A fluorometric erythrophagocytosis assay using differentiated monocytic THP-1 cells to assess the clinical significance of antibodies to red blood cells.

Vox Sanguinis
Abdelwahab Hassan Ahmed BalolaA Salama

Abstract

The significance of antibodies to red blood cells (RBCs) is variable and cannot be predicted solely by serological testing. A flow cytometry-based erythrophagocytosis assay was established using phorbol 12-myristate 13-acetate (PMA)-treated THP-1 cells and RBCs labelled with PKH26 to assess allo- and autoantibodies to RBCs. THP-1 cells were differentiated into macrophage-like cells by treatment with PMA. RBC samples coated with alloantibodies or autoantibodies were obtained from 16 patients with autoimmune haemolytic anaemia of warm type (wAIHA) as well as from five pregnant women with warm autoantibodies. RBCs from healthy blood donors were used as controls. RBCs were labelled with the red lipophilic fluorescent dye PKH26 and incubated with PMA-treated THP-1 cells. After removal of nonadherent RBCs by washing and haemolysis of adherent RBCs, erythrophagocytosis was quantified by flow cytometry. We observed significant phagocytosis of RBCs coated with clinically relevant alloantibodies (i.e. anti-D and anti-K) or autoantibodies from patients with active wAIHA, but not of those coated with alloantibodies (anti-Ch) or autoantibodies from patients and pregnant women without haemolysis. The flow cytometry-based erythrophagocytosis ...Continue Reading

References

Jan 5, 2002·Transfusion·B HoppeA Salama
Oct 13, 2006·Annual Review of Immunology·James N ArnoldRaymond A Dwek
Mar 10, 2007·Journal of Immunological Methods·Geraldine HealeyRosemary L Sparrow
Jan 1, 2008·Transfusion Medicine and Hemotherapy : Offizielles Organ Der Deutschen Gesellschaft Fur̈ Transfusionsmedizin Und Immunham̈atologie·Erwin Strobel
Aug 16, 2011·Advances in Experimental Medicine and Biology·Anja Lux, Falk Nimmerjahn
Jan 25, 2012·Blood Purification·Florian Lang, Syed M Qadri
Jun 28, 2014·Transfusion Medicine Reviews·Donald M MockRobert S Franco
Dec 24, 2015·Transfusion Medicine and Hemotherapy : Offizielles Organ Der Deutschen Gesellschaft Fur̈ Transfusionsmedizin Und Immunham̈atologie·Abdulgabar Salama
Dec 24, 2015·Transfusion Medicine and Hemotherapy : Offizielles Organ Der Deutschen Gesellschaft Fur̈ Transfusionsmedizin Und Immunham̈atologie·Gülüstan SürücüAbdulgabar Salama
Oct 25, 2016·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Etheresia PretoriusJanette Bester
Sep 30, 2017·European Journal of Haematology·Thilo BartolmäsAbdulgabar Salama
Feb 28, 2018·Transfusion·Floris HelmichFedde van der Graaf
Aug 25, 2018·Transfusion Medicine Reviews·Tik Nga TongDonald R Branch

❮ Previous
Next ❯

Related Concepts

Related Feeds

Autoimmune Hemolytic Anemia

Autoimmune hemolytic anemia (AIHA) occurs when antibodies directed against the person's own red blood cells (RBCs) cause them to burst (lyse), leading to an insufficient number of oxygen-carrying red blood cells in the circulation. Discover the latest research on AIHA here.