Journal: Nucleic Acids Research

Article Title: The p400 complex promotes HIV-1 latency by suppressing viral transcription and altering the host cell state

doi: 10.1093/nar/gkaf764

Figure Lengend Snippet: EP400 depletion results in increased expression of activation-associated proteins in T cells. ( A ) Expression of CD2, CD69, and HIV in J-Lat 10.6 cells transduced with shRNAmirs targeting CD8B, CD19, or EP400 (two independent shRNAmirs) as measured by flow cytometry, three independent experiments. Upper panels: Representative histograms showing expression of the indicated proteins. Blue traces = expression of CD69 in cells transduced with control shRNAmirs. Red traces = expression of CD69 in cells transduced with EP400 shRNAmirs. Lower panels: Percentage of cells with high levels of expression of indicated proteins. * P <.05, ** P <.01, *** P <.001, **** P <.0001, NS = not significant. One-way ANOVA with multiple comparisons. ( B ) Representative flow cytometry plots showing expression of CD2, CD69, and HIV in in J-Lat 10.6 cells transduced with shRNAmirs targeting CD8B or EP400. ( C ) Representative western blot analysis of EP400, CD2, LEF-1, and GAPDH protein levels in J-Lat 10.6 cells from panel (A). ( D ) As for panel (C) but in uninfected Jurkat cells. ( E ) As for panel (C) but in primary CD4 + cells.

Article Snippet: Membranes were probed with antibodies to EP400 (BETHYL, #A300-541A), DMAP1 (Thermo Fisher, #10411-1-AP), HDAC1 (Thermo Fisher, #PA1-860), BRD4 (Proteintech, #ABE1391) LEF-1 (Cell Signaling, #76010S), CD2 (Cell Signaling, #53460S), or GAPDH (Santa Cruz, #sc-47724) followed by horseradish peroxidase-conjugated antirabbit or antimouse IgG.

Techniques: Expressing, Activation Assay, Transduction, Flow Cytometry, Control, Western Blot

Journal: BMC Immunology

Article Title: Bovine CD2 - /NKp46 + cells are fully functional natural killer cells with a high activation status

doi: 10.1186/1471-2172-7-10

Figure Lengend Snippet: Effect of anti-CD2 mAbs on bovine NK cell functions . (A) IFN-γ response from NK cells caused by cross-linking of CD2. Plastic wells were pre-coated with two different mAbs against bovine CD2, along with NKp46-mAb as positive control and CD8-mAb or medium as negative controls. rbIL-2 activated NK cells were added and incubated for 24 h in the presence or absence of rbIL-12 (0.5 U/ml), and IFN-γ content in the supernatants analyzed by an ELISA. Data shown are means ± SD of two to four individual animals. (B-C) IL-2 activated NK cell killing of the bovine target lines BL3.1 and MDBK in a 4 h 51 Cr-release assay, with addition of the indicated mAbs for blocking. CD2mAb I = BAQ95A, CD2 mAb II = MUC2A, CD8 mAb = CACT80C, NKp46 mAb = AKS1. (D) As above, using murine Fc-receptor bearing P815 target cells to obtain a redirected lysis effect. Data shown in (B-D) are from one animal representative of three to five.

Article Snippet: Selection of NK cell subsets was performed after 24 h, by applying the anti-CD2 mAb MUC2A (VMRD, Pullman, WA, USA) and MACS anti-mouse IgG MicroBeads and LS columns (both Miltenyi Biotech, Bergish Gladbach, Germany) according to the manufacturer's instructions.

Techniques: Positive Control, Incubation, Enzyme-linked Immunosorbent Assay, Release Assay, Blocking Assay, Lysis

Journal: The Journal of Clinical Investigation

Article Title: Positive and negative selection shape the human naive B cell repertoire

doi: 10.1172/JCI150985

Figure Lengend Snippet: ( A ) Thymic graft from 1 fetal donor was cotransplanted with HSCs from a different HLA-mismatched fetus into an NSG mouse. Two 50 μg doses of anti-CD2 were injected after the surgery to deplete initial thymocytes in the thymic graft. ( B ) HEp-2 reactivity of recombinant Abs cloned from single mature naive B cells from 3 HLA-mismatched NSG + thymus humanized mice were assessed by ELISA. ( C ) Summary of the frequencies of HEp-2–reactive mature naive B cells in mismatched NSG + thymus humanized mice compared with regular NSG + thymus and NSG humanized mice. Each symbol represents a mouse, and the average is shown with a bar. ( D ) Polyreactivity of recombinant Abs cloned from single mature naive B cells from 3 HLA-mismatched NSG + thymus (NSG+Thy) humanized mice was assessed by ELISA. ( E ) Summaries of the frequencies of polyreactive and antinuclear clones in mismatched NSG + thymus humanized mice compared with regular NSG + thymus and NSG humanized mice. * P < 0.01 and **** P < 0.0001, by Kruskal-Wallis test.

