d1mt  (GL Biochem)

Journal: OncoTargets and therapy

Article Title: CXCR4 antagonism in combination with IDO1 inhibition weakens immune suppression and inhibits tumor growth in mouse breast cancer bone metastases

doi: 10.2147/OTT.S200643

Figure Lengend Snippet: Combination treatment (ie, combo in the figures) with AMD3465 and D1MT significantly increased survival in mice bearing metastasized breast cancer. ( A ) The survival curves of mice bearing spontaneously metastasized breast cancer. ( B ) The survival curves of mice with intratibial injected breast cancer cells. n=10/group. * P <0.05, ** P <0.01, *** P <0.001, and **** P <0.0001.

Article Snippet: The IDO1 inhibitor D1MT was purchased from Sigma Aldrich.

Techniques: Injection

Journal: OncoTargets and therapy

Article Title: CXCR4 antagonism in combination with IDO1 inhibition weakens immune suppression and inhibits tumor growth in mouse breast cancer bone metastases

doi: 10.2147/OTT.S200643

Figure Lengend Snippet: The anticancer effects of combination treatment (combo) relied on the function of CXCR3+ CD8+ T-cells. ( A ) Combination treatment (combo) significantly prolonged survival, while CXCR3 antagonist AMG487 could abrogate these effects. ( B ) In conventional metastasis models, AMG487 could also abolish the anticancer effects of combination treatment with AMD3465 and IDO1 inhibitor. **** P <0.0001.

Article Snippet: The IDO1 inhibitor D1MT was purchased from Sigma Aldrich.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: The amino acid sensor GCN2 inhibits inflammatory responses to apoptotic cells promoting tolerance and suppressing systemic autoimmunity

doi: 10.1073/pnas.1504276112

Figure Lengend Snippet: GCN2 activation promotes apoptotic cell-driven IL-10 production in MΦs. (A and B) BM MΦs were transduced with IDO1-GFP+ or control lentivirus as described in Materials and Methods. Then, 24 h later, kynurenine and Trp in the culture supernatant mRNA for the indicated genes were measured by sqPCR. (C) At 24 h after lentiviral transduction as described in A, BM MΦs were incubated with apoptotic thymocytes at a 10:1 apoptotic cell:MΦ ratio. Then, 18 h later, cell supernatants were collected, and cytokines were measured by ELISA. (D) Western blot analysis for the proteins indicated was done on whole-cell lysates from BM MΦ cultures as described in C. (E) mRNA from BM MΦ/apoptotic cell cultures as in C was collected at the indicated time points, and cytokine message expression was assessed by sqPCR. In some groups, 20 μM D1MT was added. (F) Sucrose gradient fractionation of cellular lysates was followed by sqPCR analysis for the indicated mRNA species from 8-h BM MΦ/apoptotic cultures treated as in C. In A, B, C, and E, bars and line points represent mean ± SD values for triplicate samples. In D, Western blot images are representative of three separate experiments. In F, pooled material from triplicate samples was analyzed. *P < 0.05; **P < 0.01, Student’s t test. Experiments were repeated three times, with similar results.

Article Snippet: D1MT (Sigma-Aldrich) was prepared as described previously ( 26 ).

Techniques: Activation Assay, Transduction, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Fractionation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: The amino acid sensor GCN2 inhibits inflammatory responses to apoptotic cells promoting tolerance and suppressing systemic autoimmunity

doi: 10.1073/pnas.1504276112

Figure Lengend Snippet: IDO1 expression decreases RNA association with polysomes in vitro, and apoptotic cell challenge induces IDO1- and GCN2-dependent stress gene expression in MZ MΦs and CD8α+ DCs in vivo. (A) Polysome analysis of IDO1+ MΦs incubated with apoptotic cells as described in Fig. 1. Cell extracts were subjected to sucrose density centrifugation, and RNA association with polysomes was analyzed by measuring the absorbance at 254 nM. Ribosomal peaks at 40 s, 60 s, and 80 s are identified. (B) At 4 h after apoptotic cell challenge (107 in vivo), SignR1+ MZ MΦs and CD11c+CD8α+ DCs were sorted by FACS from the spleen, and message expression for the indicated mRNA species was determined by sqPCR. Bars represent the relative induction vs. controls value for pooled samples from four mice. In one experimental condition, mice were given the IDO1 inhibitor D1MT ad libitum in drinking water as described in SI Materials and Methods. The experiment was repeated four times, with similar results.

Article Snippet: D1MT (Sigma-Aldrich) was prepared as described previously ( 26 ).

Techniques: Expressing, In Vitro, In Vivo, Incubation, Centrifugation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: The amino acid sensor GCN2 inhibits inflammatory responses to apoptotic cells promoting tolerance and suppressing systemic autoimmunity

doi: 10.1073/pnas.1504276112

Figure Lengend Snippet: IDO1 activity at the time of apoptotic cell exposure is required for generation of stable tolerance and allograft acceptance. Female B6 mice were given the IDO1 inhibitor D1MT ad libitum in drinking water for 2 d before administration of 107 male B6 thymocytes in vivo. Mice were then immediately taken off the D1MT containing water, followed by male skin engraftment 7 d later (as described in SI Materials and Methods), and monitored for graft rejection for 50 d. The experiment was repeated twice, with similar results. n = 10 mice/group.

Article Snippet: D1MT (Sigma-Aldrich) was prepared as described previously ( 26 ).

Techniques: Activity Assay, In Vivo

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: The amino acid sensor GCN2 inhibits inflammatory responses to apoptotic cells promoting tolerance and suppressing systemic autoimmunity

doi: 10.1073/pnas.1504276112

Figure Lengend Snippet: Autoimmunity is accelerated in myeloid GCN2KO mice exposed to apoptotic cells. (A) 3-mo-old mice of the indicated genotype were injected with 107 B6 apoptotic cells once weekly (four times total). Serum was collected at the indicated time points, and concentrations of total IgG reactive against dsDNA as a marker of systemic autoimmunity were determined by ELISA. (B) R2B and R2B GCN2flLysMcre mice were given HF injections i.p. as described in SI Materials and Methods for a period of 2 mo, after which serum was collected and IgG reactivity to dsDNA was measured by ELISA. In one experimental condition, mice were given the IDO1 inhibitor D1MT ad libitum in drinking water and examined for the impact on autoimmune disease with and without HF treatment. For both experiments, n = 7 mice/group. *P < 0.05, **P < 0.01, Student’s t test. ns, not significant. Experiments were repeated three times, with similar results.

Article Snippet: D1MT (Sigma-Aldrich) was prepared as described previously ( 26 ).

Techniques: Injection, Marker, Enzyme-linked Immunosorbent Assay