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.
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 and glycophorin.
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.
(a marker of megakaryocytes), CD235a, and CMYB were clustered together because of their similar expression dynamics (Figure 3A).
TGF-[beta] has a strong immunosuppressive effect on B cells, T cells CD41
, T cells CD81, antigen-presenting cells (APC) and macrophages.7
(2c) EpCAM, CD24, CA19- 9, CLDN3, CA-125, MUC18, EGFR, and HER2, a list of putative ovarian cancer markers; CD41
and CD45 for immune host cell markers and D2-40 a mesothelial marker for exosomes profiled left in nPLEX sensor and their parental ovarian cell lines filled in right for flow cytometer, according to Im et al.
Despite this, previous studies have shown immunostaining for markers of clot formation (CD41
, von Willebrand factor), and markers to assess the degree of crypt neuroendocrine cell degeneration (chromogranin A, CD56, or synaptophysin) may be helpful.
After 10 days culture, the cells expressed the MK markers CD41
and CD61 (Figure S1 in Supplementary Material available online at https://doi.org/10.1155/2017/2320519) and we refer to them as MK progenitors (MK-PROs).
This was accompanied by the upregulation of megakaryocyte and platelet markers such as CD41
and the platelet-specific ADP receptor P2Y12, indicating that S1PR4 signaling in this context alters genetic programs to modulate cell identity.
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.
Furthermore, immunohistochemical analysis of surgical sample was performed by using glycophorin A (DAKO, clone: JC159, 1:400) for erythroblasts, CD41
(Calbiochem, clone: 283.16B7, 1:2000) for megakaryocytes, and MPO (DAKO, rabbit polyclonal, 1: 500) for granulocytes (Figure 4).