Spanning tree progression analysis of density normalized events (SPADE) identification of novel myeloid derived suppressor cells (MDSC) subsets

Autor: James E. Talmadge, Phyllis Warkentin, Holly Briitton, Lynell W. Klassen, Kathryn Cole
Rok vydání: 2018
Předmět:
Zdroj: The Journal of Immunology. 200:46.14-46.14
ISSN: 1550-6606
0022-1767
Popis: MDSC’s are a heterogeneous population of myeloid cells at various differentiation stages. The human MDSC phenotypes, are controversial and subdivided into macrophage (M-), granulocytic (G-) and immature (i-) MDSC’s. Our studies used a single staining tube with antibodies to linage markers (CD3, CD19 and CD56), HLA-DR, CD11b, CD14, CD15, CD16, CD33, CD45, PD-L1 and LOX-1. Mobilized lymphoma patient apharesis (CA) products provided a product with a high frequency of MDSCs compared to normal donor blood samples (PB). SSCA and FSA identified lymphocytes, granulocytes and monocyte populations. A total population (CD45+), MDSCs (Lin-HLA-DR-CD11b+CD14+, Lin-HLA-DR-CD11b+CD14-CD33+ and Lin-HLA-DR−CD11b+ CD15+) and non MDSC myeloid populations (Lin-HLA-DR+CD14+, Lin-HLA-DR+CD33+ and Lin-HLA-DR+ CD15+Lin-HLA-DR-11b-) were exported from FlowJo for Spade analysis. This identified 11 nodes with 4 MDSC sub-populations. Impressively, cells from the non-MDSC populations, including monocytes and granulocytes did not occur within these nodes. SPADE identified an M-MDSC, G-MDSC and two iMDSC subsets with an increased frequency in the CA samples. Increased expression of LOX-1 and CD16 occurred in CA MDSCs vs. PB. Intracellular staining for expression of NOS-2 and Arg-1 were consistent with the M and G-MDSC subsets. Node 7 mainly expressed M-MDSC’s with a low frequency of CD15 dull G-MDSC’s. The cell frequency for G-MDSC’s in node 7 was 1,334 for CA and 0 for PB with M-MDSC’s 1,079 and 43 respectively. Node 8 had a frequency (3,319) of G-MDSC’s for CA and 2,286 for PB with M-MDSC’s 188 and 15 respectively. In summary SPADE analysis can be used to cleanly identify MDSCs from monocytes and granulocytes and provide insight into novel MDSC subsets.
Databáze: OpenAIRE
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