Journal: Cell Reports Methods

Article Title: Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors

doi: 10.1016/j.crmeth.2022.100350

Figure Lengend Snippet: γδ T cell reactivity accounts for the majority of the mouse extract response and is not mediated by peptides Total MO-specific T cell responses were measured as a percentage of AIM + (CD137 + CD69 + ) CD3 + T cells after stimulation of PBMCs with MO extract, and the individual γδ and αβ T cell reactivity was assessed. (A) Representative FACS plot of total AIM + CD3 + T cells (left) further gated as function of γδ and αβ T cell reactivity (right) (B) Graph shows the relative percentage of γδ and αβ reactivity from the total MO-specific T cell responses across all donors (n = 33). (C) (Left) Representative FACS plots of AIM + (CD137 + CD69 + ) γδ (top row) or αβ (bottom row) T cells after stimulation of PBMCs with MO peptide pools (MPs), MO extract, or control (Neg; media for γδ T cells, or DMSO for αβ T cells). (D) Graphs show the percentage of AIM + reactivity in each condition for γδ or αβ T cells across all donors (n = 20). All graphs show data represented as geometric mean with SD. Each dot represents a unique individual. Pairwise comparisons were performed with the Wilcoxon test, and p values <0.05 were considered statistically significant.

Article Snippet: APCs were separated from PBMCs in the negative flow-through using the human CD3 positive selection kit (EasySepTM Human CD3 Positive Selection Kit II, Stemcell Technologies, Vancouver, BC, Canada).

Techniques: Control

Journal: Cell Reports Methods

Article Title: Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors

doi: 10.1016/j.crmeth.2022.100350

Figure Lengend Snippet: γδ T cell reactivity to MO and CR allergen extracts is TCR specific, albeit with different requirements for the presence of APCs (A–C) Allergen-specific γδ T cell responses were measured as percentage of AIM + (CD137 + CD69 + ) γδ T cells after stimulation of PBMCs with (A) MO extract, (B) CR extract, or (C) HDMAPP in the presence (+) or absence (−) of antigen-presenting cells (APCs). (D and E) PBMCs were stimulated with an MO or CR extract, HDMAPP, or α-CD3 and cultured in the absence (−) or presence (+) of a TCR blocking reagent (dasatinib). Graphs show percentage of AIM + γδ (D) or αβ (E) T cells in response to the different stimuli and conditions (MO, n = 8; CR, n = 8; HDM, n = 16; TG, n = 16). Each dot represents a unique individual. Data are represented as geometric mean and SD. Kruskal-Wallis or Wilcoxon paired tests were performed, and p values are indicated. p < 0.05 was considered statistically significant.

Article Snippet: APCs were separated from PBMCs in the negative flow-through using the human CD3 positive selection kit (EasySepTM Human CD3 Positive Selection Kit II, Stemcell Technologies, Vancouver, BC, Canada).

Techniques: Cell Culture, Blocking Assay

Journal: Cell Reports Methods

Article Title: Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors

doi: 10.1016/j.crmeth.2022.100350

Figure Lengend Snippet:

Article Snippet: APCs were separated from PBMCs in the negative flow-through using the human CD3 positive selection kit (EasySepTM Human CD3 Positive Selection Kit II, Stemcell Technologies, Vancouver, BC, Canada).

Techniques: Recombinant, Software

Journal: Transfusion

Article Title: The feasibility and efficacy of subcutaneous plerixafor for mobilization of peripheral blood stem cells in allogeneic HLA–identical sibling donors: results of the HOVON‐107 study

doi: 10.1111/trf.15037

Figure Lengend Snippet: Composition of the grafts obtained from healthy adult sibling donors

Article Snippet: T lymphocytes were purified from peripheral blood using a human whole blood CD3 positive selection kit (STEMCELL Technologies, Inc.) according to the manufacturer's instructions.

Techniques:

Journal: Transfusion

Article Title: The feasibility and efficacy of subcutaneous plerixafor for mobilization of peripheral blood stem cells in allogeneic HLA–identical sibling donors: results of the HOVON‐107 study

doi: 10.1111/trf.15037

Figure Lengend Snippet: Chimerism in peripheral blood (PB), bone marrow (BM), and CD3 selected cells at 3, 6, and 12 months after transplant

Article Snippet: T lymphocytes were purified from peripheral blood using a human whole blood CD3 positive selection kit (STEMCELL Technologies, Inc.) according to the manufacturer's instructions.

Techniques:

Journal: Transfusion

Article Title: The feasibility and efficacy of subcutaneous plerixafor for mobilization of peripheral blood stem cells in allogeneic HLA–identical sibling donors: results of the HOVON‐107 study

doi: 10.1111/trf.15037

Figure Lengend Snippet: Immunologic reconstitution. Numbers of CD3+; CD3+/CD4+; CD3 + CD8+; CD19+, and NK cells in the peripheral blood at 3, 6, 12, and 24 months after transplant. At 12 months: CD3 + cells: median, 0.84 × 10 9 /L (0.33–1.65); normal range, 0.66–2.10. CD3+/CD4+ cells: median, 0.30 × 10 9 /L (0.17–0.75); normal range, 0. 32–1.36. CD3+/CD8+ cells: median, 0.45 × 10 9 /L (0.11–1.24); normal range, 0.15–0.88. CD3–/CD16+/CD56+ NK cells: median, 0.20 × 10 9 /L (0.13–0.52); normal range, 0.09–0.62. CD19+ cells: median, 0.27 × 10 9 /L (0.09–0.95); normal range, 0.10–0.5. Each box is drawn from the 25th percentile to the 75th percentile of the cell counts, and the horizontal bar within the box indicates the median cell count. The line up from the top of the box goes to the upper adjacent value as defined. The dots indicate cell counts larger than the upper adjacent value. [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: T lymphocytes were purified from peripheral blood using a human whole blood CD3 positive selection kit (STEMCELL Technologies, Inc.) according to the manufacturer's instructions.

Techniques: Cell Counting