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Timothy Sheahan

Publications and source records attributed to Timothy Sheahan.

5 recordsLinked to original sources
PubMed2006

Vaccine efficacy in senescent mice challenged with recombinant SARS-CoV bearing epidemic and zoonotic spike variants.

BACKGROUND: In 2003, severe acute respiratory syndrome coronavirus (SARS-CoV) was identified as the etiological agent of severe acute respiratory syndrome, a disease characterized by severe pneumonia that sometimes results in death. SARS-CoV is a zoonotic virus that crossed the species barrier, most likely originating from bats or from other species including civets, raccoon dogs, domestic cats, swine, and rodents. A SARS-CoV vaccine should confer long-term protection, especially in vulnerable senescent populations, against both the 2003 epidemic strains and zoonotic strains that may yet emerge from animal reservoirs. We report the comprehensive investigation of SARS vaccine efficacy in young and senescent mice following homologous and heterologous challenge. METHODS AND FINDINGS: Using Venezuelan equine encephalitis virus replicon particles (VRP) expressing the 2003 epidemic Urbani SARS-CoV strain spike (S) glycoprotein (VRP-S) or the nucleocapsid (N) protein from the same strain (VRP-N), we demonstrate that VRP-S, but not VRP-N vaccines provide complete short- and long-term protection against homologous strain challenge in young and senescent mice. To test VRP vaccine efficacy against a heterologous SARS-CoV, we used phylogenetic analyses, synthetic biology, and reverse genetics to construct a chimeric virus (icGDO3-S) encoding a synthetic S glycoprotein gene of the most genetically divergent human strain, GDO3, which clusters among the zoonotic SARS-CoV. icGD03-S replicated efficiently in human airway epithelial cells and in the lungs of young and senescent mice, and was highly resistant to neutralization with antisera directed against the Urbani strain. Although VRP-S vaccines provided complete short-term protection against heterologous icGD03-S challenge in young mice, only limited protection was seen in vaccinated senescent animals. VRP-N vaccines not only failed to protect from homologous or heterologous challenge, but resulted in enhanced immunopathology with eosinophilic infiltrates within the lungs of SARS-CoV-challenged mice. VRP-N-induced pathology presented at day 4, peaked around day 7, and persisted through day 14, and was likely mediated by cellular immune responses. CONCLUSIONS: This study identifies gaps and challenges in vaccine design for controlling future SARS-CoV zoonosis, especially in vulnerable elderly populations. The availability of a SARS-CoV virus bearing heterologous S glycoproteins provides a robust challenge inoculum for evaluating vaccine efficacy against zoonotic strains, the most likely source of future outbreaks.

Aging↗
PubMed2003

Biologic activity of nerve growth factor slowly released from microspheres.

Efficient and sustained delivery of neurotrophic factors to the regenerating nerve in biologically active form presents a challenge. Protein delivery in biodegradable microsphere vehicles has been difficult, based on destabilization and breakdown during both the loading and release phases. This study examines the extravasation and stability of Nerve Growth Factor (NGF) in polylactic-co-glycolic acid (PLGA) microspheres, via both ELISA and PC-12 bioassays. PLGA microspheres co-loaded with bovine serum albumin (BSA) and NGF were prepared by a water-in-oil-in-water (W/O/W) technique, using chloroform for the organic phase and 1 percent polyvinyl alcohol (PVA) for the emulsion step. Aliquots of lyophilized microspheres were incubated in double distilled water (dd H2O) at 37 degrees C, and the supernatants assayed over time for NGF activity. ELISA was utilized for quantitative determination of NGF concentration, and a PC-12 cell neurite outgrowth assay assessed biologic activity. Both ELISA and PC-12 assays demonstrated the extravasation of NGF from microspheres. Over time, the predicted concentration of NGF via the two assays differed, suggesting possible preservation of recognizable epitopes for ELISA, but loss of biologic activity. NGF can be stored and released from microspheres. Extravasation studies should include biologically relevant assays for activity.

Animals↗
PubMedDate not supplied

Baiting the cross-face nerve graft with temporary hypoglossal hookup.

BACKGROUND: Cross-face nerve grafting yields inconsistent neural regeneration, and methods that promote more robust axonal traversing of the graft would expand the indications for this procedure. OBJECTIVE: To test the hypothesis that hooking a cross-face nerve graft distally to a source of denervated muscle, rather than leaving it in the subcutaneous space, would positively affect neural ingrowth across the graft, based on elaboration of neurotrophins from the musculature. METHODS: Twenty-four rats underwent cross-face nerve grafting in which the right facial nerve buccal branch was transected and coapted to the graft. The graft was placed across the neck and into the left side of the face. The distal end of the graft was placed either in the left subcutaneous space, coapted to the marginal mandibular branch of the left facial nerve, or coapted to the distal stump of the transected left hypoglossal nerve. Eight control animals underwent right buccal branch transection and placement of a cross-face nerve graft without any proximal and distal hookup. After 12 weeks, all experimental groups underwent hookup of the distal nerve graft to the left facial nerve buccal branch. Vibrissal function was assessed during the ensuing 12 weeks, and then the graft was harvested for histomorphometric analysis. RESULTS: After 12 weeks, there was a significant difference in axon counts between the group coapted distally to the tongue (hypoglossal hookup) and that coapted to the facial musculature (marginal hookup). Twelve weeks later, after distal cross-face nerve graft hookup, this difference was not statistically significant, although the hypoglossally baited group demonstrated statistically significantly greater fiber maturity. Recovery of vibrissal movement did not differ among treatment groups. CONCLUSION: Baiting the cross-face nerve graft via temporary hookup to the distal hypoglossal nerve and tongue musculature appears to improve nerve ingrowth through a nerve graft across the face, although a corresponding improvement in facial muscle function was not observed.

Animals↗