158 BOVINE HERPES VIRUS 4 (BoHV4) INHIBITS BOVINE ENDOMETRIAL EPITHELIAL CELL (bEEC) PROLIFERATION
M. Chanrot A D , Y. Z. Guo A , G. Blomqvist B , M. Juremalm B , P. Reinaud C , G. Charpigny C , O. Sandra C , P. Chantaraprateep D , R. Båge A , G. Donofrio E , J. F. Valarcher B and P. Humblot AA Department of Clinical Sciences, SLU, Uppsala, Sweden;
B Department of Virology, Immunobiolgy and Parasitology, SVA, Uppsala, Sweden;
C INRA, UMR1198, Biology of Development & Reproduction, Jouy en Josas, France;
D Faculty of Veterinary Science, Rajamangala University of Technology, Nakhonsithammarat, Thailand;
E Department of Medical Veterinary Science, University of Parma, Parma,Italy
Reproduction, Fertility and Development 27(1) 170-170 https://doi.org/10.1071/RDv27n1Ab158
Published: 4 December 2014
Abstract
BoHV4 is a double-stranded DNA virus which has been associated to endometritis, metritis, and abortions in dairy cow. The objective of this study was to characterise its cytopathic effects on bovine endometrial epithelial cells (bEEC). Bovine uteri were collected from slaughter house and bEEC separated and cultivated as previously described (Guo et al. 2014 Reprod. Fertil. Dev. 26, 165–166). In Experiment 1 (Exp 1), bEEC (passage 5) from 3 cows were cultivated for 6 days without virus or following exposure to serial dilutions (10–4, 10–3, 10–2) of virus. Living cells were counted for each group at start of the experiment and by Day 6. Proliferation or inhibition of proliferation was calculated by (Number of cells Day 6 – Number of cells Day 0)/Number of cells Day 0. In Experiment 2 (Exp 2) cells were challenged with a single dosage of virus (MOI 0.01; 1 virus: 100 cells) and culture performed during 1, 2, 3, 4, 5, 6, or 7 days. Cells were counted at Day 0 and each day, proliferation of cells was calculated as (number of cells by Day X – number of cells Day 0)/number of cells Day 0. The effects of the dilution of virus, cow and their interaction (Exp 1) or effects of time, cow, viral exposure, and second-order interactions (Exp 2) on cow cell proliferation were analysed by ANOVA (SAS 9.2, proc GLM; SAS Institute, Inc., Cary, NC, USA). In Exp 1, the amount of living cells by Day 6 was very significantly increased in controls when compared to Day 0 (+172.6 ± 24%; P < 0.0001). A linear inhibition of proliferation was observed with increasing dilutions of virus. The number of living cells for the highest concentration of virus is not different from Day 0 numbers (–26.7 ± 24.6%). Pattern of proliferation differed between cows as evidenced by a significant interaction between cow and virus dilution (P < 0.001). In Exp 2, we observed a very strong increase of proliferation from Day 0 to Day 7 in controls (+1000 ± 87%; P < 0.0001). From Day 1 to 4, the increase in number of cells was very similar for cells exposed to BoHV4 and in controls. However, after Day 4, cells exposed to virus had a limited proliferation or expressed cell death as the number of living cells by Day 7 were not different from these observed by Day 0 (50 ± 87%; NS). These results show that both time and dose of BoHV4 affect the proliferation of bovine EEC. These results will be used to investigate further the molecular mechanisms by which BoHV4 induces cell death and their sequence.
Research was partly funded by RMUTSV.
