Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Enhanced dendritic cell maturation by TNF-alpha or cytidine-phosphate-guanosine DNA drives T cell activation in vitro and therapeutic anti-tumor immune responses in vivo.

doi: 10.4049/jimmunol.165.11.6278

Figure Lengend Snippet: FIGURE 5. Tumor specifity of vaccination with DC after coculture with tumor cells. DC were cocultured with irradiated Colon-26 cells (a and d) or Renca cells (b and c) for 4 days in the presence of 200 U/ml GM-CSF and 20 ng/ml IL-4 and additionally stimulated for 48 h with 6 mg/ml ODN-1826. Renca cells (5 3 105) were injected 7 days after vaccination with ODN-1826-stimulated DC cocultured with Colon-26 cells F, (a, p 5 0.290) or in mice vaccinated with ODN-1826-stimulated DC cocultured with Renca cells (b, f, p , 0.001). Tumor growth after injection of 5 3 105 Renca cells in unvaccinated mice (control group ¿) was rapid, leading to death at day 23. In addition, 5 3 105 Colon-26 cells were injected 7 days after vaccination with ODN-1826-stimulated DC cocultured with Renca cells (c, l, p 5 0.250) or ODN-1826-stimulated DC cocultured with Colon-26 cells (d, Œ, p , 0.001). One representative experiment of two (n 5 60) is shown. Each curve represents one mouse except for the top curve (¿), which is the mean 6 SEM of untreated control mice (n 5 5).

Article Snippet: The murine, BALB/c-derived Colon-26 carcinoma line as well as the murine Renca cells (Cell Lines Service, Heidelberg, Germany) were maintained in DMEM supplemented with 10% FCS, 1% L-glutamine, and antibiotics.

Techniques: Irradiation, Injection, Control

Journal: Frontiers in Oncology

Article Title: Alphataxin, a Small-Molecule Drug That Elevates Tumor-Infiltrating CD4 + T Cells, in Combination With Anti-PD-1 Therapy, Suppresses Murine Renal Cancer and Metastasis

doi: 10.3389/fonc.2021.739080

Figure Lengend Snippet: Tumor Growth Curves of Subcutaneously Implanted Tumor Cells. Renca cells (2 x 10 5 ) were subcutaneously implanted into 8 female BALB/c mice per group and allowed to grow for 2 weeks before the mice remained untreated (●, n = 7) or were treated with an anti-PD-1 antibody ( , 7 mg/kg, n = 6) via IP injection twice per week or with Alphataxin via daily oral gavage at a dose of 2.5 mg/kg (●), 5 mg/kg ( ), or 10 mg/kg ( ) (n = 8/arm). In combination treatment, mice were treated with anti-PD-1 (7mg/kg) via IP injection twice per week and with Alphataxin via daily oral gavage at a dose of 2.5 mg/kg ( ), 5 mg/kg ( ), or 10 mg/kg ( ) (n = 8/arm). Tumor volume was calculated periodically using calipers. Alphataxin (10 mg/kg) significantly suppressed tumor growth as compared with Alphataxin (5 mg/kg) or Alphataxin (2.5 mg/kg) (P < 0.001, n = 8/arm). Data were normalized to baseline tumor volume and are presented as the means and standard deviations.

Article Snippet: For a study conducted by AJES Life Sciences (IACUC# 764665-4), orthotopic implantation was achieved by injecting 5 x 10 2 Renca cells in 100 μl of HBSS mixed 1:1 with Matrigel (BD Biosciences) into the left kidney of 8-wk-old female BALB/c mice, and treatment was initiated 7 days after implantation.

Techniques: Injection

Journal: Frontiers in Oncology

Article Title: Alphataxin, a Small-Molecule Drug That Elevates Tumor-Infiltrating CD4 + T Cells, in Combination With Anti-PD-1 Therapy, Suppresses Murine Renal Cancer and Metastasis