Article Snippet: For HLA-mismatched NSG + thymus humanized mice, 50 μg anti-CD2 mAb (LSBio) was injected i.p. over a 2-week period (1 dose/week) to eliminate preexisting thymocytes and mature T cells from the thymic graft ( ).

Techniques: Injection, Recombinant, Clone Assay, Enzyme-linked Immunosorbent Assay

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Safety and pharmacodynamics of anti-CD2 monoclonal antibody treatment in cynomolgus macaques

doi: 10.1111/tri.13524

Figure Lengend Snippet: Animal characteristics and treatment regimens

Article Snippet: The test substances were manufactured to target the CD2 antigen, and included two modifications of the same anti-CD2 mAb: a rat anti-primate CD2 IgG2b (Immerge BioTherapeutics) (hereafter termed RT-CD2), and the further modified rhesus recombinant anti-primate CD2 IgG1 (hereafter termed RH-CD2).

Techniques:

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Safety and pharmacodynamics of anti-CD2 monoclonal antibody treatment in cynomolgus macaques

doi: 10.1111/tri.13524

Figure Lengend Snippet: A) WBC counts in all treated animals, groups A-C (n.s.). B, C) Showing treatment impact on hematocrit (Hct%) or platelet levels (n.s.). D) ALT and AST levels pre- and post-anti-CD2 infusion, group A. E, F) Post-treatment levels of leukocyte subsets. G) Showing bimodal CD2 and CD3 T cell expression in naïve cynomolgus monkeys, with memory cells having a higher CD2hi and CD3lo expression. Shown is a representative gate defining the bimodal distribution, and the three subsets of T cells (TN, TEM, TCM) gated for CD2 and CD3. Predominantly, TN are CD3hiCD2lo and memory cells CD3loCD2hi. CCR7/CD45RA gating step not shown. H) Average median fluorescence intensities (MFI) of cell surface CD2 on CD4+-, CD8+-, NK-, B-, and regulatory T cells from naïve animals (n=4). I) Showing partial competitive binding of RH-CD2 and CD2-BV786 antibodies. J, K) RH-CD2 effectively depletes memory cells, while relatively sparing naïve lymphocytes in group A. L) Illustrating the CD8/CD4 ratio before and after RH-CD2 in group A. M) Showing a 695% CD25hiFoxP3+ cell enrichment from day 0 to day 7 after RH-CD2 infusion (p=0.052) in group A. Data is presented as average (±SEM). Abbreviations: WBC, white blood cell count; CD, cluster of differentiation; CM, central memory; EM, effector memory; NK cell, natural killer cell.

Article Snippet: The test substances were manufactured to target the CD2 antigen, and included two modifications of the same anti-CD2 mAb: a rat anti-primate CD2 IgG2b (Immerge BioTherapeutics) (hereafter termed RT-CD2), and the further modified rhesus recombinant anti-primate CD2 IgG1 (hereafter termed RH-CD2).

Techniques: Expressing, Fluorescence, Binding Assay, Cell Counting

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Safety and pharmacodynamics of anti-CD2 monoclonal antibody treatment in cynomolgus macaques

doi: 10.1111/tri.13524

Figure Lengend Snippet: The allogeneic MLRs were inhibited by varying concentrations of RT- and RH-CD2, respectively, at the onset of the host effector cell cultures. After four days, MLRs were pulsed for 24 hours with 1μCi 3H-thymidine per well. The cellular radioactivity was measured in triplicates samples. Statistical analysis were performed of antibody and no antibody treatments using one-way ANOVA. (A) RT-CD2 (B) RH-CD2.

Article Snippet: The test substances were manufactured to target the CD2 antigen, and included two modifications of the same anti-CD2 mAb: a rat anti-primate CD2 IgG2b (Immerge BioTherapeutics) (hereafter termed RT-CD2), and the further modified rhesus recombinant anti-primate CD2 IgG1 (hereafter termed RH-CD2).

Techniques: Radioactivity

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Safety and pharmacodynamics of anti-CD2 monoclonal antibody treatment in cynomolgus macaques

doi: 10.1111/tri.13524

Figure Lengend Snippet: Effector cells stably transfected with A) FcγRI (n=3), B) FcγRIIA (n=3) or C) FcγRIIIA (n=2) and a luciferase reporter gene were incubated with serial dilutions of antibody. Upon binding of a target-bound IgG antibody (the reporter cells expressed CD2) luciferase expression was induced. Luminescence values were normalized to the highest concentration of the positive control. RH-CD2 displayed clear dose-dependent FcγR-mediated signaling but RT-CD2 did not. D) Both antibodies induced dose-dependent CDC of lymphocytes relative to untreated controls (n=4).

Article Snippet: The test substances were manufactured to target the CD2 antigen, and included two modifications of the same anti-CD2 mAb: a rat anti-primate CD2 IgG2b (Immerge BioTherapeutics) (hereafter termed RT-CD2), and the further modified rhesus recombinant anti-primate CD2 IgG1 (hereafter termed RH-CD2).

Techniques: Stable Transfection, Transfection, Luciferase, Incubation, Binding Assay, Expressing, Concentration Assay, Positive Control