human b lymphoblastoid cell line tk6  (ATCC)

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Clinical and Experimental Medicine

Article Title: Identification of plasma exosome circRNA as potential novel biomarkers for DLBCL and circrna-miRNA-mRNA network analysis

doi: 10.1007/s10238-025-02019-w

Figure Lengend Snippet: Candidate circRNA Screening and Validation a: Plasma exosomal expression profiles of candidate circRNAs in the pilot validation cohort ( n = 16 per group). Eight circRNAs exhibited consistent trends with microarray data, with hsa_circRNA_400230 ( P = 0.003) and hsa_circRNA_001393 ( P = 0.007) showing statistically significant differential expression. Two circRNAs (hsa_circRNA_100994 and hsa_circRNA_404762) were excluded due to low abundance (Ct > 35). b ༚Differential expression of candidate circRNAs in DLBCL (OCI-LY3) and normal B-lymphoblastoid (Hmy2.CIR) cell lines. hsa_circRNA_001393 ( P = 0.011) and hsa_circRNA_404447 ( P = 0.005) were significantly upregulated in OCI-LY3 cells, aligning with plasma exosomal trends. Remaining candidates were excluded due to discordant expression or high Ct values. Data are presented as mean ± SD.* p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human B-lymphoblastoid cell line Hmy2.CIR (Procell Life Science & Technology Co., Ltd., China) and the diffuse large B-cell lymphoma (DLBCL) cell line OCI-LY3 (BeNa Culture Collection, China) were cultured in complete growth media (Hmy2.CIR: Iscove’s Modified Dulbecco’s Medium [IMDM] supplemented with 20% fetal bovine serum [FBS]; OCI-LY3: Roswell Park Memorial Institute 1640 [RPMI-1640] medium containing 20% FBS).

Techniques: Biomarker Discovery, Clinical Proteomics, Expressing, Microarray, Quantitative Proteomics

Journal: Cell Reports Methods

Article Title: MR1-ligand cross-linking identifies vitamin B6 metabolites as TCR-reactive antigens

doi: 10.1016/j.crmeth.2025.101120

Figure Lengend Snippet: The B6 vitamers pyridoxal and PLP activate Jurkat cells expressing the A-F7 MAIT TCR and the MC.7.G5 TCR, as well as primary CD8 + T cells expressing the A-F7 MAIT TCR (A) Jurkat cells with no TCR or transduced with A-F7 MAIT TCR were co-incubated overnight with A549 WT and A549 MR1 KO cell lines treated with pyridoxal at 100, 10, and 1 μg/mL or loaded with M. smegmatis (MOI: 1:300). Cells were stained for CD69 expression with mean fluorescence intensity (MFI) displayed. Background MFI of Jurkat cells with A549 WT or MR1 KO alone with no pyridoxal or M. smegmatis was subtracted. Jurkat cells with A-F7 were gated on co-marker rCD2 + . Data display duplicate conditions ( E). (B) Jurkat cells with no TCR or transduced with A-F7 MAIT TCR were co-incubated overnight with A549 WT and the following compounds: 5-A-RU (converts to MAIT ligand 5-OP-RU in cells and was added in the absence of exogenously applied methylglyoxal, which increases potency), pyridoxal and PLP at 100, 10, 1, 0.1, 1 × 10 −2 , 1 × 10 −3 , and 1 × 10 −4 μg/mL. Cells were stained for CD69 expression with MFI displayed. Background MFI of Jurkat cells with A549 cells alone with no pyridoxal was subtracted. Jurkat cells expressing the A-F7 TCR were gated on the rCD2 co-marker. Assay was performed in triplicate ( F), and curves were fitted using a four-parameter logistic model. Points indicate mean values, with error bars depicting standard deviation. EC 50 values with a 95% confidence interval (CI) and R 2 are indicated, with the results reproducible over two assays ( G). (C) Primary CD8 + T cells from three healthy donors with no TCR transduction or expression of the A-F7 TCR to generate TCR-T cells, were co-incubated for 4 h with A549 WT cells ± pre-treatment with 100 μg/mL of pyridoxal, and then reactivity measured via T107 assay. T cells were also incubated alone or with CD3/CD28 Dynabeads, with the latter acting as a positive control. Cells were gated on lymphocytes, viable CD3 + , single cells, rCD2 + /CD8 + (or CD8 + for the untransduced), and then TNF + versus CD107a + for reactivity. For the pyridoxal conditions, background reactivity toward A549 cell lines with no pyridoxal has been subtracted. For reactivity toward CD3/CD28 Dynabeads, the reactivity for the T cell-alone condition has been subtracted ( H). (D) Jurkat cells with no TCR or transduced with MC.7.G5 TCR were co-incubated overnight with C1R cells ± pyridoxal at 100, 10, 1, 0.1, and 1 × 10 −2 μg/mL. Cells were stained for CD69 expression with MFI displayed. Background MFI of Jurkat cells alone with no pyridoxal was subtracted. Jurkat cells with MC.7.G5 TCR were gated on co-marker rCD2 + . Assay was performed in triplicate ( I), and curves were fitted using a four-parameter logistic model. Points indicate mean values, with error bars depicting standard deviation. EC 50 values with a 95% CI and R 2 are indicated ( J). See also and .

