Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP inhibited cancer cell proliferation. ( A ) Schematic illustration of the synthesis of CPP-IL-12. ( B ) Size distribution of CPP. ( C ) The average zeta potentials of CPP. ( D ) Transmission Electron Microscope images of CPP. Scale bars=200 nm. ( E ) The chemical composition was analyzed by Fourier transform infrared spectroscopy. ( F ) Heating curves of different concentrations of CPP under 808 nm (2 W/cm 2 ) laser irradiation. ( G ) The photothermal conversion cycling test of CPP during three cycles of laser on/off. The data are presented as the mean ± SD of three independent experiments. ( H ) Electrophoretic analysis of pEGFP plasmid in the complexes of CPP under various N/P ratios. ( I ) B16F10 cells incubated with CPP@pEGFP complexes (N/P from 10:1 to 40:1); green colour, EGFP expression. Scale bars=125 μm. ( J ) Analyzing the cytotoxicity of CPP, ANOVA. ( K ) ROS detection. CPP treatment increased ROS levels, and 2.0 W/cm 2 of laser further increased ROS levels. ** P <0.01, ANOVA. ( L ) Cell proliferation analysis. B16-F10 cell proliferation was suppressed at 3 min to 9 min after laser treatment, ** P <0.01, ANOVA.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Transmission Assay, Microscopy, Fourier Transform Infrared Spectroscopy, Spectroscopy, Irradiation, Plasmid Preparation, Incubation, Expressing

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP transferred IL-12 into B16F10 cells and CPP-IL-12 induced immunogenic cell death. ( A ) The structure of IL-12 vector. ( B ) Western blot detection of IL-12 expression. ** P < 0.01, Student’s t -test. ( C ) ELISA. CPP-IL-12 increased IL-12 levels in the supernatant of B16-F10 cells, and 2.0 W/cm 2 of laser further increased IL-12 levels. ** P < 0.01, ANOVA. ( D ) Live/dead staining assay. Scale bars=100 μm. ( E ) The number of dead cells. CPP-IL-12 increased and 2.0 W/cm 2 of laser further increased the number of dead cells. ** P < 0.01, ANOVA. ( F ) Cell apoptosis analysis. CPP-IL-12 increased and laser treatment further increased the number of apoptotic cells. ( G ) Immunofluorescence. Green color, CALR expression. Scale bars=2 μm. ( H, I ) ELISA analysis of extracellular and intracellular HMGB1 levels, respectively. * P < 0.05, ** P < 0.01, ANOVA. (J) ATP levels. The ATP levels decreased in CPP-IL-12-treated cells, and laser treatment further reduced ATP content. ** P < 0.01, ANOVA. ( K ) CPP increased CD80 + /CD86 + expression of DC.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Plasmid Preparation, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP-IL-12 suppressed melanoma cell growth in vivo. ( A ) Schematic illustration of in vivo study. ( B ) The xenografts dissected from mice. ( C, D ) The weights and volumes of xenografts were analyzed. Data were presented as the mean ± SD or median (interquartile range). n=4 or 5, ** P < 0.01, * P < 0.05; Student’s t -test or Mann–Whitney U -test. ( E ) HE staining demonstrated that CPP-IL-12 suppressed and laser further inhibited the growth of tumor xenografts in vivo. ( F ) The levels of IL-12 in tumor xenografts analyzed by immunohistochemistry staining. The brown signal indicated the expression of IL-12 in the tissue, the blue signal was nuclear of cells. There was more IL-12 expression in CPP-IL-12 treated groups. Scale bars=125 μm.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: In Vivo, MANN-WHITNEY, Staining, Immunohistochemistry, Expressing

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP-IL-12 promoted systemic antitumor immunity. ( A ) Analyzing CALR expression in tissues by immunofluorescence. Green color, CALR expression. Scale bars=20 μm. ( B ) CPP-IL-12 increased CD80 + /CD86 + expression of DC in lymph glands. ( C ) CPP-IL-12 increased the number of CD3 + /CD8 + T cells from spleen. ( D ) CPP-IL-12 increased the number of CD3 + /CD4 + T cells from spleen.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Expressing, Immunofluorescence

