PLoS Pathogens最新文献

Acute hepatopancreatic necrosis disease (AHPND) is a disease that has caused significant losses to shrimp farming since 2009. The primary mechanism of this disease involves the binary toxins PirAvp and PirBvp, which are produced by specific strains of Vibrio parahaemolyticus, and which lead to significant damage to the hepatopancreatic cells of shrimps. Recent studies on the pathology of AHPND have also highlighted the role of the Vibrio quorum sensing (QS) system, which affects growth, virulence, and biofilm regulation in Vibrio species. For example, deletion of the qseC gene reduces the virulence of the AHPND-causative V. parahaemolyticus. Most importantly, the QS regulators LuxOvp and AphBvp have been implicated as they control the growth-phase-dependent expression of the pirAvp/pirBvp genes. Additionally, given the growing problem of antibiotic resistance, this article reviews several alternative control strategies targeting the QS system, including QS inhibition using natural products, biofloc technology, and the development of small-molecule inhibitors against AphBvp. Finally, we also discussed the potential of using probiotics to enhance shrimp disease resistance through QS inhibition, highlighting the feasibility of targeting the QS system for AHPND control.

Surface adhesion is critical to the survival of pathogenic bacteria both in natural niches and during infections, often via forming matrix-embedded communities called biofilms. Vibrio cholerae, the causal agent of pandemic cholera, is capable of forming biofilms adhering to both biotic and abiotic surfaces and the biofilm lifestyle has been implicated in promoting the survival of V. cholerae both in the natural reservoir and during host colonization. Previously, a 57-amino acid loop in the biofilm-specific adhesin Bap1 (Bap1-57aa) has been identified as a key contributor to the adhesion of V. cholerae biofilms to various surfaces including lipid membranes. However, the mechanism underlying its interaction with lipids, as well as its secondary structures, remain unresolved. Here, we combined biophysical, computational, and genetic approaches to elucidate the molecular mechanism of how this adhesive peptide interacts with lipids and lipid-coated surfaces. We found that a central aromatic-rich motif anchors the peptide to lipid bilayers while peripheral pseudo repeats enhance binding through avidity. Surprisingly, the core motif undergoes a lipid-induced conformational transition into a β-hairpin, enabling robust membrane insertion. We confirmed these findings both in vitro and in the biofilm context. Moreover, we demonstrated that the adhesive peptide can adhere to model host surfaces and is sensitive to membrane curvature. Finally, we show that the biofilm-derived peptide is found in several other Vibrio species, and its sequence is well-conserved. Our results provide molecular insight into biofilm adhesion and may lead to new strategies for targeted biofilm removal, as well as the design of bioinspired underwater adhesives.

[This corrects the article DOI: 10.1371/journal.ppat.1010211.].

New SARS-CoV-2 variants pose an ongoing threat due to persistent immune escape of natural and vaccine-induced immunity. The emergence of BA.1 (Omicron) produced a large antigenic shift in the spike protein, rendering many antibodies ineffective with concomitant loss of Emergency Use Authorization (EUA) status. While strains have evolved far from BA.1, re-emergence of variants from branches closer to BA.1 are of recent concern. Here, we engineered a self-assembling nanoparticle displaying RBD 4mut g5.1, an immunogen developed using structure-guided design to focus antibody responses to the receptor binding site (RBS) epitope and promote cross-reactivity by inclusion of four rationally selected BA.1 mutations in the RBS. Unlike multi-component RBD approaches, we demonstrate a single, rationally designed component is sufficient for generating broad immunity. We demonstrate that in both naïve and antigen-experienced mice, RBD 4mut g5.1 nanoparticle induced cross-reactive and durable antibody responses capable of potent neutralization of ancestral SARS-CoV-2 and many Omicron variants. RBD 4mut g5.1 provided heterologous protection at a memory timepoint. By showcasing how subtle changes in an epitope can trigger a diversified antibody response, this study offers a promising new avenue for developing vaccines that can more effectively tackle the ever-evolving threat of immune escape, not only against SARS-CoV-2 but potentially against a range of variable pathogens.

The RIG-I-like receptor (RLR) signaling pathway plays a critical role in the host defense against RNA virus infection. Among the RLR family members, retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5) are key cytosolic sensors that initiate type I interferon (IFN-I) responses. Their controllable expression, activation, and degradation are essential for maintaining immune homeostasis. However, the precise regulatory mechanisms governing RIG-I and MDA5 function during viral infection remain unclear. Here, we uncover that the E3 ubiquitin ligase RNF20 exerts dual regulatory roles in RLR signaling by modulating the expression and promoting the degradation of RIG-I and MDA5 in a nucleocytoplasmic translocation-dependent manner during viral infection. Under resting conditions, RNF20 resides in the nucleus, where it maintains immune readiness by regulating the basal and inducible transcription of RIG-I and MDA5. Upon RNA virus infection, RNF20 translocates to the cytoplasm via the export receptor CRM1. There, it recognizes the degron motifs of RIG-I and MDA5 through its coiled-coil domain and catalyzes their K27-linked ubiquitination and degradation, thereby preventing excessive antiviral signaling. These findings shed light on the significant and dual regulatory roles of RNF20 in maintaining innate immune homeostasis.

