Journal: Cell reports

Article Title: Chronic stress physically spares but functionally impairs innate-like invariant T cells

doi: 10.1016/j.celrep.2021.108979

Figure Lengend Snippet:

Article Snippet: Mouse: B16-F10-Red-FLuc (B16-FLuc) melanoma cells , PerkinElmer , Cat # BW124734.

Techniques: Control, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, In Situ, Selection, cDNA Synthesis, Software

Journal: Acta Pharmaceutica Sinica. B

Article Title: Boosting 5-ALA-based photodynamic therapy by a liposomal nanomedicine through intracellular iron ion regulation

doi: 10.1016/j.apsb.2021.03.017

Figure Lengend Snippet: Intracellular iron ion regulating mediated PpIX accumulation and DNA repair enzyme inhibition. (A) The membrance fusion of CLs@DiO and MFLs@DiO (scale bar = 10 μm), the distribution of MFLs@DiI in B16–F10 cells (B) (scale bar = 10 μm). (C) The detection of Fe 2+ after incubating with B16–F10 cells for 4 h (scale bar = 25 μm). (D, E) The transformation of 5-ALA analyzed after incubating with B16–F10 cells for 4 h by CLSM (scale bar = 10 μm) and flow cytometry, respectively. (F) DFO inhibits ALKBH2 repair of m1A in dsDNA by using the DpnII digestion assay and semi-quantitative analysis (G), 1: maker, 2: dsDNA (m1A), 3: dsDNA (m1A)+DpnII, 4: dsDNA (m1A)+ALKBH2+Fe 2+ +DFO + DpnII, 5: dsDNA (m1A)+ALKBH2+Fe 2+ +DpnII. Data are presented as means ± SD ( n = 3). ∗∗∗∗ P < 0.0001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The B16–F10 cells were seeded into confocal dish (2 × 10 5 cells per dish) for 24 h of incubation and then treated with control, 5-ALA, DFO, 5-ALA + DFO, CLs@5-ALA/DFO, MFLs@5-ALA/DFO (with the same dose of 5-ALA and DFO: 1 mmol/L and 100 μmol/L respectively) for 4 h. After that, FerroOrange working solution (1 mL, 1 μmol/L) was added to each dish and incubated for a certain period of time, and then the samples were observed directly by a confocal laser-scanning microscopy (CLSM, Leica TCS SP8).

Techniques: Enzyme Inhibition Assay, Transformation Assay, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Boosting 5-ALA-based photodynamic therapy by a liposomal nanomedicine through intracellular iron ion regulation

doi: 10.1016/j.apsb.2021.03.017

Figure Lengend Snippet: In vitro enhanced PDT of MFLs@5-ALA/DFO. (A) The production of reactive oxygen species (ROS) after 532 nm laser irradiation by fluorescence microscope (scale bar = 200 μm). (B, C) The analyzed of DNA damage by western blotting and comet assay (scale bar = 200 μm). (D) Cells cytotoxicity of 5-ALA, 5-ALA + DFO, MFLs@5-ALA/DFO. (E) Living and dead cell staining and apoptosis test by Calcein-AM and Propidium Iodide (Calcein-AM/PI) (scale bar = 200 μm). (F) Annexin V/PI staining assays of B16–F10 cells. All of them were incubated with different preparations for 4 h and irradiated with 532 nm laser. Data are presented as means ± SD ( n = 3). ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The B16–F10 cells were seeded into confocal dish (2 × 10 5 cells per dish) for 24 h of incubation and then treated with control, 5-ALA, DFO, 5-ALA + DFO, CLs@5-ALA/DFO, MFLs@5-ALA/DFO (with the same dose of 5-ALA and DFO: 1 mmol/L and 100 μmol/L respectively) for 4 h. After that, FerroOrange working solution (1 mL, 1 μmol/L) was added to each dish and incubated for a certain period of time, and then the samples were observed directly by a confocal laser-scanning microscopy (CLSM, Leica TCS SP8).

