An M protein coiled coil unfurls and exposes its hydrophobic core to capture LL-37

Surface-associated, coiled-coil M proteins of Streptococcus pyogenes (Strep A) disable human immunity through interaction with select proteins. However, coiled coils lack features typical of protein-protein interaction sites, and it is therefore challenging to understand how M proteins achieve specific binding, for example, with the human antimicrobial peptide LL-37, which results in its neutralization. The crystal structure of a complex of LL-37 with M87 protein, an antigenic variant from a strain that is an emerging threat, revealed a novel interaction mode. The M87 coiled coil unfurled and asymmetrically exposed its hydrophobic core to capture LL-37. A single LL-37 molecule bound M87 in the crystal, but in solution recruited additional LL-37 molecules, consistent with a protein trap neutralization mechanism. The interaction mode visualized crystallographically was verified to contribute significantly to LL-37 resistance in an M87 Strep A strain, and was identified to be conserved in a number of other M protein types that are prevalent in human populations. Our results provide specific detail for therapeutic inhibition of LL-37 neutralization by M proteins.

Genetic Variability among Swine Influenza Viruses in Italy: Data Analysis of the Period 2017–2020

Swine play an important role in the ecology of influenza A viruses (IAVs), acting as mixing vessels. Swine (sw) IAVs of H1N1 (including H1N1pdm09), H3N2, and H1N2 subtypes are enzootic in pigs globally, with different geographic distributions. This study investigated the genetic diversity of swIAVs detected during passive surveillance of pig farms in Northern Italy between 2017 and 2020. A total of 672 samples, IAV-positive according to RT-PCR, were subtyped by multiplex RT-PCR. A selection of strains was fully sequenced. High genotypic diversity was detected among the H1N1 and H1N2 strains, while the H3N2 strains showed a stable genetic pattern. The hemagglutinin of the H1Nx swIAVs belonged to HA-1A, HA-1B, and HA-1C lineages. Increasing variability was found in HA-1C strains with the circulation of HA-1C.2, HA-1C.2.1 and HA-1C.2.2 sublineages. Amino acid deletions in the HA-1C receptor binding site were observed and antigenic drift was confirmed. HA-1B strains were mostly represented by the Δ146-147 Italian lineage HA-1B.1.2.2, in combination with the 1990s human-derived NA gene. One antigenic variant cluster in HA-1A strains was identified in 2020. SwIAV circulation in pigs must be monitored continuously since the IAVs’ evolution could generate strains with zoonotic potential.

Retrospective Genotyping and Whole Genome Sequencing of a Canine Parvovirus Outbreak in Bangladesh

Canine parvovirus 2 (CPV-2) outbreaks in close quarters such as kennels or shelters can cause substantial case fatality. Thirteen dead Labradors from a secluded kennel of security dogs presented with typical clinical signs and gross pathology of parvovirus infection. Whole genome shotgun sequencing from tissue-extracted genomic DNA detected new CPV-2a as the contributing antigenic variant. Further genotyping using polymerase chain reaction coupled with high-resolution melt assays (PCR-HRM) confirmed new CPV-2a infection in all deceased dogs. PCR-HRM of additional thirty-four clinically suspected dogs suggested that this variant is in wider community circulation, at least in the southeastern part of Bangladesh. We present complete genome sequence of the new CPV-2a variant circulating in the domestic canine population of Bangladesh.

Biological Characterization and Evolutionary Dynamics of Pigeon Paramyxovirus Type 1 in China

Pigeon paramyxovirus type 1 (PPMV-1) is considered as an antigenic variant of Newcastle disease virus (NDV) which has an obvious host preference for pigeons and has caused significant economic losses to the global poultry industry. The evolutionary dynamics of PPMV-1 in China, however, are poorly understood. In this study, we characterized seven PPMV-1 isolates from diseased pigeons collected in Jiangsu, Anhui, and Henan provinces during 2020. Phylogenetic analysis revealed that seven isolates belonged to sub-genotype VI.2.1.1.2.2. Biological characterization indicated that seven isolates were mesogenic based on the mean death time (69.6–91.2 h) and intracerebral pathogenicity index (1.19–1.40) and had similar growth kinetics in chicken embryos and CEFs. Furthermore, the four representative viruses (AH/01/20/Pi, JS/06/20/Pi, HN/01/20/Pi, and HN/02/20/Pi) could result in marked cytopathic effects (CPE) in CEFs and induced syncytium formation in Vero cells. Our Bayesian phylogenetic analysis showed that PPMV-1 might first emerge in East China in 1974 and East China had the highest genotypic diversity of PPMV-1. Besides, phylogeographic analysis indicated that East China and South China were probably the major epicenters of dissemination of PPMV-1 in China. Selection pressure analysis and amino acid substitutions analysis revealed that the viral replication complex (NP, P, and L proteins) was likely related with the host preference of PPMV-1. Collectively, this study uncovered the epidemiology and evolutionary dynamics of PPMV-1 circulating in China, emphasizing the importance of strengthening the monitoring of PPMV-1 in East China and South China and providing significant clues for further studies on the molecular mechanism underlying host preference of PPMV-1.

