A rapid response vaccine development strategy for newcastle disease in poultry

Poultry is a major source of protein in sub-Saharan Africa and many other lower-income regions. Newcastle disease virus (NCDV) comprises a significant threat toward poultry production. While NCDV vaccines are routinely used in developed countries, those used in sub-Saharan Africa are mostly imported and are not specific to locally circulating strains. Indeed, the lack of rapid, field-based NCDV detection and the absence of cost-effective production methods for pure, strain-specific vaccines hampers efficient poultry production throughout these regions. This remains a major problem for both subsistence and commercial farming.The aim for this study was firstly, to develop a field-based isothermal PCR assay for NCDV detection that employed a portable instrument and real-time data transfer application. Secondly,to use the nucleic acid sequence data obtained from field isolates to develop a protocol compatible with rapid emergency vaccine production for NCDV.To achieve this, the isothermal PCR detection assay was applied to field isolates from suspected NCDV outbreaks on commercial poultry farms in KwaZulu-Natal, South Africa, while for the vaccine development, the NCDV matrix gene of one of the isolates was sequenced and used to design primers for the recombinant cloning of this antigen into an adenoviral vector.This‘vaccine vector’ and a control adenoviral vector were each amplified in 293T cells and then used to infect both 293T cells as a production cell line and chicken embryo fibroblasts (CEF) as a preliminary model of the target host. Western blotting confirmed the successful expression of the V5epitopetagby the control vector in both cell lines, which established the compatibility of the adenovirus vector as an appropriate carrier of the target antigen. Mass spectrometry confirmed expression of the NCDV matrix protein by the vaccine vector in both cell lines. In conclusion, the improved turnaround time from detection to the production of the vaccine antigen was under6weeks.The approach described here provides a rapid and cost-effective protocol for both the pathogen detection on-site and the production of pure vaccine antigens specific to an emerging field strain of NCDV within lower-income regions.</jats:p

Galactosemia, a single gene disorder with epigenetic consequences

Long-term outcomes of classic galactosemia (GAL) remain disappointing. It is unclear if the complications result mainly from prenatal-neonatal toxicity or persistent glycoprotein and glycolipid synthesis abnormalities. We performed gene expression profiling (T transcriptome) to characterize key-altered genes and gene clusters of four patients with GAL with variable outcomes maintained on a galactose-restricted diet, compared with controls. Significant perturbations of multiple cell signaling pathways were observed including mitogen-activated protein kinase (MAPK) signaling, regulation of the actin cytoskeleton, focal adhesion, and ubiquitin mediated proteolysis. A number of genes significantly altered were further investigated in the GAL cohort including SPARC (osteonectin) and S100A8 (S100 calcium-binding protein). The whole serum N-glycan profile and IgG glycosylation status of 10 treated patients with GAL were compared with healthy control serum and IgG using a quantitative high-throughput analytical HPLC platform. Increased levels of agalactosylated and monogalactosylated structures and decreases in certain digalactosylated structures were identified in the patients. The persistent abnormal glycosylation of serum glycoproteins seen with the microarray data indicates persisting metabolic dyshomeostasis and gene dysregulation in “treated” GAL. Strict restriction of dietary galactose is clearly life saving in the neonatal period; long-term severe galactose restriction may contribute to ongoing systemic abnormalities