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(American Journal of Pathology. 2002;161:1695-1700.)
© 2002 American Society for Investigative Pathology

Differentiation between High- and Low-Grade Astrocytoma Using a Human Recombinant Antibody to the Extra Domain-B of Fibronectin

Patrizia Castellani*, Laura Borsi*, Barbara Carnemolla*, Attila Birò*, Alessandra Dorcaratto{dagger}, Giuseppe L. Viale{dagger}, Dario Neri{ddagger} and Luciano Zardi*

From the Laboratory of Cell Biology,* Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy; the Department of Surgery,{dagger} Division of Neurosurgery, Department of Surgical Sciences, Anesthesiology, and Organ Transplantation, University of Genoa, Italy; and the Department of Applied BioScience,{ddagger} Swiss Federal Institute of Technology, Zürich, Switzerland

Different fibronectin (FN) isoforms are generated by the alternative splicing of the primary FN transcript. We previously demonstrated that the isoform containing the extra domain B sequence of fibronectin (B-FN), a complete type-III-homology repeat, is a marker of angiogenesis that accumulates around neovasculature only during angiogenic processes. We produced a single-chain human recombinant antibody (scFv), L19, which reacts specifically with B-FN and selectively targets tumor vasculature in vivo. We used this scFv and an antibody against a pan-endothelial marker (Factor VIII) in a double-staining procedure on specimens of low- and high-grade astrocytomas to determine the percentage of B-FN-positive vessels, (denominating the resulting value angiogenic index [AI]). Compared to vascular density and proliferative activity (evaluated using antibodies to Factor VIII and Ki67, respectively), AI correlated better with tumor grade (1.6 ± 2.6% and 92.0 ± 8.7% of B-FN-positive vessels in low- and high-grade astrocytomas, respectively) and was a more precise diagnostic tool than either of the two conventional methods. In fact, discriminating analysis using these three parameters showed that only AI accurately classified 100% of the cases studied, compared to 64% and 89% correctly diagnosed by vascular density and of proliferating cells, respectively.




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