Journal: Journal for Immunotherapy of Cancer

Article Title: Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation

doi: 10.1136/jitc-2020-001223

Figure Lengend Snippet: Identification of further technical variables in MDSC immunomonitoring. Participating centers stained for MDSC subsets in healthy donor controls. Beforehand every participant received the identical batch of blood collection tubes. From the same donor sodium citrate-, EDTA and heparin blood was collected and PBMC were stained for MDSC subsets in PBMC. Lymphocyte separation medium was free of choice. MDSC frequency was determined in the core lab as described for . Influence of anticoagulants (A) and lymphocyte separation medium (B) on total frequency of putative ‘MDSC subsets’ in healthy individuals with mean and SD is shown. Kruskal-Wallis was used for statistical analysis. Results were considered significant at *p≤0.05, **p≤0.001 and ***p≤0.0001. M-MDSC, monocytic myeloid-derived suppressor cell; PBMC, peripheral blood mononuclear cell; PMN, polymorphonuclear.

Article Snippet: Before separation blood was admixed with the same volume of phosphate-buffered saline (PBS) and overlayed on 1.077 g/mL separation medium (Biocoll, Merck Millipore; Ficoll-Paque, GE healthcare; Pancoll, PAN-Biotech; Lymphoprep, Stemcell Technologies).

Techniques: Staining, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation

doi: 10.1136/jitc-2020-001223

Figure Lengend Snippet: Standardization of anticoagulant and separation medium reduces intercenter variability of PMN-MDSC frequencies. All participants used sodium citrate as anticoagulant and Biocoll as separation medium and determined the frequency of PMN-MDSC in healthy donor controls. Data were compared with data obtained for . (A) Frequency of PMN-MDSC in Step1 ( , free choice of anticoagulant and separation medium) and step 2 (sodium citrate and Biocoll) from all healthy blood donors in the five centers is shown (plus mean and SD). (B) The mean frequency was determined for each of the five participating centers and (C) % cv was calculated. In all panels the comparison to step 1 (data from ) is shown. F test was used for statistical analysis. Results were considered significant at *p≤0.05 and ****p≤0.0001. PBMC; peripheral blood mononuclear cell.

Article Snippet: Before separation blood was admixed with the same volume of phosphate-buffered saline (PBS) and overlayed on 1.077 g/mL separation medium (Biocoll, Merck Millipore; Ficoll-Paque, GE healthcare; Pancoll, PAN-Biotech; Lymphoprep, Stemcell Technologies).

Techniques: Comparison

Journal: Journal for Immunotherapy of Cancer

Article Title: Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation

doi: 10.1136/jitc-2020-001223

Figure Lengend Snippet: Expression of molecules associated with T cell suppression expression of PD-L1 and LOX-1 on MDSC subsets in patients and healthy donor controls was determined using the harmonized flow cytometry labeling protocol, Biocoll as standard separation medium and sodium citrate as standard anticoagulant. Flow cytometry analysis was performed in the core lab to ensure standardized gating. Staining intensity of PD-L1 (A) and LOX-1 (B) on MDSC-subsets was determined in five different disease settings. Delta median (median signal intensity of antibody minus median signal intensity of isotype control) is shown. Data are depicted as mean and SD is shown. CVD, cardiovascular disease; M-MDSC, monocytic myeloid-derived suppressor cells; PD-L1, programmed death-ligand 1; PMN, polymorphonuclear.

Article Snippet: Before separation blood was admixed with the same volume of phosphate-buffered saline (PBS) and overlayed on 1.077 g/mL separation medium (Biocoll, Merck Millipore; Ficoll-Paque, GE healthcare; Pancoll, PAN-Biotech; Lymphoprep, Stemcell Technologies).

Techniques: Expressing, Flow Cytometry, Labeling, Staining, Control, Derivative Assay

Journal: Cell reports

Article Title: Systematic Single Cell Pathway Analysis (SCPA) to characterize early T cell activation

doi: 10.1016/j.celrep.2022.111697

Figure Lengend Snippet: Key resources table

Article Snippet: Human bulk CD4 + or CD8 + T cells were isolated from PBMCs of one donor (male aged 39 years old) by centrifugation using Lymphoprep separation medium (Corning, Vienna, VA) and subsequent use of either the MACS Human CD8 + T cell Isolation Kit (Miltenyi Biotech), or the Negative Selection EasySep CD4 + T cell kit (Stemcell Technologies) according to the manufacturers’ instructions.

Techniques: Generated, Virus, Recombinant, Lysis, Enzyme-linked Immunosorbent Assay, Cell Isolation, Staining, CRISPR, Control, Software

Journal: Cell reports

Article Title: Systematic Single Cell Pathway Analysis (SCPA) to characterize early T cell activation

doi: 10.1016/j.celrep.2022.111697

Figure Lengend Snippet: Key resources table

Article Snippet: Human bulk CD4 + or CD8 + T cells were isolated from PBMCs of one donor (male aged 39 years old) by centrifugation using Lymphoprep separation medium (Corning, Vienna, VA) and subsequent use of either the MACS Human CD8 + T cell Isolation Kit (Miltenyi Biotech), or the Negative Selection EasySep CD4 + T cell kit (Stemcell Technologies) according to the manufacturers’ instructions.

Techniques: Generated, Virus, Recombinant, Lysis, Enzyme-linked Immunosorbent Assay, Cell Isolation, Staining, CRISPR, Control, Software