Journal: Bone Marrow Transplantation

Article Title: Brief ex vivo Fas-ligand incubation attenuates GvHD without compromising stem cell graft performance

doi: 10.1038/s41409-020-0941-2

Figure Lengend Snippet: T-lymphocytes isolated from FasL-treated and control MPBCs were stimulated using anti-CD3/CD28 activation beads. a Activated T helper (CD4 + CD25 + ) and b T cytotoxic (CD8 + CD25 + ) cells were quantified 24 and 48 h post stimulation using flow cytometry. c Concentration of IFN-γ secreted to the medium of the cultured T-cells was measured 24, 48 and 72 h post stimulation using ELISA. d The percentages of CD4 + and CD8 + IFN-γ secreting (IFN-γ + ) cells were quantified by flow cytometry 48 h post stimulation. e Percent of TH1 (CD4 + CXCR3 + ) and f TC1 (CD8 + CXCR3 + ) cells were quantified 24 and 48 h post stimulation using flow cytometry. g The percentages of CD4 + and CD8 + IL17 secreting (IL17 + ) cells were quantified by flow cytometry 48 h after stimulation. h TH17 (CD4 + CCR6 + ) T cells populations were quantified 24 and 48 h post stimulation using flow cytometry. i Regulatory CD4 + and CD8 + T cells (CD25 + FoxP3 + ) were quantified 72 h post stimulation using flow cytometry. Representative results of one independent study out of two is presented. Data presented as Mean+SD, n = 3. Statistical analysis was performed using unpaired, parametric t-test; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: Intracellular staining was performed using either Inside Stain Kit or FoxP3 Staining Buffer Set (130-090-477 or 130-093-142, respectively, Miltenyi Biotech, Bergisch Gladbach, Germany) according to manufacturer’s instructions.

Techniques: Isolation, Control, Activation Assay, Flow Cytometry, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay