PLoS Neglected Tropical Diseases最新文献

Background: Machupo virus (MACV) is a New World mammarenavirus (hereafter referred to as "arenavirus") and the etiologic agent of Bolivian hemorrhagic fever (BHF). No vaccine or antiviral therapy exists for BHF, which causes up to 35% mortality in humans. New World arenaviruses evolve separately in different locations. To date, up to eight lineages of MACV have been identified in Bolivia. While the prototype MACV Carvallo strain belongs to lineage I discovered in the Memore Province in the 1960, the MACV lineage II strains have become the dominantly-circulating lineage in the same province since 1993.

Methods: We report the development of a reverse genetics system for the MACV lineage II Chicava strain, using a pRF42 plasmid encoding the L and S segment genomic RNA under the transcriptional control of a murine DNA-dependent RNA polymerase I promoter sequence. Rescue of the recombinant MACV Chicava strain (rMACV-Chicava) was accomplished by expression of the L protein and nucleoprotein genes of the MACV Carvallo strain in trans in transfected baby hamster kidney (BHK-21) cells. We characterized the multiplication kinetics of rMACV-Chicava in African green monkey kidney epithelial Vero cells, followed by determining the virulence phenotype in outbred Hartley guinea pigs.

Principal findings: We demonstrated that the multiplication kinetics in Vero cells, virulence phenotype in guinea pigs, and neutralizing antibody titers are indistinguishable between rMACV-Chicava and the wild-type parental virus.

Conclusion and significance: We conclude that rMACV-Chicava provides a useful model system to investigate the emergence of MACV lineage II strains and the guinea pig model has utility for the development of candidate vaccines and therapeutic antibodies for BHF.

Chikungunya virus (CHIKV) is primarily associated with non-human-primates (NHPs) in Africa, which also infect humans. Since its introduction to Brazil in 2014, CHIKV has predominantly thrived in urban cycles, involving Aedes aegypti mosquitoes. Limited knowledge exists regarding CHIKV occurrence and implications in rural and sylvatic cycles where neotropical NHPs are potential hosts, from which we highlight Leontopithecus chrysomelas (Kuhl, 1820), the golden-headed lion tamarin (GHLT), an endangered species endemic to the Atlantic Forest (AF) in Southern Bahia State, Brazil. The present study investigated wild GHLT groups across two municipalities, Ilhéus and Una, Bahia. Surveys were conducted in three groups within cocoa agroforests (cabrucas) in Ilhéus, and four groups in anthropized forest and agroforestry fragments in Una, between 2021 and 2022. Thirty-two GHLT specimens were captured and chemically immobilized, examined and submitted to blood sample collection; nine specimens were later recaptured in 2022, totaling 41 samples. CHIKV viremia was not detected in any specimens (as assayed by RT-qPCR). Plaque reduction neutralization test (PRNT90) detected CHIKV antibodies in two (6.3%) GHLTs, with 10-20 antibody titers. Seroprevalence in 2021 was 5.6% and in 2022 was 8.7% with an incidence of 4.5%, whereas, a male adult tested seropositive in both years, suggesting either natural re-exposure and antibody maintenance over time. All samples tested seronegative for Mayaro Virus. Eight mosquito species from the Culicidae family were collected, identified and assayed for CHIKV genomes, showing negative results. This study provides the first evidence of natural CHIKV exposure among free-living GHLTs in Brazil, emphasizing their susceptibility and potential role as reservoirs. These findings underscore the possible consequences of anthropic disturbances in the Brazilian AF, without a seroprevalence difference between non-protected forest formations, agroforest fragments and various mosaic farming landscapes in South Bahia, and highlight the importance of conservation efforts for this endemic and endangered primate species.

Background: Schistosoma haematobium is the causative pathogen for urogenital schistosomiasis. To achieve progress towards schistosomiasis elimination, there is a critical need for developing highly sensitive and specific tools to monitor transmission in near-elimination settings. Although antibody detection is a promising approach, it is usually unable to discriminate active infections from past ones. Moreover, crude antigens such as soluble egg antigen (SEA) show cross-reactivity with other parasitic infections, and it is difficult to formulate the standard preparations. To resolve these issues, the performances of recombinant antigens have been evaluated. The antibody responses against recombinant S. haematobium serine-protease inhibitor (ShSerpin) and RP26 were previously shown to reflect active schistosome infection in humans. Furthermore, antibody detection using multiple recombinant antigens has been reported to improve the accuracy of antibody-based assays compared to single-target assays. Therefore, we examined the performances of ShSerpin, RP26 and the mixture of these antigens for detecting S. haematobium low-intensity infection and assessed the potential for transmission monitoring.

Methodology/principal findings: We collected urine and plasma samples from school-aged children in Kwale, Kenya and evaluated S. haematobium prevalence by number of eggs in urine and worm-derived circulating anodic antigen (CAA) in plasma. Among 269 pupils, 50.2% were CAA-positive by the lateral flow test utilizing up-converting phosphor particles (UCP-LF CAA), while only 14.1% were egg-positive. IgG levels to S. haematobium SEA (ShSEA), ShSerpin, RP26, and the mixture of ShSerpin and RP26 were measured by ELISA. The mixture of ShSerpin and RP26 showed the highest sensitivity, 88.7%(125/141)among the four antigens in considering indecisive UCP-LF CAA results as negative.

