In vivo behaviour of rat band 3 cross-linked carrier erythrocytes.

Rat band 3 cross-linked carrier erythrocytes have been prepared. Iodinated carbonic anhydrase has been encapsulated into rat erythrocytes. Then, carrier erythrocytes were labeled with 51chromium. Eventually, these doubly labeled rat RBCs were treated with a band 3 cross-linking reagent, namely bis(sulfosuccinimidyl)suberate (BS3). 51Chromium labeling and 125I CA showed to have cytosolic localization in cross-linked carrier erythrocytes. Estimation of the band 3 cross-linking induced by BS3 on rat carrier erythrocytes has been done rendering values around 25% of band 3 monomer reduction. BS3-cross-linked carrier erythrocytes when injected into rats are mainly targeted to liver as shown by chromium labeling localization. Also, encapsulated CA radioactivity carried by cross-linked carrier rat erythrocytes when injected into rats is localized predominantly in liver as shown by in vivo experiments. Accordingly, cross-linked carrier erythrocytes are highly recognized by peritoneal macrophages as detected by in vitro analyses of macrophage recognition. Thus, our data revealed a targeting of carrier rat erythrocytes induced by cross-linking of band 3 protein by BS3. These results support claims in favor of this animal model as a feasible system to analyze cross-linked carrier erythrocytes survival and targeting as well as the in vivo efficacy of targeting of loaded compounds to liver.