A next-generation intranasal trivalent MMS vaccine induces durable and broad protection against SARS-CoV-2 variants of concern.

As SARS-CoV-2 variants of concern (VoCs) that evade immunity continue to emerge, next-generation adaptable COVID-19 vaccines which protect the respiratory tract and provide broader, more effective, and durable protection are urgently needed. Here, we have developed one such approach, a highly efficacious, intranasally delivered, trivalent measles-mumps-SARS-CoV-2 spike (S) protein (MMS) vaccine candidate that induces robust systemic and mucosal immunity with broad protection. This vaccine candidate is based on three components of the MMR vaccine, a measles virus Edmonston and the two mumps virus strains [Jeryl Lynn 1 (JL1) and JL2] that are known to provide safe, effective, and long-lasting protective immunity. The six proline-stabilized prefusion S protein (preS-6P) genes for ancestral SARS-CoV-2 WA1 and two important SARS-CoV-2 VoCs (Delta and Omicron BA.1) were each inserted into one of these three viruses which were then combined into a trivalent "MMS" candidate vaccine. Intranasal immunization of MMS in IFNAR1-/- mice induced a strong SARS-CoV-2-specific serum IgG response, cross-variant neutralizing antibodies, mucosal IgA, and systemic and tissue-resident T cells. Immunization of golden Syrian hamsters with MMS vaccine induced similarly high levels of antibodies that efficiently neutralized SARS-CoV-2 VoCs and provided broad and complete protection against challenge with any of these VoCs. This MMS vaccine is an efficacious, broadly protective next-generation COVID-19 vaccine candidate, which is readily adaptable to new variants, built on a platform with a 50-y safety record that also protects against measles and mumps.

Independent evolution of porcine reproductive and respiratory syndrome virus 2 with genetic heterogeneity in antigenic regions of structural proteins in Korea.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen that affects the global swine industry. The continuous evolution of this virus has made control and prevention difficult, which emphasizes the importance of monitoring currently circulating PRRSV strains. In this study, we investigated the genetic characteristics of whole structural genes of 35 PRRSV-2 isolates that circulated between 2012 and 2017 in Korea. Genetic and phylogenetic analysis demonstrated that a recently identified PRRSV-2 shared a relatively low level of nucleotide sequence identity that ranged from 86.2% to 92.8%; however, they were clustered into four distinct Korean field clades, except KU-N1702, in ORF2-7-based phylogeny. KU-N1702 was closely related to the NADC30-like strains that were identified in the USA and China. Amino acid sequence analysis showed that the GP5 neutralizing epitope was conserved among the KU viruses. In contrast, the viruses had genetic mutations in key residues for viral neutralization within GP5 and M. For minor structural proteins, neutralizing epitopes, aa 41-55 of GP2, 61-75 of GP3, and 51-65 of GP4, were variable among the KU viruses. Bioinformatics demonstrated diversifying evolution within the GP2 and GP4 neutralizing epitopes and the emergence of a novel glycosylation site within the GP3 and GP4 neutralizing epitopes. Taken together, these data provide evidence that Korean PRRSV-2 evolved independently in Korea, with genetic heterogeneity in antigenic regions of structural proteins.

Classical Swine Fever Outbreak after Modified Live LOM Strain Vaccination in Naive Pigs, South Korea.

We report classical swine fever outbreaks occurring in naive pig herds on Jeju Island, South Korea, after the introduction of the LOM vaccine strain. Two isolates from sick pigs had >99% identity with the vaccine stain. LOM strain does not appear safe; its use in the vaccine should be reconsidered.

Three SARS-CoV-2 spike protein variants delivered intranasally by measles and mumps vaccines are broadly protective.

As the new SARS-CoV-2 Omicron variants and subvariants emerge, there is an urgency to develop intranasal, broadly protective vaccines. Here, we developed highly efficacious, intranasal trivalent SARS-CoV-2 vaccine candidates (TVC) based on three components of the MMR vaccine: measles virus (MeV), mumps virus (MuV) Jeryl Lynn (JL1) strain, and MuV JL2 strain. Specifically, MeV, MuV-JL1, and MuV-JL2 vaccine strains, each expressing prefusion spike (preS-6P) from a different variant of concern (VoC), were combined to generate TVCs. Intranasal immunization of IFNAR1-/- mice and female hamsters with TVCs generated high levels of S-specific serum IgG antibodies, broad neutralizing antibodies, and mucosal IgA antibodies as well as tissue-resident memory T cells in the lungs. The immunized female hamsters were protected from challenge with SARS-CoV-2 original WA1, B.1.617.2, and B.1.1.529 strains. The preexisting MeV and MuV immunity does not significantly interfere with the efficacy of TVC. Thus, the trivalent platform is a promising next-generation SARS-CoV-2 vaccine candidate.