Conclusion/significance: IgG detection against the ShSerpin-RP26 mixture demonstrated better sensitivity for detection of active S. haematobium infection. This recombinant antigen mixture is simpler to produce with higher reproducibility and can potentially replace ShSEA in monitoring transmission under near-elimination settings.

Understanding host utilization by mosquito vectors is essential to assess the risk of vector-borne diseases. Many studies have investigated the feeding patterns of Culex mosquitoes by molecular analysis of blood-meals from field collected mosquitoes. However, these individual small-scale studies only provide a limited understanding of the complex host-vector interactions when considered in isolation. Here, we analyse the Culex blood-feeding data from 109 publications over the last 15 years to provide a global insight into the feeding patterns of Culex mosquitoes, with particular reference to vectors of currently emerging Culex-borne viruses such as West Nile and Usutu. Data on 29990 blood-meals from 70 different Culex species were extracted from published literature. The percentage of blood-meals on amphibian, avian, human, non-human mammalian, and reptilian hosts was determined for each Culex species. Our analysis showed that feeding patterns were not significantly explained by mosquito species-level phylogeny, indicating that external factors play an important role in determining mosquito feeding patterns. For Cx. quinquefasciatus, 'Cx. pipiens pooled', and Cx. tritaeniorhynchus, feeding patterns were compared across the world's seven biogeographical realms. Culex tritaeniorhynchus, 'Cx. pipiens pooled' and Cx. quinquefasciatus all had significantly varied feeding patterns between realms. These results demonstrate that feeding patterns of Culex mosquitoes vary between species but can also vary between geographically distinct populations of the same species, indicating that regional or population-level adaptations are major drivers of host utilization. Ultimately, these findings support the surveillance of vector-borne diseases by specifying which host groups are most likely to be at risk.

The global priorities in the field of infectious diseases are constantly changing. While emerging viral infections have regularly dominated public health attention, which has only intensified after the COVID-19 pandemic, numerous bacterial diseases have previously caused, and continue to cause, significant morbidity and mortality-deserving equal attention. Three potentially life-threatening endemic bacterial diseases (leptospirosis, melioidosis, and rickettsioses) are a huge public health concern especially in low- and middle-income countries. Despite their continued threat, these diseases do not receive proportionate attention from global health organizations and are not even included on the WHO list of neglected tropical diseases (NTDs). This, in turn, has led to a vicious circle of neglect with continued, yet conceivably preventable, hospitalizations and deaths each year especially in the vulnerable population. This is a call from a group of multi-institutional experts on the urgent need to directly address the circle of neglect and raise support in terms of funding, research, surveillance, diagnostics, and therapeutics to alleviate the burden of these 3 diseases.

Background: Intestinal larva invasion is a crucial step of Trichinella spiralis infection. Intestinal infective larvae (IIL) and their excretory/secretory proteins (ESP) interact with gut epithelium, which often results in gut epithelium barrier injuries. Previous studies showed when T. spiralis invaded intestinal epithelium cells, the IIL ESP disrupted the tight junctions (TJs) of Caco-2 monolayer, but the mechanism is not clear. The IIL ESP might cause gut epithelial apoptosis, weaken the gut barrier and aid the larval invasion. The aim of this study was to investigate whether T. spiralis IIL ESP participate in enterocyte apoptosis and disrupt gut epithelial barrier to promote the larval invasion.

Methodology/principal findings: Cell viability was assessed by CCK-8 assay and the results showed that 200 μg/ml of IIL ESP incubated with Caco-2 cells for 18 h inhibited the Caco-2 cell viability. The results of trans-epithelial electrical resistance (TEER) and FITC-dextran showed that IIL ESP decreased the TEER, increased FITC-dextran flux in Caco-2 monolayer. qPCR, Western blot and immunofluorescence test (IFT) showed that IIL ESP decreased the mRNA and protein expression of TJs (ZO-1, E-cad, Occludin and Claudin-1). The IIL ESP-induced Caco-2 cell apoptosis was observed by DAPI, Hoechst 33358, TUNEL and Annexin V/PI staining. Besides, flow cytometry revealed an increasing apoptosis rate in Caco-2 cells after the IIL ESP treatment. qPCR and Western blot analysis indicated that IIL ESP activated caspases (Caspase 3, Caspase 9 and Caspase 8), up-regulated the pro-apoptotic factors (Bax and Cytochrome c) and down-regulated the anti-apoptosis molecule Bcl-2. Interestingly, pretreatment of Caco-2 cells with apoptosis inhibitor Z-VAD-FMK abrogated and recovered the barrier function of Caco-2 monolayer destroyed by IIL ESP. Furthermore, the Z-VAD-FMK pretreatment also impeded the in vitro larva invasion of Caco-2 monolayer.

