title: A second generation transgenic mouse model expressing both hemoglobin S (HbS) and HbS-Antilles results in increased phenotypic severity.
abstract: We report on a second generation of transgenic mice produced by crossing a transgenic mouse line expressing high levels of human alpha and beta S chains (alpha H beta S [beta MDD]) with a line expressing human alpha and beta S-Antilles (beta SAnt). We hypothesized that mice expressing both hemoglobins (Hbs) would have a more severe phenotype because the reduced oxygen affinity and solubility of the beta S-Antilles might enhance the rate and extent of polymer formation. We obtained mice that expressed both beta S and beta S-Antilles. The doubly transgenic mice that are heterozygous for deletion of mouse beta Major (beta MD) occurred with reduced frequency and those that are homozygous for deletion of mouse beta Major (beta MDD) occurred at a much reduced frequency and suffered early mortality. Human alpha was 58% of all alpha globin for all animals, whereas beta S and beta S-Antilles were 34% and 28% of all beta globins for beta MD mice and 42% and 36% for beta MDD mice. Hematocrit, Hb, and mean corpuscular Hb were normal for all transgenic mice, but reticulocyte levels were higher for the doubly transgenic mice versus alpha H beta S [beta MDD] mice older than 30 days (10.0% +/- 1.0% v 4.3% +/- 0.4%; P < .001, mean +/- SE, n = 20 and n = 10, respectively) and control mice (3.9% +/- 0.4%). Reticulocytosis was more severe in mice less than 30 days old ( > 20% for alpha H beta S beta S-Ant[beta MDD] mice). The median mean corpuscular hemoglobin concentration of doubly transgenic mice was higher than that of alpha H beta S[beta MDD] mice with a variable number of very dense cells. Delay times for polymerization of Hb in red blood cells from alpha H beta S beta S-Ant[beta MDD] mice were shorter than those of alpha H beta S[beta MDD] mice, and there were fewer cells with delay times greater than 100 seconds. Urine-concentrating ability in control mice under ambient conditions is 2,846 +/- 294 mOsm and was reduced 30% to 1,958 +/- 240 mOsm, P < 4 x 10(-8) in all mice expressing both transgenes. We conclude that doubly transgenic mice have a more severe phenotype than either of the two parental lines. These mice may be suitable for validating therapeutic intervention in sickle cell disease.
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: M E
authors.initials: ME
authors.lastname: Fabry
authors.forename: A
authors.initials: A
authors.lastname: Sengupta
authors.forename: S M
authors.initials: SM
authors.lastname: Suzuka
authors.forename: F
authors.initials: F
authors.lastname: Costantini
authors.forename: E M
authors.initials: EM
authors.lastname: Rubin
authors.forename: J
authors.initials: J
authors.lastname: Hofrichter
authors.forename: G
authors.initials: G
authors.lastname: Christoph
authors.forename: E
authors.initials: E
authors.lastname: Manci
authors.forename: D
authors.initials: D
authors.lastname: Culberson
authors.forename: S M
authors.initials: SM
authors.lastname: Factor
authors.forename: R L
authors.initials: RL
authors.lastname: Nagel
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood / Q000235:genetics* / Q000473:pathology; D000818:Animals; D001921:Brain / Q000473:pathology; D002499:Centrifugation, Density Gradient; D003433:Crosses, Genetic; D004195:Disease Models, Animal; D004906:Erythrocyte Count; D004913:Erythrocytes, Abnormal; D005914:Globins / Q000235:genetics*; D006451:Hemoglobin, Sickle / Q000096:biosynthesis / Q000235:genetics*; D006454:Hemoglobins / Q000032:analysis; D006455:Hemoglobins, Abnormal / Q000235:genetics; D006579:Heterozygote; D006801:Humans; D007668:Kidney / Q000473:pathology; D008099:Liver / Q000473:pathology; D008168:Lung / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic / Q000097:blood / Q000235:genetics*; D009929:Organ Size; D009994:Osmolar Concentration; D010641:Phenotype; D017354:Point Mutation; D011994:Recombinant Proteins / Q000235:genetics; D012156:Reticulocytes; D012720:Severity of Illness Index; D013154:Spleen / Q000473:pathology; D014556:Urine / Q000737:chemistry
chemical_list: D006451:Hemoglobin, Sickle; D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D011994:Recombinant Proteins; C055657:hemoglobin S-Antilles; D005914:Globins; C100003:deoxyhemoglobin; C026237:hemoglobin D Punjab