Genomic characteristics and pathogenicity of natural recombinant porcine reproductive and respiratory syndrome virus 2 harboring genes of a Korean field strain and VR-2332-like strain.

Porcine reproductive and respiratory syndrome (PRRS), an economically-important disease caused by PRRS virus (PRRSV), has become endemic to most pig-producing countries. Point mutation and recombination are responsible for genetic heterogeneity, resulting in circulation of genetically-diverse strains. However, no natural recombinant PRRSV has yet been identified in Korea. Here, we successfully isolated natural recombinant PRRSV-2 (KU-N1202) using cell culture, investigated its genomic characteristics, and further evaluated its pathogenicity. KU-N1202 is a recombinant strain between Korean MN184-like and VR-2332-like strains. Specifically, ORF5 to partial ORF7 of the VR-2332-like strain was inserted into the backbone of a CP07-626-2-like strain. KU-N1202 induced mild-to-moderate clinical signs and mild histopathological changes with low viral loads in challenged pigs. Contact pigs showed minimal clinical signs and lower viral loads than those in the challenge group. This study demonstrates the genomic characteristics and pathogenicity of natural recombinant PRRSV-2, illustrating the potential importance of recombination in the field.

Differential evolution of antigenic regions of porcine reproductive and respiratory syndrome virus 1 before and after vaccine introduction.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a widespread viral pathogen that has caused tremendous economic losses throughout most pig-producing countries. Nowadays, both PRRSV-1 and PRRSV-2 co-circulate in Korean pig populations, and commercial modified live vaccine (MLV) is predominantly used to control PRRS. Specifically, control strategy using only PRRSV-2 MLV that was used since 1995 cannot prevent the spread of PRRSV-1 and damage from its infection, which led to the first introduction of two additional PRRSV-1 vaccines in 2014. Despite the wide implementation with PRRSV-1 vaccines, there is a lack of knowledge about the currently circulating Korean PRRSV-1 strains. Whole structural genes of PRRSV-1 before (11) and after (17) the introduction of vaccine were compared to determine the genetic evolutionary features of PRRSV. Genetic and phylogenetic analysis indicated that Korean PRRSV-1 shared 91.5 ± 1.7% nucleotide identity but formed a unique clade based on ORF2-7 phylogeny. Bioinformatics showed increased genetic heterogeneity, enhanced diversifying selection, and the emergence of novel glycosylation sites within neutralizing epitopes of minor structural proteins after vaccine introduction. Taken together, our data provide novel insight into the evolution of minor structural proteins of PRRSV-1 in the vaccination era.

Challenges of influenza A viruses in humans and animals and current animal vaccines as an effective control measure.

Influenza A viruses (IAVs) are genetically diverse and variable pathogens that share various hosts including human, swine, and domestic poultry. Interspecies and intercontinental viral spreads make the ecology of IAV more complex. Beside endemic IAV infections, human has been exposed to pandemic and zoonotic threats from avian and swine influenza viruses. Animal health also has been threatened by high pathogenic avian influenza viruses (in domestic poultry) and reverse zoonosis (in swine). Considering its dynamic interplay between species, prevention and control against IAV should be conducted effectively in both humans and animal sectors. Vaccination is one of the most efficient tools against IAV. Numerous vaccines against animal IAVs have been developed by a variety of vaccine technologies and some of them are currently commercially available. We summarize several challenges in control of IAVs faced by human and animals and discuss IAV vaccines for animal use with those application in susceptible populations.

Complete Genomic Characterization of Korean Porcine Epidemic Diarrhea Virus Strain KUPE21.

Nationwide porcine epidemic diarrhea virus (PEDV) outbreaks occurred in late 2013 in the Republic of Korea, resulting in an expansion of genomic data for Korean PEDVs. However, the data available for Korean PEDVs before 2013 are insufficient. Therefore, we sequenced and analyzed the complete genome of a Korean PEDV strain, KUPE21, which was isolated in the early 2000s.

Genetic identification and serological evaluation of commercial inactivated foot-and-mouth disease virus vaccine in pigs.

