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: Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, In Situ, Selection, 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

Journal: Life Science Alliance

Article Title: The HDL particle composition determines its antitumor activity in pancreatic cancer

doi: 10.26508/lsa.202101317

Figure Lengend Snippet: WT or Apoa1 KO C57Bl6/J mice were inoculated with 0.5 × 10 6 syngeneic tumor cells either subcutaneously in the right flank (Panc02, B16F10, lung carcinoma cell) or orthotopically in the second mammary fat pad (E0771). (A) Tumor growth kinetics and tumor weight at experimental end stage of the Panc02 pancreatic-cancer model (n = 7/7). (B) Tumor growth kinetics and tumor weight at experimental end stage of the B16F10 melanoma model (n = 13/10 WT and Apoa1 KO mice, respectively). (C) Tumor growth kinetics and tumor weight at experimental end stage of the lung carcinoma cell lung cancer model (n = 9/8). (D) Tumor growth kinetics and tumor weight at experimental end stage of the E0771 breast cancer model (n = 10/13; * P < 0.05; unpaired t test for A, B, C, D).

Article Snippet: The murine melanoma cell line B16F10, Lewis LLCs, and the breast adenocarcinoma cell line E0771 were a kind gift of Prof. Dr. Peter Carmeliet and Prof. Massimiliano Mazzone (VIB Vesalius Research Center, KU Leuven) and maintained in DMEM medium supplemented with 10% FCS, 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin (complete DMEM).

Techniques: