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Fig. 2 HCMV secretome mediates EC wound healing. Wound healing activity of the HCMV secretome. ECs were grown to confluence on ECIS arrays and exposed to test supernatants prior to electrical wounding. Wound healing, as indicated by increasing resistance, is plotted as a function of time. Healing traces for duplicate wells are shown for the HSV-1 Secretome (yellow), mock (green), HCMV (red), or UV-inactivated HCMV (light blue). Control traces include a negative control (SSFM; pink), a positive control (Complete SFM; dark blue) and an unwounded control (black). Cells exposed to the HCMV secretome show wound repair within 6-10 h, whereas cells exposed to the HSV-1 or mock secretomes repopulate the wound inefficiently, indicating that the production of wound healing factors is specific for HCMV infections o -,-,-,-,-

Time (hrs)

Fig. 2 HCMV secretome mediates EC wound healing. Wound healing activity of the HCMV secretome. ECs were grown to confluence on ECIS arrays and exposed to test supernatants prior to electrical wounding. Wound healing, as indicated by increasing resistance, is plotted as a function of time. Healing traces for duplicate wells are shown for the HSV-1 Secretome (yellow), mock (green), HCMV (red), or UV-inactivated HCMV (light blue). Control traces include a negative control (SSFM; pink), a positive control (Complete SFM; dark blue) and an unwounded control (black). Cells exposed to the HCMV secretome show wound repair within 6-10 h, whereas cells exposed to the HSV-1 or mock secretomes repopulate the wound inefficiently, indicating that the production of wound healing factors is specific for HCMV infections cells cultured in the presence of the HCMV secretome were able to repopulate the wound with the same kinetics as with complete medium, although steady state resistance levels were slightly lower. In contrast, cells cultured in SFM lacking growth factors, the mock secretome, or the secretomes from HCMV UV- or foscarnet-treated cells all were unable to repopulate the electrode, even over the total 20 h measured. Similarly, cells incubated with a secretome derived from HSV-1 infected cells also failed to mediate WH, suggesting that the WH effects are specific for CMV. Importantly, these data clearly show that the HCMV secretome contains factors that promote cell migration into a mechanical wound. The ability of the HCMV secretome to mediate WH is due to active viral replication since both UV-treated virions and foscarnet-treated cells did not promote WH. Since foscarnet treatment of infected cells resulted in an inactive secretome, this observation suggests that a late kinetic class of HCMV gene(s) is involved in the generation of secretome WH factors. These observations correlate with studies in human heart transplant patients as well as our own observations in heart transplants in rats in which ganciclovir treatment prolongs graft survival.

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