry

Journal: The Journal of Experimental Medicine

Article Title: Activating receptors promote NK cell expansion for maintenance, IL-10 production, and CD8 T cell regulation during viral infection

doi: 10.1084/jem.20082387

Figure Lengend Snippet: Effect of Ly49H on CD8 T cell responses in MCMV-infected Prf1 −/− mice. (A) Splenic leukocytes from day 0 and 7 MCMV-infected wt , Prf1 −/− , and Ly49h −/− Prf1 −/− mice were analyzed for frequencies of NK cells and CD8 T cells, and expression of IFN-γ and granzyme B in CD8 T cells. (B) Percentages and absolute numbers of CD8α + IFN-γ + in total splenic leukocytes isolated from day 0 and 7 MCMV-infected wt , Prf1 −/− , and Ly49h −/− Prf1 −/− mice were determined. (C) Virus titers in spleens, and IFN-γ, TNF-α, and IL-10 in serum samples from day 0 and 7 MCMV-infected wt , Prf1 −/− , and Ly49h −/− Prf1 −/− mice were measured. The numbers indicate the percentages of cells in each area. Data are presented as means ± SD of three to six mice. Statistical significances between groups are indicated (*, P < 0.05; **, P < 0.01). Results are representative of at least two independent experiments with at least three mice per group. φ, not detected.

Article Snippet: For CD8 T cell blocking, 1 mg anti-CD8α antibody (clone 53.6.72; Bio-Express) was administered by i.p. injection at day 5.

Techniques: Infection, Expressing, Isolation, Virus

Journal: The Journal of Experimental Medicine

Article Title: Activating receptors promote NK cell expansion for maintenance, IL-10 production, and CD8 T cell regulation during viral infection

doi: 10.1084/jem.20082387

Figure Lengend Snippet: Effect of blocking of IL-10 on CD8 T cell responses in and survival of MCMV-infected Prf1 −/− mice. (A) Splenic leukocytes from day 0 and 7 MCMV-infected Prf1 −/− mice treated with either control antibody or anti–IL-10 antibody were analyzed for frequencies of NK cells and CD8 T cells, and expression of IFN-γ and granzyme B in CD8 T cells. (B) Percentages and absolute numbers of CD8α + IFN-γ + in total splenic leukocytes from day 0 and 7 MCMV-infected Prf1 −/− mice treated with either control antibody or anti–IL-10 antibody are shown. (C) Virus titers in spleens, and IFN-γ, TNF-α, and IL-10 levels in serum samples from day 0 and 7 MCMV-infected Prf1 −/− mice treated with either control antibody or anti–IL-10 antibody were measured. The numbers indicate the percentages of cells in each area. Data are presented as means ± SD of three to six mice. Statistical significances between groups are indicated (**, P < 0.01). Results are representative of at least two independent experiments with at least three mice per group. (D) Survivals of Prf1 −/− mice infected with 5,000 PFU and treated with either control antibody or anti–IL-10 antibody were evaluated. Data were compiled from two independent experiments, and the p-value was determined by the log-rank survival test. φ, not detected.

Article Snippet: For CD8 T cell blocking, 1 mg anti-CD8α antibody (clone 53.6.72; Bio-Express) was administered by i.p. injection at day 5.

Techniques: Blocking Assay, Infection, Control, Expressing, Virus

Journal: The Journal of Experimental Medicine

Article Title: Activating receptors promote NK cell expansion for maintenance, IL-10 production, and CD8 T cell regulation during viral infection

doi: 10.1084/jem.20082387

Figure Lengend Snippet: Effect of blocking CD8 T cell responses during MCMV infection of Ly49h −/− Prf1 −/− mice. (A) Virus titers in spleens, and IFN-γ, TNF-α, and IL-10 levels in serum samples from day 0 and 7 MCMV-infected Ly49h −/− Prf1 −/− mice treated with either control antibody or anti-CD8α antibody were measured. Symbols provide results from individual mice. Horizontal line data presented are means, and the vertical lines indicate ± SD of five mice. Statistical significances between groups are indicated (*, P < 0.05; **, P < 0.01). Results are representative of at least two independent experiments. (B) Survivals of Ly49h −/− Prf1 −/− mice MCMV infected with 5,000 PFU and treated with either control antibody or anti-CD8α antibody were evaluated. Data were compiled from two independent experiments, and the p-value was determined by the log-rank survival test.

Article Snippet: For CD8 T cell blocking, 1 mg anti-CD8α antibody (clone 53.6.72; Bio-Express) was administered by i.p. injection at day 5.

Techniques: Blocking Assay, Infection, Virus, Control

Journal: The Journal of Experimental Medicine

Article Title: Activating receptors promote NK cell expansion for maintenance, IL-10 production, and CD8 T cell regulation during viral infection

doi: 10.1084/jem.20082387

Figure Lengend Snippet: Model for sustaining NK cells for regulation of adaptive immune responses during viral infections. (A) In the absence of stimulation through activating receptors, NK cells decrease during extended viral infections. As a result, they are not available to contribute antiviral and/or immune regulatory functions for extended periods. If the virus is unchecked, unregulated downstream adaptive immune responses can lead to immune pathology and death. (B) In the presence of stimulation through activating receptors, NK cell proliferation is supported and the cells are available to contribute a wide range of functions for extended periods. The studies in this paper demonstrate that during profound viral infections, the conditions can result in induction of NK cell IL-10 production to regulate the magnitude of adaptive CD8 T cell responses and protect from CD8 T cell–dependent death.

Article Snippet: For CD8 T cell blocking, 1 mg anti-CD8α antibody (clone 53.6.72; Bio-Express) was administered by i.p. injection at day 5.

Techniques: Virus