human t lymphoblast cell line  (ATCC)

Journal: bioRxiv

Article Title: Beta-2-microglobulin stimulates neutrophil phagocytosis of bacteria and apoptotic cells

doi: 10.1101/2025.10.30.685267

Figure Lengend Snippet: Efferocytosis of apoptotic Jurkat cells labeled with CellTrace™ Violet by CD66b-positive granulocytes stained with CellTrace™ Far Red was examined under various conditions. (A) PMNs were exposed to varying concentrations of β2m or dK58β2m in the presence of apoptotic Jurkat cells. (B) PMNs were treated with apoptotic cells alone or in combination with 50 µg/ml β2m, 50 µg/ml dK58β2m, 100 ng/ml GM-CSF, or combinations thereof. (C-D) PMNs were treated with apoptotic Jurkat cells with 50 µg/ml β2m (C) or dK58β2m (D) combined with 10 or 25% human AB serum. In all experiments, a 4:1 apoptotic cell-to-PMN ratio was used. Treatment with 100 ng/ml GM-CSF served as a positive control, and pre-incubation with 10 µg/ml cytochalasin D (cyto D) served as a negative control. Results are shown as mean ± SD of double-positive PMNs. Sample sizes were n=6 or n=4 (A), n=10 (B), and n=8 or 3 donors (C and D). Group comparisons were analyzed using mixed-effects analysis and Tukey’s multiple comparisons test (A, C, and D) or repeated-measures ANOVA and Tukey’s multiplecomparisons test(B). Statistical significance is indicated by *P < 0.05 and **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: The human T lymphoblast cell line Jurkat (TIB-152TM, ATCC®) was cultured in RPMI 1640 medium supplemented with 1% L-glutamine, 1% penicillin and streptomycin, and 10% fetal bovine serum (FBS; Corning, #35–079-CV) at 37°C with 5% CO 2 .

Techniques: Labeling, Staining, Positive Control, Incubation, Negative Control

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Flow cytometry analysis of T lymphocytes identified by the CD3 surface marker. Total leukocytes were obtained using density gradient separation with Lymphoprep and then resuspended in 1 ml of PBS. Magnetic nanoparticles conjugated with anti-CD3 were used to purify T lymphocytes using AUTOMACS equipment from Miltenyi Biotec, Inc. Total leukocytes, purified T lymphocytes and Jurkat cells were incubated with an anti-CD3 monoclonal antibody conjugated with the PerCP fluorochrome to detect the CD3 surface marker. (A) Prior to magnetic separation, 25.2% of the total leukocytes were found to be positive for the CD3 marker. (B) Following purification, the average purity of the T lymphocytes was 98.36%. (C) Jurkat cells labeled with a PerCP-conjugated anti-CD3 antibody were 84.89% positive for the CD3 surface marker.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: Flow Cytometry, Marker, Purification, Incubation, Labeling

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: One-step RT-PCR analysis of ACHE mRNA transcripts in Jurkat E6-1 leukemia cells. The total RNA isolated from Jurkat E6-1 leukemia cells was used for one-step RT-PCR with specific primers for AChE-H (487 bp), AChE-T (444 bp) and AChE-R (333 bp) variants, along with β-actin (500 bp) as a housekeeping gene control. The PCR products were separated on a 2% (w/v) agarose gel containing 0.5 µg/ml ethidium bromide and visualized under UV light. The analysis revealed the presence of AChE-H (487 bp) and AChE-T (444 bp) transcripts, whereas AChE-R was not detected. RT-PCR, reverse transcription PCR; AChE, acetylcholinesterase.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: One Step RT-PCR, Isolation, Control, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: One-step RT-PCR analysis of ACHE mRNA transcripts in human T lymphocytes and cell lines. Total RNA isolated from normal human T lymphocytes, HepG2 hepatoma cells and Jurkat E6-1 leukemia cells and was used for one-step RT-PCR. Specific primers were used for AChE-H (487 bp), AChE-T (444 bp) and AChE-R (574 bp) variants, along with β-actin (500 bp) as a housekeeping gene control. The PCR products were separated on a 2% (w/v) agarose gel containing 0.5 µg/ml ethidium bromide and visualized under UV light. Normal T lymphocytes expressed only the AChE-H transcript (487 bp), whereas Jurkat cells expressed both AChE-H (487 bp) and AChE-T (444 bp) transcripts. RNA from the HepG2 cell line was used as a positive control for AChE-R amplification, resulting in a band at 574 bp with the same primer set used for AChE-H (487 bp). β-Actin bands for each cell line were included for normalization purposes. RT-PCR, reverse transcription PCR; AChE, acetylcholinesterase.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: One Step RT-PCR, Isolation, Control, Agarose Gel Electrophoresis, Positive Control, Amplification, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Relative intensity of mRNA splicing isoforms for the ACHE gene.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques:

