human cd34 hematopoietic stem cells  (Jackson Laboratory)

Journal: Blood Advances

Article Title: CAR T cells targeting CCR4 selectively deplete human Tregs ex vivo and in vivo

doi: 10.1182/bloodadvances.2025017573

Figure Lengend Snippet: Tregs are depleted by the CCR4-CAR in a humanized mouse model. (A) Experimental design. NSG-SGM3-IL15 engrafted with CD34 + hematopoietic stem cells were injected with 1 million CAR + CCR4-CARTs IV. Blood was collected on days 0, 3, 5, and 8, and mice were euthanized on day 11. (B) Representative flow plots showing the proportion of human CD45 (hCD45) and mCD45 leukocytes at baseline. (C) Proportion of Tregs, CD4 + non-Treg, and CD4 − cells of hCD45 percent at baseline. (D) Percentage of Tregs, non-Treg, and CD4 − cells that are CCR4 + at baseline. (E) Representative flow plots showing the CCR4 + and FOXP3 + expression on the CD4 + population before and after CART administration gated on CD4 + cells. (F-K) Proportions of Tregs, CD4 + non-Tregs, and CD4 − cells over time. Significance was determined using t tests corrected for multiple comparisons, with comparison to baseline indicated on graph; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. M1, Mouse 1; M2, Mouse 2; M3, Mouse 3; mCD45, mouse CD45.

Article Snippet: Mice were engrafted with human cord blood–derived CD34 + hematopoietic stem cells, and females were available for use at age 13 to 18 weeks after evaluation of engrafted human cell populations by flow cytometry at The Jackson Laboratory.

Techniques: Injection, Expressing, Comparison

Journal: PLOS Pathogens

Article Title: Mechanistic insights and in vivo HIV suppression by the BRD4-targeting small molecule ZL0580

doi: 10.1371/journal.ppat.1013449

Figure Lengend Snippet: (A) Experimental design and timeline for the in vivo study using HIV-infected CD34 + Hu-mice. At week 0, 12 Hu-mice were intravenously inoculated with a single dose of HIV-1 JR-CSF (10 ng HIV p24 antigen per animal). Plasma samples were collected at weeks 2 and 3 after HIV inoculation for confirmation of HIV infection by RT-qPCR and HIV p24 ELISA. At week 3, 11 out of the 12 Hu-mice were confirmed with systemic HIV infection (the one without infection was removed from the study). The 11 Hu-mice with successful HIV infection were randomized into four treatment groups at week 3: Empty Vehicle (EV) (n = 2), ART (n = 3), ZL0580 (n = 3), and ART + ZL0580 (n = 3). Treatment regimens were initiated at week 3 and administered daily by intraperitoneal until week 7. Plasma viral loads were closely monitored during the treatments at weeks 5, 6, 7. From week 7, all animals were subjected to treatment interruption (TI) and plasma viral loads were closely monitored once every 2 weeks until week 15. (B) Kinetics of plasma viral loads (HIV RNA copies/ml) post HIV inoculation for each mouse in EV (black circles), ART (orange triangles), ZL0580 (blue squares), and ART + ZL0580 (red diamonds) treatment groups. The shaded areas denote the phase of establishing systemic HIV infection (orange), treatment phase (blue), and treatment interruption (TI) monitoring phase (green). RT-qPCR was performed in duplicate and the mean of viral RNA copies for each sample at different time points was shown. (C) Summary data for kinetics of plasma viral loads over weeks post HIV inoculation for all animals in the four treatment groups: EV (black circles), ART (orange triangles), ZL0580 (blue squares), and ART + ZL0580 (red diamonds). Data are presented as mean ± SEM. Statistical comparison of viral loads among the four groups at each time point was performed using one-way ANNOVA. * at week 7 and *** at week 13 denote significant difference of the individual treatment group (ART, ZL0580, or ART + ZL0580) as compared to the EV group. The dashed line indicates the limit of detection (LOD). (D) Comparison of area under the curve (AUC) among the four groups after treatment interruption (TI) between weeks 7 and 15. Statistical comparison was performed using one-way ANNOVA. (E) Percentage change in body weight over days after treatment for all treatment groups. Data are presented as mean ± SEM. In this figure: * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: To assess in vivo HIV-suppressive activity of ZL0580, we utilized a humanized mouse model (Hu-mice) generated by engrafting human CD34 + hematopoietic stem cells (HSCs) into the immunodeficient NOD SCID gamma (NSG) mice (the Jackson Laboratory).

Techniques: In Vivo, Infection, Clinical Proteomics, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Comparison