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. 2007 Jun 18;96(12):1862-70.
doi: 10.1038/sj.bjc.6603806. Epub 2007 May 22.

Characterisation and radioimmunotherapy of L19-SIP, an anti-angiogenic antibody against the extra domain B of fibronectin, in colorectal tumour models

E El-Emir  1 , J L J Dearling, A Huhalov, M P Robson, G Boxer, D Neri, G A M S van Dongen, E Trachsel, R H J Begent, R B Pedley
Affiliations

Characterisation and radioimmunotherapy of L19-SIP, an anti-angiogenic antibody against the extra domain B of fibronectin, in colorectal tumour models

E El-Emir et al. Br J Cancer. .

Abstract

Angiogenesis is a characteristic feature of tumours and other disorders. The human monoclonal antibody L19- SIP targets the extra domain B of fibronectin, a marker of angiogenesis expressed in a range of tumours. The aim of this study was to investigate whole body distribution, tumour localisation and the potential of radioimmunotherapy with the L19-small immunoprotein (SIP) in colorectal tumours. Two colorectal tumour models with highly different morphologies, the SW1222 and LS174T xenografts, were used in this study. Localisation and retention of the L19-SIP antibody at tumour vessels was demonstrated using immunohistochemistry and Cy3-labelled L19-SIP. Whole body biodistribution studies in both tumour models were carried out with (125)I-labelled L19-SIP. Finally, (131)I-labelled antibody was used to investigate the potential of radioimmunotherapy in SW1222 tumours. Using immunohistochemistry, we confirmed extra domain B expression in the tumour vasculature. Immunofluorescence demonstrated localisation and retention of injected Cy3-labelled L19-SIP at the abluminal side of tumour vessels. Biodistribution studies using a (125)I-labelled antibody showed selective tumour uptake in both models. Higher recorded values for localisation were found in the SW1222 tumours than in the LS174T (7.9 vs 6.6 %ID g(-1)), with comparable blood clearance for both models. Based on these results, a radioimmunotherapy study was performed in the SW1222 xenograft using (131)I-Labelled L19-SIP (55.5 MBq), which showed selective tumour uptake, tumour growth inhibition and improved survival. Radio- and fluorescence-labelled L19-SIP showed selective localisation and retention at vessels of two colorectal xenografts. Furthermore, (131)I-L19-SIP shows potential as a novel treatment of colorectal tumours, and provides the foundation to investigate combined therapies in the same tumour models.

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Figures

Figure 1
Figure 1
Immunohistochemical and immunofluorescence staining with L19-SIP antibody in SW1222 and LS174T xenograft-bearing mice. (A) Immunohistochemical staining of SW1222 xenograft lines with L19-SIP antibody. (B–D) Multiple digital fluorescence images of an LS174T tumour injected with Cy3-labelled L19-SIP antibody for 6 h demonstrating (B) perfusion, (C) blood vessel staining and (D) L19-SIP in the same tumour section. (E) Triple fluorescence staining of CD31 (green), L19-SIP (red) and DAPI (blue) in LS174T tumours. (F) High-power image of CD31 (green), L19-SIP (red) and Hoechst (blue) staining in LS174T (arrow indicating abluminal localisation of the L19-SIP antibody). All images are at × 20 magnification except (F) which is at × 40 magnification.
Figure 2
Figure 2
Biodistribution of intravenously injected 125I-L19-SIP in SW1222 xenograft-bearing mice at 1, 3, 6 and 24 h after injection. The mean±s.d. of four different mice are shown.
Figure 3
Figure 3
Biodistribution of intravenously injected 125I-L19-SIP in LS174T xenograft-bearing mice at 1, 3, 6 and 24 h after injection. The mean±s.d. of four different mice are shown.
Figure 4
Figure 4
Blood stability data. Antigen binding of 125I-L19-SIP in blood of LS174T tumour-bearing mice at 1, 3, 6 and 24 h post-injection. The mean±s.d. of four different mice are shown. Binding of the labelled protein before injection was 84.11%.
Figure 5
Figure 5
Biodistribution of intravenously injected 131I-L19-SIP in SW1222 xenograft-bearing mice at 3, 24 and 72 h after injection. The mean±s.d. of four different mice are shown.
Figure 6
Figure 6
Tumour volumes of control and treated SW1222 xenografts following a single injection of 55.5 MBq of 131I-L19-SIP. The mean±s.e.m. of six mice for the control and four mice for the treated groups are shown.
Figure 7
Figure 7
Survival of SW1222 xenograft-bearing nude mice after a single injection of 55.5 MBq of 131I-L19-SIP.
Figure 8
Figure 8
Individual body weights of (A) control and (B) treated mice bearing SW1222 xenografts, after a single injection of 55.5 MBq of 131I-L19-SIP.
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