Vaccination is considered a frequently used tool to prevent and control foot-and-mouth disease (FMD). However, the effectiveness of conventional FMD virus (FMDV) vaccines in pigs has been controversial because the massive prophylactic vaccination could not elicit proper immune response nor prevent the broad spread of FMD outbreak, mainly in pig farms, in South Korea during outbreaks of 2014. In addition, there has been little information on the efficacy of inactivated, high potency, multivalent, oil-based FMDV vaccine in pigs, because an evaluation of FMDV vaccines had been mainly carried out using cattle. In this study, we evaluated the genetic identification of commercial inactivated FMDV vaccine and monitored the immune responses in pigs under the field condition. Results implied that it contained three different serotypes with a high level of antigen payload. However, serological results showed low mean percentage of inhibition, and positive rate reached its peak at 6-week post-vaccination, indicating current FMDV vaccine need to improve for a prophylactic vaccination policy in pigs. Therefore, there is an imperative need to develop FMDV vaccine that can provide rapid and long-lasting protective immunity in pigs.

Prevalence of novel porcine circovirus 3 in Korean pig populations.

Porcine circovirus 3 (PCV3) is a novel porcine circovirus that was identified in pigs with porcine dermatitis and nephropathy syndrome, reproductive failure, and multi-systemic inflammation. However, the distribution and genetic characteristics of emerging PCV3 in Korea remains unclear. In this study, we determined the nationwide prevalence and genetic characteristics of PCV3 using pen-based oral fluid samples. The total prevalence of PCV3 in individual samples and at the farm level was 44.2% (159/360) and 72.6% (53/73), respectively. Korean PCV3 shared 99.2±0.2% (98.9-99.8%) and 98.6±0.5% (97.9-99.8%) nucleotide identity in the complete genome and ORF2, respectively, when compared to those of US strains. These data suggested that PCV3 is widely distributed throughout Korean pig populations.

Genotypic diversity of porcine circovirus type 2 (PCV2) and genotype shift to PCV2d in Korean pig population.

Porcine circovirus type 2 (PCV2) is a causative agent of PCV2-associated disease (PCVAD), which leads to enormous economic losses in the swine industry worldwide. A high nucleotide substitution rate allows for the continuous evolution of PCV2 and the emergence of novel PCV2 strains. However, the distribution of emerging PCV2 genotypes and the co-existence of multiple genotypes in Korea have not been elucidated. The objective of this study was to determine the genetic diversity of PCV2 in Korean pig herds between 2009 and 2016. The overall prevalence of PCV2, from various samples originating from commercial pigs, was 53.8% (325/604). Two cases of a genotype shift to PCV2d at the farm level showed that the genotype shift started before 2012. In addition, genotype-specific PCR, on pen-based oral fluid samples for nationwide PCV2 surveillance in 2016, indicated that the infection pattern of PCV2 genotypes at the farm level was as follows; none (6/69), PCV2a (6/69), PCV2b (2/69), PCV2d (33/69), PCV2a/b (2/69), PCV2a/d (4/69), PCV2b/d (11/69), and PCV2a/b/d (2/69), respectively. This suggests that the genotype shift to PCV2d occurred on a nationwide scale and that the co-existence of different genotypes is common in Korean pig herds. In addition, seven sites on the capsid protein of Korean PCV2 were identified as being under positive selection pressure, all of which are related to the epitope region and neutralization activity. These data provide evidence of increased genetic diversity and shifts among Korean PCV2 isolates.

Genetic diversity of ORF 4-6 of type 1 porcine reproductive and respiratory syndrome virus in naturally infected pigs.

Genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) has been highly prevalent throughout Korea since the virus was first detected in 2005. However, genetic analyses of genotype 1 PRRSV in Korea have been limited to ORF5 and/or ORF7. In the present study, we determined 10 representative sequence covering ORF4 to ORF6 and each individual ORFs of genotype 1 PRRSV in Korea, and performed molecular analyses. The most variable gene among the individual ORFs of field strains was ORF4, and this gene exhibited only 74.5-87.3% sequence homology compared with strains reported elsewhere. However, the strains showed analogous sequence arrangements with each other. In the phylogenetic analysis, the sequences of Korean field strains formed a distinct cluster with some Austrian and German strains compared to genotype 1 PRRSV strains available in GenBank. In the amino acid analysis, the putative antigenic region of GP4 was highly variable, whereas the predicted epitope regions of ORF5 and ORF6 were relatively conserved. The hydropathy plots of GP4 showed a highly variable pattern in the antigenic region. The non-synonymous and synonymous substitution analysis suggested that ORF4 presumably had more immunogenic pressure compare with the other ORFs. According to these findings, genotype 1 PRRSV in Korea has been diversified and indigenized in Korea, and these strains might have multifarious immunological and genetic properties. This study provides novel insights into genotype 1 PRRSV in a geographically remote area and contributes to the information for further research on the evolution of type 1 PRRSV in the Korean peninsula.

