Journal: AIDS Research and Human Retroviruses

Article Title: Purging Exhausted Virus-Specific CD8 T Cell Phenotypes by Somatic Cell Reprogramming

doi: 10.1089/aid.2017.0161

Figure Lengend Snippet: Establishment of B6/129OKSM mouse line. (A) Schematic illustration of B6/129OKSM mouse generation. We first established a DOX-reprogrammable fibroblast-like cell line originated from AinV15 mouse ESCs by transduction of STEM-CCA lentiviral vector encoding four DOX-inducible mouse reprogramming genes (Oct4, Klf4, Sox2, and c-Myc). Chimeric mice were generated by the injection of 8–12 established iPSCs into a B6 mouse blastocyst. The mice were bred with B6 mice to obtain F1 hybrids derived from 129P2/OlaHsd strain (agouti coat). F1 hybrids were then crossbred for >10 generations until we established a stable mouse line carrying a single copy of a reprogramming gene cassette introduced by STEM-CCA lentiviral vector homozygously and enabling induction of ESC-like colony formation from peripheral blood cells with high efficiency in the presence of DOX. DOX: doxycycline. Diff.: differentiation. (B) Fibroblasts were isolated from tail-tip biopsies of chimeric mice and exposed to 1 μg/ml of DOX for 2° iPSC induction. No 2° iPSC formation was seen with no DOX addition. (C) Expression of two pluripotent stem cell markers (Nanog and SSEA1) in 2° iPSCs. (D) Embryoid bodies generated from 2° iPSCs. iPSC, induced pluripotent stem cell.

Article Snippet: AinV15 mouse embryonic stem cells (ESCs) were purchased from American Type Culture Collection (ATCC SCRC-1029, Manassas, VA).

Techniques: Transduction, Plasmid Preparation, Generated, Injection, Derivative Assay, Isolation, Expressing

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Structure and in vitro activities of BKI-1748 against N. caninum (Nc-β-gal) and T. gondii (Tg-β-gal). (A) molecular structure of BKI-1748, MW = molecular weight; EC 50 = concentration of half maximal proliferation inhibition; *toxicity levels against human foreskin fibroblasts (HFF) and zebrafish embryos were determined previously [22]. (B,C) Dose-response curves for Neospora and Toxoplasma tachyzoites grown in HFF.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: In Vitro, Molecular Weight, Concentration Assay, Inhibition

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Litter size, parasite burden, neonatal and postnatal mortality rates of N. caninum infected mice treated with BKI-1748.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: Infection

Journal: Molecular cancer research : MCR

Article Title: A Functional Precision Oncology Approach to Identify Treatment Strategies for Myxofibrosarcoma Patients

doi: 10.1158/1541-7786.MCR-21-0255

Figure Lengend Snippet: (A, B) Ex vivo drug testing of the novel patient derived cell model (WCM197) together with the OH931, NMFH-1 and HFF (benign human fibroblasts) using abemaciclib a second line CDK4/6-inhibitor and pralatrexate an antifolate inhibitor in a 6–8-point dilution dose response log scale to determine the IC50 values, highest dose was 10uM and lowest dose was 4.7 pM with an assay time of 96 hours. Greater cytostatic and cytotoxic effect was seen in the cells with the CDKN2A/B and MTAP null status (WCM197 and OH931) compared to CDKN2A/B and MTAP wild type status (NMFH-1 and HFF). (C) The combination of pralatrexate (IC30) and abemaciclib killed all our patient cells within 48 hours compared to the vehicle and abemaciclib and pralatrexate mono-therapy. (D) WCM197 sarco-sphere formation is completely inhibited with the combination of abemaciclib and pralatrexate. (E) Drug combination of abemaciclib and pralatrexate causes massive cell swellings and cytoplasmatic vacuole formation. (F) Cleaved caspase 3 and cleaved PARP at 24, 48 and 72 hours were detected with the combination of abemaciclib and pralatrexate indicating the induction of cell death in the tumor cells after treatment.

Article Snippet: The Human Foreskin Fibroblast (HFF) cells were provided by Dr. Carla Grandori, SEngine Precision Medicine, Seattle, WA, USA as a control specimen for high throughput drug screening ( 22 ).

