Journal: eLife

Article Title: Seroconversion stages COVID19 into distinct pathophysiological states

doi: 10.7554/eLife.65508

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-Human CD33 , Fluidigm , Cat# 3163023B, RRID: AB_2687857 , Monoclonal-Clone: WM53 Dilution: 1/100.

Techniques: Staining, Biomarker Discovery, Software

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: Cytotoxic IC 50 values of HAA 2020 , the eleven inhibitors and their combinations in HL60 cells (24 h, nM).

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques:

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: Cell cycle analysis in HL60 cells. Cells were treated (+) for 24 h with either 100 nM doxorubicin (Dox), 500 nM HAA 2020 , 10 nM dinaciclib or a combination of HAA 2020 and dinaciclib (500 nM and 10 nM, respectively). Data are represented as mean ± SEM ( n = 2, two independent experiments) for each of the phases of the cell cycle, sub-G 1 ( A ), G 0 /G 1 ( B ), S ( C ) and G 2 /M ( D ). Statistical differences, compared with the untreated control cells (-), were assessed by a one-way ANOVA with the Tukey’s post-hoc multiple comparison test (GraphPad Prism). p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****) were taken as significant.

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques: Cell Cycle Assay, Control, Comparison

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: Detection of apoptosis in HL60 cells. Cells were treated for 6 h, 12 h and 24 h with either HAA 2020 (500 nM), dinaciclib (10 nM) or their combination. Following treatment, the cells were stained with annexin V FITC/PI. A total of 20,000 single-cell events were acquired on a BC-500 flow cytometer and analyzed by the Expo 32 software. Data are represented as mean ± SEM ( n = 3, two independent experiments) for each of the cell staining statuses: live cells (annexin V-/PI-), early apoptotic cells (annexin V+/PI-), late apoptotic cells (annexin V+/PI+) and necrotic cells (annexin V-/PI+).

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques: Staining, Flow Cytometry, Software

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: mRNA expression levels of TNF-α and caspase-7 ( A ), Hsp90-α and EGFR ( B ), compared with the control. HL60 cells were treated for 6 h with either HAA 2020 (500 nM), dinaciclib (10 nM) or their combination. Data are represented as mean ± SEM ( n = 3, two independent experiments). The results were expressed as fold-change compared with the untreated group. Statistical differences, compared with the untreated control cells (dashed line), were assessed by a one-way ANOVA with the Tukey’s post-hoc multiple comparison test. p < 0.05 (*), p < 0.01 (**) and p < 0.001 (***) were taken as significant.

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques: Expressing, Control, Comparison

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: Expression of p27, cyclin D3 and CDK2 in HL60 cells treated with either HAA 2020 , dinaciclib or their combination as above for 6 h. Data are represented as mean ± SEM ( n = 2, two independent experiments). The result was normalized as fold-change compared with the untreated group. Ratio above 1 (dashed line) shows the amount of mRNA for the three genes in the untreated cells (control = 1). Statistical differences, compared with the untreated control cells, were assessed by a one-way ANOVA with the Tukey’s post-hoc multiple comparison test. p < 0.05 (*), p < 0.01 (**) and p < 0.001 (***) were taken as significant. The Image J software was used for densitometry.

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques: Expressing, Control, Comparison, Software

Journal: Molecules

Article Title: Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors

doi: 10.3390/molecules25092220

Figure Lengend Snippet: The effect of the HAA 2020 and dinaciclib combination on apoptosis, cell proliferation and survival of HL60 cells. Created in BioRender.com.

Article Snippet: HL60 suspension cells (Hematopoietic neoplasm, Human acute myeloid leukemia: AML, Myc+, CLL1, CD33, CD123 and CD135) were obtained from the ATCC.