Cross-Reactivity of Porcine Immunoglobulin A Antibodies with Fecal Immunoglobulins of Wild Boar (Sus scrofa) and Other Animal Species.

Fecal samples obtained from wild boar habitats are useful for the surveillance of diseases in wild boar populations; however, it is difficult to determine the species of origin of feces collected in natural habitats. In this study, a fecal IgA ELISA was evaluated as a method for identifying the porcine species from fecal samples. Both domestic pigs (Sus scrofa domestica) and wild boars (Sus scrofa coreanus) showed significantly higher levels of fecal IgA than other animal species. Additionally, age dependent changes in the level of Ig A in wild boars and domestic pigs were identified; Titers of Ig A were highest in suckling period and lowest in weanling period.

Efficacy of commercial genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) vaccine against field isolate of genotype 2 PRRSV.

Although several recent studies have found that type 1 porcine reproductive and respiratory syndrome virus (PRRSV) modified live virus (MLV) vaccine showed appreciable levels of cross-protection against type 2 PRRSV infection, the possibility of cross-protection between two genotype of PRRSV is still controversial. To determine potential protective efficacy against hetero-genotype field strain of PRRSV and to improve understandings of the mechanisms underlying performance improvement after infection in vaccinated animals, piglets were vaccinated with type 1 PRRSV MLV vaccine and challenged with type 2 field strain of PRRSV. As a result, vaccinated animals gained on average 8.45 kg in comparison to 4.77 kg measured in non-vaccinated animals during a 3-week period after viral challenge, which shows using a certain PRRSV vaccine could be clinically effective against heterologous genotypic virus challenge. In vaccinated animals, viremia was reduced and cleared rapidly, whilst viral load was much higher and reduced more slowly, indicating rebound viremia in non-vaccinated animals. The titers of neutralizing antibody against the type 2 PRRSV did not exceed the protective level in any animal from both vaccinated and control groups. Instead, antibody avidity of vaccinated animals was much higher than in the control group clearly. Furthermore, a strong negative correlation between antibody avidity and viremia was noted in 80% of vaccinated animals. Through those results from tests evaluating degree of antibody maturation and its relevance with clearing viremia, it could be suggested that non-neutralizing antibodies induced by vaccination prior to challenge might play a key role in protection against PRRSV infection, especially in early time course.

Wild boars harboring porcine epidemic diarrhea virus (PEDV) may play an important role as a PEDV reservoir.

Porcine epidemic diarrhea virus (PEDV) is a burdensome pathogen in the swine industry. Wild boar population poses a high risk for reservoir of viral pathogen. Two hundred eighty seven samples from wild boar (Sus scrofa) collected in South Korea during 2010/11 were analyzed using RT-PCR, revealing a PEDV infection rate of 9.75% (28/287). PEDV positive samples were distributed throughout the mainland of South Korea, clustering at the northern border adjacent to the Demilitarized Zone (DMZ) and in mountainous regions. PEDV in wild boar was genetically similar to Chinese PEDV strains in phylogenetic investigations. Our results indicated that PEDV is circulating in the wild boar and provided a novel knowledge into epidemiology of PEDV infection.

A novel porcine bocavirus harbors a variant NP gene.