Evolution of Rabies in South America and Inter-Species Dynamics (2009–2018)

Rabies is listed as one of the World Health Organisation’s (WHO) Neglected Tropical Diseases Worldwide, with a significant impact in South America. This paper explores the dynamics of rabies cases in humans, pets (dogs and cats), livestock and wildlife (bats in particular) in South America during the period 2009–2018. The data used in this study were derived from the two main databases for rabies in South America: the OIE-WAHIS from the World Organisation for Animal Health (OIE) and PANAFTOSA’s Regional Information System for the Epidemiological Surveillance of Rabies (SIRVERA). Being a neglected disease with possible underreporting in some areas, the reported rabies cases may not always represent the real disease burden. The analysis focuses on the evolution of the number of cases in time and their spatial distribution, as well as on the main source of infections in humans, determined by laboratory assays of the antigenic variant or through epidemiological investigations. Additionally, Generalised Linear Mixed Models (GLMM) were used to evaluate the risk factors associated with the occurrence of human cases. Our results show that the highest impact of the disease in terms of number of cases was reported on livestock, while the overall number of cases (in animals and humans) progressively decreased along the study period. The spatial distribution of rabies in livestock showed two main clusters in the north-western (mainly Colombia) and in the south-eastern part of the affected area (Brazil), and a third smaller cluster in Peru. A cluster in dogs was observed in Bolivia. Out of the 192 human cases reported during the study period, 70% of them were transmitted by bats. The number of human cases reported during the study period were significantly associated with the number of rabies cases reported in livestock, pets and wildlife. Despite the overall decreasing case report rate, the disease still represents a major animal and public health concern in South America, and new strategies for compiling systematic information, networking and education are needed, as well as the education and training of veterinary staff.

Recombinant pseudorabies virus with gI/gE deletion generated by overlapping polymerase chain reaction and homologous recombination technology induces protection against the PRV variant PRV-GD2013

Background Since 2011, numerous highly virulent and antigenic variant viral strains have been reported in pigs that were vaccinated against the swine pseudorabies virus. These infections have led to substantial economic losses in the Chinese swine industry. Results This study, constructed a novel recombinant vaccine strain with gI/gE deletion (PRV-GD2013-ΔgI/gE) by overlapping PCR and homologous recombination technology. The growth curves and plaque morphology of the recombinant virus were similar to those of the parental strain. However, PRV-GD2013-ΔgI/gE infection was significantly attenuated in mice compared with that of PRV-GD2013. Two-week-old piglets had normal rectal temperatures and displayed no clinical symptoms after being inoculated with 105 TCID50 PRV-GD2013-ΔgI/gE, indicating that the recombinant virus was avirulent in piglets. Piglets were immunized with different doses of PRV-GD2013-ΔgI/gE, or a single dose of Bartha-K61 or DMEM, and infected with PRV-GD2013 at 14 days post-vaccination. Piglets given high doses of PRV-GD2013-ΔgI/gE showed no obvious clinical symptoms, and their antibody levels were higher than those of other groups, indicating that the piglets were completely protected from PRV-GD2013. Conclusions The PRV-GD2013-ΔgI/gE vaccine strain could be effective for immunizing Chinese swine herds against the pseudorabies virus (PRV) strain.

Evaluation of the Pathogenicity and the Escape from Vaccine Protection of a New Antigenic Variant Derived from the European Human-Like Reassortant Swine H1N2 Influenza Virus

The surveillance of swine influenza A viruses in France revealed the emergence of an antigenic variant following deletions and mutations that are fixed in the HA-encoding gene of the European human-like reassortant swine H1N2 lineage. In this study, we compared the outcomes of the parental (H1huN2) and variant (H1huN2Δ14–147) virus infections in experimentally-inoculated piglets. Moreover, we assessed and compared the protection that was conferred by an inactivated vaccine currently licensed in Europe. Three groups of five unvaccinated or vaccinated piglets were inoculated with H1huN2 or H1huN2Δ14–147 or mock-inoculated, respectively. In unvaccinated piglets, the variant strain induced greater clinical signs than the parental virus, in relation to a higher inflammatory response that involves TNF-α production and a huge afflux of granulocytes into the lung. However, both infections led to similar levels of virus excretion and adaptive (humoral and cellular) immune responses in blood. The vaccinated animals were clinically protected from both infectious challenges and did not exhibit any inflammatory responses, regardless the inoculated virus. However, whereas vaccination prevented virus shedding in H1huN2-infected animals, it did not completely inhibit the multiplication of the variant strain, since live virus particles were detected in nasal secretions that were taken from H1huN2Δ14–147-inoculated vaccinated piglets. This difference in the level of vaccine protection was probably related to the poorer ability of the post-vaccine antibodies to neutralize the variant virus than the parental virus, even though post-vaccine cellular immunity appeared to be equally effective against both viruses. These results suggest that vaccine antigens would potentially need to be updated if this variant becomes established in Europe.

Genomic epidemiology and evolutionary dynamics of respiratory syncytial virus group B in Kilifi, Kenya, 2015–17

Respiratory syncytial virus (RSV) circulates worldwide, occurring seasonally in communities, and is a leading cause of acute respiratory illness in young children. There is paucity of genomic data from purposively sampled populations by which to investigate evolutionary dynamics and transmission patterns of RSV. Here we present an analysis of 295 RSV group B (RSVB) genomes from Kilifi, coastal Kenya, sampled from individuals seeking outpatient care in nine health facilities across a defined geographical area (∼890 km2), over two RSV epidemics between 2015 and 2017. RSVB diversity was characterized by multiple virus introductions into the area and co-circulation of distinct genetic clusters, which transmitted and diversified locally with varying frequency. Increase in relative genetic diversity paralleled seasonal virus incidence. Importantly, we identified a cluster of viruses that emerged in the 2016/17 epidemic, carrying distinct amino-acid signatures including a novel nonsynonymous change (K68Q) in antigenic site ∅ in the Fusion protein. RSVB diversity was additionally marked by signature nonsynonymous substitutions that were unique to particular genomic clusters, some under diversifying selection. Our findings provide insights into recent evolutionary and epidemiological behaviors of RSVB, and highlight possible emergence of a novel antigenic variant, which has implications on current prophylactic strategies in development.