Deletion of the Vaccinia Virus Gene A46R, Encoding for an Inhibitor of TLR Signalling, Is an Effective Approach to Enhance the Immunogenicity in Mice of the HIV/AIDS Vaccine Candidate NYVAC-C.

Viruses have developed strategies to counteract signalling through Toll-like receptors (TLRs) that are involved in the detection of viruses and induction of proinflammatory cytokines and IFNs. Vaccinia virus (VACV) encodes A46 protein which disrupts TLR signalling by interfering with TLR: adaptor interactions. Since the innate immune response to viruses is critical to induce protective immunity, we studied whether deletion of A46R gene in a NYVAC vector expressing HIV-1 Env, Gag, Pol and Nef antigens (NYVAC-C) improves immune responses against HIV-1 antigens. This question was examined in human macrophages and in mice infected with a single A46R deletion mutant of the vaccine candidate NYVAC-C (NYVAC-C-ΔA46R). The viral gene A46R is not required for virus replication in primary chicken embryo fibroblast (CEF) cells and its deletion in NYVAC-C markedly increases TNF, IL-6 and IL-8 secretion by human macrophages. Analysis of the immune responses elicited in BALB/c mice after DNA prime/NYVAC boost immunization shows that deletion of A46R improves the magnitude of the HIV-1-specific CD4 and CD8 T cell immune responses during adaptive and memory phases, maintains the functional profile observed with the parental NYVAC-C and enhances anti-gp120 humoral response during the memory phase. These findings establish the immunological role of VACV A46R on innate immune responses of macrophages in vitro and antigen-specific T and B cell immune responses in vivo and suggest that deletion of viral inhibitors of TLR signalling is a useful approach for the improvement of poxvirus-based vaccine candidates.

A Candidate HIV/AIDS Vaccine (MVA-B) that Enhances the Magnitude and Polyfunctionality of Memory HIV-1-Specific T-Cell Responses

Background: The poxvirus vector Modified Vaccinia Virus Ankara (MVA) expressing HIV-1 Env, Gag, Pol and Nef antigens from clade B (MVA-B) is currently used as a HIV/AIDS vaccine candidate. A general strategy to try to improve the immunogenicity of poxvirus HIV-1 vaccine candidates is the deletion of known or suggested immunomodulatory vaccinia virus (VACV) genes.Methods: We have generated and characterized the innate immune sensing and the immunogenicity profile of a new HIV-1 vaccine candidate, which contains a deletion in a VACV gene.Results: We show that this VACV protein is expressed early during virus infection and localizes to the cytoplasm of infected cells. Deletion of this VACV gene from the MVA-B had no effect on virus growth kinetics; therefore this VACV protein is not essential for virus replication. The innate immune signals elicited by the MVA-B deletion mutant in human macrophages and monocyte-derived dendritic cells were characterized. In a DNA prime/MVA boost immunization protocol in mice, flow cytometry analysis revealed that the MVA-B deletion mutant enhanced the magnitude and polyfunctionality of the HIV-1-specific CD4 + and CD8 + T-cell memory immune responses, with most of the HIV-1 responses mediated by the CD8 + T-cell compartment with an effector phenotype. Significantly, while MVA-B induced preferentially Env- and Gag-specific CD8 + T-cell responses, the MVA-B deletion mutant induced more GPN-specific CD8 + T-cell responses. Furthermore, the MVA-B deletion mutant enhanced the levels of antibodies against Env in comparison with MVA-B.Conclusion: These findings revealed that this new VACV protein can be considered as an immunomodulator and that deleting this gene in MVA-B confers an immunological benefit by inducing innate immune responses and increasing the magnitude and quality of the T-cell memory immune responses to HIV-1 antigens. Our observations are relevant for the improvement of MVA vectors as HIV-1 vaccines.

