Biotechnology and Applied Biochemistry (1998) 27, (133–137) (Printed in Great Britain)
Differential induction of macrophage recognition of carrier erythrocytes by treatment with band 3 cross-linkers
José A. Jordán, F. Javier Alvarez, L. Alfredo Lotero, Gemma Olmos, Paloma Calleja, M. Cristina Tejedor and José C. Díez
Departamento de Bioquímica y Biología Molecular, Campus Universitario, Universidad de Alcalá, 28871 Alcalá de Henares (Madrid), Spain
Abbreviations: BS3, bis(sulphosuccinimidyl)suberate;
CA, carbonic anhydrase;
DTSP, 3,3 -dithiobis(sulphosuccinimidyl propionate).
Correspondence: José C. Díez, Departamento de
Bioquímica y Biología
Molecular, Campus Universitario, Universidad de Alcalá, 28871 Alcalá de Henares
(Madrid), Spain
Mouse native and hypotonically loaded erythrocytes were treated with
two cross-linking reagents: bis(sulphosuccinimidyl)suberate
(BS3)- and 3,3-
dithiobis(sulphosuccinimidyl propionate) (DTSP), excluding
clustering agents. Microscopic analyses revealed that band 3 cross-linked native
and hypotonically loaded erythrocytes are more strongly recognized by peritoneal
macrophages than native and loaded erythrocytes as a result of
the cross-linking of band 3 protein in accordance with studies
in vivo. Macrophage-recognition analyses of
51Cr-labelled erythrocytes also demonstrated increased
recognition of cross-linked and cross-linked loaded erythrocytes. This shows that
the only action of these two band 3 cross-linkers on
mouse erythrocytes promotes recognition by macrophages without requiring the use
of clustering agents. The extent of recognition of BS3 cross-linked
and cross-linked loaded erythrocytes by macrophages is dependent on the
presence or absence of homologous serum or immunoglobulins. In contrast,
the presence of serum factors or IgG in the incubation
medium did not seem to influence the recognition of DTSP-modified
erythrocytes by macrophages. These results seem to indicate a different
mechanism of recognition for the erythrocytes modified with either one
or the other band 3 cross-linker. In summary, the unique
use of both band 3 cross-linkers procedures can be used
to target carrier erythrocytes conveying active compounds to macrophages, with
possible therapeutical applications. Different mechanisms of
induction of macrophage recognition
by these band 3 cross-linkers could reveal differential actions on
erythrocytes or the involvement of different factors in the recognition
process.
Received 29 September 1997/20
November 1997; accepted 24 November 1997
© 1998 The International Union of Biochemistry and Molecular Biology
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