doi: 10.3389/fonc.2021.739080

Figure Lengend Snippet: Long-term Remission Achieved by Treatment with Alphataxin and an Anti-PD-1 Antibody. Mice were orthotopically implanted with 5x 10 3 Renca-GL cells, and treatment was initiated the following day. (A) Kidneys were excised at the time of euthanization and stained with H&E. Images were captured at identical magnification, and representative images at 1X are shown. (B) Longitudinal dorsal images of the orthotopic kidney tumor growth of mice. A representative animal that showed similar bioluminescence with at least 2 other animals within the same treatment arm after 25 days of treatment was selected from each treatment arm. The mice in the combination treatment arm were bifurcated into 2 groups, those with tumors (“Combination +”, 4 th row, n = 4) and those without tumors (“Combination −”, 5 th row, n = 2). The luminescence color scale represents photons/sec/cm 2 /steradian x 10 9 with minimum 4.48e6 and maximum 19.4e9. (C) As compared with untreated mice and monotherapy, after 25 days of treatment the tumor volumes (BLI) were significantly smaller in mice receiving combination treatment with tumors ( P = 0.05, n = 4) and without tumors ( P = 0.02, n = 2). After 25 days of treatment, there were no significant differences between untreated and Alphataxin-treated mice ( P = 0.24, n = 6/arm) or between untreated and anti-PD-1-treated mice ( P = 0.06, n = 6) although the difference between untreated and anti-PD-1-treated mice have practical statistical importance. Means and standard deviations are depicted. (D) Median longitudinal tumor growth demonstrates that as compared with untreated animals ( ), Alphataxin ( ) and anti-PD-1( ) individually impeded tumor growth and in combination + animals ( ), substantially impeded tumor growth while in combination – animals ( ), tumor growth regressed. Asterisks designate statistically significant differences (*P < 0.05).

Article Snippet: For a study conducted by AJES Life Sciences (IACUC# 764665-4), orthotopic implantation was achieved by injecting 5 x 10 2 Renca cells in 100 μl of HBSS mixed 1:1 with Matrigel (BD Biosciences) into the left kidney of 8-wk-old female BALB/c mice, and treatment was initiated 7 days after implantation.

Techniques: Staining

Journal: Frontiers in Oncology

Article Title: Alphataxin, a Small-Molecule Drug That Elevates Tumor-Infiltrating CD4 + T Cells, in Combination With Anti-PD-1 Therapy, Suppresses Murine Renal Cancer and Metastasis

doi: 10.3389/fonc.2021.739080

Figure Lengend Snippet: Increases in the Tumor-infiltrating CD4/CD8 Ratio Achieved by Treatment with Alphataxin or an Anti-PD-1 Antibody. Excised kidneys from mice in each treatment arm of (5 x 10 2 orthotopically implanted Renca cells and 5 x 10 3 orthotopically implanted Renca-GL cells) were immunohistochemically stained to determine the numbers of CD4 + and CD8 + TILs which were counted in all the tumor fields of the stained slides. Because the tumors varied in size, yielding large variation in the number of TILs, the CD4/CD8 ratio is depicted to produce more accurate comparisons. (A) In the Renca cell orthotopic study, after 21 days of treatment, the tumor-infiltrating CD4/CD8 ratio in Alphataxin-treated animals was significantly greater than that in untreated and anti-PD-1 antibody-treated mice ( P = 0.04). (B) In the Renca-GL study, tumor-infiltrating CD4/CD8 ratio in Alphataxin-treated mice was significantly greater than that in untreated ( P < 0.001, n = 3) and anti-PD-1-treated mice ( P < 0.001, n = 3). In anti-PD-1 treatment, CD4/CD8 ratio was significantly greater than in untreated animals ( P = 0.02, n = 3). Combination treatment is not depicted due to the lack of suitable kidney sectioning. Means and standard deviations are depicted. The CD4/CD8 ratios were not normally distributed and were compared by the Mann-Whitney rank- sum test. (C) In the Renca-GL study, there were more CD4 + TILs (green bars) in the tumors of Alphataxin-treated mice than in those of anti-PD-1 antibody-treated mice ( P = 0.086, n = 3), which demonstrates practical statistical importance. The numbers of CD8 + TILs (blue bars) were not different among the treatment arms ( P = 0.94), n =3). Note that the scales for the CD4/CD8 ratio and TIL counts are not equivalent in different graphs. TILs were normally distributed and were tested by ANOVA. Asterisks designate statistically significant differences (*P < 0.05, ***P < 0.001).