Parvovirus B19 (B19V) is a prevalent human pathogen that can cross the placenta by a mechanism that remains unknown, posing a risk of severe fetal complications, particularly during the first trimester of pregnancy. We investigated the expression of B19V-specific receptors in the three trophoblast cell types, cytotrophoblasts (CTBs), syncytiotrophoblasts (STBs), and extravillous trophoblasts (EVTs), and assessed their susceptibility to infection. VP1uR, the receptor that mediates viral uptake and infection in erythroid progenitor cells, is expressed in CTBs and STBs, but not in EVTs. Globoside, a glycosphingolipid that is essential for the escape of the virus from endosomes, is also expressed in these cells, except for choriocarcinoma-derived CTBs. In the latter, the absence of globoside can be overcome by promoting endosomal leakage with polyethyleneimine. While erythropoietin receptor (EpoR) signaling is associated with the strict erythroid tropism of B19V, it is not required for infection in trophoblasts. Transfection experiments revealed that highly proliferative first-trimester CTBs are more susceptible to B19V infection than the low-proliferative CTBs from term placenta. These findings demonstrate that B19V targets specific trophoblast cells, where viral entry and replication are collectively mediated by VP1uR, globoside, and high cellular proliferative activity, but are independent of EpoR signaling.

Porcine astrovirus (PAstV) is globally prevalent in swine and is associated with diarrhea and encephalitis in piglets, posing a threat to porcine health. However, its pathogenic mechanisms remain poorly understood. This study used the PAstV1-GX1 strain to infect PK-15 cells, revealing that the virus induces significant apoptosis, with late apoptotic cells reaching 41.2% at 24 hours post-infection. The infection activates caspase-9 and caspase-3, but not caspase-8, and causes mitochondrial damage, indicating apoptosis via the mitochondrial pathway. The apoptosis inhibitor Z-VAD-FMK reduced viral replication, while apoptosis inducer ABT-263 enhanced it at later stages. The nsP1a/3 protein, which interacts with MAVS and localizes to mitochondria, was identified as key in inducing apoptosis. Its 3C-like serine protease domain likely mediates this interaction. Knocking down MAVS reduced apoptosis and increased early-stage replication but decreased it later. Overexpressing MAVS increased apoptosis and decreased replication. Furthermore, we observed that the expression of nsP1a/3 resulted in the cleavage of MAVS and the suppression of the type I interferon (IFN) response. Notably, treatment with Z-VAD-FMK did not influence nsP1a/3-mediated MAVS cleavage or type I IFN inhibition, suggesting that the induction of apoptosis and MAVS cleavage are distinct processes. By employing site-directed mutagenesis to substitute alanine for the catalytic triad residues (His459, Asp487, and Ser549) of the 3C-like serine protease, we significantly reduced the ability of nsP1a/3 to induce apoptosis, cleave MAVS, and suppress the type I IFN response, underscoring the essential role of protease activity in these functions. Furthermore, the use of a serine protease inhibitor markedly decreased PAstV replication. These findings provide significant insights into the pathogenesis of PAstV and establish a foundation for the development of novel antiviral therapies.

Rift Valley fever virus (RVFV) poses a continued threat to human health and animal husbandry. Two neutralizing and protective human monoclonal antibodies (mAbs), RVFV-268 and RVFV-379, exhibit similar affinities and epitope footprints on the Gn glycoprotein component of the RVFV Gn-Gc capsomeric lattice. Here, we define fine details of the biophysical determinants of Gn recognition used by RVFV human monoclonal antibodies through studying an antibody encoded by a set of recombined genes not previously identified in RVFV antibodies. We find that RVFV-379 exhibits a larger footprint than that observed for RVFV-268 and other antibodies targeting the same region, which involves major contributions of both the light and heavy chains. RVFV-379 also uses an oblique angle of approach towards the virion surface that contrasts with the perpendicular angle of engagement observed for some other potently neutralizing human mAbs. Further, consistent with amino acid sequence variation within and proximal to the RVFV-379 epitope, in vitro neutralization screening reveals a limited degree of neutralization breadth across prevalent RVFV strains, suggesting that RVFV has fewer functional constraints at this region of the virus envelope. By dissecting the molecular determinants of mAb recognition of Gn, this integrated analysis refines strategies needed for the rational design of vaccines that can elicit a potent and species-wide protective antibody immune response to this important re-emerging pathogen.

The Mab21/cGAS protein family has diversified across metazoans to regulate development and innate immunity. In vertebrates, cGAS detects cytosolic DNA and synthesizes 2'3'-cGAMP to activate STING-TBK1-IRF signaling, while invertebrate cGAS-like receptors (cGLRs) recognize RNA or DNA and generate non-canonical cyclic dinucleotides. However, whether shrimp Mab21 proteins function as canonical nucleic acid sensors remains unresolved. Here, we identified three Mab21 proteins from Litopenaeus vannamei-LvMab21-1, LvMab21-2, and LvMab21-3. Although they are phylogenetically related to cGAS-like proteins, none bound dsDNA or dsRNA or synthesized cGAMP in response to ISD or poly(I:C). Instead, all three interacted directly with the TBK1 homolog LvIKKε, promoted its phosphorylation at serine 175, and thereby activated the downstream IRF-Vago4 signaling axis. This mechanism defines a non-canonical nucleic acid sensing paradigm, whereby Mab21 proteins act as protein-based enhancers of kinase activation rather than as nucleic acid-dependent CDN synthases. We further show that these proteins display tissue-specific antiviral functions: all three act in hemocytes, LvMab21-1 predominates in hepatopancreas, LvMab21-2 and LvMab21-3 are most critical in gills, and LvMab21-1 and LvMab21-3 cooperate in intestine. Silencing any Mab21 paralog reduced survival and increased white spot syndrome virus (WSSV) burden, underscoring their physiological relevance. Together, our findings expand the known repertoire of innate immune strategies within the Mab21 family, highlight a previously unrecognized non-canonical mechanism of interferon-like activation, and reveal tissue-specific specialization that tailors antiviral responses across shrimp organs. These insights provide both evolutionary context and candidate targets for breeding disease-resistant shrimp.