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Blood, 1 May 2006, Vol. 107, No. 9, pp. 3745-3752.
Prepublished online as a Blood First Edition Paper on December 29, 2005; DOI 10.1182/blood-2005-10-4094.


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RED CELLS

Ex vivo perfusion of human spleens maintains clearing and processing functions

Pierre A. Buffet, Geneviève Milon, Valentine Brousse, Jean-Michel Correas, Bertrand Dousset, Anne Couvelard, Reza Kianmanesh, Olivier Farges, Alain Sauvanet, François Paye, Marie-Noëlle Ungeheuer, Catherine Ottone, Huot Khun, Laurence Fiette, Ghislaine Guigon, Michel Huerre, Odile Mercereau-Puijalon, and Peter H. David

From the Biomedical Research Team, Medical Center, the Molecular Immunology of Parasites Unit, Centre National de la Recherche Scientifique, Unité de Recherche Associée (CNRS URA) 2581, Parasitology Department, the Immunophysiology and Intracellular Parasitism Unit, Parasitology Department, the Unite de Recherche et d'Expertise (URE) Histotechnology and Pathology, and the Public Health Platform, Institut Pasteur, Paris, France; and Assistance Publique-Hôpitaux de Paris, Paris, France.

The spleen plays a central role in the pathophysiology of several potentially severe diseases such as inherited red cell membrane disorders, hemolytic anemias, and malaria. Research on these diseases is hampered by ethical constraints that limit human spleen tissue explorations. We identified a surgical situation—left splenopancreatectomy for benign pancreas tumors—allowing spleen retrieval at no risk for patients. Ex vivo perfusion of retrieved intact spleens for 4 to 6 hours maintained a preserved parenchymal structure, vascular flow, and metabolic activity. Function preservation was assessed by testing the ability of isolated-perfused spleens to retain Plasmodium falciparum-infected erythrocytes preexposed to the antimalarial drug artesunate (Art-iRBCs). More than 95% of Art-iRBCs were cleared from the perfusate in 2 hours. At each transit through isolated-perfused spleens, parasite remnants were removed from 0.2% to 0.23% of Art-iRBCs, a proportion consistent with the 0.02% to 1% pitting rate previously established in artesunate-treated patients. Histologic analysis showed that more than 90% of Art-iRBCs were retained and processed in the red pulp, providing the first direct evidence of a zone-dependent parasite clearance by the human spleen. Human-specific physiologic or pathophysiologic mechanisms involving clearing or processing functions of the spleen can now be experimentally explored in a human tissue context.


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