jurkat cells tib 152  (ATCC)

Journal: Frontiers in Immunology

Article Title: Split intein-mediated reconstitution of regulatable CAR T-cells

doi: 10.3389/fimmu.2026.1844972

Figure Lengend Snippet: IMPDH.1 N -CD1a scFv is functional and competent for PTS. (A) CD1a + MOLT-4 cells were incubated with IMPDH.1 N -CD1a scFv, and polypeptide attachment was detected by staining with an anti-His-tag antibody. (B) PTS activity of IMPDH.1 N -CD1a scFv was initially assessed using a model Int C partner designed to transfer a variable heavy domain of a heavy chain (VHH) onto the CD1a scFv upon successful trans-splicing. Upper: Schematic representation of the reaction: His-tagged IMPDH.1 N -CD1a scFv (MW = 39,6 kDa) + small ubiquitin-like modifier (SUMO)-His-IMPDH.1 C -VHH (MW = 33 kDa) → CD1a scFv-VHH (MW = 40,5 kDa; red asterisk) + His-tagged IMPDH.1 N (MW = 13,2 kDa; black asterisk) + SUMO-His-IMPDH.1 C (MW = 19 kDa; black asterisk). Reactants (green box); products (blue boxes). Scheme designed with FigureLabs™. To specifically identify the trans-splicing product (TSP), the reaction mixture was applied to a Ni resin (IMAC). Because the TSP lacks a His-tag, it does not bind the resin, like the other reaction components do. Bottom: Coomassie-stained SDS-PAGE gel showing reaction products obtained from the PTS assay: I N : His-tagged IMPDH.1 N -CD1a scFv; I C : SUMO-His-IMPDH.1 C -VHH; t 0 : initial time; 1h and 3h refer to the reaction time; FT: flow-through; R: Ni resin-bound proteins. The anticipated molecular weights (MW) are specified in parentheses. (C) JURKAT-cells transduced with IMPDH.1 C - t -CAR (J-IMPDH.1 C , upper ) or NRDJ.1 C - t -CAR (J-NRDJ.1 C , bottom ). Transduction efficacy was determined by GFP reporter and anti-His-tag antibody. (percentage shown in the inset). (D, E) JurkaT-cells were used for preliminary technical optimization of the conditions required for split-intein-mediated protein trans-splicing. These experiments were exploratory and intended to identify conditions for subsequent validation in primary human T-cells; therefore, the Jurkat optimization data were not subjected to inferential statistical analysis. Cells were subjected to PTS with IMPDH.1 N -CD1a scFv under reducing conditions (+DTT). PTS occurred efficiently in J-IMPDH.1 C cells (D) but not in J-NRDJ.1 C cells (E) , in a concentration-dependent manner after 3h of incubation. Green denotes the CD1a scFv-negative cell population, whereas cyan blue represents the CD1a scFv-positive population detected using anti-scFv Ab (percentage shown in the inset).

Article Snippet: The human T-ALL cell lines JURKAT and MOLT-4 cells were purchased from the DSMZ cell line bank (Braunschweig, Germany) and cultured in RPMI-1640 supplemented with 10% heat-inactivated FBS.

Techniques: Functional Assay, Incubation, Staining, Activity Assay, Ubiquitin Proteomics, SDS Page, Transduction, Biomarker Discovery, Analysis, Concentration Assay

Journal: Toxicological Sciences

Article Title: Nonclinical safety assessment of SBT777101, a CAR-engineered regulatory T cell therapy for autoimmune diseases

doi: 10.1093/toxsci/kfag125

Figure Lengend Snippet: Selective activation of SBT777101 Jurkat reporter cells by citrullinated proteins. Representative dose–response curves showing fold-increase in luminescence ( y -axis) relative to baseline following incubation with increasing concentrations (log scale, x -axis) of citrullinated (red) or noncitrullinated (black) protein targets. Data points represent the mean ± SD of 3 replicates.

Article Snippet: NFAT-luciferase reporter Jurkat cells (BPS Bioscience) were stably transduced with the SBT777101 CAR.

Techniques: Activation Assay, Incubation