The primary panel of monoclonal antibodies (mAbs) used were CD45 (for blast identification), CD13, CD33, CD34, HLA-DR and for AML M4/M5 CD14, for AML M6 anti-glycophorin A and for AML-M7, CD61
, CD41 and CD42 were used.
Lineage-specific markers, including myeloid (CD13, CD33, CD117, MPO), monocytic (CD11b, CD14, CD64), megakaryocytic (CD61
), B-cell (CD19, CD10, CD20) and T-cell (CD3) antigens, were negative.
Eight-color MFC was performed in all patients as a routine clinical test on BM samples that were obtained as part of baseline assessment at the time of diagnosis, the end of the first and second courses of chemotherapy, as well as pre-HSCT., A panel of eight antibody combinations that recognize CD7, CD11b, CD13, CD14, CD16, CD19, CD33, CD34, CD38, CD41, CD45, CD56, CD61
, CD64, CD71, CD117, CD123, and HLA-DR was used for MRD detection, and 0.2-1 million events per tube were acquired on a Fluorescence activated cell sorter (FACS Canto II) (BD Co., USA).
Wherever indicated the primary panel was extended up to (secondary panel) antibodies against cytoplasmic (cyt) CD3, CD4, CD8, myeloperoxidase (MPO), terminal deoxynucleotidyl transferase (TdT), CD79, CD41, CD61
Immunophenotyping revealed 23% blast cells, positive for megakaryocyte markers (CD42b, CD41, CD61
), myeloid markers (CD33), progenitor cell markers (CD117, CD34) and T cell marker--CD7 positive.
Using flow cytometry to detect epithelial cell adhesion molecule positive ([EpCAM.sup.+]) EVs from MCF7 cell culture and integrin /33 positive (CD61
) EVs from human plasma, we compared the performance of the most promising generic markers, namely, calcein AM, calcein violet, CFSE, di-8-ANEPPS, and lactadherin, and used side scatter as a reference.
Less than 1% of cells contained within water fibres expressed CD235a and CD61
, whereas DMSO fibres contained a higher number of cells expressing the platelet marker CD61
It was reported that elevated EVs (Annexin V-binding EV, CD61
, P-selectin-exposing EV, and E-selectin-exposing EV) in women were related to the menstrual luteal cycle .
HLA-DR, CD58, CD99, CD56, CD38, cy-MPO, CD11b, CD13, CD33, CD65, CD64, CD117, CD36, CD61
, CD4, and 7.1 were positive, and CD14, CD15, CD19, CD10, CD20, CD3, and CD7 were negative.
Followup platelet immunophenotyping using flow cytometry demonstrated decreased expression of CD41 (a marker of platelet membrane GPIIb) and normal expression of CD61
(a marker of GPIIIa) and CD42b (a marker of GPIb), suggesting a specific deficiency in GPIIb and confirming the diagnosis of GT.
Modulation of monocyte Fc[gamma]R balance by TPO-ra was also found in a murine model of ITP established by transferring splenocytes from immunized CD61
knockout mice into CD611 severe combined immunodeficient mice.
In brief, samples of fixed blood were incubated with fluorescent antibodies to the platelet specific marker CD61
(used to identify platelets) and CD62P (P-selectin).