Starting with human whole blood, the CD3+ cell content of the isolated fraction typically ranges from 92.4 - 99.8% (as assessed by staining the start and isolated fractions with anti-CD3 or anti-CD2 antibodies, respectively). In the above example, the purities of the start and the final isolated fractions are 18.2% and 98.1%, respectively (gated on CD45). NOTE: Red blood cells were removed from the start sample by lysis prior to flow cytometry.
Figure 2. FACS Data for Anti-Human CD2 Antibody, Clone RPA-2.10, PE-Conjugated
(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD2 Antibody, Clone RPA-2.10, PE (Catalog #60007PE) and anti-human CD45 APC. (B) Flow cytometry analysis of human PBMCs processed with the EasySep™ HLA CD3 Positive Selection Kit (Catalog #17871) and labeled with Anti-Human CD2 Antibody, Clone RPA-2.10, PE. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgG1, kappa PE isotype control antibody is shown (open histogram).
Figure 3. FACS Data for Anti-Human CD5 Antibody, Clone UCHT2, PE-Conjugated
(A) Flow cytometry analysis of human buffy coat nucleated cells labeled with Anti-Human CD5 Antibody, Clone UCHT2, PE (Catalog #60082PE) and Anti-Human CD20 Antibody, Clone 2H7, APC (Catalog #60008AZ). (B) Flow cytometry analysis of human buffy coat nucleated cells labeled with a mouse IgG1, kappa PE isotype control antibody and Anti-Human CD20 Antibody, Clone 2H7, APC. (C) Flow cytometry analysis of human buffy coat nucleated cells processed with the EasySep™ HLA CD3 Positive Selection Kit (Catalog #17871) and labeled with Anti-Human CD5 Antibody, Clone UCHT2, PE. Histograms show labeling of buffy coat nucleated cells (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgG1, kappa PE isotype control antibody is shown (solid line histogram).
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Neutrophil and Granulocytic Myeloid-Derived Suppressor Cell-Mediated T Cell Suppression Significantly Contributes to Immune Dysregulation in Common Variable Immunodeficiency Disorders. M. Vlkova et al. Journal of immunology (Baltimore, Md. : 1950) 2018 NOV
Abstract
Common variable immunodeficiency disorders (CVID) represent a group of primary immunodeficiency diseases characterized by hypogammaglobulinemia and impaired specific Ab response, resulting in recurrent infections due to dysfunctional immune response. The specific mechanisms mediating immune deficiency in CVID remain to be determined. Previous studies indicated that immune dysregulation in CVID patients is associated with chronic microbial translocation, systemic immune activation, and altered homeostasis of lymphocytic and myeloid lineages. A detailed phenotypic, functional characterization of plasma markers and immune cell populations was performed in 46 CVID patients and 44 healthy donors. CVID patients displayed significantly elevated plasma levels of a marker of neutrophil activation neutrophil gelatinase-associated lipocalin. Neutrophils from CVID patients exhibited elevated surface levels of CD11b and PD-L1 and decreased levels of CD62L, CD16, and CD80, consistent with a phenotype of activated neutrophils with suppressive properties. Neutrophils from CVID patients actively suppressed T cell activation and release of IFN-$\gamma$ via the production of reactive oxygen species. Furthermore, CVID was associated with an increased frequency of low-density neutrophils (LDNs)/granulocytic myeloid-derived suppressor cells. LDN/granulocytic myeloid-derived suppressor cell frequency in CVID patients correlated with reduced T cell responsiveness. Exogenous stimulation of whole blood with bacterial LPS emulated some but not all of the phenotypic changes observed on neutrophils from CVID patients and induced neutrophil population with LDN phenotype. The presented data demonstrate that neutrophils in the blood of CVID patients acquire an activated phenotype and exert potent T cell suppressive activity. Specific targeting of myeloid cell-derived suppressor activity represents a novel potential therapeutic strategy for CVID.
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Species
Human
Magnet Compatibility
• “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
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