Journal: Cell Reports Medicine

Article Title: METTL3 drives NAFLD-related hepatocellular carcinoma and is a therapeutic target for boosting immunotherapy

doi: 10.1016/j.xcrm.2023.101144

Figure Lengend Snippet:

Article Snippet: Human CD8 + T cells were co-cultured with conditional medium collecting from different NASH-HCC cells, with human T-activation CD3/CD28 Dynabeads (Invitrogen) and human IL-2 (R&D Systems, 10ng/ml) for 72 hours, with or without cholesterol (Sigma), cholesterol ester (Sigma) or β- cyclodextrin (Sigma).

Techniques: Control, Recombinant, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Cell Isolation, Magnetic Beads, Transfection, Red Blood Cell Lysis, Staining, Reporter Assay, RNA Sequencing, Gene Expression, Plasmid Preparation, Software

Journal: Cell chemical biology

Article Title: Dual Modifications of an α-Galactosyl Ceramide Synergize to Promote Activation of Human Invariant Natural Killer T Cells and Stimulate Anti-Tumor Immunity

doi: 10.1016/j.chembiol.2018.02.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Besra, University of Birmingham, UK https://avantilipids.com/product/867000/ AH10-7 glycolipid This paper N/A AH03-1 glycolipid Ndonye et al., 2005 N/A AH15-1 glycolipid This paper N/A DB03-4 glycolipid Dr. G. S. Besra, University of Birmingham, UK N/A PBS-44 glycolipid Dr. P. Savage, Brigham Young University, USA N/A Insect-XPRESSTM Medium Lonza http://www.lonza.com/products-services/bio-research/cell-culture-products/specialty-media/insect-xpress-protein-free-insect-cell-medium-with-l-glutamine.aspx Fetal Calf Serum Atlanta Biologicals Cat# {"type":"entrez-protein","attrs":{"text":"S11150","term_id":"98016","term_text":"pir||S11150"}} S11150 Triton X-100 Sigma Cat# T8787 Tween-20 ThermoFisher Cat# BP337-500 Tyloxapol Sigma Cat# T8761-50G Dimethyl sulfoxide (DMSO) Sigma Cat# D8418-100ML Recombinant human IL-7 R&D Systems Cat# 207-IL Recombinant human IL-2 Peprotech Cat# 200-02 Recombinant human IFNγ ThermoFisher Scientific Cat# RIFNG50 Recombinant mouse IL-2 R&D Systems Cat# 402-ML-100 Recombinant 2C12 NKT TCR This paper N/A Recombinant mouse CD1d (mCD1d) This paper N/A HisPurTM Ni-NTA affinity resin ThermoFisher Scientific Cat# 88223 Superdex 200 16/60 size exclusion column GE Healthcare Cat# 28989335 MonoQ GL anion exchange column GE Healthcare Cat# 17-5166-01 PEG 3350 Sigma-Aldrich Cat# 202444 Tacsimate Hampton Research Cat# HR2-825 Carboxyfluorescein succinimidyl ester (CFSE) ThermoFisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C34554","term_id":"2370695","term_text":"C34554"}} C34554 Mouse CD1d tetramer, αGalCer-loaded This paper N/A Critical Commercial Assays Mouse CBA Th1/Th2/Th17 Cytokine kit BD Biosciences Cat# 560485 Deposited Data Crystal structure 2C12 TCR-CD1d-AH10-7 This paper –structure deposited PDB code: 6BNL Crystal structure 2C12 TCR-CD1d-‹-GalCer This paper – structure deposited PDB code: 6BNK Crystal structure mouse CD1d-NU-alpha-GalCer-iNKT TCR complex Aspeslagh et al., 2011 PDB code: 3QUZ Crystal structure human NKT TCR-CD1d-4′deoxy-alpha- galactosylceramide ternary complex Wun et al., 2012 PDB code: 3VWK Crystal structure of CD1d-lipid-antigen complexed with Beta-2-Microglobulin, NKT15 Alpha-Chain and NKT15 Beta-Chain Borg et al., 2007 PDB code: 2PO6 Structure of the mouse CD1d-4ClPhC-alpha-GalCer- iNKT TCR complex Aspeslagh et al., 2013 PDB code: 4IRJ Experimental Models: Cell Lines SF9 insect cells ThermoFisher Scientific Cat# 12659017 HighFiveTM insect cells ThermoFisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"B85502","term_id":"2926634","term_text":"B85502"}} B85502 DN3A4-1.2 mouse iNKT cell hybridoma Dr. M. Kronenberg N/A BW5147α-β-, mouse thymoma line Dr. S. Behar, University of Massachusetts, USA Letourneur F & Malissen B. Eur J Immunol.