Techniques:

Journal: Nature biotechnology

Article Title: Development and function of human innate immune cells in a humanized mouse model

doi: 10.1038/nbt.2858

Figure Lengend Snippet: a , Percentages of human myeloid cells (hCD33 + ) among human hematopoietic cells (hCD45 + ) in the blood of the indicated recipient mice, engrafted as newborns by intra-hepatic injection of fetal liver CD34 + cells after X-ray preconditioning. Each symbol represents an individual mouse and the red bars indicate mean values (n=20-113; statistical analysis is shown in ). b , Composition of human white blood cells in the same mice as in a (n=20-113 mice/group; n=8 human donors; error bars indicate SEM). c , Immunohistological staining of human myeloid cells (hCD68 + ) in non-lymphoid tissues of the indicated recipient mice. The black bar represents 20 μm, and the images shown are representative of at least three mice analyzed per group. d-e , Representative flow cytometry analysis ( d ) and frequencies ( e ) of human monocyte subsets, identified by expression of CD14 and CD16 among hCD45 + CD33 + cells in the blood of recipient mice (n=8-12 mice/group; error bars indicate SEM). Dot plots in d are gated on CD33 hi SSC lo cells to show the subset distribution among monocytic cells. f , Human monocytes in the blood of MISTRG recipients and human monocytes from a human donor were stained with the indicated antibodies. Staining with isotype control antibodies is shown in red and specific antibodies in blue.

Article Snippet: Human CD33 + cells were enriched by magnetic isolation (EasySep CD33 selection kit, StemCell Technologies).

Techniques: Injection, Staining, Flow Cytometry, Expressing, Control

Journal: Oncology Reports

Article Title: Kevetrin induces apoptosis in TP53 wild-type and mutant acute myeloid leukemia cells

doi: 10.3892/or.2020.7730

Figure Lengend Snippet: Effect of continuous kevetrin treatment on viability and apoptosis induction in AML cell lines and primary samples. (A and B) Viability of TP53 -wt OCI-AML3 and MOLM-13 and TP53 -mut KASUMI-1 and NOMO-1 cell lines treated with different concentrations of kevetrin (85, 170 and 340 µM) for 24 and 48 h. (C and D) Quantification of Annexin V + cells in AML cell lines treated with kevetrin at different concentrations (85, 170 and 340 µM) for 24 and 48 h. (E and F) Cell cycle analysis of cell lines treated with 340 µM kevetrin for 24 and 48 h. Values represent the mean ± standard deviation of three biological replicates. (G) Cell viability and (H) percentage of Annexin V + cells in bone marrow mononuclear cells from AML patients exposed to increasing concentrations of kevetrin. (I) Percentage of AML blasts relative to control. Values represent the mean ± standard deviation. Symbols indicate samples from AML patients (no. 01: TP53 -mut AML; nos. 02, 03 and 04: TP53 -wt AML). (J) Percentage of Annexin V − cells in AML blasts, monocytes and lymphocytes. Blasts were defined as CD33 + CD14 − cells in the CD45/side scatter ‘blast gate’, monocytes as CD33 + CD14 + , B lymphocytes as CD19 + and T lymphocytes as CD3 + . *P<0.05, **P<0.01, ***P<0.001. AML, acute myeloid leukemia; KEV, kevetrin; CTRL, control; wt, wild-type.

Article Snippet: In primary samples, Annexin V staining was combined with surface markers using the following antibodies: CD45-APC Vio770 (cat. no. 130-110-635), CD33-APC (cat. no. 130-111-020), CD14-PerCP Vio 700 (cat. no. 130-110-523), CD3-PE (cat. no. 130-113-139) (all from Miltenyi Biotec GmbH, dilution 1:50) and CD19-PE Cy7 (cat. no. 302216, 1:10, BioLegend, Inc.).

Techniques: Cell Cycle Assay, Standard Deviation, Control

Journal: Biomacromolecules

Article Title: Development of CD33-Targeted Dual Drug-Loaded Nanoparticles for the Treatment of Pediatric Acute Myeloid Leukemia

doi: 10.1021/acs.biomac.4c00672

Figure Lengend Snippet: Preparation and characterization of CD33 NPs. (A) Schematic overview of the antibody–nanoparticle conjugation process, using maleimide–thiol chemistry. (B) Table summarizing conjugated NPs characteristics in terms of drug loading, amount of antibody conjugated obtained via Micro BCA, DLS-measured hydrodynamic diameter (nm), and polydispersity index (PdI) values, and PALS-measured zeta potential values. Data are presented as mean ± SD, from measurements performed in triplicate and averaged from at least n = 3. (C) TEM images of CD33 NPs, representative of two independent experiments.

