Paracrine delivery of secreted protein by transgenized EADSCs in vivo. A) Western immunoblotting at 24, 48 and 72 h of gE2-EADSCs conditioned medium and compared to 72 h conditioned medium coming from mock-transfected and selected EADSCs. B) Humoral immune responses of 3 horses (1, 2 and 3) inoculated with gE2-EADSCs. Preimmune sera (time 0), sera at 2 weeks post gE2-EADSCs inoculation (time 2) and sera at 4 weeks post gE2-EADSCs inoculation (time 4 or time 2 from the second post gE2-EADSCs inoculation). Anti BVDV antibodies were detected by serum neutralization (SN) test. SN antibodies were expressed as the reciprocal of the highest dilution of the serum that inhibited the development of virus-induced CPE in MDBK cells. Virus neutralization (VN) titers of >1 (log2) were considered to be positive. Each value represents the response of each horse. Multiwells plate image with respective horse sera dilution, where crystal violet staining allows macroscopic evaluation of the integrity (violet wells) or the destruction (clear transparent wells) of the cell monolayer. The test was repeated three times and the same result was obtained. Controls were established in the absence of sera (-S) or with sera coming from BVDV infected animals.
Donofrio et al. BMC Cell Biology 2010 11:73 doi:10.1186/1471-2121-11-73