Techniques: Enzyme-linked Immunosorbent Assay, Virus, Recombinant, Transfection, Plasmid Preparation, Software

Journal:

Article Title: Induction of T Helper Type 1 Responses by a Polysaccharide Deacetylase from Cryptococcus neoformans

doi: 10.1128/IAI.71.9.5412-5417.2003

Figure Lengend Snippet: rd25-induced IFN-γ and IL-2 production in cultures of splenocytes from rd25-immunized or C. neoformans-infected mice. Spleens were collected at 7, 14, and 28 days after immunization or infection (see the legend to Fig. ​Fig.1),1), and splenocytes (5 × 106/ml) were stimulated with CneF (10 μg/ml), rd25 (10 μg/ml), or PBS. Cytokine levels were assayed at 24 h (IL-2) or 48 h (IFN-γ) after the addition of the stimuli. Data are expressed as means ± SEM for three experiments, each performed in triplicate. Asterisk, P < 0.05, relative to results with cells from PBS-CFA-injected mice by one-way analysis of variance and the Student-Keuls-Newman test.

Article Snippet: To assess the role of IL-2, groups of d25-immunized and nonimmunized mice were treated with polyclonal goat anti-mouse IL-2 IgG (R&D Systems) or normal goat IgG and challenged with C. neoformans (7.5 × 10 3 CFU).

Techniques: Infection, Injection

Journal:

Article Title: Induction of T Helper Type 1 Responses by a Polysaccharide Deacetylase from Cryptococcus neoformans

doi: 10.1128/IAI.71.9.5412-5417.2003

Figure Lengend Snippet: rd25-induced IFN-γ and IL-2 production in cultures of CD4+ or CD8+ cells from C. neoformans-infected mice. Spleens were collected at 14 days after infection, and CD4+ or CD8+ T cells (5 × 106/ml), separated by means of a panning procedure (16), were cultured in the presence of splenic adherent cells (5 × 105/ml) and CneF (10 μg/ml), d25 (10 μg/ml), or PBS. Cytokine levels were assayed as indicated in the legend to Fig. ​Fig.2.2. Data are expressed as means ± SEM for three experiments, each performed in triplicate. Asterisk, P < 0.05, relative to results with cells from PBS-CFA-injected mice by one-way analysis of variance and the Student-Keuls-Newman test.

Article Snippet: To assess the role of IL-2, groups of d25-immunized and nonimmunized mice were treated with polyclonal goat anti-mouse IL-2 IgG (R&D Systems) or normal goat IgG and challenged with C. neoformans (7.5 × 10 3 CFU).

Techniques: Infection, Cell Culture, Injection

Journal:

Article Title: Induction of T Helper Type 1 Responses by a Polysaccharide Deacetylase from Cryptococcus neoformans

doi: 10.1128/IAI.71.9.5412-5417.2003

Figure Lengend Snippet: Role of IL-12 and TNF-α in the development of cytokine responses to d25. Spleens were collected at 7 and 14 days after immunization with rd25 (100 μg per mouse in CFA), and cultures of splenocytes (5 × 106/ml) were stimulated with rd25 (10 μg/ml). Cytokine levels were assayed at 24 h (IL-2) or 48 h (IFN-γ) after the addition of the stimulus. Experimental groups included immunized mice that were injected i.p. with 0.2 mg of goat polyclonal anti-mouse TNF-α IgG, goat anti-IL-12 IgG, or normal goat IgG at 1 h prior to infection. Data are expressed as means ± SEM for three experiments, each performed in triplicate. Asterisk, significantly (P < 0.05) different from normal IgG control group by one-way analysis of variance and the Student-Keuls-Newman test.

Article Snippet: To assess the role of IL-2, groups of d25-immunized and nonimmunized mice were treated with polyclonal goat anti-mouse IL-2 IgG (R&D Systems) or normal goat IgG and challenged with C. neoformans (7.5 × 10 3 CFU).

Techniques: Injection, Infection

Journal:

Article Title: Induction of T Helper Type 1 Responses by a Polysaccharide Deacetylase from Cryptococcus neoformans

doi: 10.1128/IAI.71.9.5412-5417.2003

Figure Lengend Snippet: Role of IL-2 and IFN-γ in the protective effects of d25 immunization. Panels A and B show the effects of, respectively, IL-2 blockade and IFN-γ deficiency on C. neoformans-induced lethality in d25-immunized and unimmunized mice. (A) Eight-week-old female BALB/c mice were immunized s.c. with rd25 (100 μg per mouse in CFA) or injected with PBS-CFA. Mice belonging to each of these two subgroups were injected i.p. at the time of immunization and 15 days later with 0.5 mg of goat polyclonal anti-mouse IL-2 IgG or normal goat IgG. All mice were challenged i.v. with 7.5 × 103 viable C. neoformans cells (strain H99) at 1 week after immunization, and lethality was observed daily for 30 days. (B) Eight-week-old female IFN-γ−/− mice on a C57BL/6 background and control WT C57BL/6 mice were immunized with d25 in CFA or injected with PBS-CFA as described above. After 1 week, mice were challenged i.v. with 3 × 104 viable C. neoformans cells (strain H99), and lethality was observed daily for 30 days. Survival data were analyzed with Kaplan-Meier survival plots followed by the log rank test (JMP Software; SAS Institute, Cary, N.C.) on an Apple Macintosh computer.

Article Snippet: To assess the role of IL-2, groups of d25-immunized and nonimmunized mice were treated with polyclonal goat anti-mouse IL-2 IgG (R&D Systems) or normal goat IgG and challenged with C. neoformans (7.5 × 10 3 CFU).

Techniques: Injection, Software