Flow cytofluorimetric analysis of young and senescent human erythrocytes probed with lectins. Evidence that sialic acids control their life span
- Cite this article as:
- Bratosin, D., Mazurier, J., Debray, H. et al. Glycoconjugate J (1995) 12: 258. doi:10.1007/BF00731328
Analysis by flow cytofluorimetry of the binding of fluorescein isothiocyanate labelled lectins specific for sialic acids shows that the aged erythrocytes bind less WGA, LPA, SNA and MAA than young erythrocytes. The binding of DSA and LCA is not modified. On the contrary, the number of binding sites of UEA-I specific for O antigen and of AAA decreases significantly. PNA and GNA do not bind to erythrocytes.
RCA120 as well asErythrina cristagalli andErythrina corallodendron lectins specific for terminal β-galactose residues lead to unexpected and unexplained results with a decrease in the number of lectin binding sites associated with increasing desialylation.
The glycoconjugates from the old erythrocytes incorporate more sialic acid than the young cells. This observation results from the determination of the rate of transfer by α-2,6-sialyltransferase of fluorescent or radioactiveN-acetylneuraminic acid, using as donors CMP-9-fluoresceinyl-NeuAc and CMP-[14C]-NeuAc, respectively.
Microscopy shows that the old erythrocytes are captured preferentially by the macrophages relative to the young ones. Fixation of erythrocytes by the macrophage membrane is inhibited by lactose, thus demonstrating the involvement of a terminal β-galactose specific macrophage lectin.
Comparative study of the binding of WGA, LPA, SNA and MAA to the aged erythrocytes and to thein vitro enzymatically desialylated erythrocytes shows that the desialylation rate of aged cells is low but sufficient to lead to their capture by the macrophages
KeywordsSenescent erythrocyteslectinsflow cytofluorimetrysialic aciderythrophagocytosismacrophagesendogeneous galactolectin
bovine serum albumin
cell sialylation buffer
ethylene diamine tetraacetic acid
polyacrylamide gel electrophoresis
Dulbecco's phosphate buffer saline solution
red blood cells
Senescent Cell Antigen
sodium dodecyl sulfate
senescent factor glycopeptides
Aleuria aurantia agglutinin
Datura stramonium agglutinin
Erythrina cristagalli agglutinin
Galanthus nivalis agglutinin
Lens culinaris agglutinin
Limax flavus agglutinin
Limulus polyphemus agglutinin
Maackia amurensis agglutinin
Ricinus communis agglutinin
Sambucus nigra agglutinin
Ulex europeus agglutinin-I
Wheat germ agglutinin