What is FLAER flow cytometry?
What is FLAER flow cytometry?
Fluorescein-labeled proaerolysin (FLAER) is used in a flow cytometric assay to diagnose paroxysmal nocturnal hemoglobinuria (PNH). Proaerolysin binds to the glycophosphatidylinositol(GPI) anchor in the plasma membrane of cells. Cells affected by PNH lack GPI anchoring proteins, and thus are not bound by proaerolysin.
What is FLAER?
FLAER is an Alexa488-labeled inactive vari- ant of aerolysin that does not cause lysis of cells. Our goals were to develop a FLAER-based assay to diagnose and monitor patients with PNH and to improve detection of minor populations of PNH clones in other hematologic disorders.
Why is it called paroxysmal nocturnal hemoglobinuria?
The condition gets its name from one of its symptoms: dark or bright red blood in your urine at night or in the morning. “Paroxysmal” means “sudden,” “nocturnal” means “at night,” and “hemoglobinuria” means “blood in the urine.” It happens in up to 50% of people with PNH.
What is PNH blood disease?
Paroxysmal nocturnal hemoglobinuria (PNH) is a rare acquired, life-threatening disease of the blood. The disease is characterized by destruction of red blood cells (hemolytic anemia), blood clots (thrombosis), and impaired bone marrow function (not making enough of the three blood components).
Which of the following antibodies are used for the diagnosis of PNH using flow cytometry?
To improve the resolution of PNH detection, the present study formulated an 8-color combination for the detection of PNH white blood cells (WBCs), which included lineage-gating antibodies (CD64 for monocytes and CD15 for granulocytes) together with fluorescein-labeled proaerolysin (FLAER) and three GPI-linked markers.
What is CD55 and CD59?
CD55 and CD59 are glycosylphosphatidylinositol (GPI)-anchored proteins with complement inhibitory properties. Congenital deficiency of the expression of these proteins, as in paroxysmal nocturnal hemoglobinuria, lead to augmented sensitivity to complement mediated lysis, which results in hemolytic anemia.
How do you test for PNH?
The main diagnostic test for individuals with suspected PNH is flow cytometry, a blood test that can identify PNH cells (blood cells that are missing GPI-anchored proteins).
What is MDS and PNH?
Bone marrow failure happens when the marrow does not produce enough red cells, white cells or platelets, or the blood cells that are produced are damaged or defective. This means the body can not supply itself with the blood it needs. PNH, along with aplastic anemia and MDS, are bone marrow failure diseases.
Is PNH treatable?
PNH is a debilitating, fatal but treatable disease: same disease, different clinical presentations.
How is aerolysin used in the treatment of parasites?
Aerolysin has also been shown to be useful for the purification of parasites such as trypanosomes and Leishmania from tissue or blood. These parasites are completely insensitive to the toxin, in contrast to the host blood cells, thereby allowing lysis of the mammalian cells and purification of the parasite [116].
How is proaerolysin used in flow cytometric assay?
Fluorescein-labeled proaerolysin. Fluorescein-labeled proaerolysin ( FLAER) is used in a flow cytometric assay to diagnose paroxysmal nocturnal hemoglobinuria (PNH). The assay takes advantage of the action of proaerolysin, a prototoxin of aerolysin, a virulence factor of the bacterium Aeromonas hydrophila.
How does aerolysin affect the permeability of blood cells?
Aerolysins are proteins capable of altering the permeability of blood cells, as well as other eukaryotic cells (and consequently promoting osmotic lysis) due to polymerization of the structure induced by binding to a membrane-specific glycoprotein site [101]. Ioan Iacovache,
What is the structure of the aerolysin monomer?
Aerolysin family of β-PFPs. (a) Structure of aerolysin (1PRE) monomer showing the four structural domains of the protein (D1, orange; D2, red; D3, light blue; and D4, dark blue).
