Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: PE Mouse monoclonal anti human CD45RB (clone MT4 (6B6)) , BD PharMingen , Cat# 555904; RRID:AB_396215.

Techniques: Negative Control, Recombinant, Purification, Staining, Expressing, Lysis, Immunoprecipitation, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Cell Isolation, Software, Microscopy

Journal: Blood

Article Title: ORP4L is a prerequisite for the induction of T-cell leukemogenesis associated with human T-cell leukemia virus 1

doi: 10.1182/blood.2021013579

Figure Lengend Snippet: Tax triggers ORP4L expression and T-cell malignant transformation via suppression of miR-31. (A-B) qPCR (A) and western blot (B) analyses confirming the effects of miR-31 inhibitor on ORP4L expression in HeLa cells and the effects of miR-31 mimic on ORP4L expression in MT-4 cells. Mean ± standard deviation (SD; n = 3 experimental repeat; Student t test). (C) qPCR analysis of miR-31 expression in T cells of patients with ATL (n = 37), as compared with normal T cells (n = 5). (D) Pearson’s correlation coefficient of miR-31 and ORP4L expression within human ATL specimens (n = 37). (E) Multicolor fluorescence in situ hybridization analysis of the genomic status of miR-31 encoding (green probe) and its host gene (origin probe) regions in T cells of 2-month-old WT and LCK/R26 Tax mice. The chromosome 4 centromere (red probe) is included as a control. Scale bar, 5 μm. (F) miR-31 expression in 2-month-old WT and LCK/R26 Tax mouse T cells analyzed by qPCR. Mean ± SD (n = 3 mice of each group; Student t test). (G) Western blot analysis of the expression of ORP4L in LCK/R26 Tax T cells or LCK/R26 Tax T cells overexpressing miR-31. The cells were isolated and infected with lentivirus carrying control RNA or miR-31 and cultured for 96 hours in vitro. (H) Western blot analysis of the expression of ORP4L in human normal T cells or human ATL T cells overexpressing miR-31. The cells were isolated and infected with lentivirus carrying control RNA or miR-31 and cultured for 96 hours in vitro. (I) Kaplan-Meier comparative survival analysis of B-NDG mouse recipients of WT T cells or LCK/R26 Tax T cells overexpressing miR-31 or overexpressing both miR-31 and ORP4L. The LCK/R26 Tax T cells were coinfected with the lentivirus carrying control RNA miR-31, empty vector (Emp.vec.), or ORP4L as follows: control RNA+Emp.vec., miR-31+Emp.vec., miR-31+ORP4L, and cultured for 96 hours in vitro before transplantation (n = 7; mice of each group, log-rank test). (J) Representative images of peripheral blood smears from B-NDG mice treated as in panel I. The number of abnormal lymphocytes on smears in each cohort are indicated below the images. Scale bar, 10 μm. (K) The percentage of CD3 + CD4 + , CD4 + CD8 + , CD44 + CD25 + , and CD3 + c-Kit + cells in peripheral blood of B-NDG mice treated as in panel I. Mean ± SD (n = 7 mice of each group; Student t test). (L) Representative histologic hematoxylin and eosin– and anti-CD3–stained sections showing T-cell infiltration in spleen and liver of B-NDG mice treated as in panel I. Scale bars, 100 μm. (M) Kaplan-Meier comparative survival analysis of B-NDG mouse recipients of T cells of patients with ATL (ATL#21). The cells were isolated and coinfected with lentivirus carrying control RNA, miR-31, Emp.vec. or ORP4L as follows: control RNA+Emp.vec., miR-31+Emp.vec., control RNA+ORP4L, and miR-31+ORP4L and cultured for 96 hours in vitro before transplantation (n = 8 mice each group, log-rank test). (N) The percentage of human CD45 + CD3 + ATL T cells in peripheral blood of B-NDG mice treated as in panel M. Mean ± SD (n = 8 mice in each group; Student t test). *** P < .001.

