Freeze preservation of sickle erythrocytes

Castro, Oswaldo; Hardy, Kathryn Phillips; Winter, William P.; Hornblower, M.; Meryman, Harold T.

doi:10.1002/ajh.2830100309

Cited by 20 publications

(11 citation statements)

References 10 publications

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“…The solubility of an equimolar mixture of Hb S plus Hb C measured in low-ionicstrength phosphate buffer (,u < 0.2) is, however, lower than that of a similar mixture of Hb S plus Hb A (39). The isosolubility of deoxy-Hb F or deoxy-Hb A2 in a 50:50 mixture with deoxy-Hb S confirms an earlier report to this effect (4) but is discrepant with others that report Hb A2 to be either less (7) or more (10) effective than Hb F in enhancing the solubility of unliganded Hb S. In any case, our results, in which buffer composition, pH, and temperature were care- (38) (29) at 37°C, the measured solubilities more closely mimic those ofthe intact cell. These studies assess the influence of non-S Hbs on the solubility of Hb S as a function of ligand saturation under conditions that approximate those of the sickling disorders.…”

Section: Discussioncontrasting

confidence: 67%

Sparing effect of hemoglobin F and hemoglobin A2 on the polymerization of hemoglobin S at physiologic ligand saturations.

Poillon

Kim

Rodgers

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

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Recent interest in therapies for sickle cell anemia based on elevating fetal Hb has made accurate estimates of the sparing effect offetal Hb (Hb F) and other non-sickle Hbs on sickle Hb (Hb S) polymerization essential. We have developed a technique, using HbCO as surrogate for HbO2, that enables us to assess the solubility of Hb S as a function of ligand saturation under conditions that mimic those of the sickling disorders. Equimolar mixtures of unliganded Hb S with Hb F or normal Hb A2 were isosoluble. Solubilities for equimolar mixtures with normal (Hb A) or abnormal (Hb C) Hbs were also identical but were lower than in the prior case. Thus, the sparing effect of both Hb F and Hb A2 should be considered in therapeutic strategies designed to modify Hb S polymerization. Hemolysates, stripped of 2,3-bisphosphoglycerate, from sickle cell disease patients with Hb (F + A2) levels varying from 6 to 25%, as well as from a sickle trait individual, were used to evaluate equilibrium solubiity as a function of ligand saturation over the range of pathophysiologic interest (25-70%). Our results show that the sparing effect of Hb (F + A2) increases relative to that of Hb A as ligand saturation increases, and that in the absence of ligand, '30% Hb (F + A2) is essentially isosoluble with the 60% Hb A of sickle trait. Although detailed knowledge of expected therapeutic benefits is confounded by the heterogeneity of Hb F distribution and other variables, these data should provide a framework for estimating likely clinical benefit from pharmacologic efforts to modulate globin gene expression.Sickle cell anemia exists in individuals who are homozygous for a point mutation (GAG --GTG) at codon 6 of the 3-globin gene, which results in a Glu-6 -* Val substitution in P-globin. are, thus, said to have a "sparing" effect on intracellular Hb S polymerization (3, 4). For Hbs F and A2, this solubilization can be described well with a model that assumes complete exclusion of either the homotetramer (a2-y2 or a2&2) or the hybrid heterotetramer (a238y or a2138) from the nascent polymer (5-7). For Hbs A and C, the data are compatible with exclusion of only homotetramers (a2I3A and a2K) from the polymer. Hybrid heterotetrameric forms (a2f3Sf3A and a2138S,c) appear to be incorporated into the polymer, but with a copolymerization probability roughly one-half that of Hb S (8-10). These copolymerization tendencies explain why the solubility enhancement induced by Hb F or A2 in admixture with Hb S is greater than that for Hb A or C.The solubilizing properties of various non-S Hbs have been established by use of a chemical reducing agent (dithionite) to completely deoxygenate mixtures of oxy-Hb S and the oxy form of other hemoglobins (Hb X), followed by centrifugation to fractionate the resulting semisolid gel into a tightly packed polymer pellet and a supernatant consisting of soluble Hb S and Hb X. The concentration of all Hb species in the supernatant is by definition the saturation concentration (csat), or equilibrium solubility, under g...

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Section: Discussioncontrasting

confidence: 67%

Sparing effect of hemoglobin F and hemoglobin A2 on the polymerization of hemoglobin S at physiologic ligand saturations.

