Red blood cell membrane and density changes under ambient and hypoxic conditions in transgenic mice producing human...

Red blood cells (RBC) from patients with sickle cell disease (SCD) are characterized by membrane lesions caused by cell sickling and oxidative damage due to denatured hemichromes. We have developed three lines of transgenic human sickle hemoglobin (Hb S) mice, which produce 30, 50, and 80% human beta sickle globin (h beta S), by crossing transgenic progeny with nonthalassemic, heterozygous, or homozygous beta-thalassemic mice, respectively. Transgenic mice that produce Hb A, developed in a similar fashion, were used as controls. RBC from each transgenic line were examined for pathologic changes. RBC from 50 and 80% h beta S mice sickle upon deoxygenation in vitro while RBC from 30% h beta S mice and all Hb A mice do not. Density gradients of RBC from each Hb S line, including those from 30% h beta S that do not sickle, show broad distributions with increased dense fractions, similar to those of patients with SCD. RBC from Hb S lines exhibit elevated membrane-associated denatured hemoglobin (MADH) levels (0.250 +/- 0.080%) when compared to RBC from nontransgenic (0.073 +/- 0.021%) and transgenic Hb A (0.062 +/- 0.033%) mice. Elevated MADH levels in RBC from the 30% h beta S line suggest that membrane changes occur even though these cells do not sickle. These Hb S-dependent pathologic changes suggest that transgenic Hb S lines may be useful for the study of not only RBC sickling in vivo but also membrane oxidative damage and other chronic changes attributed to abnormal properties of both oxygenated and deoxygenated Hb S.