MDSC Phenotyping in PBMC

PBMC were analyzed for the presence of MDSC as previously described [21 (link)]. MDSC were defined as cells positive for CD33, CD11b and lacking HLA-DR with subsets expressing CD15 or CD14 representing granulocytic and monocytic MDSC, respectively (Fig. 1). Notably, the current method for phenotyping M-MDSC (CD33+/HLADR-/CD14+/CD11b+) compares favorably to methods employed by other investigators (e.g., CD14+/HLADRlow/−) with respect to the percentages of M-MDSC obtained [22 (link)]. Specific antibodies included CD15-FITC, CD33-APC, HLA-DR-PC7, CD11b-PE (all Beckman Coulter), and CD14-V450 (BD Biosciences). All samples were run on a BD LSR-II flow cytometer and data was analyzed with FlowJo software (Tree Star, Inc.).

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Wesolowski R., Duggan M.C., Stiff A., Markowitz J., Trikha P., Levine K.M., Schoenfield L., Abdel-Rasoul M., Layman R., Ramaswamy B., Macrae E.R., Lustberg M.B., Reinbolt R.E., Mrozek E., Byrd J.C., Caligiuri M.A., Mace T.A, & Carson WE I.I.I. (2017). Circulating myeloid derived suppressor cells increase in patients undergoing neo-adjuvant chemotherapy for breast cancer. Cancer immunology, immunotherapy : CII, 66(11), 1437-1447.

Publication 2017

CD15-FITC CD33-APC HLA-DR-PC7 CD11b-PE CD14-V450

Antibodies Cd11b Cells Fitc Granulocytic Hla dr Mdsc Monocytic Tree

Corresponding Organization : The Ohio State University

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Variable analysis

independent variables

MDSC phenotyping method (CD33+/HLADR-/CD14+/CD11b+ vs CD14+/HLADRlow/−)

dependent variables

Percentage of M-MDSC

control variables

Specific antibodies used: CD15-FITC, CD33-APC, HLA-DR-PC7, CD11b-PE, CD14-V450
Flow cytometry instrument: BD LSR-II
Data analysis software: FlowJo

controls

No positive or negative controls were explicitly mentioned.

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