Journal: Cell Death & Disease

Article Title: Colorectal cancer patients-derived immunity-organoid platform unveils cancer-specific tissue markers associated with immunotherapy resistance

doi: 10.1038/s41419-024-07266-5

Figure Lengend Snippet: A Immunohistochemical staining of CD3, CD4, CD8, CD68 and Ki-67 in CRC tissues showed the presence of T-cells populations (CD4 + , CD8 + ) and macrophages (CD68 + ) in MSI and MSS patients. Ki-67 was used as a control proliferation marker of tumor cells; ( B ) Quantification of positive cells/area was performed with QuPath Software. Significance shown refers to Wilcoxon test p -values *< 0.05, **< 0.01, ***< 0.001.

Article Snippet: T-cells and Myeloid Derived Suppressor Cells (MDSCs) were isolated from patient peripheral blood mononuclear cells (PBMCs) by CD3 + immunomagnetic negative selection (#17951, StemCell Technologies) and CD33 + immunomagnetic positive selection (#17876, StemCell Technologies) following positive selection with CD3 + immunomagnetic positive selection (#17851, StemCell Technologies), respectively.

Techniques: Immunohistochemical staining, Staining, Control, Marker, Software

Journal: Cell reports

Article Title: DNA Damage Baseline Predicts Resilience to Space Radiation and Radiotherapy

doi: 10.1016/j.celrep.2020.108434

Figure Lengend Snippet: (A) Baseline number of foci per nucleus as a function of age in CD3 + lymphocytes extracted from finger prick collection in 339 healthy donors (***p < 0.0001, deviation from zero of the slope of linear regression). Four donors in this cohort were considered healthy but were in remission from cancer at the time of the finger prick (shown as red dots). See also . (B) Distribution of spontaneous foci per nucleus after bead-based extraction of CD3 + lymphocytes from finger prick samples (finger prick), Ficoll-based extraction of PBMCs from buffy coat samples (PBMC baseline), and a further freeze-thaw cycle and plating in the cell culture media (PBMC 0 Gy). ***p < 0.0001, one-way ANOVA, Tukey’s post hoc test for multiple comparisons among all three groups: 1–2, 2–3, and 1–3. Boxplots show median ± quartiles ± min/max values.

Article Snippet: After incubation, 4.5 μL of the CD3 Positive Selection Cocktail (StemCell Tech Cat# 18081C.1) was added to each well and incubated for 15 min at room temperature.

Techniques: Extraction, Cell Culture

Journal: Cell reports

Article Title: DNA Damage Baseline Predicts Resilience to Space Radiation and Radiotherapy

doi: 10.1016/j.celrep.2020.108434

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: After incubation, 4.5 μL of the CD3 Positive Selection Cocktail (StemCell Tech Cat# 18081C.1) was added to each well and incubated for 15 min at room temperature.

Techniques: Recombinant, Red Blood Cell Lysis, Selection, Flow Cytometry, Multiplex Assay, Software

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