hcd28  (Cytiva Europe)

Journal: Gene Therapy

Article Title: Baboon endogenous retrovirus (ERV) envelope pseudotyped lentiviral vectors outperform human ERV lentivectors for transduction of T, B, NK and HSPCs

doi: 10.1038/s41434-025-00587-w

Figure Lengend Snippet: A Schematic representation of the different WT or cytoplasmic domain-modified viral envelope glycoproteins (gp) for pseudotyping of LVs. MLV, murine leukemia virus; HERV-W, human endogenous retrovirus V-W, HERV-Cyt16, HERV- Cyt31: cytoplasmic tail deletion mutants, HERVTR, HERV gp mutant carrying the cytoplasmic tail and R-peptide of the MLV gp; HERVRLess, HERV gp mutant lacking the R-peptide, BaEV baboon endogenous retrovirus gp, BaEVTR, BaEV gp carrying the cytoplasmic tail and R-peptide of the MLV gp, BaEVRless is a BaEV gp mutant lacking the R-peptide. B Titer of the different vector pseudotypes carrying a GFP reporter gene determined on 293 T cells by serial vector dilutions. Titer was analysed by FACS at day 3 post-transduction for GFP expression (IU/ml; mean ± SD; n = 6; two-way Anova, ** p < 0.01, *** p < 0.001,**** p < 0.0001).

Article Snippet: Peripheral blood (PB) T lymphocytes were prestimulated for 3 days with T cell human hCD3/hCD28 TransAct beads (Miltenyi Biotec; #130-111-160,) supplemented with IL-2 (100 ng/ml) (Miltenyi Biotec; #130-097-743,) in RPMI or were prestimulated with recombinant 10 ng/mL human rIL-7 (rhIL-7; Miltenyi Biotec, #130-093-937) and 10 ng/ml rhIL-15 (Miltenyi Biotec 130-093-955,) for 3 days.

Techniques: Modification, Virus, Mutagenesis, Plasmid Preparation, Transduction, Expressing

Journal: Gene Therapy

Article Title: Baboon endogenous retrovirus (ERV) envelope pseudotyped lentiviral vectors outperform human ERV lentivectors for transduction of T, B, NK and HSPCs

doi: 10.1038/s41434-025-00587-w

Figure Lengend Snippet: A , C Human T cells were pre-stimulated by IL-7 and IL-15 for 3 days and subsequently transduced with GFP encoding LVs pseudotyped with HERV-W or BaEVRless gps. The multiplicities of infections (MOI) are indicated. Representative FACS plots are shown in ( A ) and data are summarized in ( C , left histograms). (mean ± SD; BaEVRless, n = 3; HERV-W, n = 4 two-way Anova,**** p < 0.0001). B , C Human T cells were pre-stimulated by anti-CD3/anti-CD28 and IL-2 for 3 days and subsequently transduced with GFP encoding LVs pseudotyped with HERV-W or BaEVRless gps. The multiplicities of infection (MOI) are indicated. Representative FACS plots are shown in ( B ) and data are summarized in ( C , right histograms). (mean ± SD; BaEVRless, n = 4; HERV-W, n = 5; two-way Anova, *** p < 0.001,**** p < 0.0001).

Article Snippet: Peripheral blood (PB) T lymphocytes were prestimulated for 3 days with T cell human hCD3/hCD28 TransAct beads (Miltenyi Biotec; #130-111-160,) supplemented with IL-2 (100 ng/ml) (Miltenyi Biotec; #130-097-743,) in RPMI or were prestimulated with recombinant 10 ng/mL human rIL-7 (rhIL-7; Miltenyi Biotec, #130-093-937) and 10 ng/ml rhIL-15 (Miltenyi Biotec 130-093-955,) for 3 days.

