Volume 13, Issue 4, Pages 716-728 (April 2006) Biological Response Determinants in HSV-tk + Ganciclovir Gene Therapy for Prostate Cancer  Gustavo Ayala, Takefumi Satoh, Rile Li, Moshe Shalev, Yehoshua Gdor, Estuardo Aguilar-Cordova, Anna Frolov, Thomas M. Wheeler, Brian J. Miles, Kate Rauen, Bin S. Teh, E. Brian Butler, Timothy C. Thompson, Dov Kadmon  Molecular Therapy  Volume 13, Issue 4, Pages 716-728 (April 2006) DOI: 10.1016/j.ymthe.2005.11.022 Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 1 Whole-mount sections of prostates. (A) Close-up view of a single prostate whole-mount section. The tumor is present within the black line. Within the tumor, the areas with cytopathic effect are stained green (yellow arrows), while those with no effect remain pink. Each image represents a different patient. (B) Large prostatic carcinoma showing large areas showing cytopathic effect. (C) Small prostatic carcinoma with very little cytopathic effect. (D) Large bilateral tumor showing cytopathic effect on both sides of the prostate. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 2 Graphical representation of the increased effect of HSV-tk with increased tumor size. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 3 Local immune response results. (A) CD8+ cells are increased significantly in the tumors of patients treated with HSV-tk (GT) compared to controls (C). (B) CD20+ cells. (C) CD68+ cells (macrophages) are increased in the nonneoplastic tissues. In each graph, the first two bars on the left represent benign nonneoplastic prostate (B) and the two on the right the tumor (T). Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 4 Bimodal increase in IL-12 is seen at 1 week and at the time of radical prostatectomy. TGF-β1 values trend in the opposite direction. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 5 Detection by FACS of total CD8+ and DR+CD8+ T cells in peripheral blood after vector injection. Note the significant increase of both. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 6 NK cell levels decreased and reached a nadir at the end of the ganciclovir treatment, rose slightly on the day of surgery, and again at 2–6 months, but remained significantly lower than injection day levels. B cell levels remained almost unchanged, except for at 2–6 months. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

FIG. 7 CAR expression is membranous and is present in (C) HGPIN and (A, B) cancer at differing levels. (D) The membranous staining (yellow arrow) is broken into granules in areas of cytopathic effect (red arrowhead). (E, F) Whole-mount sections of prostates showing the tumors delineated in black and the areas of cytopathic effect shaded in green (red arrowhead). The left image shows areas with high expression of CAR in red and lower expression in blue. No expression has no shading. Note that the areas with cytopathic effect seem to have greater expression of CAR. Molecular Therapy 2006 13, 716-728DOI: (10.1016/j.ymthe.2005.11.022) Copyright © 2006 The American Society of Gene Therapy Terms and Conditions