West 1 St Street/ IA 57 from Highland Drive to Center Street/Franklin Street, Blackhawk County, Iowa, Project # STP-57-2(28)--2C-07, Environmental Assessment and Section 4(f) De Minimis Impact Finding, 2015

From Proposed Action: "Iowa Northland Regional Council of Governments (INRCOG) and the City of Cedar Falls, in coordination with the Iowa Department of Transportation (Iowa DOT) and the Federal Highway Administration (FHWA), are proposing to upgrade and modernize an approximate 4,900-foot segment of Iowa Highway 57 (IA 57), locally known as West 1st Street, in Cedar Falls, Black Hawk County, Iowa."

Road Safety Audit for US 6 from the ECL of Iowa City to the WCL of West Liberty in Johnson and Muscatine Counties, Iowa, 2008

Approximately 13.2 miles of US 6 in eastern Iowa extends from the east corporate limits of Iowa City, Iowa, to the west corporate limits of West Liberty, Iowa. This segment of US 6 is a service level B primary highway, with an annual daily traffic volume varying from 3,480 vehicles per day (vpd) to 5,700 vpd. According to 2001–2007 crash density data from the Iowa Department of Transportation (Iowa DOT), the corridor is currently listed among the top 5% of non-freeway Iowa DOT roads in several crash categories, including crashes involving excessive speed, impaired drivers, single-vehicle run-off-road, and multiple-vehicle crossed centerline. A road safety audit of this corridor was deemed appropriate by the Iowa Department of Transportation’s Office of Traffic and Safety. Staff and officials from the Iowa DOT, Iowa State Patrol, Governor’s Traffic Safety Bureau, Federal Highway Administration, Center for Transportation Research and Education, and several local law enforcement and transportation agencies met to review crash data and discuss potential safety improvements to this segment of US 6. This report outlines the findings and recommendations of the road safety audit team to address the safety concerns on this US 6 corridor and explains several selected mitigation strategies.

Road Safety Audit for the US 61/Harrison Street and West Locust Street Intersection in Davenport-Scott County, Iowa, 2008

On the October 7 and 8, 2008, a road safety audit was conducted for the intersection of US 61/Harrison Street and West Locust Street in Davenport, Iowa. US 61/Harrison Street is a one-way street and a principal arterial route through Davenport, with three southbound lanes. Locust Street is a four-lane, two-way minor arterial running across the city from west to east. The last major improvement at this intersection was implemented approximately 20 years ago. The Iowa Department of Transportation requested a safety audit of this intersection in response to a high incidence of crashes at the location over the past several years, in view of the fact that no major improvements are anticipated for this intersection in the immediate future. The road safety audit team discussed current conditions at the intersection and reviewed the last seven years of crash data. The team also made daytime and nighttime field visits to the intersection to examine field conditions and observe traffic flow and crossing guard operations with younger pedestrians. After discussing key issues, the road safety audit team drew conclusions and suggested possible enforcement, engineering, public information, and educational strategies for mitigation.

Road Safety Audit for Intersection of US 218 and County Road C-57 (West Cedar Wapsi Road) in Black Hawk County, Iowa, 2009

Beginning on June 22, 2009, a road safety audit was initiated for the intersection of US 218 and County Road C-57 in Black Hawk County, Iowa. Due to the traffic volumes and the number of conflicting traffic movements on these two roadways, this intersection has developed a crash history that concerns the Iowa Department of Transportation (Iowa DOT), Iowa State Patrol, and local agencies. This intersection is ranked seventh in Iowa for the highest number of at-grade expressway intersection crashes. Considering this, Black Hawk County and the Iowa DOT requested that a road safety audit be conducted to address the safety concerns and recommend possible mitigation strategies.

A novel HIV vaccine adjuvanted by IC31 induces robust and persistent humoral and cellular immunity.