Techniques: Transduction, Infection

Journal: Gene Therapy

Article Title: Baboon endogenous retrovirus (ERV) envelope pseudotyped lentiviral vectors outperform human ERV lentivectors for transduction of T, B, NK and HSPCs

doi: 10.1038/s41434-025-00587-w

Figure Lengend Snippet: A Schematic representation of the experimental set-up; CD34+ cells are isolated from cord blood, subsequently stimulated with SCF/TPO/Flk3 overnight and then transduced with HERV-W LV or BaEVRless-LV at an MOI of 20. After 24 h of transduction CD34+ cells were injected into NBSGW mice ( n = 6 for BaEVRless LVs; n = 5 for HERV-W LVs) to allow reconstitution of the mice with a human blood system for 16 weeks before sacrifice and FACS analysis of the different hematopoietic tissues. B Detection of humanization in the blood of the recipient mice reconstituted with BaEVRLess or HERV-W LV transduced CD34+ cells. C Percentage of transduced (GFP + ) per total hCD45+ cells in bone marrow, spleen, thymus and blood at mice sacrifice for BaEVRless LVs (M1 to M6) and HERV-W LVs (M1 to M5). ND thymus not detected. D Representative FACS plots for GFP+ cells in the blood for the different lymphocyte subpopulations (CD19 for B cells; CD4 and CD8 for T cells, CD14 for monocytes and CD56 for NK cells) for mice M4 in the BaEVRless LV group and the mice M3 in the HERV-W group. Distribution of different hematopoietic lineages in the blood of humanized NBSGW mice a sacrifice (left histogram); Data for the transduction of different blood cell lineages ( D , right histogram) are summarized for all the mice per vector group.

Article Snippet: Peripheral blood (PB) T lymphocytes were prestimulated for 3 days with T cell human hCD3/hCD28 TransAct beads (Miltenyi Biotec; #130-111-160,) supplemented with IL-2 (100 ng/ml) (Miltenyi Biotec; #130-097-743,) in RPMI or were prestimulated with recombinant 10 ng/mL human rIL-7 (rhIL-7; Miltenyi Biotec, #130-093-937) and 10 ng/ml rhIL-15 (Miltenyi Biotec 130-093-955,) for 3 days.

Techniques: Isolation, Transduction, Injection, Plasmid Preparation

Journal: Gene Therapy

Article Title: Baboon endogenous retrovirus (ERV) envelope pseudotyped lentiviral vectors outperform human ERV lentivectors for transduction of T, B, NK and HSPCs

doi: 10.1038/s41434-025-00587-w

Figure Lengend Snippet: The experimental set-up for the humanization is outlined in Fig. . A Representative FACS plots for GFP+ cells in the spleen for the different lymphocyte subpopulations (CD19 for immature and mature B cells; CD4 and CD8 for T cells, CD14 for monocytes and CD56 for NK cells) for mice M4 in the BaEVRless LV group and the mice M3 in the HERV-W group. B Distribution of different hematopoietic lineages in the spleen of humanized NBSGW mice a sacrifice (left histogram) and data from ( A ) are summarized for all the mice per vector group (right histogram). C Representative FACS plots for GFP+ cells in the BM for the different lymphocyte subpopulations (early progenitors (CD34 + CD19-CD10-), pro-B cells (CD34 + CD19-CD10 + ) and pre B cells (CD34l ow CD19 + CD10 + ) and CD19+ immature/mature B cells) for mice M4 in the BaEVRless LV group and the mice M3 in the HERV-W group. D Distribution of different hematopoietic lineages in the BM of humanized NBSGW mice a sacrifice (left histogram) and data from ( C ) are summarized for all the mice per vector group (right histogram).

Article Snippet: Peripheral blood (PB) T lymphocytes were prestimulated for 3 days with T cell human hCD3/hCD28 TransAct beads (Miltenyi Biotec; #130-111-160,) supplemented with IL-2 (100 ng/ml) (Miltenyi Biotec; #130-097-743,) in RPMI or were prestimulated with recombinant 10 ng/mL human rIL-7 (rhIL-7; Miltenyi Biotec, #130-093-937) and 10 ng/ml rhIL-15 (Miltenyi Biotec 130-093-955,) for 3 days.

Techniques: Plasmid Preparation

Journal: Cell reports

Article Title: Mitochondrial reactive oxygen species regulate acetyl-CoA flux between cytokine production and fatty acid synthesis in effector T cells

doi: 10.1016/j.celrep.2025.115430

Figure Lengend Snippet:

Article Snippet: anti-hCD28 antibody , Bio X Cell , Cat# BE0248.

Techniques: FLAG-tag, Virus, Recombinant, Protease Inhibitor, Adjuvant, Staining, Isolation, Citrate Assay, Luminescence Assay, Chromatin Immunoprecipitation, Sample Prep, Plasmid Preparation, Software, Imaging, Microscopy, Transmission Assay, Gene Expression, Flow Cytometry, Mass Spectrometry