Article Snippet: FLISA studies were performed as previously described, using recombinant human CD33-Fc (Sino Biological) at 1 μg/mL to coat the plate wells.

Techniques: Conjugation Assay, Zeta Potential Analyzer

Journal: Biomacromolecules

Article Title: Development of CD33-Targeted Dual Drug-Loaded Nanoparticles for the Treatment of Pediatric Acute Myeloid Leukemia

doi: 10.1021/acs.biomac.4c00672

Figure Lengend Snippet: Binding of nanoformulations to recombinant CD33-Fc and CD33-expressing cells. (A–D) Binding of rhodamine 6G-loaded NPs to recombinant CD33-Fc in FLISA assays: (A) dose-dependent binding, (B) binding of NPs (50 μg polymer/mL) ± preincubation with CD33-Fc (10 μg/mL), (C) binding of NPs (500 μg polymer/mL) ± preblock with CD33 mAb (40 μg/mL), (D) binding of NPs (500 μg polymer/mL) in competition with varying concentrations of gemtuzumab (0.00256–40 μg/mL). Data are presented as mean ± SD, n = 3. (E) Binding of nonfluorescent nanoformulations to CD33-Fc and IgG-Fc evaluated by SPR: (i) binding of the NPs at 4 mg/mL to CD33-Fc and IgG-Fc, (ii) binding of CD33 NPs to CD33-Fc at varying concentrations, with linear regression and corresponding goodness of fit ( R 2 ). Binding is presented as response relative to baseline observed 5 s before the end of the injection period. Data are presented as mean ± SD, n = 2. (F) Cells were treated with blank CD33 or nude NPs (750 μg polymer/mL) for 1 h at 4 °C. Then, cells were washed, stained with PE-labeled anti-CD33 antibody or isotype control antibody, and PE-fluorescence was analyzed by flow cytometry. Representative histograms are shown for each condition tested, (i), as well as the corresponding reduction of fluorescence compared with the positive stained control observed after treatment with the NPs for each cell line (ii). (G) Confocal microscopy images of MOLM-13 cells treated with 500 μg polymer/mL rhodamine 6G-loaded NPs for 1 h at 4 °C, followed by a washing step and a further 2 h-incubation at 37 °C. Scale bar is 50 μm, and blue and red staining denote cell nuclei and nanoparticles, respectively. Representative data from n = 2.

Article Snippet: FLISA studies were performed as previously described, using recombinant human CD33-Fc (Sino Biological) at 1 μg/mL to coat the plate wells.

Techniques: Binding Assay, Recombinant, Expressing, Fluorophore-linked Immunoabsorbent Assay, Polymer, Injection, Staining, Labeling, Control, Fluorescence, Flow Cytometry, Confocal Microscopy, Incubation

Journal: Biomacromolecules

Article Title: Development of CD33-Targeted Dual Drug-Loaded Nanoparticles for the Treatment of Pediatric Acute Myeloid Leukemia

doi: 10.1021/acs.biomac.4c00672

Figure Lengend Snippet: Targeted delivery of synergistic drug combination within dual CD33 NPs. (A) MV4-11, (B) MOLM-13, and (C) MOLM-14 cells were treated with blank or dual-loaded conjugated (CD33 NP) or nonconjugated (nude NP) nanoparticles for 1 h at 4 °C. Then, cells were washed, counted, and reseeded for 72 h of incubation at 37 °C, prior to measurement of cell viability. Where appropriate, cells were preincubated with 5 μg of free-gemtuzumab for 15 min at 4 °C and washed, prior to NP incubation. Data are presented as mean ± SD, n = 3.

Article Snippet: FLISA studies were performed as previously described, using recombinant human CD33-Fc (Sino Biological) at 1 μg/mL to coat the plate wells.

Techniques: Incubation