The HIV vaccine strategy that, to date, generated immune protection consisted of a prime-boost regimen using a canarypox vector and an HIV envelope protein with alum, as shown in the RV144 trial. Since the efficacy was weak, and previous HIV vaccine trials designed to generate antibody responses failed, we hypothesized that generation of T cell responses would result in improved protection. Thus, we tested the immunogenicity of a similar envelope-based vaccine using a mouse model, with two modifications: a clade C CN54gp140 HIV envelope protein was adjuvanted by the TLR9 agonist IC31®, and the viral vector was the vaccinia strain NYVAC-CN54 expressing HIV envelope gp120. The use of IC31® facilitated immunoglobulin isotype switching, leading to the production of Env-specific IgG2a, as compared to protein with alum alone. Boosting with NYVAC-CN54 resulted in the generation of more robust Th1 T cell responses. Moreover, gp140 prime with IC31® and alum followed by NYVAC-CN54 boost resulted in the formation and persistence of central and effector memory populations in the spleen and an effector memory population in the gut. Our data suggest that this regimen is promising and could improve the protection rate by eliciting strong and long-lasting humoral and cellular immune responses.

Road Safety Audits for County Road X-23 from IA 2 to the South Corporate Limits of West Point and for County Road W-62 from US 218 to IA 27 in Lee County, Iowa, 2010

On October 20–21, 2009, two road safety audits were conducted in Lee County, Iowa: one for a 6 mile section of County Road X-23 from IA 2 to the south corporate limits of West Point and one for a 9.7 mile section of County Road W-62 from US 218 to IA 27. Both roads have high severe crash histories for the years of 2001 through 2008. Using these crash data, the Iowa Department of Transportation (Iowa DOT) has identified County Road X-23 as being in the top 5 percent of similar roads for run-off-road crashes. The Iowa DOT lists County Road W-62 as a high-risk rural road that has above-average crash numbers and is eligible for funding under the Federal High-Risk Rural Road Program. Considering these issues, the Lee County Engineer and Iowa DOT requested that road safety audits be conducted to address the safety concerns and to suggest possible mitigation strategies.

A prospective observational safety study on MF59(®) adjuvanted cell culture-derived vaccine, Celtura(®) during the A/H1N1 (2009) influenza pandemic.

BACKGROUND: The present study was a prospective observational study to evaluate the safety profile of Celtura(®), a monovalent, cell culture-derived, inactivated subunit influenza vaccine prepared from A/California/07/2009(H1N1) with the adjuvant MF59(®). Subjects were enrolled prospectively during the H1N1 2009 influenza pandemic at medical centres in Colombia, Chile, Switzerland, and Germany during the period December 2009 to June 2010. METHODS: Subjects ages 18 and older were followed for the occurrence of adverse events (AEs) for six months after vaccination. Adverse events of special interest (AESIs) were neuritis, convulsion (seizure), anaphylaxis, encephalitis, vasculitis, Guillain-Barre syndrome, demyelinating conditions, Bell's palsy, and laboratory-confirmed vaccination failure. RESULTS: Overall, 7348 AEs were reported in 2296 of 3989 enrolled subjects (57.6%). Only two AEs were considered related to injection site reactions. No laboratory-confirmed cases of influenza were reported. There were 108 medically confirmed serious adverse events (SAEs) reported among 73 subjects with 6 such SAEs described as possibly or probably related to vaccination. Three fatal cases were reported and assessed as not related to vaccination. Two AESIs classified as convulsion were reported and assessed as not related to vaccination. Both AESIs occurred well outside the pre-specified 7 day risk window representing the likely timeframe of the occurrence of seizure following vaccination. CONCLUSIONS: The results of this study support the overall good safety profile of MF59 adjuvanted cell culture-derived influenza vaccine as administered in adults during the 2009-2010 H1N1 influenza pandemic. No concern is raised regarding the occurrence of AESIs.

