Journal:

Article Title: Influence of Glycosylation on the Efficacy of an Env-Based Vaccine against Simian Immunodeficiency Virus SIVmac239 in a Macaque AIDS Model

doi: 10.1128/JVI.79.16.10386-10396.2005

Figure Lengend Snippet: Env-specific CD4+ T-cell and CD8+ T-cell responses in 12 macaques. A: Env-specific CD8+ T cells in PBMCs were measured by ELISPOT assay for IFN-γ in three groups. B: Env-specific CD4+ T cells in PBMCs were measured by ELISPOT assay for IFN-γ in three groups. ELISPOT results are colored as follows: Δ5G SU-specific T cells (red), wt-SU-specific T cells (green), and TM-specific T cells (yellow). Arrows with a dotted line, arrows with broken line, and arrows with a solid line indicate the time of the third DNA vaccination at 8 weeks p.p., the time of the vaccine boost at 21 weeks p.p., and the time of SIVmac239 challenge at 28 weeks p.p., respectively. C: Comparison of SU-specific CD8+ T cells and CD4+ T cells in PBMCs among the wt-Env vaccine group, the Δ5G Env vaccine group, and the vector control group at 26 weeks p.p. and 4, 7, and 12 days p.i. The numbers of SFC responding to SIVmac239 SU were used to compare the effects of the two vaccines. w, weeks; d, days.

Article Snippet: PBMCs were subjected to the depletion of CD4 + cells with magnet beads coated with anti-human CD4 Ab (Dynal ASA, Oslo, Norway) or subjected to the depletion of CD8 + cells with magnet beads coated with anti-human CD8 Ab (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Enzyme-linked Immunospot, Comparison, Plasmid Preparation, Vaccines

Journal:

Article Title: Influence of Glycosylation on the Efficacy of an Env-Based Vaccine against Simian Immunodeficiency Virus SIVmac239 in a Macaque AIDS Model

doi: 10.1128/JVI.79.16.10386-10396.2005

Figure Lengend Snippet: SIV-specific CD8+ T-cell and CD4+ T-cell responses in 12 animals. A: SIV viral-protein-specific CD8+ T cells in PBMCs were measured by ELISPOT assay for IFN-γ in three groups: vector controls, wt-Env vaccine group, and Δ5G Env vaccines. B: SIV viral-protein-specific CD4+ T cells in PBMCs were measured by ELISPOT assay for IFN-γ in three groups. ELISPOT results of individual SIV proteins are colored as follows: Gag (red), Nef (green), Tat/Rev (blue), Vif/Vpr/Vpx (yellow), and Pol (pink). C: Comparison of cumulated CD8+ T cells or CD4+ T cells specific to the viral proteins Gag, Pol, Nef, Tat/Rev, and Vif/Vpr/VpX between SIV infection-controlled and uncontrolled animals. w, weeks; d, days.

Article Snippet: PBMCs were subjected to the depletion of CD4 + cells with magnet beads coated with anti-human CD4 Ab (Dynal ASA, Oslo, Norway) or subjected to the depletion of CD8 + cells with magnet beads coated with anti-human CD8 Ab (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Enzyme-linked Immunospot, Plasmid Preparation, Vaccines, Comparison, Infection

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Representative flow analysis of peripheral blood apheresis collections from the preselection, post-CD45RA depletion, and CD8+enrichment steps. Cells were stained for expression of CD4, CD8, CD45RA, and CD45RO. Plots show CD45 gated events.

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques: Staining, Expressing

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Flow cytometric analysis, cytokine expression, mixed lymphocyte reaction, and cytokine secretion. (A) Representative flow analysis of 1 experiment repeated 5 times that shows cell composition and subsets from start to finish of preselection, post–CD45RA depletion, and post–CD8+ enrichment. Cells were stained for expression of CD45RA, CD45RO, and CD62L, and plots are shown for CD4+ and CD8+ gated cells at each step of the processing procedure. The surface phenotype of the CD45RA–CD8+-enriched cells is consistent with the phenotypic CD8+ TEM subset being predominantly CD45RA–CD45RO+CD62L–. The TEM cells ranged from 81.7% to 98.1% of the final cell composition. (B) Flow cytometric analysis of cytokine expression by enriched cell subsets. Cells were activated by ionomycin and phorbol myristate acetate, treated with monensin, and stained for expression of CD45, CD4, CD8, and CD45RA. Cells were fixed, permeabilized, and stained for INF-γ, and IL-2. Plots show comparison of IL-2 and IFN-γ expression in CD8+CD45RA–, CD8+CD45RA+, and CD4+CD45RA– cells as indicated. (C) Proliferation of CD8+CD45R– and CD4+ cells activated by co-culture with or without irradiated allogeneic stimulators and assessed by 3H-thymidine uptake during the last 24 hours of culture. Mean and standard deviations are shown (n = 4). (D) Cytokine secretion assessment in CD8+CD45RA– and CD4+ cells activated by co-culture with or without irradiated allogeneic stimulators. Supernatants from 7-day cultures were analyzed by flow cytometry using cytokine bead arrays for INF-γ, IL-2, and TNF-α (n = 4). Means with standard deviations are shown. Teff, T effector cells.

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques: Expressing, Staining, Co-Culture Assay, Irradiation, Flow Cytometry

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Patient characteristics

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques:

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Patient outcomes

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques:

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Kaplan-Meier estimates of event-free survival (EFS) for the 15 patients receiving CD8+TMcell infusion.

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques:

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: Kaplan-Meier estimates of overall survival (OS) for the 15 patients receiving CD8+TMcell infusion.

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques:

Journal: Blood Advances

Article Title: Infusion of donor-derived CD8 + memory T cells for relapse following allogeneic hematopoietic cell transplantation

doi: 10.1182/bloodadvances.2017012104

Figure Lengend Snippet: CD8 + T M cell yield, purity, and infusion dose after tandem selection

Article Snippet: Thereafter, the flow-through CD45RA – cells were incubated with Miltenyi anti-CD8 antibody-conjugated beads to positively select the CD8 + cells.

Techniques: Selection, Cell Counting