Article Snippet: MT-4 and HeLa cells were purchased from American Type Culture Collection and maintained in RPMI 1640 (For MT-4 cells) or Dulbecco’s modified Eagle’s medium (for HeLa cells) containing 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin at 37°C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Transformation Assay, Western Blot, Standard Deviation, Fluorescence, In Situ Hybridization, Control, Isolation, Infection, Cell Culture, In Vitro, Plasmid Preparation, Transplantation Assay, Staining

Journal: Viruses

Article Title: Inhibition of Human Immunodeficiency Virus Type 1 Entry by a Keggin Polyoxometalate

doi: 10.3390/v10050265

Figure Lengend Snippet: The anti-human immunodeficiency virus (HIV) activity in activated peripheral blood mononuclear cells (PBMCs) and toxicity of the Keggin polyoxometalate PT-1. ConA stimulated PBMCs were incubated with the HIV-1 NL4-3 strain and increasing doses of PT-1( A ) or zidovudine (AZT) ( B ), HIV-1 replication was evaluated by HIV-1 p24 enzyme-linked immunosorbent assay (ELISA) on days 3, 5, 7, and 9 after infection, and the data were expressed as the means ± standard deviations (SD); ( C ) The cytotoxicity of PT-1 were measured using TZM-bl, MT4, and PBMCs with Cell counting kit -8 (CCK-8) assay; ( D ) Representative plots of PBMC apoptosis measured by Annexin V-FITC/PI staining and flow cytometry (FACS) analysis. Early apoptotic cells (Annexin-V positive-PI negative) and late apoptotic cells (Annexin-V-PI positive) are shown in the lower right and upper right, respectively; ( E ) Representative image of mice bone marrow cells smears from vehicle control ( a ), 200 mg/kg PT-1 ( b ), 600 mg/kg PT-1 ( c ), 1800 mg/kg PT-1 ( d ), and cyclophosphamide positive control group ( e ). Cells were fixed and stained with Giemsa stain, magnification 1000× polychromatic erythrocytes (PCEs) with one or more nuclei (white arrow) scored. All data represent two independent experiments.

Article Snippet: HEK-293T cells and the human T-cell line MT4 was purchased from American Type Culture Collection (ATCC) and maintained in RPMI 1640 (Gibco) supplemented with 10% ( v / v ) fetal bovine serum, 200 μg/mL l -glutamine and 100 U/mL penicillin-streptomycin.

Techniques: Virus, Activity Assay, Incubation, Enzyme-linked Immunosorbent Assay, Infection, Cell Counting, CCK-8 Assay, Staining, Flow Cytometry, Control, Positive Control, Giemsa Stain

Journal: Viruses

Article Title: Inhibition of Human Immunodeficiency Virus Type 1 Entry by a Keggin Polyoxometalate

doi: 10.3390/v10050265

Figure Lengend Snippet: PT-1 inhibited gp120/CD4 binding. ( A ) Surface plasmon resonance (SPR) signals of gp120/CD4 binding. Recombinant gp120 protein was immobilized on a CM5 sensor chip, followed by injection of soluble CD4 (0.11 μM), PT-1 (2 μM), or a pre-incubated mixture of CD4 (0.11 μM) and PT-1 (0.7 μM); ( B ) SPR sensorgram of PT-1 binding to CD4 binding. CD4 protein was immobilized on a CM5 sensor chip, followed by injection of a serial dilution of PT-1 (0.7–140.81 μM). Gp120 was used as a positive control; ( C ) FACS profiles of PT-1 binding to the CD4 receptor on MT4 cells. MT4 cells were incubated PT-1 (0.7, 3.5, 17.6 μM) or gp120 (positive control) for 30 min. Cells were then stained with PE-conjugated anti-CD4 RPA-T4 antibodies (D1 domain) for FACS analysis.

Article Snippet: HEK-293T cells and the human T-cell line MT4 was purchased from American Type Culture Collection (ATCC) and maintained in RPMI 1640 (Gibco) supplemented with 10% ( v / v ) fetal bovine serum, 200 μg/mL l -glutamine and 100 U/mL penicillin-streptomycin.

Techniques: Binding Assay, SPR Assay, Recombinant, Injection, Incubation, Serial Dilution, Positive Control, Staining