1245-02IJ West Tarkio Final Report

Initiated in 2001, the West Tarkio Creek Watershed Project has a proven track record of implementing an enormous amount of structural conservation practices. To date, over $925,000 has been spent to build 69 miles of terraces on 63 cooperators' land. The success of the Project was due in large part to the conservation ethic of the landowners to improve their farms, preserve the productivity of the land, and protect West Tarkio Creek. This has been made possible through funding from DSC Watershed Protection Funds (WSPF) which has provided $1,362,592 in cost share funds since 2001 but is has been severely limited in recent years due to shortages within the State’s budget. The original project goals called for the construction of 750,000 feet (142 miles) of terraces to effectively treat the watershed. In order to meet these goals and bring the project to a successful endpoint, another 153,000 feet (29 miles) remain to be constructed by the landowners with the help of the SWCD staff. Severe rain events in recent years have caused an enormous amount of damage throughout the region, these storms were helpful in identifying where watershed work remains to be completed. Scars on the landscape in the aftermath of the storms clearly etched out the specific location where additional practices are needed in addition to those proposed in the original project work plan. Project supporters are confident that the WIRB Program can unlock this potential and pave the way for what can become known as one of the most effective land treatment projects in Iowa.

Improved NYVAC-based vaccine vectors.

While as yet there is no vaccine against HIV/AIDS, the results of the phase III Thai trial (RV144) have been encouraging and suggest that further improvements of the prime/boost vaccine combination of a poxvirus and protein are needed. With this aim, in this investigation we have generated derivatives of the candidate vaccinia virus vaccine vector NYVAC with potentially improved functions. This has been achieved by the re-incorporation into the virus genome of two host range genes, K1L and C7L, in conjunction with the removal of the immunomodulatory viral molecule B19, an antagonist of type I interferon action. These novel virus vectors, referred to as NYVAC-C-KC and NYVAC-C-KC-ΔB19R, have acquired relevant biological characteristics, giving higher levels of antigen expression in infected cells, replication-competency in human keratinocytes and dermal fibroblasts, activation of selective host cell signal transduction pathways, and limited virus spread in tissues. Importantly, these replication-competent viruses have been demonstrated to maintain a highly attenuated phenotype.

Health consultation : Buchanan Bulk Oil -- Ma & Pa Stores West 6th Avenue and South F Street, Oskaloosa, Mahaska County, Iowa (2006)

The Iowa Department of Natural Resources (IDNR) has requested the Iowa Department of Public Health (IDPH) Hazardous Waste Site Health Assessment Program to evaluate the potential health impacts of the future development at the Buchanan Bulk Oil – Ma & Pa Stores site. A Targeted Brownfields Assessment was completed by the IDNR at this site to measure existing on-site contaminants. Assistance was sought from the IDPH to determine potential health risks if the site was developed for residential use. This health consultation addresses potential health risks to people from exposure to the contaminants found in the soil and groundwater within the property boundary. The information in this health consultation was current at the time of writing. Data that emerges later could alter this document’s conclusions and recommendations.

Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI).

The Cancer Vaccine Consortium of the Sabin Vaccine Institute (CVC/SVI) is conducting an ongoing large-scale immune monitoring harmonization program through its members and affiliated associations. This effort was brought to life as an external validation program by conducting an international Elispot proficiency panel with 36 laboratories in 2005, and was followed by a second panel with 29 participating laboratories in 2006 allowing for application of learnings from the first panel. Critical protocol choices, as well as standardization and validation practices among laboratories were assessed through detailed surveys. Although panel participants had to follow general guidelines in order to allow comparison of results, each laboratory was able to use its own protocols, materials and reagents. The second panel recorded an overall significantly improved performance, as measured by the ability to detect all predefined responses correctly. Protocol choices and laboratory practices, which can have a dramatic effect on the overall assay outcome, were identified and lead to the following recommendations: (A) Establish a laboratory SOP for Elispot testing procedures including (A1) a counting method for apoptotic cells for determining adequate cell dilution for plating, and (A2) overnight rest of cells prior to plating and incubation, (B) Use only pre-tested serum optimized for low background: high signal ratio, (C) Establish a laboratory SOP for plate reading including (C1) human auditing during the reading process and (C2) adequate adjustments for technical artifacts, and (D) Only allow trained personnel, which is certified per laboratory SOPs to conduct assays. Recommendations described under (A) were found to make a statistically significant difference in assay performance, while the remaining recommendations are based on practical experiences confirmed by the panel results, which could not be statistically tested. These results provide initial harmonization guidelines to optimize Elispot assay performance to the immunotherapy community. Further optimization is in process with ongoing panels.