title: Systemic lupus erythematosus serum deposits C4d on red blood cells, decreases red blood cell membrane deformability, and promotes nitric oxide production.
abstract: OBJECTIVE
  Systemic lupus erythematosus (SLE) is characterized by intravascular activation of the complement system and deposition of complement fragments (C3 and C4) on plasma membranes of circulating cells, including red blood cells (RBCs). The aim of this study was to address whether this process affects the biophysical properties of RBCs.
  
  
  METHODS
  Serum and RBCs were isolated from patients with SLE and healthy controls. RBCs from healthy universal donors (type O, Rh negative) were incubated with SLE or control serum. We used flow cytometry to assess complement fragment deposition on RBCs. RBC membrane deformability was measured using 2-dimensional microchannel arrays. Protein phosphorylation levels were quantified by Western blotting.
  
  
  RESULTS
  Incubation of healthy universal donor RBCs with sera from patients with SLE, but not with control sera, led to deposition of C4d fragments on the RBCs. Complement-decorated RBCs exhibited significant decreases in both membrane deformability and flickering. Sera from SLE patients triggered a transitory Ca(++) influx in RBCs that was associated with decreased phosphorylation of β-spectrin and with increased phosphorylation of band 3, two key proteins of RBC cytoskeleton. Finally, incubation with SLE sera led to the production of nitric oxide by RBCs, whereas this did not occur with control sera.
  
  
  CONCLUSIONS
  Our data suggest that complement activation in patients with SLE leads to calcium-dependent cytosketeletal changes in RBCs that render them less deformable, probably impairing their flow through capillaries. This phenomenon may negatively affect the delivery of oxygen to the tissues.
authors.affiliation: Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Ionita C
authors.initials: IC
authors.lastname: Ghiran
authors.forename: Mark L
authors.initials: ML
authors.lastname: Zeidel
authors.forename: Sergey S
authors.initials: SS
authors.lastname: Shevkoplyas
authors.forename: Jennie M
authors.initials: JM
authors.lastname: Burns
authors.forename: George C
authors.initials: GC
authors.lastname: Tsokos
authors.forename: Vasileios C
authors.initials: VC
authors.lastname: Kyttaris
mesh_terms: D000017:ABO Blood-Group System; D000328:Adult; D001782:Blood Donors / Q000145:classification; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D015935:Complement C4b / Q000276:immunology / Q000378:metabolism*; D004907:Erythrocyte Deformability / Q000502:physiology*; D004910:Erythrocyte Membrane / Q000276:immunology / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology / Q000378:metabolism* / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D009569:Nitric Oxide / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D044967:Serum / Q000378:metabolism*; D012720:Severity of Illness Index; D013049:Spectrin / Q000378:metabolism; D055815:Young Adult
chemical_list: D000017:ABO Blood-Group System; D010446:Peptide Fragments; D013049:Spectrin; D009569:Nitric Oxide; D015935:Complement C4b; C032261:complement C4d; D002118:Calcium