BACKGROUND: Porcine bocavirus is classified within the genus Bocaparvovirus, family Parvoviridae. Unlike other parvoviruses, the members of genus Bocaparvovirus (bocaparvoviruses) encode an additional open reading frame (NP1). Many strains of PBoVs have been identified in domestic pigs and recognized as a potential emerging pathogen causing respiratory and gastrointestinal disease. FINDINGS: A new strain of porcine bocavirus (PBoV) that harbored the shortest NP1 gene among all currently characterized PBoVs (provisionally named as 'PBoV-KU14') was detected in domestic pigs. Almost the complete genome sequence was obtained, approximately 4,630 nucleotides in lengths with putative NS1, NP1, and VP1/2 genes of 1,908, 600, 1,851 bp, respectively. Phylogenetic and comparative analysis was performed using protein and nucleotide sequences. It was revealed that PBoV-KU14 belongs to the genus Bocaparvovirus and species Ungulate bocaparvovirus 4. However, phylogenetic incongruence was observed among species classifications based on the NS1, NP1 and VP1/2 proteins, which indicates a probability of crossover recombination. Conserved protein domains unique for genus Bocaparvovirus in NP1, VP1 protein were also detected. CONCLUSION: NP1 gene truncation supposed to be caused by cross over recombination was detected in a new strain of PBoV (PBoV-KU14). Considering high rates of substitution and recombination in parvovirus, periodic surveillance study to monitor genomic variation and find new strainsof PBoVs seems to be needed.

Cost effectiveness of telecare management for pain and depression in patients with cancer: results from a randomized trial.

OBJECTIVE: Pain and depression are prevalent and treatable symptoms among patients with cancer, yet they are often undetected and undertreated. The Indiana Cancer Pain and Depression (INCPAD) trial demonstrated that telecare management can improve pain and depression outcomes. This article investigates the incremental cost effectiveness of the INCPAD intervention. METHODS: The INCPAD trial was conducted in 16 community-based urban and rural oncology practices in Indiana. Of the 405 participants, 202 were randomized to the intervention group and 203 to the usual-care group. Intervention costs were determined, and effectiveness outcomes were depression-free days and quality-adjusted life years. RESULTS: The intervention group was associated with a yearly increase of 60.3 depression-free days (S.E. = 15.4; P < 0.01) and an increase of between 0.033 and 0.066 quality-adjusted life years compared to the usual care group. Total cost of the intervention per patient was US$1189, which included physician, nurse care manager and automated monitoring set-up and maintenance costs. Incremental cost per depression-free day was US$19.72, which yields a range of US$18,018 to US$36,035 per quality-adjusted life year when converted to that metric. When measured directly, the incremental cost per quality-adjusted life year ranged from US$10,826 based on the modified EQ-5D to US$73,286.92 based on the SF-12. CONCLUSION: Centralized telecare management, coupled with automated symptom monitoring, appears to be a cost effective intervention for managing pain and depression in cancer patients.

Family out-of-pocket health care burden and children's unmet needs or delayed health care.

OBJECTIVE: To assess the relationship between family members' out-of-pocket (OOP) health care spending and unmet needs or delayed health care due to cost for children with and without special health care needs (SHCN). METHODS: Data come from the Medical Expenditure Panel Survey, 2002-2009, and include 63,462 observations representing 41,748 unique children. The primary outcome was having any unmet needs/delayed care as a result of the cost of medical care, dental care, or prescription drugs. We also examined having unmet needs/delayed care due to cost for each service separately. Key explanatory variables were OOP spending on the index child and OOP spending on other family members. We estimated multivariate instrumental variable models to adjust the results for potential bias from any unobserved factors that might influence both other family OOP costs and the outcome variable. RESULTS: An increase of other family OOP costs from $500 (50th percentile) to $3000 (90th percentile) was associated with a higher adjusted rate of any unmet need/delayed care due to cost (1.39% to 5.62%, P < .001, among children without SHCN; 3.17% to 7.87%, P = .01, among those with SHCN). Among children without SHCN, higher OOP costs among other family members were associated with higher levels of unmet needs or delays in medical, prescription drug, and dental care, while among children with SHCN, higher OOP costs among other family members was primarily associated with unmet or delayed dental care. CONCLUSIONS: Programs and policies that reduce the OOP costs of family members other than the child may improve the child's access to care.

Recession led to a decline in out-of-pocket spending for children with special health care needs.

The 2007-09 recession led to an overall slowing in health care spending growth, but it is unclear whether the slowed spending growth had different impacts on adults and children. Although most children are healthy, forgoing routine health care could have long-term adverse implications for public health. Furthermore, children with special health care needs are at risk of adverse outcomes if they do not receive adequate care. Focusing on privately insured families with children, we investigated how out-of-pocket spending trends changed before and during the recession. Medical Expenditure Panel Survey data from the period 2001-09 revealed that the recession did not affect out-of-pocket spending for most children, but it led to a decline in spending for children with special needs, who had much higher out-of-pocket spending at baseline. Adults had significantly lower out-of-pocket spending during the recession, which suggests that parents may reduce their own medical care in difficult economic times to meet their children's health care needs.