Super-permissiveness does not correlate with cytotoxicity It has been reported in CNN in 2005, in work done by Craig Meyers, that AAV preferentially kills cancer cellshttp://​www.​cnn.​com/​2005/​HEALTH/​06/​22/​cancer.​virus/​. This reported cancer cell killing may be related to parvovirus replication as certain parvoviruses have been reported to preferentially replicate in malignant cells [44]. Thus we tested the high and low AAV-permissive cells for their sensitivity

to killing by AAV infection. The results are shown in Figure4and demonstrate that PT3 was not preferentially sensitive to killing by AAV2 infection compared to other AZD1480 squamous cells. Figure 4 Lack of cytotoxicity by AAV. Various squamous cell isolates were grown in culture and infected with AAV2 virus as indicated. Note that increasing mois of AAV2 did not result in increased cell toxicity of PT3, and had only minimal effects on the cell growth of the S63845 nmr other cells. Shown is a representative experiment of three done. Discussion Earlier studies

by Niet aland Nashet alidentified a number of cellular components which are required forin vitroAAV DNA replication using both adenovirus-infected and uninfected cell extracts [41,42]. These cellular components, found to be critical, include PCNA, RFC, LY2606368 order RPA and DNA polymerase delta (POLD1). This study demonstrates that the PT3 primary cervical cancer cell isolate, which is super-permissive for AAV replication [40], over-expresses all four of these components, when compared with PT1/NK. Thus, the data presented here are fully consistent

with the earlierin vitrostudies, but now extend these studies into the context of the living cell. These data also further characterize the primary cervical cancer isolate PT3 and confirms the ability of AAV to replicate in SSE, now including malignant cells [34–36]. It is also confirmed that AAV2 variably replicates in multiple cervical cancer isolates [40]. Thus far, to our knowledge, Tacrolimus (FK506) only the PT3 isolate has been described as super-permissive for AAV replication, this being when compared to a variety of cells of squamous origin. Both the Affymetrix DNA microarray data and real-time quantitative PCR results demonstrated that all four of these cellular components were over expressed in PT3 cells. POLD1 and PCNA were strongly over-expressed in PT3. Moreover, multiple RFC and RPA family members were over-expressed in PT3. Thus, these data support the unusual phenotype of PT3 cells and suggest their use as a unique reagent for identifying critical genes involved in AAV replication. This phenotype also suggests the possibility that PT3, itself, may be useful as a platform for rAAV production. One issue against this idea is that AAV replication in PT3 takes place during cellular differentiation (induced by air interface and calcium).