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Solubilized acetylcholinesterase activity of normal T lymphocytes and Jurkat cells.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: Activity Assay

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Lectin affinity chromatography analysis of AChE glycosylation in human T lymphocytes and Jurkat cells. Solubilized AChE protein extracts (S 1 +S 2 fractions) from normal human T lymphocytes and Jurkat E6-1 leukemia cells were incubated overnight at 4˚C with Sepharose-4B beads (control for nonspecific binding) or lectin-conjugated Sepharose-4B beads specific for ConA, recognizing mannose residues, LCA recognizing D-mannose bound to fucose residues, RCA, recognizing galactose or terminal sialic acid residues and WGA recognizing N-acetyl-glucosamine residues. Unbound AChE activity in the supernatants was measured via a microplate assay and expressed as a percentage of the total AChE activity in the Sepharose-4B control (100%). T lymphocytes: AChE displayed decreased binding to ConA (33%), moderate binding to LCA (66%) and WGA (63%) and increased binding to RCA (75%). Jurkat cells: AChE exhibited increased binding to ConA (66%) and LCA (72%) compared with T-lymphocytes, suggesting increased levels of mannose and fucose-associated mannose residues. Conversely, Jurkat cells showed decreased binding to RCA (67%) compared with T lymphocytes, indicating potentially reduced levels of galactose or sialic acid residues. Binding to WGA (67%) remained statistically similar (P<0.05) between both cell lines. AChE, acetylcholinesterase; ConA, Concanavalin A; LCA, Lens culinaris agglutinin; RCA, Ricinus communis agglutinin; WGA, wheat germ agglutinin.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: Affinity Chromatography, Glycoproteomics, Incubation, Control, Binding Assay, Activity Assay

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Sucrose density gradient sedimentation analysis of the AChE molecular forms in human T lymphocytes and Jurkat cells. The molecular forms of AChE were identified using gradient centrifugation with 5 and 20% sucrose solutions in Tris-HCl buffer containing NaCl, MgCl 2 and Triton X-100. Bovine liver catalase (11.4S) and bovine intestine phosphatase (6.1S) served as sedimentation standards. The gradients were subsequently centrifuged at 200,000 x g for 18 h at 4˚C. Fractions 37-40 were collected following centrifugation and sedimentation coefficients were calculated from the distance traveled by AChE compared with that traveled by standard proteins. The following marker proteins were used to determine sedimentation coefficients: Alkaline phosphatase (6.1S, F) and catalase (11.4S, C). (A) AChE protein fractions of T lymphocytes displayed sedimentation coefficients corresponding to the amphiphilic dimer G 2 A (5.2S) and the amphiphilic monomer G 1 A (3.5S). (B) AChE protein fractions of Jurkat cells revealed the presence of all three forms identified in T lymphocytes (G 2 A , 5.2S and G 1 A , 3.5S) along with an additional higher-molecular-weight form consistent with hydrophilic tetramers G 4 H (10.6S). The presence of G 4 H suggested the expression of the AChE-T transcript variant in Jurkat cells. AChE, acetylcholinesterase; A.U., arbitrary units.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: Sedimentation, Gradient Centrifugation, Centrifugation, Marker, Molecular Weight, Expressing, Variant Assay

Journal: Biomedical Reports

Article Title: Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells

doi: 10.3892/br.2024.1846

Figure Lengend Snippet: Schematic model of ACh-mediated induction of cell proliferation in Jurkat leukemia cells compared with normal cells. Normal cells : Functional AChE maintains steady-state levels of ACh via hydrolysis. ACh interacts nAChRs and mAChRs receptors at physiological levels, leading to balanced regulation of intracellular signaling pathways. Jurkat cells: Deficient AChE activity leads to increased ACh accumulation. Elevated ACh levels overstimulate nAChRs and mAChRs, resulting in the excessive activation of ChAT and PLC. PLC activation increases the Ca² + release from the endoplasmic reticulum. Increased Ca² + signaling promotes the expression of the proto-oncogene c-fos, which upregulates genes involved in cell proliferation. ACh, acetylcholine; AChE, acetylcholinesterase; nAChRs, nicotinic ACh receptor; mAChRs, muscarinic ACh receptor; ChAT, choline acetyltransferase; PLC, phospholipase C; PIP 2 , phosphatidylinositol bisphosphate; DAG, diacylglycerol; PKC, protein kinase C; IP 3 , inositol triphosphate.

Article Snippet: The human T lymphoblast cell line Jurkat E6-1 (TIB-152) was obtained from the American Type Culture Collection.

Techniques: Functional Assay, Protein-Protein interactions, Activity Assay, Activation Assay, Expressing