Journal: Immunotherapy Advances

Article Title: Characterization of sabatolimab, a novel immunotherapy with immuno-myeloid activity directed against TIM-3 receptor

doi: 10.1093/immadv/ltac019

Figure Lengend Snippet: ADCP activity of sabatolimab in vitro . (A) THP-1 cells (TIM-3 sufficient, control, or TIM-3 deficient, KO) were cultured for 1 hour with Raji TIM-3 o.e. cells and treated with different concentrations of sabatolimab or hIgG4 isotype control in duplicates. ADCP was determined by flow cytometric analysis as % of CFSE + CD11c + cells. Plots depict means ± SD for each condition. (B) THP-1 cells (TIM-3 sufficient, control, or TIM-3 deficient, KO) cultured as in A with with Raji hTIM-3 o.e. cells were treated with sabatolimab or hIgG4 isotype control, with Latrunculin A or Fc-blocking antibody added to some of the wells in duplicates. ADCP was determined by flow cytometric analysis as % of CFSE + CD11c + cells. Plots depict means ± SD for each condition. (C,D) Macrophages differentiated from PBMC-isolated monocytes were cultured for 4 hours with CFSE-labeled leukemia cells (HNT-34, in C or SKM-1, in D) and treated with different concentrations of sabatolimab or hIgG4 isotype control in duplicates. ADCP was determined by flow cytometric analysis as % of CFSE + CD11c + cells. Plots depict means ± SD for each condition. Two separate donors are shown for each cell line. Asterisks mark statistically significant P values between sabatolimab- and isotype control-treated samples ( T test * P < 0.05, ** P < 0.005, *** P < 0.001).

Article Snippet: The Raji human lymphoblast-like cell line, established from a Burkitt’s lymphoma of the left maxilla of an 11-year-old black male [ ], was obtained from Deutsche Sammlung von Mikrooganismen Zellkulturen (DSMZ, Braunschweig, Germany) and engineered to stably express the human TIM-3 at Trenzyme (Konstanz, Germany).

Techniques: Activity Assay, In Vitro, Control, Cell Culture, Blocking Assay, Isolation, Labeling

Journal: Genes and Environment

Article Title: Weight of evidence approach using a TK gene mutation assay with human TK6 cells for follow-up of positive results in Ames tests: a collaborative study by MMS/JEMS

doi: 10.1186/s41021-021-00179-1

Figure Lengend Snippet: Differentially expressed proteins (DEP) identified in TK6 cells exposed to 4-nitroanthranilic acid. a A total of 1078 DEPs that differentially expressed compared to control groups were identified based on a log 2 fold change of ≥2. Effects of treatment time are greater than that of dose. b A total of 168 specific DEPs that differentially expressed compared to other groups were further extracted from the 420 DEPs in 24 h 800 μg/mL group based on a log 2 fold change of ≥1

Article Snippet: The human lymphoblast cell line TK6 was purchased from the Japanese Collection of Research Bioresources cell bank and the American Type Culture Collection.

Techniques: Control

Journal: Genes and Environment

Article Title: Weight of evidence approach using a TK gene mutation assay with human TK6 cells for follow-up of positive results in Ames tests: a collaborative study by MMS/JEMS

doi: 10.1186/s41021-021-00179-1

Figure Lengend Snippet: Target gene ontology analysis for TK6 cells exposed to 4-nitroanthranilic acid for 24 h. Whereas no significance was found in GO:0006974 (cellular response to DNA damage stimulus), GO:0006281 (DNA repair), and GO:0006979 (response to oxidative stress), statistically significance was found in GO:0008631 ~ intrinsic apoptotic signaling pathway in response to oxidative stress among DEPs in 24 h 800 μg/mL group and specific DEPs in 24 h 800 μg/mL group, indicating involvement with oxidative stress

Article Snippet: The human lymphoblast cell line TK6 was purchased from the Japanese Collection of Research Bioresources cell bank and the American Type Culture Collection.

Techniques:

Journal: Genes and Environment

Article Title: Weight of evidence approach using a TK gene mutation assay with human TK6 cells for follow-up of positive results in Ames tests: a collaborative study by MMS/JEMS

doi: 10.1186/s41021-021-00179-1

Figure Lengend Snippet: Possible mechanisms of oxidative stress produced by the redox reaction of amino- and nitro compounds in human TK6 cells

Article Snippet: The human lymphoblast cell line TK6 was purchased from the Japanese Collection of Research Bioresources cell bank and the American Type Culture Collection.

Techniques: Produced

Journal: Nucleic Acids Research

Article Title: Antisense oligonucleotide-mediated correction of CFTR splicing improves chloride secretion in cystic fibrosis patient-derived bronchial epithelial cells

doi: 10.1093/nar/gkaa490

Figure Lengend Snippet: CFTR c.3718-2477C>T aberrant splicing and correction with a splice-switching antisense oligonucleotide. ( A ) Schematic of the aberrant splicing caused by the CFTR c.3718-2477C>T mutation in intron 22. The C>T mutation creates a de novo 5′ splice site resulting in the insertion into the mRNA of an 84 nucleotide pseudoexon (ψ-Ex) that has an in-frame stop codon. An ASO designed to mask the CFTR c.3718-2477C>T de novo splice site redirects splicing to the canonical splice site, restoring wild-type (WT) mRNA. ( B ) Sequence alignment of ASO-ψ to the ψ-Ex 5′ splice site found in CFTR c.3718-2477C>T. ( C ) RT-PCR analysis of CFTR splicing in a lymphoblast cell line from a CF patient homozygous for the CFTR c.3718-2477C>T mutation following transfection with vehicle only (–), a control, non-targeted ASO (ASO-C) or ASO-ψ targeting the ψ-Ex 5′ splice site. Cells were untreated (−) or treated (+) with puromycin before RNA isolation. GAPDH was analyzed as a control for total cellular RNA abundance. Splicing was quantified as the percent of RNA transcripts with ψ-Ex [(ψ-Ex/(ψ-Ex+WT))x100] and is shown below each lane.

Article Snippet: The human patient lymphoblast cell line (GM11860; Coriell Institute) was grown in RPMI media supplemented with 15% fetal bovine serum (FBS).

Techniques: Mutagenesis, Sequencing, Reverse Transcription Polymerase Chain Reaction, Transfection, Control, Isolation