Effects of Adaptation of Infectious Bronchitis Virus Arkansas Attenuated Vaccine to Embryonic Kidney Cells.

The population structure of an embryo-attenuated infectious bronchitis virus (IBV) Arkansas (Ark) Delmarva Poultry Industry (DPI)-derived vaccine was characterized during serial passages in chicken embryo kidney (CEK) cells and after back-passage in embryonated chicken eggs (ECE) and in chickens. Both conventional and deep-sequencing results consistently showed population changes occurred during adaptation to CEK cells. Specifically, 13 amino acid (aa) positions seemed to be targets of selection when comparing the vaccine genome prior to and after seven passages in CEK (CEKp7). Amino acid changes occurred at four positions in the spike (S) gene and, at two positions in the S gene, large shifts in frequencies of aa encoding were observed. CEK adaptation shifted the virus population towards homogeneity in S. The changes achieved in the S1 gene in CEKp7 were maintained after a back-passage in ECE. Outside the S gene, aa changes at three positions and large shifts in frequencies at four positions were observed. Synonymous nucleotide changes and changes in noncoding regions of the genome were observed at eight genome positions. Inoculation of early CEK passages into chickens induced higher antibody levels and CEKp4 induced increased respiratory signs compared to CEKp7. From an applied perspective, the fact that CEK adaptation of embryo-attenuated Ark vaccines reduces population heterogeneity, and that changes do not revert after one replication cycle in ECE or in chickens, provides an opportunity to improve commercial ArkDPI-derived vaccines.

Timing of ICAM-I Expression in a Canine Model of Post-Haemorrhagic Cerebral Vasospasm.

SUMMARY: Temporal alterations in endothelial intercellular adhesion molecule I (ICAM-I) expression during the course of post-haemorrhagic cerebral vasospasm (PHCV) are correlated with angiographic and histologic changes in the canine basilar artery. Angiography was performed in six dogs to obtain baseline measurements of basilar artery diameter. In three dogs subarachnoid haemorrhage (SAH) was created by performing percutaneous puncture of the cisterna magna, and replacing 7 ml of cerebrospinal fluid with 7 ml of arterial blood. The remaining three dogs were used as controls. Daily angiography was performed on all dogs to determine the percent reduction in basilar artery diameter (%RBAD). One dog from each group was sacrificed after 24 hours. The remaining two dogs in each group were sacrificed after 48 hours. Each basilar artery was perfusion fixed and subjected to histologic, and immunohistochemical analysis. In the SAH group, the average %RBAD was 4 (+/- 3) at 24 hours, and 36 (+/- 1) at 48 hours. In the control group, the average %RBAD was - 1 (+/- 1) at 24 hours, and 0 (+/- 2) at 48 hours. Endothelial edema and endothelial expression of ICAM-I were found at 24 hours.At 48 hours post-SAH there was widespread endothelial desquamation, but no evidence of ICAM-I expression. In the control group, histology was normal and no ICAM-I expression was found at 24 or 48 hours. The results suggest that a brief window of therapeutic efficacy exists during the first postictal 24 hours where ICAM-I antagonists may be useful in suppressing the pathogenesis of PHCV.