Article Snippet: For a study conducted by AJES Life Sciences (IACUC# 764665-4), orthotopic implantation was achieved by injecting 5 x 10 2 Renca cells in 100 μl of HBSS mixed 1:1 with Matrigel (BD Biosciences) into the left kidney of 8-wk-old female BALB/c mice, and treatment was initiated 7 days after implantation.

Techniques: Staining, MANN-WHITNEY

Journal: Journal of Nanobiotechnology

Article Title: A multifunctional PEGylated liposomal-encapsulated sunitinib enhancing autophagy, immunomodulation, and safety in renal cell carcinoma

doi: 10.1186/s12951-024-02664-5

Figure Lengend Snippet: The biodistribution pattern of lipo-sunitinib in RCC models. A Lipo-sunitinib was labeled with the lipophilic tracers DiI or DiR to assess biodistribution and observation in vitro and in vivo. B A498 and RENCA cells were subjected to IF to evaluate the observation of DiI-labeled lipo-sunitinib. DAPI and DiI were used to represent nuclei and observed lipo-sunitinib, respectively. C, D Lipo-sut DiR and free DiR were intravenously injected into RCC orthotopic models, and the distribution of DiR signals was tracked over a time period of 0–48 h. E The primary organs were harvested for ex vivo detection of DiR signals to analyze the biodistribution of lipo-sut-DiR and free DiR groups (T=tumor, K=kidney, B=brain, H/LG=heart/lungs, L=livers, S=spleen). F The DiR signals from RCC kidneys were compared with those from normal kidneys. G RCC orthotopic tumors were harvested from DiI-labeled lipo-sunitinib, and DiI fluorescence signals were detected with excitation and emission wavelengths of 555 and 584 nm, respectively (T = tumor tissue, N = normal tissue). Scale bar, 100 μm

Article Snippet: RENCA cells and A498 cells were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu City, Taiwan) and maintained in Roswell Park Memorial Institute (RPMI) 1640 and Eagle’s Minimum Essential (MEM) medium contenting 10% fetal bovine serum (FBS) and 1 × penicillin/streptomycin (PS), then incubated with 5% CO 2 and 95% air mixture at 37 °C humanity incubator.

Techniques: Labeling, In Vitro, In Vivo, Injection, Ex Vivo, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: A multifunctional PEGylated liposomal-encapsulated sunitinib enhancing autophagy, immunomodulation, and safety in renal cell carcinoma

doi: 10.1186/s12951-024-02664-5

Figure Lengend Snippet: Lipo-sunitinib strongly upregulates cytotoxicity through autophagy induction in RCC. A , B MTT assays were conducted to compare the cytotoxicity of sunitinib and lipo-sunitinib in RENCA and A498 cell lines. C RENCA cells treated with lipo-sunitinib underwent RNA-seq analysis, revealing significantly upregulated pathways compared to the non-treated group. D RNA-seq data for lipo-sunitinib were subjected to GSEA analysis with a focus on macrophagy and autophagosome pathways. E IF analysis of RENCA and A498 cells included detection of LC3B and p62 expression using GFP and RFP channels, with DAPI staining representing nuclei. F In addition, LC3B protein expression in RENCA and A498 cells were assessed using western blotting assays. G The subcutaneous RCC xenograft models were constructed using BALB/c mice with RENCA cells subcutaneously injection. Vehicle and sunitinib groups were administrated 0.1% DMSO and 30 mg/kg sunitinib through daily gavage, respectively, and the H tumor volume of RCC was measured twice daily. Mice are sacrificed on day 14, and all tumors were isolated from mice for I weighting and J imaging. K Body weights of mice were also measured twice daily after treatment started. L Several organs of the subcutaneous RCC xenograft model (liver, heart, and kidneys) were subjected to H&E staining. Scale bar, 50 μm

Article Snippet: RENCA cells and A498 cells were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu City, Taiwan) and maintained in Roswell Park Memorial Institute (RPMI) 1640 and Eagle’s Minimum Essential (MEM) medium contenting 10% fetal bovine serum (FBS) and 1 × penicillin/streptomycin (PS), then incubated with 5% CO 2 and 95% air mixture at 37 °C humanity incubator.

Techniques: RNA Sequencing, Expressing, Staining, Western Blot, Construct, Injection, Isolation, Imaging