Techniques: In Vitro, Irradiation, Fluorescence, Microscopy, Western Blot, Single Cell Gel Electrophoresis, Staining, Incubation

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Nexavant recruits diverse immune cells to the draining lymph nodes near the injection sites. C57BL/6 mice ( n = 4/group) were injected subcutaneously with 25 μg of Nexavant. At 1, 4, 12, 24, and 48 h post-injection, the mice were monitored for ( A ) immune cell profile and DC activation in the inguinal lymph nodes and ( B ) IFN-β, IL-6, and IL-12 cytokine levels in the blood. C57BL/6 mice ( n = 5/group) inoculated subcutaneously with 5 × 10 5 B16F10 cells were treated with 100 μg of Nexavant or Poly(I:C) on days 8, 11, 14, and 17 post-inoculation, and ( C ) tumor growth was measured. ( D ) Blood levels of IL-6 and IFN-β cytokines were measured 4 h after the first injection of Nexavant or Poly(I:C) ( n = 6/group). ( E ) The immune cell profile of mouse tumors was analyzed on day 15 ( n = 4, 4, 5). * p < 0.05, ** p < 0.01, and **** p < 0.0001; n.s., not significant; two-way ANOVA with Dunnett’s test ( C ) and one-way ANOVA with Dunnett’s test ( D , E ).

Article Snippet: The B16F10 murine melanoma cell line was purchased from Korean Cell Line Bank (Seoul, Korea), and the CT26 murine colon cancer cell line, the LL/2 murine lung cancer cell line, and the 4T-1 murine breast cancer cell line were purchased from ATCC.

Techniques: Injection, Activation Assay

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Intratumoral administration of Nexavant suppressed tumor growth in various syngeneic models. ( A ) C57BL/6 mice ( n = 4/group) inoculated subcutaneously with 5 × 10 5 LL/2, CT26, or 4T-1 cells were treated with Nexavant (25 μg) (LL/2 and CT26 models on days 7, 9, 11, and 13 post-inoculation; the 4T-1 model on days 6, 8, 10, and 12 post-inoculation). ( B ) C57BL/6 mice ( n = 5/group) were inoculated subcutaneously in both flanks with 5 × 10 5 B16F10 cells. Nexavant (25, 50, 75, 100, and 150 ugs) was injected into the right flank tumor 7-, 11-, and 14-days post-inoculation. ( C ) C57BL/6 mice ( n = 4/group) inoculated subcutaneously with 5 × 10 5 B16F10 cells were treated with Nexavant (100 μg and 500 μg) on days 22, 25, 29, and 32 post-inoculation. ** p < 0.01, and **** p < 0.0001; n.s., not significant; two-way ANOVA with Dunnett’s test ( A – C ).

Article Snippet: The B16F10 murine melanoma cell line was purchased from Korean Cell Line Bank (Seoul, Korea), and the CT26 murine colon cancer cell line, the LL/2 murine lung cancer cell line, and the 4T-1 murine breast cancer cell line were purchased from ATCC.

Techniques: Injection

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Combination therapy with anti-PD-1 antibodies and Nexavant shows an improved anti-tumor effect. ( A ) C57BL/6 mice ( n = 5/group) were inoculated subcutaneously in their right flanks with 5 × 10 5 B16F10 tumor cells and treated with 25 μg of Nexavant (intratumoral) and 100 μg of anti-PD-1 antibodies (intraperitoneal) on days 6, 8, 10 and 12 post-inoculation. The mice were inoculated again, this time in their left flanks, with 5 × 10 5 B16F10 tumor cells on day 24 after the first challenge (corresponding to day 0 of the rechallenge). Since all individuals in the PBS group died before day 24, naïve mice were used as a control group (CTL) for rechallenge tumors. The volume of injected and non-injected tumors was measured at the indicated time points. ( B ) The immune cell profiles of the B16F10 tumors were assessed on day 25 using flow cytometry ( n = 6/group). ( C ) C57BL/6 mice ( n = 5/group) were subcutaneously inoculated in their right flanks with 5 × 10 5 K1735 tumor cells and immunized with Nexavant and anti-PD-1 antibodies on days 17, 19, 21, 23, and 25 post-inoculation. ( D ) BALB/c mice ( n = 5/group) were inoculated in their mammary fat pads with 5 × 10 5 EMT6 tumor cells and then immunized with Nexavant and anti-PD-1 antibodies on days 7, 9, 11, 13, and 15 post-inoculation. ( E ) C57BL/6 mice ( n = 5/group) were inoculated in their mammary fat pads with 5 × 10 5 EO771 tumor cells and then immunized with Nexavant and anti-PD-1 antibodies on days 11, 15, 18, 22, 25, 29, and 32 post-inoculation. Tumor volumes were measured at indicated time points. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; n.s., not significant; two-way ANOVA with Dunnett’s test ( A , C – E ) and one-way ANOVA with Dunnett’s test ( B ).

Article Snippet: The B16F10 murine melanoma cell line was purchased from Korean Cell Line Bank (Seoul, Korea), and the CT26 murine colon cancer cell line, the LL/2 murine lung cancer cell line, and the 4T-1 murine breast cancer cell line were purchased from ATCC.

Techniques: Injection, Flow Cytometry

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Improved anti-cancer efficacy of the combined treatment with Nexavant and nivolumab (a human anti-PD-1 antibody) was tested in the transgenic mice expressing a chimeric PD-1 with the extracellular domain of human PD-1 and the transmembrane and cytoplasmic domains of mouse PD-1 compared with the respective monotherapies. C57BL/6 and BALB/c mice with human PD-1 were inoculated with 5 × 10 5 B16F10 melanoma cells and EMT6 TNBC cells, respectively (day 0). ( A , B ) Tumor growth inhibition and improved survival in the group administered the combination therapy compared with the groups administered monotherapies in ( A ) B16F10 melanoma and ( B ) EMT6 breast cancer models ( n = 6/group). B16F10-bearing mice were treated with Nexavant and anti-PD-1 antibodies on days 8, 11, 14, 18, 21, and 24 post-inoculation, and EMT6-bearing mice were treated on days 7, 11, 14, 18, 21, and 25 post-inoculation. Tumor volumes and survivals were measured at indicated time points. * p < 0.05, ** p < 0.01, and **** p < 0.0001; n.s., not significant; two-way ANOVA with Dunnett’s test ( A , B ).

Article Snippet: The B16F10 murine melanoma cell line was purchased from Korean Cell Line Bank (Seoul, Korea), and the CT26 murine colon cancer cell line, the LL/2 murine lung cancer cell line, and the 4T-1 murine breast cancer cell line were purchased from ATCC.

Techniques: Transgenic Assay, Expressing, Inhibition

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Intranasal administration of Nexavant has a potent antitumor effect but there is no additive effect when it is combined with anti-PD-1 antibodies. ( A ) In all, 25 μg of Nexavant was slowly instilled through the nostrils ( n = 4/group). After 24 h, bronchoalveolar lavage fluid (BALF) was collected and analyzed using flow cytometry. ( B , C ) C57BL/6 mice were injected with 5 × 10 5 B16F10 cells via the tail vein. Next, 25 μg of Nexavant or Poly(I:C) was slowly instilled through the nostrils every two days starting on day 8 or 14. Tumor-bearing lungs (( B ); n = 8, 6, 8/group) were harvested on day 18, and overall survival (( C ); n = 8, 8, 7) was monitored. ( D , E ) C57BL/6 mice were injected with 5 × 10 5 B16F10 cells via the tail vein. Next, 25 ug of Nexavant was slowly instilled through the nostrils every two days starting on days 2, 8, or 14 ( n = 8, 8, 8, 6). ( E ) On day 15, the lungs were harvested from B16F10 lung metastases, and the tumor nodules were counted ( n = 6, 5). ( F ) C57BL/6 mice ( n = 6, 5, 5, 4) were injected with 5 × 10 5 B16F10 cells via the tail vein. Next, 25 μg of Nexavant was instilled via the nostrils, and 200 μg of anti-PD-1 antibodies was injected intraperitoneally on days 2, 4, 6, 8, and 10. On day 18, B16F10 lung metastasis mice were sacrificed and the lungs were harvested. The gross photograph of a lung with tumor burden was taken from a certain distance. Scale bar = 1 cm; ** p < 0.01, and *** p < 0.001; n.s., not significant; one-way ANOVA with Dunnett’s test ( A , E ). The uncropped blots are shown in .

Article Snippet: The B16F10 murine melanoma cell line was purchased from Korean Cell Line Bank (Seoul, Korea), and the CT26 murine colon cancer cell line, the LL/2 murine lung cancer cell line, and the 4T-1 murine breast cancer cell line were purchased from ATCC.

Techniques: Flow Cytometry, Injection