Poillon

Kim

Rodgers

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

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“…Although there are reports of gelling after thawing of cryopreserved red blood cell products obtained from donors with SCT, 13,15,19 no such gelling was observed in our study, regardless of whether or not products from SCT donors underwent red cell removal before cryopreservation. However, special processing requirements of products obtained from SCT subjects were identified.…”

Section: Discussioncontrasting

confidence: 46%

“…Finally, there have been reports of red cell gelling and clumping, 13,14 requiring special processing of blood products obtained from SCT donors. 13,15 Because there is no urgency with regard to the timing of transplantation for patients with SCA, it should be possible to collect and preserve PBSCs before transplantation.…”

Section: Introductionmentioning

confidence: 99%

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Mobilization, collection, and processing of peripheral blood stem cells in individuals with sickle cell trait

Kang

Areman

David-Ocampo

et al. 2002

Blood

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Mobilized peripheral blood is increasingly used as the source of hematopoietic stem cells for allogeneic transplantation, currently the only curative approach for sickle cell anemia. However, the safety and feasibility of stem cell mobilization in individuals with sickle cell trait (SCT) has not been documented. This study is a prospective controlled trial to evaluate the safety and feasibility of peripheral blood stem cell (PBSC) mobilization in 8 SCT subjects and 8 control subjects matched for age and race. Mobilization with filgrastim 10 g/kg subcutaneous daily for 5 days was followed by 12-L apheresis on the fifth day. Filgrastim administration was accompanied by similar symptoms in all subjects; no untoward adverse events occurred in either group, including sickle cell crises. CD34 ؉ cell mobilization response was not significantly different between SCT and control subjects. Median CD34 ؉ cell content was also similar in PBSCs collected from SCT versus control subjects, 6.8 versus 3.9 ؋10 6 CD34 ؉ cells/70 kg, P ‫؍‬ .165. Red cell depletion from SCT products was not possible by using hydroxyethyl starch sedimentation but was achievable with ammonium chloride lysis. There was no evidence of gelling of SCT products after thaw, and no difference in cell recovery was seen among red cell-depleted versus nondepleted products. Cryopreservation in 5% dimethyl sulfoxide/6% pentastarch was associated with superior cell recovery (both SCT and control subjects) compared with 10% dimethyl sulfoxide (P ‫؍‬ .001). The study concluded that filgrastim mobilization, large volume apheresis, processing, and cryopreservation appears to be safe in donors with SCT, allowing PBSC use for transplantation in patients with sickle cell anemia. (Blood. 2002;99:850-855)

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“…There are no naturally occurring animal models for the disease. For this reason, transfusions of human sickle erythrocytes to laboratory animals such as rats have been used to investigate the disease's pathophysiology [2][3][4][5] and the in vivo effectiveness of agents that inhibit red cell sickling [6][7][8]. However, because of their anatomic and physiologic differences from man, small laboratory animals do not represent optimal models for the circulation and function of human red cells.…”

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confidence: 99%

Human Sickle Erythrocytes: Survival in Chimpanzees

Castro¹,

Ww²,

Moor‐Jankowski³

1982

J Med Primatol

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The survival characteristics of human sickle (SS) erythrocytes (RBCs) transfused to intact chimpanzees were determined. The mean post-transfusion recovery of^51 Crlabelled SS RBCs in four chimpanzees was 30.5 % ±15.2 SD, and the half-life survival was 4.2 h ± 0.8 SD. The recovery of control (hemoglobin AA) human red cells in five chimpanzees was complete and their mean intravascular T 1/2 was 22.3 h. Shorter survival of sickle erythrocytes was also shown by transfusing chimpanzees with mixtures of human cells such as 51Cr AA RBCs and 59Fe SS RBCs, or 51Cr SS RBCs and non-labelled fetal (cord blood) erythrocytes. The difference in survival of AA and SS RBCs resembles that in human recipients and was probably caused by sickling of SS cells in the chimpanzee circulation. These primate animals could, therefore, be used as a model for in vivo studies of sickle cell disease.

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Freeze preservation of sickle erythrocytes

Cited by 20 publications

References 10 publications

Sparing effect of hemoglobin F and hemoglobin A2 on the polymerization of hemoglobin S at physiologic ligand saturations.

Sparing effect of hemoglobin F and hemoglobin A2 on the polymerization of hemoglobin S at physiologic ligand saturations.

Mobilization, collection, and processing of peripheral blood stem cells in individuals with sickle cell trait

Human Sickle Erythrocytes: Survival in Chimpanzees

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