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP-IL-12 increased the expression of CD4, CD8, granzyme B, IFN-γ and TNF-α. ( A ) CPP-IL-12 increased IL-12 levels in tumor xenografts. Scale bars =125 μm. ( B ) The levels of CD8 were increased in tumor xenografts analyzed by immunohistochemistry staining. Scale bars =125 μm. ( C ) CPP-IL-12 increased CD4 expression in tumor xenografts. Scale bars=125 μm. ( D ) ELISA analysis showed that Granzyme B levels increased in CPP-IL-12-treated tumor xenografts. ** P < 0.01, ANOVA. ( E ) IFN-γ levels increased in CPP-IL-12-treated tumor xenografts. ** P < 0.01, ANOVA. ( F ) TNF-α levels increased in CPP-IL-12-treated tumor xenografts. ** P < 0.01, ANOVA. ( G ) CPP-IL-12 treatment increased granzyme B levels in mouse serum. ** P < 0.01, ANOVA. ( H ) CPP-IL-12 treatment increased IFN-γ levels in mouse serum. ** P < 0.01, ANOVA. ( I ) CPP-IL-12 treatment increased TNF-α levels in mouse serum. ** P < 0.01, ANOVA.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Expressing, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP-IL-12 suppressed the growth of local and distant xenografts. ( A ) Schematic description and the mechanism of the effect of local treatment to tumor on distant untreated one. This image was made by using Figdraw software. ( B – D ) CPP-IL-12 with laser treatment inhibited the growth, weights and volumes of local xenografts, respectively. ** P < 0.01, ANOVA. ( E – G ) CPP-IL-12 with laser treatment suppressed the growth, weights and volumes of distant tumors, respectively. ** P < 0.01, ANOVA. ( H – J ) ELISA analysis showed that IFN-γ, TNF-α, and granzyme B levels were increased in the serum of CPP-IL-12-treated mice, respectively. ** P < 0.01, ANOVA. ( K ) CD80 + /CD86 + expression of DC was increased in the lymph glands of CPP-IL-12-treated mice. ( L ) CD4 expression of T cells was increased in the spleens of CPP-IL-12-treated mice. ( M ) CD8 expression was increased in the spleens of CPP-IL-12-treated mice. ( N, O ) The CD8 and CD4 levels were analyzed by immunohistochemistry staining, respectively. Scale bars=125 μm.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Software, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Staining

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: Analyzing the distribution and toxicity of CPP. ( A ) HE staining demonstrated CPP did not lead to significant damage to the organs of mice. Scale bars=125 μm. ( B, C ) The blood compatibility of nanoparticles was evaluated by Hemolysis test. ( D ) The distribution of CPP in mice. ( E ) CPP-IL-12 nanoparticles could significantly suppress melanoma cell proliferation. On one hand, IL-12 delivery by CPP-IL-12 involves immunotherapy and PTT, which induced IL-12 expression and ICD in tumor environment. On the other hand, the ICD would further activate dendritic cells, NK cells, and T cells to promote antitumor immunity.

Article Snippet: The endogenous peroxidase activity in tissues was inactivated by 3% hydrogen peroxide for 15 min. Then, the sections were washed with PBS and incubated in 10% normal goat serum (BeiJingZhongShan Golden Bridge Technology Co, Ltd., Beijing) for 20 min. Next, the sections were incubated with rabbit anti-IL-12 primary antibodies (1:300, Bioworld Technology, Ltd.) at 4°C overnight.

Techniques: Staining, Expressing

Journal: Inflammatory Intestinal Diseases

Article Title: Mucosal Healing in Crohn's Disease: Bull's Eye or Bust? The “Relative” Con Position

doi: 10.1159/000519731

Figure Lengend Snippet: Failed clinical trials in Crohn's disease

Article Snippet: Briakinumab , Anti-IL-12/23 p40 monoclonal antibody , Primary endpoint (clinical remission) at week 6 not met.

Techniques: Clinical Proteomics, Inhibition