Journal: Cell metabolism

Article Title: CD36-mediated ferroptosis dampens intratumoral CD8 + T-cell effector function and impairs their antitumor ability

doi: 10.1016/j.cmet.2021.02.015

Figure Lengend Snippet: (A) IPA analysis of canonical pathway changes and (B) CD36 expression in tumor-infiltrating CD8+ T cells from long- and short-survival melanoma patients. (C) CD8+ T cells from MM and MGUS patient’s bone marrow analyzed for the expression of CD36. (D) CD36 expression of CD8+ T cells from MM patient’s bone marrow and blood. (E) Tumor volume of the mice and CD36 expression on tumor-infiltrating CD8+ T cells at days 7 and 14 after tumor injection. (F) Tumor burden of the mice and CD36 expression on tumor-infiltrating CD8+ T cells after Vk*MYC MM cell injection. (G and H) CD36 expression on cultured CD8+ T cells with tumor tissue (20 or 50 μl at 1 mg/ml), βCD, or cholesterol treatment. (I and J) Tumor growth curves, tumor burden at week 3, and survival of B16 bearing WT and CD36−/− B6 mice. (K and L) WT and CD36−/− B6 mice were injected with anti-CD8 antibodies 1 day before B16 tumor injection and every 3 days for a total of 5 injections. Tumor growth curves, tumor volumes at week 4, and mouse survival are shown. Differences in survival curves between the groups were analyzed by log-rank (Mantel–Cox) test. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. See also Figure S1.

Article Snippet: Cell purification and culture Murine B16 cell line was purchased from ATCC and maintained in the indicated medium.

Techniques: Expressing, Injection, Cell Culture

Journal: Cell metabolism

Article Title: CD36-mediated ferroptosis dampens intratumoral CD8 + T-cell effector function and impairs their antitumor ability

doi: 10.1016/j.cmet.2021.02.015

Figure Lengend Snippet: (A) Correlation between CD36 and cytotoxic cytokine expressions in melanoma patient’s tumor-infiltrating CD8+ T cells. (B) Expression of IFNγ and TNFα by tumor-infiltrating CD8+ T cells at days 7 and 14 after B16 tumor injection. (C-E) Tumor-infiltrating (C), spleen (D), draining lymph node and lymph node (E) CD8+ T cells were analyzed for the expression of IFNγ or TNFα at day 14 after tumor injection in WT and CD36−/− B6 mice. (F-H) Tumor-infiltrating lung (F), spleen (G), draining lymph node and lymph node (H) CD8+ T cells were analyzed for the expression of IFNγ and TNFα at day 14 after tumor injection in WT and CD36−/− B6 mice. (I) Tumor foci in the lung were counted after 2 weeks of B16 tumor injection in WT and CD36−/− B6 mice. (J) Expression of IFNγ and TNFα at day 28 by tumor-infiltrating CD8+ T cells after Vk*MYC cell injection in WT or CD36−/− B6 mice. (K) Tumor development at day 28 in WT and CD36−/− B6 mice after Vk*MYC cell injection. (L) At day 16 after lung B16 tumor inoculation, adoptively transferred WT or CD36−/− CD8+ Pmel-1 T cells in tumor, lymph nodes, and spleen were analyzed for IFNγ expression. (M) Adoptively transferred WT or CD36−/− CD8+ Pmel-1 T cells in s.c. B16 tumor were analyzed for IFNγ and TNFα expression at day 17 after tumor inoculation. (N) B16 tumor burden and survival of mice treated with WT or CD36−/− CD8+ Pmel-1 T cells. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. See also Figure S2.

Article Snippet: Cell purification and culture Murine B16 cell line was purchased from ATCC and maintained in the indicated medium.

Techniques: Expressing, Injection

Journal: Cell metabolism

Article Title: CD36-mediated ferroptosis dampens intratumoral CD8 + T-cell effector function and impairs their antitumor ability

doi: 10.1016/j.cmet.2021.02.015

Figure Lengend Snippet: Fatty acid (5 μl), RSL-3 (10 μM), or ferrostatin-1 (5 μM) was added during T cell differentiation. (A-D) Relative numbers of transferred T cells measured by flow cytometry on day 7 (A) or 14 (B) after T cell transfer. Lipid peroxidation and 7-AAD of transferred T cells were measured by flow cytometry on day 14 (B). IFNγ or Ki67 expression of transferred T cells were measured by flow cytometry on day 7 (C and D). (E and F) B16 tumor growth and survival of mice treated with adoptive transfer of control CD8+ Pmel-1 T cells, fatty acid-, RSL-3-, or ferrostatin-1- treated Pmel-1 T cells (E), and tumor growth and survival of mice treated with IgG control, anti-PD-1 antibodies, CD8+ Pmel-1 T cells, CD36−/− CD8+ Pmel-1 T cells, CD8+ Pmel-1 T cells in combination with anti-PD-1 antibodies, or CD36−/− CD8+ Pmel-1 T cells in combination with anti-PD-1antibodies (F). (G) Schematic diagram showing the mechanism underlying fatty acid-induced dysfunction in CD8+ effector T cells in TME. FA: fatty acid; Ferro: ferrostatin-1. Differences in survival curves between the groups were analyzed by log-rank (Mantel–Cox) test. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Cell purification and culture Murine B16 cell line was purchased from ATCC and maintained in the indicated medium.

Techniques: Cell Differentiation, Flow Cytometry, Expressing, Adoptive Transfer Assay, Control

Journal: Cell metabolism

Article Title: CD36-mediated ferroptosis dampens intratumoral CD8 + T-cell effector function and impairs their antitumor ability

doi: 10.1016/j.cmet.2021.02.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell purification and culture Murine B16 cell line was purchased from ATCC and maintained in the indicated medium.

Techniques: Control, Virus, Recombinant, Cell Isolation, Activation Assay, Amplex Red Cholesterol Assay, Quantitation Assay, Peroxidation Assay, Sequencing, Software