Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: IDO shRNA and D-1MT treatment inhibits LLC1 tumor growth and causes increasing number of tumor-infiltrating neutrophils in a subcutaneous lung tumor model. ( a ) Experimental design. 2 × 10 5 LLC1 cells were subcutaneously injected into male inbred C57BL/6JNarl mice. Scramble IDO shRNA (Scr IDOsh), IDO shRNA (IDO sh), and 1-methyl-D-tryptophan (D-1MT) was treated on 7 days after LLC1 injection. ( b ) Tumor growth curve. *A statistically significant difference when compared with scramble IDO shRNA group on 16 days after LLC1 injection ( P < 0.05). ( c ) Number of tumor-infiltrated CD11b + Ly6G + neutrophils. *A statistically significant difference when compared with scramble IDO shRNA group ( P < 0.05). ( d ) Number of CD11b + Ly6G + neutrophils in spleen at 16 days after LLC1 injection. IDO, Indoleamine 2,3-dioxygenase; shRNA, short hairpin RNA.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Injection

Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: IDO shRNA and D-1MT treatment induced Th1-bias immune responses. Spleens were collected at 16 days after LLC1 injection. Splenocytes and culture medium were collected after 24 hours culture. ( a ) mRNA expression of IDO. ( b ) mRNA expression of IFN-γ and IL-10. ( c ) Cytokines concentration of IFN-γ, TNF-α, IL-4 and IL-10. * or **A statistically significant difference when compared with scramble IDO shRNA group (* P < 0.05; ** P < 0.01). D-1MT, 1-methyl-D-tryptophan; IDO, Indoleamine 2,3-dioxygenase; IFN-γ, interferon- γ; IL-4, interlukein-4; shRNA, short hairpin RNA; TNF-α, tumor necrosis factor- α.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Injection, Expressing, Concentration Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: IDO shRNA and D-1MT did not significantly affect the CD4 + CD25 + regulatory T cells. ( a ) Spleens were collected at 16 days after LLC1 injection and the population of CD4 + CD25 + Treg cells were determined on a flow cytometry. Data are the representative of at least three independent experiments. ( b ) The quantitative results. D-1MT, 1-methyl-D-tryptophan; IDO, Indoleamine 2,3-dioxygenase; shRNA, short hairpin RNA.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Injection, Flow Cytometry

Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: The level of tryptophan transfer RNA was induced by IDO shRNA treatment in total splenocytes. Total splenocytes were collected at 16 days after LLC1 injection. The levels of tryptophan transfer RNA (tRNA Trp ) and initiator methionine transfer RNA (tRNA i Met ) was measured. *A statistically significant difference when compared with scramble IDO shRNA group (* P < 0.05). IDO, Indoleamine 2,3-dioxygenase; shRNA, short hairpin RNA.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Injection

Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: Tumor-infiltrating CD11b + Ly6G + neutrophils expressed high level of TNF- α and TGF- β in IDO shRNA treated mice. ( a ) Enriched CD11b + Ly6G + neutrophils from tumor-infiltrated cells. ( b ) The mRNA expression of tumor-infiltrating neutrophils which was collected on day 16 post LLC1 injection. ( c ) Tumor volume at day 23. ( d ) The mRNA expression of tumor-infiltrating neutrophils which was collected at day 23 post LLC1 injection. * or **A statistically significant difference when compared with scramble IDO shRNA group (* P < 0.05; ** P < 0.01). IDO, Indoleamine 2,3-dioxygenase; shRNA, short hairpin RNA; TGF-β, Transforming growth factor-β. TNF-α, tumor necrosis factor-α.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Expressing, Injection

Journal: Molecular Therapy. Nucleic Acids

Article Title: Neutrophils are Essential in Short Hairpin RNA of Indoleamine 2,3- Dioxygenase Mediated-antitumor Efficiency

doi: 10.1038/mtna.2016.105

Figure Lengend Snippet: Neutrophils depletion abolished the antitumor effect of IDO shRNA. ( a ) Experimental design. 300 µg of anti-Ly6G antibody or IgG control was intraperitoneally injected at day 7, 10, and 14 after LLC1 cells injection. All mice were sacrificed at day 17. ( b ) Evaluation of depletion efficiency. The Ly6G + cells were detected in total splenocytes after injection of depletion antibodies. ( c ) Tumor volume was measured at day 17. *A statistically significant difference between IgG and Ly6G groups (* P < 0.05). IDO, Indoleamine 2,3-dioxygenase; IgG, immunoglobulin G; shRNA, short hairpin RNA.

Article Snippet: Mouse Lewis lung carcinoma cell line, LLC1 (LL/2), was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was maintained in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD) which was supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/0.1 mg/ml) (Life Technologies, Grand Island, NY).

Techniques: shRNA, Control, Injection