Dapsone hydroxylamine induces premature removal of human erythrocytes by membrane reorganization and antibody binding.
OBJECTIVE
N-hydroxylation of dapsone leads to the formation of the toxic hydroxylamines responsible for the clinical methaemoglobinaemia associated with dapsone therapy. Dapsone has been associated with decreased lifespan of erythrocytes, with consequences such as anaemia and morbidity in patients treated with dapsone for malaria. Here, we investigated how dapsone and/or its hydroxylamine derivative (DDS-NHOH) induced erythrocyte membrane alterations that could lead to premature cell removal.


METHODS
Erythrocytes from healthy donors were subjected to incubation with dapsone and DDS-NHOH for varying times and the band 3 protein tyrosine-phosphorylation process, band 3 aggregation, membrane alteration and IgG binding were all examined and compared with erythrocytes from two patients receiving dapsone therapy.


RESULTS
The hydroxylamine derivative, but not dapsone (the parent sulphone) altered membrane protein interactions, leading both to aggregation of band 3 protein and to circulating autologous antibody binding, shown in erythrocytes from patients receiving dapsone therapy. The band 3 tyrosine-phosphorylation process can be used as a diagnostic system to monitor membrane alterations both in vitro, assessing concentration and time-dependent effects of DDS-NHOH treatment, and in vivo, evaluating erythrocytes from dapsone-treated patients, in resting or oxidatively stimulated conditions.


CONCLUSIONS
DDS-NHOH-induced alterations of human erythrocytes can be directly monitored in vitro by tyrosine-phosphorylation level and formation of band 3 protein aggregates. The latter, together with antibody-mediated labelling of erythrocytes, also observed after clinical use of dapsone, may lead to shortening of erythrocyte lifespan.
Bordin
Luciana
L
Department of Biological Chemistry, University of Padova, Viale G. Colombo 3, Padua, Italy. luciana.bordin@unipd.it
Fiore
Cristina
C
Zen
Francesco
F
Coleman
Michael D
MD
Ragazzi
Eugenio
E
Clari
Giulio
G
D001457:Anion Exchange Protein 1, Erythrocyte / Q000187:drug effects / Q000378:metabolism; D000890:Anti-Infective Agents / Q000008:administration & dosage / Q000633:toxicity*; D000906:Antibodies / Q000378:metabolism; D003622:Dapsone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010766:Phosphorylation / Q000187:drug effects; D013997:Time Factors; D014443:Tyrosine / Q000187:drug effects / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D000890:Anti-Infective Agents; D000906:Antibodies; D007074:Immunoglobulin G; D014443:Tyrosine; D003622:Dapsone; C018209:4-amino-4'-hydroxylaminodiphenylsulfone