Article Snippet: B-cell lymphoblastoid cell line C1R (ATCC information, CRL-1993) were sourced locally and engineered to overexpress MR1∗01 using lentivirus.

Techniques: Expressing, Transduction, Incubation, Staining, Fluorescence, Marker, Standard Deviation, Positive Control

Journal: The Journal of Experimental Medicine

Article Title: A CARMIL2 gain-of-function mutation suffices to trigger most CD28 costimulatory functions in vivo

doi: 10.1084/jem.20250339

Figure Lengend Snippet: Effects of the CARMIL2 QE mutation on physiological Jurkat T cell activation. (A) WT, CARMIL2 OST , and CARMIL2 QE-OST Jurkat T cells were analyzed by flow cytometry for the expression of CD3 and CD28 (shaded histograms). Dashed line curves correspond to isotype-matched control antibodies (negative controls), and data are representative of two independent experiments. (B) WT, CARMIL2 OST , and CARMIL2 QE-OST Jurkat T cells were left untreated (−) or stimulated (+) with anti-CD3 plus anti-CD28 for 2 and 5 min at 37°C. Immunoblot analysis of equal amounts of TL of the specified cells probed with anti-CARMIL2, anti-CARD11, and an anti-ZAP70 loading control. Data are representative of two independent experiments. Left margin, molecular size in kilodaltons. (C) Quantitation of the immunoblot analysis shown in B. Bars represent normalized CARMIL2-ZAP-70 and CARD11-ZAP70 ratios (see Materials and methods). Data in C and E are presented as the mean ± SE. (D) WT, CARMIL2 OST , and CARMIL2 QE-OST Jurkat T cells were activated as in B, and immunoblot analysis of equal amounts of lysates from the specified cells subjected to AP on Strep-Tactin Sepharose beads, followed by elution of proteins with D-biotin, and probed with anti-CARMIL2 or anti-CARD11. Data are representative of two independent experiments. Left margin, molecular size in kilodaltons. (E) Quantitation of the immunoblot analysis shown in D. Bars represent normalized CARD11-CARMIL2 ratios. (F) CARMIL2 OST and CARMIL2 QE-OST Jurkat T cells were left untreated (−) or stimulated with anti-CD3 (+) in the presence (+) or absence (−) of anti-CD28 for 2 min at 37°C. Immunoblot analysis of equal amounts of lysates from the specified cells subjected to AP on Strep-Tactin Sepharose beads, followed by elution of proteins with D-biotin, and probed with anti-CARMIL2 or anti-CARD11. Data are representative of two independent experiments. Left margin, molecular size in kilodaltons. (G) WT, CARMIL2 OST , and CARMIL2 QE-OST Jurkat cells were stimulated with Raji cells that were preincubated in the absence (0) or presence of the specified concentrations of SEE. For each condition, the MFI of CD69 + cells was measured by flow cytometry 24 h after stimulation. Numbers on the y axis correspond to the MFI of CD69 + cells. Error bars correspond to the mean and SD. Data are representative of two independent experiments. (H) IL-2 production by CARMIL2 OST and CARMIL 2 QE-OST Jurkat T cell clones stimulated with Raji cells alone (0) or in the presence of 0.05, 0.1, and 0.5 and 1 ng/ml SEE. Analysis of IL-2 production was performed 24 h after stimulation. The expression of IL-2 (pg/ml) is shown using boxplot with the median, boxed interquartile range, and whiskers extending to the most extreme point up to 1.5 times the interquartile range. Data are representative of two independent experiments, involving eight independent clones of Carmil2 OST and Carmil2 Q539E- OST Jurkat cells. Each dot corresponds to a clone of the specified Jurkat T cells. **P < 0.01, ***P ≤ 0.001, and ns, nonsignificant; unpaired Student’s t test. AP, affinity purification; TL, total lysates; MFI, mean fluorescence intensity. Source data are available for this figure: SourceData F1 .

Article Snippet: The Jurkat human leukemic T cell line and Raji lymphoblastoid B cell line originated from the American Type Culture Collection and were provided by A. Weiss (University of California, San Francisco, San Francisco, CA, USA).

Techniques: Mutagenesis, Activation Assay, Flow Cytometry, Expressing, Control, Western Blot, Quantitation Assay, Clone Assay, Affinity Purification, Fluorescence