Volume 48, Issue 1, Pages 153-161 (July 2005) Virus Specific Immune Responses after Human Neoadjuvant Adenovirus-mediated Suicide Gene Therapy for Prostate Cancer  R.R.M. van der Linden, B.L. Haagmans, P. Mongiat-Artus, G.J. van Doornum, R. Kraaij, D. Kadmon, E. Aguilar-Cordova, A.D.M.E. Osterhaus, T.H. van der Kwast, C.H. Bangma  European Urology  Volume 48, Issue 1, Pages 153-161 (July 2005) DOI: 10.1016/j.eururo.2005.02.013 Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 1 Serum PSA response during the gene therapy protocol (A) and follow up (B). The insert (A) shows PSA levels between day 0 (day of entry protocol) and day 21 (day of radical prostatectomy). Patients A-D were treated with a low dose of Adv-HSV-tk (2×1010vp) and patients E-H were treated with a higher dose of Adv-HSV-tk (2×1011vp). European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 2 Anti-Adv neutralizing (A, B), IgA (C, D) and IgG (E,F) levels after treatment with Adv-HSV-tk. Data are presented as titer (A,B) or a percentage of the pre-treatment level (day 0, C-F). Patients A-D (A, C, E) were treated with a low dose of Adv-HSV-tk (2×1010vp) and patients E-H (B, D, F) were treated with a higher dose of Adv-HSV-tk (2×1011vp); legends as in Fig. 1. European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 3 Stimulation index (SI) of PBMC isolated at several time-points during therapy. PBMC were stimulated for 6 days with 5000 MOI Adv-HSV-tkfollowed by determination of 3H-thymidine incorporation. Data are normalized for the proliferation index of the negative control (stimulation with human serum). Patients A-D (A) were treated with a low dose of Adv-HSV-tk (2×1010vp) and patients E-H (B) were treated with a higher dose of Adv-HSV-tk (2×1011vp); legends as in Fig. 1. DL=detection limit. European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 4 IFN-γ production of PBMC isolated at day 0, day 7 and day 21. PBMC were stimulated for 6 days with 5000MOI Adv-HSV-tk (tk) or 100MOI Influenzae vaccine (i). Data are normalised for IFN-γ production of the negative control (stimulation with human serum) and expressed as a percentage of IFN-γ production at day 0. Patients A-D were treated with a low dose of Adv-HSV-tk (2×1010vp) and patients E-H were treated with a higher dose of Adv-HSV-tk (2×1011vp). European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 5 Illustration of B (A,B) and T (C,D) lymphocyte infiltration in tumoral (D) and non-tumoral (C) areas of prostate specimens after ADV-HSV-tk gene therapy (40× magnification). B lymphocytes and T lymphocytes were visualized by immunohistochemistry using monoclonal antibodies directed against CD20 and CD3, respectively. European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions

Fig. 6 Quantitation of B-lymphocyte infiltration (A), expressed as the mean number of B lymphocyte foci per cm2 counted in 36 slides per specimen. B lymphocytes were visualized by immunohistochemistry using a monoclonal antibody directed against CD20 and counted at 400× magnification. Lymphocyte infiltration in tumoral and non-tumoral tissue, expressed as the absolute number of T lymphocytes (B) counted in 36 slides per specimen. T lymphocytes were visualized by immunohistochemistry using a monoclonal antibody directed against CD3 and counted at 400× magnification. Patients A-D were treated with a low dose of Adv-HSV-tk (2×1010vp) and patients E-H were treated with a higher dose of Adv-HSV-tk (2×1011vp). European Urology 2005 48, 153-161DOI: (10.1016/j.eururo.2005.02.013) Copyright © 2005 Elsevier B.V. Terms and Conditions