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To determine whether the immunomodulatory effect of PDE5 inhibition affected T cell activation within the tumor microenvironment, we examined IL-2 production by TILs using a transgenic mouse in which expression of GFP is under an IL-2 promoter (BALB/c–IL-2p/GFP) (25). In this model, T cell stimulation activates the IL-2 promoter and results in expression of the reporter transgene, GFP, which is easily detectable by flow cytometry. C26GM-primed BALB/c–IL-2p/GFP splenocytes were adoptively transferred into tumor-bearing recipients that were either left untreated or treated with sildenafil for 9 d. Single cell suspensions of the tumor-infiltrating CD8+ T cells were analyzed by flow cytometry for GFP expression. Adoptively transferred, vaccine-primed T cells were activated in the tumor microenvironment only in the presence of PDE5 inhibition, whereas in its absence they produced no IL-2 and, hence, were bona fide anergic T cells (Fig. 4 D). To further prove that these effects were dependent on CD8+ T cells, mice were challenged with C26GM and were (a) left untreated, (b) given sildenafil, (c) given an anti-CD8+ depleting antibody, or (d) given both sildenafil and the CD8+ depleting antibody. Sildenafil treatment again demonstrated a statistically significant reduction in tumor outgrowth, an effect completely abrogated by CD8+ depletion (Fig. 4 E). These experiments demonstrate that PDE5 inhibition enhances the tumor-specific T cell response, increases intratumoral T cell infiltration and activation, and underscores the role of CD8+ T cells in sildenafil-mediated antitumor responses.
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Acknowledgments We thank Sergio Martinez for constructing Fig. 3.
Erectile Dysfunction and Gene Therapy
No reports proving benefit
The patient's flaccid penis was somewhat turgid at the onset of physical examination, but became soft after a few minutes. The remainder of the examination was unremarkable except for a tender area in the left lateral aspect of the perineum. There was no evidence of ecchymosis or any other signs of acute trauma. An ultrasound revealed no abnormality in the pendulous portion of the penis. The patient developed 70% penile rigidity after intracavernous injection of 0.3 ml of a mixture of papaverine and phentolamine and self-genital stimulation, consistent with the erections he had at home. Color duplex ultrasound revealed peak flow velocities of 35 cm/s and 40 cm/s in the right and left cavernous arteries respectively, both within the normal range. His end-diastolic flow velocity was >5 cm/s. At this point, the patient was told that he had venogenic erectile dysfunction.
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