Conclusions: T. spiralis IIL ESP induced gut epithelial apoptosis, reduced the TJs expression, damaged gut epithelial integrity and barrier function, and promoted larval invasion. These findings provided a basis of further understanding the interaction mechanism between T. spiralis and host gut epithelium, and they were valuable to the development new prevention and therapeutic strategy of early T. spiralis infection.

Background: Riverine communities face various health problems, which involve geographical and cultural barriers to accessing care, in addition to a lack of financial investments in services aimed at these communities, resulting in a process of invisibility for the population living in these regions. In this scenario, the significant burden of snakebite envenoming (SBE) highlights the need for participatory research to address ways to minimize this situation. Thus, this study aimed to describe the priority health problems identified by this population and the ranking of SBEs in that context, mapping solutions according to the local reality.

Methodology/principal findings: This study was conducted in Limeira, a riverine community located in Tabatinga, in the extreme Western Brazilian Amazonia, on the borders with Peru and Colombia. The research lasted approximately one year, from 2021 to 2022.It is a participatory study that followed three steps: baseline assessment of the community, community assembly, and final data analysis. The study included a total of 42 participants in the sociodemographic survey, which served as the basis for the subsequent stages of data collection. Of these 42 individuals, 32 participated in the qualitative interviews, and 20 took part in the community assembly. Participants emphasized snakebite envenoming as a significant health issue, though not the only one, and reported frequent encounters with snakes, underscoring its severity as a concern. The qualitative analysis identified three main themes: Snakebites in the Community, which focused on personal experiences with snakes; Common Health Problems, which addressed other health issues faced by community members; and Community Defining Solutions, which discussed strategies and solutions proposed by the community to address these challenges.

Conclusions/significance: Improvements in health care delivery to populations living in Amazonian communities are possible with the judicious use of tested integrated interventions, particularly when the community identifies various concurrent health problems. SBE control programs in remote areas of the Brazilian Amazon should be planned with a multidisciplinary and intercultural approach, preferably integrated with broader interventions that address the population's needs for a range of health issues.

Background: C-type lectin (CTL) plays an important act in parasite adhesion, host's cell invasion and immune escape. Our previous studies showed that recombinant Trichinella spiralis C-type lectin (rTsCTL) mediated larval invasion of enteral mucosal epithelium. The aim of this study was to investigate protective immunity produced by vaccination with rTsCTL and its effect on gut epithelial barrier function in a mouse model.

Methodology/principal finding: The ELISA results showed that subcutaneous vaccination of mice with rTsCTL elicited a systemic humoral response (high levels of serum IgG, IgG1/IgG2a and IgA) and significant gut mucosal sIgA responses. The levels of Th1/Th2 cytokines (IFN-γ/IL-4) secreted from spleen, mesenteric lymph nodes and Peyer's patches were distinctly increased at 6 weeks following vaccination (P < 0.05). At one week after challenge, the numbers of goblet cells and expression level of Muc2, Muc5ac and pro-inflammatory cytokines (TNF-α and IL-1β) in gut tissues of vaccinated mice were obviously decreased, while expression of anti-inflammatory cytokines (IL-4 and IL-10) was evidently increased, compared to the infected PBS group. It is interesting that expression levels of gut epithelial tight junctions (TJs; occludin, claudin-1 and E-cad) were prominently elevated and intestinal permeability was interestingly declined in vaccinated mice. The rTsCTL-vaccinated mice exhibited a 51.69 and 48.19% reduction of intestinal adult and muscle larva burdens, respectively. The female fecundity in rTsCTL vaccinated mice was reduced by 40.51%. These findings indicated that rTsCTL vaccination impeded larval invasion and improved gut epithelial integrity and barrier function, reduced worm burdens, and relieved gut and muscle inflammation.

Conclusions: Vaccination of mice with rTsCTL elicited an obvious protective immunity against larval challenge, impeded larval invasion of gut mucosa, enhanced gut epithelial integrity and barrier function, reduced worm burdens; it also alleviated gut and muscle inflammation. TsCTL might be a novel candidate target molecule for anti-Trichinella vaccines.

Scrub typhus is caused by Orientia tsutsugamushi infection and occurs frequently in an area called the Tsutsugamushi Triangle. Currently, there is no vaccine for O. tsutsugamushi, and its infection is treated with antibiotics such as doxycycline. Scrub typhus responds to effective treatment, and early treatment shortens the course of the disease, reduces mortality, and accelerates recovery. Therefore, it is important to rapidly diagnose O. tsutsugamushi infection to ensure successful outcomes. Here, we developed a CRISPR-Cas12a-based diagnostic method targeting the bacterial 16S rRNA to detect O. tsutsugamushi infection of all known genotypes. To reduce the possibility of contamination and increase field applicability, we designed the one-pot assay system in addition to conventional two-pot assay system. Using this method, we successfully detected up to 100 copies of in vitro transcribed O. tsutsugamushi 16S rRNA within 1 hour under isothermal conditions. In blood samples from patients confirmed to be infected with O. tsutsugamushi by nested PCR, the developed method exhibited a clinical sensitivity of 98% and high specificity. These data demonstrate that the presented method is applicable for the rapid and universal diagnosis of scrub typhus to facilitate timely and appropriate treatment.