Exploring the immunogenicity of an insect-specific virus vectored Zika vaccine candidate.

Zika virus (ZIKV) is an important re-emerging flavivirus that presents a significant threat to human health worldwide. Despite its importance, no vaccines are approved for use in humans. Insect-specific flaviviruses (ISFVs) have recently garnered attention as an antigen presentation platform for vaccine development and diagnostic applications. Here, we further explore the safety, immunogenicity, and efficacy of a chimeric ISFV-Zika vaccine candidate, designated Aripo-Zika (ARPV/ZIKV). Our results show a near-linear relationship between increased dose and immunogenicity, with 1011 genome copies (i.e., 108 focus forming units) being the minimum dose required for protection from ZIKV-induced morbidity and mortality in mice. Including boosters did not significantly increase the short-term efficacy of ARPV/ZIKV-vaccinated mice. We also show that weanling mice derived from ARPV/ZIKV-vaccinated dams were completely protected from ZIKV-induced morbidity and mortality upon challenge, suggesting efficient transfer of maternally-derived protective antibodies. Finally, in vitro coinfection studies of ZIKV with Aripo virus (ARPV) and ARPV/ZIKV in African green monkey kidney cells (i.e., Vero-76) showed that ARPV and ARPV/ZIKV remain incapable of replication in vertebrate cells, despite the presence of active ZIKV replication. Altogether, our data continue to support ISFV-based vaccines, and specifically the ARPV backbone is a safe, immunogenic and effective vaccine strategy for flaviviruses.

Novel murine models for studying Cache Valley virus pathogenesis and in utero transmission.

Cache Valley virus (CVV) is a prevalent emerging pathogen of significant importance to agricultural and human health in North America. Emergence in livestock can result in substantial agroeconomic losses resulting from the severe embryonic lethality associated with infection during pregnancy. Although CVV pathogenesis has been well described in ruminants, small animal models are still unavailable, which limits our ability to study its pathogenesis and perform preclinical testing of therapeutics. Herein, we explored CVV pathogenesis, tissue tropism, and disease outcomes in a variety of murine models, including immune -competent and -compromised animals. Our results show that development of CVV disease in mice is dependent on innate immune responses, and type I interferon signalling is essential for preventing infection in mice. IFN-αßR-/- mice infected with CVV present with significant disease and lethal infections, with minimal differences in age-dependent pathogenesis, suggesting this model is appropriate for pathogenesis-related, and short- and long-term therapeutic studies. We also developed a novel CVV in utero transmission model that showed high rates of transmission, spontaneous abortions, and congenital malformations during infection. CVV infection presents a wide tissue tropism, with significant amplification in liver, spleen, and placenta tissues. Immune-competent mice are generally resistant to infection, and only show disease in an age dependent manner. Given the high seropositivity rates in regions of North America, and the continuing geographic expansion of competent mosquito vectors, the risk of epidemic and epizootic emergence of CVV is high, and interventions are needed for this important pathogen.

Comparison of hip fracture risk prediction by femoral aBMD to experimentally measured factor of risk.

Areal BMD (aBMD) derived from DXA is currently the gold standard for diagnosis of osteoporosis. A biomechanical approach to fracture risk assessment comparing the ratio of applied load to bone strength, termed the factor of risk (Phi), may be useful to better identify patients at risk for fracture. We obtained 73 human cadaveric femurs (48 women and 25 men, aged 74.2+/-8.7 years, range 55-98 years), measured femoral neck (FN) aBMD by DXA, and mechanically tested the femurs to failure in a sideways fall configuration. The force applied to the hip during a sideways fall was estimated from height and weight, and accounted for trochanteric soft tissue thickness. Compared to men, women had significantly lower FN aBMD and femoral strength, and tended to have higher factor of risk for hip fracture Phi. Fifty-three of 54 (98%) specimens that had a FN aBMD T-score below -2.5 also had a Phi>1. However, 10/19 (53%) specimens with FN aBMD T-score above -2.5 also had Phi>1. These data indicate that whereas an aBMD-based diagnosis of osteoporosis is highly associated with fracture risk as assessed by the factor of risk, about 50% of individuals not designated as osteoporotic by aBMD testing would be at high risk for hip fracture should they experience a sideways fall. These findings strongly support the investigation of new biomechanically-based methods of fracture risk prediction.

Collagen-platelet rich plasma hydrogel enhances primary repair of the porcine anterior cruciate ligament.

The anterior cruciate ligament (ACL) fails to heal after suture repair. One hypothesis for this failure is the premature loss of the fibrin clot, or provisional scaffolding, between the two ligament ends in the joint environment. To test this hypothesis, a substitute provisional scaffold of collagen-platelet rich plasma (PRP) hydrogel was used to fill the ACL wound site at the time of suture repair and the structural properties of the healing ACLs evaluated 4 weeks after surgery. Bilateral ACL transections were performed in five 30-kg Yorkshire pigs and treated with suture repair. In each animal, one of the repairs was augmented with placement of a collagen-PRP hydrogel at the ACL transection site, while the contralateral knee had suture repair alone. In addition, six control knees with intact ACLs from three additional animals were used as a control group. No postoperative immobilization was used. After 4 weeks the animals underwent in vivo magnetic resonance imaging to assess the size of the healing ACL, followed by biomechanical testing to determine tensile properties. The supplementation of suture repair with a collagen-PRP hydrogel resulted in significant improvements in load at yield, maximum load, and linear stiffness at 4 weeks. We conclude that use of a stabilized provisional scaffold, such as a collagen-PRP hydrogel, to supplement primary repair of the ACL can result in improved biomechanical properties at an early time point. Further studies to determine the long-term effect of primary repair enhancement are needed.