Detection of red blood cell antibodies in mitogen‐stimulated cultures from patients with hereditary spherocytosis

• DOI: 10.1111/trf.13257
• Publication Date: 2015-08-11T04:37:32Z
• Authors: Anna Zaninoni; Cristina Vercellati; Francesca G. Impe...; Anna P. Marcello; Bruno Fattizzo; Elisa Fermo; Paola Bianchi; Claudia Grossi; Alessandra Cattaneo; Agostino Cortelezzi; Alberto Zanella; Wilma Barcellini
• Keywords: Hereditary spherocytosis; Spherocytosis

BACKGROUND Hereditary spherocytosis (HS) is a congenital hemolytic anemia caused by defects in red blood cell (RBC) membrane proteins leading to premature RBC clearance the spleen. The presence of autoantibodies has never been extensively investigated HS. STUDY DESIGN AND METHODS antibody–bound immunoglobulin (Ig)G was 91 consecutive HS patients mitogen‐stimulated direct antiglobulin test (MS‐DAT), sensitive method able magnify latent antibody autoimmunity and related with variables, previous splenectomy, type defect. RESULTS A total 61% cases had antibodies MS‐DAT (29 Band 3, 17 spectrin deficiency, nine no defined defect). amount RBC‐bound IgG greater compared controls (236 ± 192 ng/mL vs. 52 29 ng/mL, p < 0.0001), although lower than that observed autoimmune (AIHA; 634 371 236 0.0001). Western blot experiments showed purified fraction from MS‐DAT–positive bind α‐ β‐spectrin, 4.9. Positive displayed increased reticulocytosis slightly reduced hemoglobin (Hb) values negative ones. Patients displaying more 250 (the positive threshold AIHA) number spherocytes mainly deficiency. free Hb over time after storage at 4°C, surrogate ex vivo aging, evidently controls, particularly 3 CONCLUSION were detected half patients. evident pattern suggesting pathogenic role these opsonization splenic removal.