cd38 cells  (Elabscience Biotechnology)

Journal: Molecular Therapy Oncology

Article Title: A measles virus encoding a CD19/CD3 bispecific T cell engager shows enhanced preclinical anti-BCP-ALL efficacy without significant toxicity

doi: 10.1016/j.omton.2026.201127

Figure Lengend Snippet: MV-Blina infected cells secrete target-binding secBlina, which initiates ALL cell kill more efficiently than CD19/CD3-bsTE (A) Vero cells secrete secBlina. secBlina was harvested from the supernatant of infected Vero cells, followed by purification and concentration. Eluate (2 μg protein) was detected via His-tag by Western Blot in comparison to the commercial CD19/CD3-bsTE (1.8 μg). (B) Purified and concentrated secBlina binds specifically to target cells. Binding to target cells was confirmed by flow cytometry using CD3 − CD19 + REH, CD3 + CD19 - Jurkat, and CD3 − CD19 - K562 cell lines incubated with secBlina. Binding of secBlina to target cells was detected by flow cytometry via a His-tag antibody. (C) secBlina strongly binds to REH target cells. The REH cell line was first incubated with 1 μg secBlina. Increasing concentrations of CD19/CD3-bsTE were then added to displace cell-bound secBlina. Cell-bound secBlina was detected via an HA tag antibody by flow cytometry. (D) T cells in human PBMCs are activated by secBlina. Pooled PBMCs were stimulated with an (E) T cells are activated by secBlina during co-culture of PBMCs with REH cells. Pooled PBMCs were co-cultured with REH cells in a 1:1-ratio for 24 h while being treated with secBlina or the CD19/CD3-bsTE. Activated CD2 + CD69 + T cells were detected by flow cytometry. (F) MV-Blina infected leukemia cells produce and secrete secBlina. REH and Jurkat cells are shown 72 h post MV infection or Opti-MEM control. secBlina was detected by His-tag IF staining. Scale bars 50 μm. (G) secBlina effectively induces REH target cell kill compared to CD19/CD3-bsTE, whereas K562 control cells are unaffected. Target (T) cells REH (left panels) and negative control cells K562 (right panels) were incubated with secBlina (green) or CD19/CD3-bsTE (blue) for 24 h at indicated concentrations in the presence of PBMCs (effector; E) at different E:T ratios. Specific cell kill was determined by flow cytometry. Results in A, B, C, and F are shown as representatives of three independent experiments. Data are shown as mean ± SD of n = 3 (D, E) or n = 5 (G) independent experiments. Statistical analysis was performed using two-way ANOVA with Dunnett’s correction. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; and ∗∗∗∗, p < 0.0001.

Article Snippet: For the displacement assay, 1 × 10 6 REH cells were incubated with 1 μg secBlina for 30 min, followed by increasing concentrations of a research-grade analog of the CD19/CD3-bsTE blinatumomab (BPS Bioscience) for an additional 30 min, all at 4°C. secBlina bound to target cells was detected using rabbit anti-HA antibody (clone EPR22819-101, 1:600, ab256483, Abcam) for 30 min at 4°C, followed by a goat-anti-rabbit IgG (H&L) AF488 (1:2000, ab150077, Abcam).

Techniques: Infection, Binding Assay, Purification, Concentration Assay, Western Blot, Comparison, Flow Cytometry, Incubation, Co-Culture Assay, Cell Culture, Control, Staining, Negative Control

Journal: Nature Immunology

Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

doi: 10.1038/s41590-025-02265-w

Figure Lengend Snippet: a , Schematic of the experimental design for de novo generation of human T-ALL. Sublethally irradiated NSG mice were subjected to i.v. injection with human cord blood (CB) CD34 + HPCs (1–1.5 × 10 5 cells per mouse) retrovirally transduced with either ICN1 along with GFP, or GFP alone as the control. b , Representative flow cytometry expression of CD4, CD8, CD3, TCRαβ and TCRγδ by ICN1 + human T-ALL cells infiltrating the bone marrow of diseased mice in a at 27 weeks after transplant. c , Representative CD3 versus TCRαβ expression of human ICN1 + T-ALL cells recovered from diseased mice in a (primary), or from NSG mice after serial transplantation with bone marrow from primary mice (1st transfer and 2nd transfer). Results correspond to T-ALL cells isolated from the bone marrow at 39, 8 and 9 weeks after transplant, respectively. d , Mean percentages ± s.e.m. of human CD8 + CD4 + double-positive, CD3 lo TCRαβ neg ICN1 + cells infiltrating the bone marrow of either primary diseased mice, or serially transplanted mice (nine mice per group from two independent experiments) as in c . Data were analyzed by one-way analysis of variance (ANOVA) with Kruskal–Wallis Dunn’s multiple-comparisons test. **** P < 0.0001.

Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

Techniques: Irradiation, Injection, Transduction, Control, Flow Cytometry, Expressing, Transplantation Assay, Isolation

Journal: Nature Immunology

Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

doi: 10.1038/s41590-025-02265-w

Figure Lengend Snippet: ( A ) Representative pseudocolour plots of CD3 vs pTα expression in pre-TCR-expressing SupT1, and TCRαβ-expressing HPB-ALL and Jurkat human cell lines from three independent experiments, as assessed by reactivity with anti-CD3ε and anti-pTα monoclonal antibodies (left biparametric histograms). Representative monoparametric histograms on the right show surface pre-TCR expression levels analysed with anti-pTα monoclonal antibody plus goat anti-mouse FITC. Numbers indicate percentages of positive cells. Shaded histograms show background staining with an irrelevant isotype-matched antibody. ( B ) Representative CD3 vs TCRαβ expression of either primary human thymocytes from a single 17-weeks-old fetal thymus sample (upper left), or developing T cells derived either from two independent human CB HPC samples after 70 days of culture onto OP9-Jag2 cells (middle left), or from a single CD34 + human ETP sample after 18 days of culture in ATOs (bottom left). Pre-TCR expression levels of electronically gated CD3 lo TCRαβ neg , TCRαβ + and CD3 neg cells analysed with anti-pTα mAb plus biotin-coupled anti-mouse (Fab’)2 plus BV421-coupled streptavidin are shown in the monoparametric histograms on the right. Numbers indicate percentages of positive cells relative to background staining with an isotype-matched mAb.

Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

Techniques: Expressing, Bioprocessing, Staining, Derivative Assay

Journal: Nature Immunology

Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

doi: 10.1038/s41590-025-02265-w

Figure Lengend Snippet: ( A ) Immunoblot analysis of CMS expression in SupT1 cells that were either transfected with several shRNAs (shCMS1-5) specific for CD2AP , the gene encoding CMS, or left untransfected (NT) (left panel). Relative CMS expression normalized to α-tubulin expression used as loading control is shown in the right panel. ( B ) Immunoblot analysis of CMS expression (upper) and PLCγ tyrosine (Tyr) phosphorylation (bottom) in JR.pTα pre-TCR + cells transduced either with shCMS4 or with shSC as control, prior to activation with an anti-CD3ε monoclonal antibody for the indicated times. PLCγ Tyr-phosphorylation was analysed after immunoprecipitation with anti-PLCγ1 monoclonal antibody by probing with an anti-Y783-PLCγ1 antibody. Tubulin expression was analysed as loading control. ( C ) Relative NFAT activity of JR.pTα cells transduced with either shCMS4 or shSC as control, analysed upon stimulation with an anti-CD3ε monoclonal antibody by a luciferase assay. Data are shown as mean percentages ± SEM of NFAT activity relative to activated shSC cells analysed by two-tailed unpaired t test (n = 3). **** P < 0.0001. ( D ) Immunoblot analysis of CMS in human CD34 + early thymic progenitors (ETPs) transduced with a lentivirus encoding either shCMS4 and GFP or shSC and GFP as control. ( E ) Absolute numbers of shCMS- or shSC-transduced ETP-derived human cells reconstituting the thymus of RAG-2 −/− γc −/− mice at the indicated weeks post-transplant (n = 6 week 3, n = 4 week 5; *** P = 0.0002). ETP transduction efficiencies with shCMS and shSC were 24,5% ± 4,5% and 21,5% ± 3,5%, respectively. ( F ) Absolute numbers of shCMS- or shSC-transduced human thymocytes in ( E ) expressing the double positive CD3 lo TCRαβ − phenotype at 3- and 5-weeks post-transplant (left panel; * P = 0.0222) or the post-β selected DP CD3 + TCRαβ + phenotype at 5-weeks post-transplant (right panel; *P = 0.0146). Data in ( E, F ) are shown as mean numbers ± s.e.m. of transduced (GFP + ) cells normalized to 10 5 transduced input cells from two independent experiments, analysed by two-tailed unpaired t test.

Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

Techniques: Western Blot, Expressing, Transfection, Control, Phospho-proteomics, Activation Assay, Immunoprecipitation, Activity Assay, Transduction, Luciferase, Two Tailed Test, Derivative Assay