Techniques: Ex Vivo, Derivative Assay

Journal: Molecular cancer research : MCR

Article Title: A Functional Precision Oncology Approach to Identify Treatment Strategies for Myxofibrosarcoma Patients

doi: 10.1158/1541-7786.MCR-21-0255

Figure Lengend Snippet: (A) High throughput drug screening of all 4 cell strains (HFF, OH931, WCM197, NMFH-1) shown as a heat map (blue is sensitive to red resistant). RB1 wt and CDKN2A/B/MTAP null cells (WCM197 and OH931) show higher sensitivity to CDK4/6 inhibititors such as abemaciclib (indicated by the yellow arrows) and the antifolate pralatrexate and methotrexate (indicated by the green arrows) compared to the CDKN2A/B/MTAP wild type lines (NMFH-1 and HFF). Selective sensitivity to PLK-1 inhibitors was detected in WCM197 and OH931 cells compared to the NMFH-1 and the control line HFF, shown in orange arrows. (B) Graphs show the response of the sarcoma cells to each compound in the library as area under the curve (AUC) compared to the benign human foreskin fibroblast line (HFF) as a normal control. (C-D) Ex vivo drug validation of the two PLK-1 inhibitors (volasertib and rigosertib) confirms high sensitivity for WCM197 and OH931 cells. (E, F) WCM197 3D sarco-spheres are completely inhibited in growth under volasertib treatment and show cell death indicated by cleaved caspase 3, PARP and cleaved PARP. Cell death was monitored with cleaved caspase 3 and cleaved PARP over 96 hours with a peak at 24–72 hours.

Article Snippet: The Human Foreskin Fibroblast (HFF) cells were provided by Dr. Carla Grandori, SEngine Precision Medicine, Seattle, WA, USA as a control specimen for high throughput drug screening ( 22 ).

Techniques: High Throughput Screening Assay, Drug discovery, Control, Ex Vivo, Biomarker Discovery

Journal: Nature Communications

Article Title: Adoptive γδ T cell therapy controls cytomegalovirus infection in preclinical transplantation models

doi: 10.1038/s41467-026-69538-2

Figure Lengend Snippet: a Analysis of DOT cell degranulation in response to CMV infection. Mean percentages of CD107a+ cells are shown, from cocultures with fibroblasts ( n = 6 donors) and macrophages ( n = 7 donors) conducted in technical duplicates. b DOT cells were cultured alone (medium) or cocultured with mock- or CMV-infected fibroblasts ( n = 27 donors), macrophages ( n = 12 donors) or endothelial cells ( n = 10 donors), in the presence of IL-18 and in technical triplicates. Means of IFNγ secretion are shown. a , b Lines connect conditions from the same donor. Statistical analysis was performed using a repeated-measures one-way ANOVA in ( a ) or a mixed-effects model in ( b ), both with Geisser-Greenhouse correction followed by Tukey’s multiple comparisons test. c Comparison of IFNγ secretion by DOT cells expanded from CMVneg ( n = 13) versus CMVpos ( n = 14) donors in fibroblast cultures. Shown are means ± SD, compared using two-tailed Mann–Whitney tests. d Fibroblasts were mock-infected or infected with CMV TB42/E, CMV Merlin, HSV or VZV, then cultured alone (white) or with DOT cells (gray) in the presence of IL-18. IFNγ secretion is represented as a fold change compared to a DOT cell-alone control. Box plots show the median (centre line), 25th–75th percentile (box) and minimum-maximum values (whiskers) of six independent donors (technical triplicates). Statistical analysis was performed using two-tailed Wilcoxon matched-pairs signed rank tests. e Schematic of the CMV dissemination assay. f Fibroblasts infected with CMV strain Merlin UL32-GFP were cultured alone or with DOT cells at various E:T ratios. Shown are percentages of GFP-positive fibroblasts, normalized to the infected monoculture control (set to 100%). Each bar represents the mean ± SD of 5 (mock and E:T 0.05:1 conditions) or 7 (other conditions) independent experiments using different DOT donors, each performed in technical duplicates. Statistical comparisons used a mixed-effects model with Geisser-Greenhouse correction and Dunnett’s two-tailed multiple comparisons test, comparing each condition to the infected monoculture control. Source data is provided as a Source Data file. CMV cytomegalovirus, DOT Delta One T, E:T effector to target, GFP green fluorescent protein, HFF human foreskin fibroblasts, HSV herpes simplex virus, IFN interferon, IL interleukin, Mϕ macrophages, neg negative, pos positive, VZV varicella-zoster virus.

Article Snippet: Human foreskin fibroblasts (HFF) were provided by Dr. Hamid-Reza Rezvani (INSERM, U1035, Bordeaux).

Techniques: Infection, Cell Culture, Comparison, Two Tailed Test, MANN-WHITNEY, Control, Virus