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