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Senescence of canine biotinylated erythrocytes: increased autologous
immunoglobulin binding occurs on erythrocytes aged in vivo for 104 to 110
days
JA Christian, AH Rebar, GD Boon and PS Low
Department of Chemistry, Purdue University, West Lafayette, IN 47907.
We have evaluated senescence related changes in canine red blood cells
(RBCs) using the biotinylation system, where RBCs are labeled in vivo with
biotin at the beginning of their life span, and retrieved from circulation
on immobilized avidin at the end of their life span. This approach avoids
the controversial use of density gradient centrifugation to collect
presumably old RBCs. Furthermore, the dog is an appropriate model for human
RBC senescence because it has a low degree of random RBC loss and a
similarly long RBC life span (approximately 110 days). Two dogs had 97% to
100% of their circulating RBCs biotinylated by infusion of
N-hydroxysuccinimido biotin (Clontech, Palo Alto, CA; Calbiochem, La Jolla,
CA) dissolved in dimethyl sulfoxide. At postbiotinylation days 104 and 107
for one dog and day 110 for the other dog, biotinylated RBCs were isolated
by magnetic cell sorting and analyzed for the presence of autologous IgG
using 125I- labeled sheep-antidog IgG (SAD IgG). On all 3 days, there were
at least three times more SAD IgG molecules per RBC on senescent
biotinylated RBCs than on control (unfractionated) RBCs (day 104: 11,677 v
3,399; day 107: 6,710 v 2,115; day 110: 6,042 v 1,838 molecules of SAD IgG
per senescent v control RBC). Furthermore, it is unlikely that an immune
response to the conjugated biotin had been elicited, because fresh in vitro
biotinylated RBCs that were incubated in autologous plasma (taken after
exposure to circulating biotinylated RBCs for 113 days) and then exposed to
the SAD IgG showed no increase in antibody binding over control
(non-biotinylated) RBCs (1,431 v 1,378 cpm/10(8) biotinylated v control
RBCs; P > .20). These results suggest that senescence of canine
biotinylated RBCs is characterized by binding of autologous IgG and that
antibiotin antibodies do not contribute to this process.
Volume 82,
Issue 11,
pp. 3469-3473,
12/01/1993
Copyright © 1993 by The American Society of Hematology
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