Filterability of sickle cells as a function of pO2: role of physico-chemical factors.

A rigidity index (RI) related to red blood cell deformability was measured by using the hemorheometre. The RI for 13 patients homozygous for sickle cell disease was 109 +/- 44 at 37 degrees C and at atmospheric pO2. The filtration time curve as a function of pO2 is biphasic for sickle cell suspensions. The pO2 at which filtration time is maximum, pO2max., correlated with the rigidity index measured at atmospheric pO2. This pO2max. value was very sensitive to small changes in physico-chemical parameters such as osmolality, pH, temperature, hematocrit, and cell density. Conditions which reduced the Hb S polymerization induced a leftward shift of pO2max.. The experimental curves are in agreement with theoretical models based on the presence of two abnormal cell types: filtrable "slow cells" and infiltrable "sickled cells".

Inhibition of erythrocyte sickling by thiol reagents.

The antisickling effects of eight thiol reagents that cross the red cell membrane and then react with the cysteine beta 93, the only accessible thiol group of hemoglobin, have been investigated at various pO2 values. In spite of completely reacted hemoglobins, the potent antisickling effect varied from one compound to the other and was partially related to the extent of the increased oxygen affinity of intact sickle cells induced by these compounds. The formation of methemoglobin upon the incubation of red blood cells with some disulfides had only a small effect on the sickling process.

[Use of a new rheometer for the study of the filtrability of a suspension of sickled red cells as a function of PO2]. / Utilisation d'un nouveau rhéomètre pour étudier la filtrabilité d'une suspension de globules rouges drepanocytaires en fonction de la PO2.

The filtration time of a small volume (0.1 ml) of red cell suspension from normal (AA), heterozygous (AS) and homozygous (SS) subjects for sickle cell disease was investigated as a function of PO2 The curve of filtration time of AS and SS red cell suspensions was biphasic. At high values of PO2, the progressive reduction of filtrability of sickle cell suspensions with decreasing PO2 occurred without new change in morphology of most of the cells. In contrast, at lower PO2 the apparent filtrability was improved and the cells were sickled." However the red blood cells were retained by the filter and the "solvent" filtrability was improved because rigid and highly deformed sickled cells did not clogged completely the pores of the filter. This study allowed to distinguish a new concept of apparent filtrability for red blood cells in sickle cell disease.

Hemoglobin Bougardirey-Mali beta 119 (GH2) Gly replaced by Val. An electrophoretically silent variant migrating in isoelectrofocusing as Hb F.

Hemoglobin Bougardirey-Mali was detected by isoelectrofocusing during a screening in a 32 years old African, a native of Mali. This abnormal Hb, representing 35% of the total, exhibited the same pI as that of Hb F. In contrast, it was indistinguishable from Hb A in all the electrophoretic systems tested, and equally by its resistance to alkaline denaturation. Structural studies have shown that the abnormality was localized on the beta chain. A fingerprint of the tryptic digest of the aminoethylated beta chain indicated the absence of the beta T12 b. The presence of an abnormal beta T12 b was suspected in the T14-15 spot, as indicated by the intensity of staining and its amino acid composition. beta T12 b was isolated by chromatography on PA 35. Its sequential analysis by manual Edman-dansyl degradation showed that glycine 119 was replaced by a valine residue. This mutation is localized in a alpha 1 beta 1 contact, which makes the molecules slightly unstable. The clinical consequences of this mutation seem to be minor; similar observations have been reported for the other Hb mutated at the same locus, i.e. Hb Fannin-Lubbock beta 119 Gly leads to Asp.

Haemoglobin Saki alpha 2 beta 2 14 Leu-Pro(a11) structure and function.

The characterization of haemoglobin Saki alpha 2 beta 2 14 Leu-Pro(a11) is described. This new mutation is unique since it only induces modification of the stability of the molecule. In vitro precipitation of haemoglobin Saki upon heat or in the presence of chemicals is compared to the stability of haemoglobin A and haemoglobin S.