Differentiation between high- and low-grade astrocytoma using a human recombinant antibody to the extra domain-B of fibronectin - PubMed (original) (raw)

Comparative Study

Differentiation between high- and low-grade astrocytoma using a human recombinant antibody to the extra domain-B of fibronectin

Patrizia Castellani et al. Am J Pathol. 2002 Nov.

Abstract

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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Figures

Figure 1.

Figure 1.

Serial sections of a glioblastoma (A and B), normal brain (C and D), and a pilocytic astrocytoma (E and F) stained with the scFv L-19 specific for B-FN (A, C, E) and with the anti-factor VIII mAb (B, D, F). Scale bars, 10 μm.

Figure 2.

Figure 2.

Serial sections of a anaplastic astrocytoma showing neoplastic and normal tissue (A and B) stained with the anti-factor VIII mAb and with scFv L-19, respectively. Only the vessels within the tumors were stained by the scFv L-19. Two different magnifications (C and D) of a glioblastoma multiforme specimen showing neoplastic and normal tissue double stained with the scFv L-19 (brown) and with anti-factor VIII mAb (red). Two different magnifications (E and F) of an in situ hybridization experiment using human glioblastoma cryostat sections with the DIG-labeled cRNA ED-B repeat probe. A positive signal was visible in the endothelial cells. Scale bars, 10 μm.

Figure 3.

Figure 3.

Left: Number of Ki-67-positive cells calculated in five different areas of each specimen of high- and low-grade astrocytomas using the mAb specific for proliferating cells. Middle: Number of vessels calculated in five different areas of each specimen of high- and low-grade astrocytomas using the mAb anti-factor VIII. Right: Percentage of B-FN-positive vessels calculated in five different areas of each specimen of high- and low-grade astrocytomas double stained using the scFv L-19 and the mAb anti-factor VIII. Although considerable overlapping of the results of high- and low-grade astrocytomas for proliferating cells and vascular density was observed, no overlapping was seen when measuring the percentage of B-FN-positive vessels.

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