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Exhausting carbon-based gases from building construction or comfortable household supplies have caused the main pollution gas species for the indoor air environment. Given the variety of indoor activities, many reports have been challenged to find room-temperature operational and harmless materials. Here, we report that the activating MoO vibration mode in Na,Sb doped-NiMoO4 micro-sized powder reveals the high response of CO and VOC, occupying multiple bonds at room temperature. We performed Na and Sb doping in the α-NiMoO4 crystal structure, such as introducing the Sb in Ni and interstitial doping of Na in around 4 Å of free space in the NiMoO4 crystal structure that manipulates the partial β-NiMoO4 phase. The Na,Sb-codoped NiMoO4 generates the higher MoO vibration mode from FT-IR measurement and enables carbon-based gas detection with 38 responses to CO and nearly 20 responses to VOC under 20 ppm of each analyte gases environment. Furthermore, the n-type gas sensing behavior of Na,Sb doped-NiMoO4 chemiresistor exhibits an immediate sensing response instead of a none-of response than intrinsic or single element doped NiMoO4. These results suggest that Na,Sb codoped-NiMoO4 is the applicable material for emergent warning against carbon-based gas exposure in the indoor environment. In addition, activating the electrochemical reaction site of MoO4–2 in NiMoO4 is tailorable by a simple doping process.

In this study, non-oriented Au-decorated SnO2 nanospheres (NSs) were synthesized using hydrothermal and microwave methods. The SnO2 spheres ranged from hundreds of nanometers to a microscale. The oxygen vacancy on the surface increased after Au adsorption on the surface of the SnO2 NSs, ultimately showing a synergistic effect with the spillover effect of the existing Au catalyst. Specifically, at 100 °C and 200 °C, the response to 10 ppm H2S gas improved to 16.14 and 46.81, respectively. These gas sensing effects were approached individually by being divided into two disadvantages (oxygen adsorption and homojunction) and five advantages (oxygen vacancies, spill-over, surface area, H2S + SnO2 reaction, and H2S + O2 reaction). Based on this reference, we investigated SnO2 NSs of various sizes and functions by adjusting the process variables.

The demand for gas-sensing operations with lower electrical power and guaranteed sensitivity has increased over the decades due to worsening indoor air pollution. In this report, we develop room-temperature operational NH3 gas-sensing materials, which are activated through electron doping and crystal structure distortion effect in Fe0.2Ni0.8WO4. The base material, synthesized through solid-state synthesis, involves Fe cations substitutionally located at the Ni sites of the NiWO4 crystal structure and shows no gas-sensing response at room temperature. However, doping Na into the interstitial sites of Fe0.2Ni0.8WO4 activates gas adsorption on the surface via electron donation to the cations. Additionally, the hydrothermal method used to achieve a more than 70-fold increase in the surface area of structure-distorted Na-doped Fe0.2Ni0.8WO4 powder significantly enhances gas sensitivity, resulting in a 4-times increase in NH3 gas response (Rg/Ra). Photoluminescence and XPS results indicate negligible oxygen vacancies, demonstrating that cation contributions are crucial for gas-sensing activities in Na-doped Fe0.2Ni0.8WO4. This suggests the potential for modulating gas sensitivity through carrier concentration and crystal structure distortion. These findings can be applied to the development of room-temperature operational gas-sensing materials based on the cations.

After forming the double layer of SnO2 and Au, the compound of Sn and O with multiple band gaps was formed in a short time period via flame chemical vapor deposition (FCVD). The Sn component of SnO2 has fluidity and is the only one moving to the discrete Au side and leaving O behind. The capillarity of Sn allows it to form a unique structure with Au-Sn-O nanoparticles and amorphous Sn-O nanotubes of multiple compositions in a single process. For verification, the gradual morphological, crystallographic, compositional, and optical changes in the material were sequentially presented as per the degree of FCVD.

WO3 nanostructures with different morphologies and dimensions were fabricated via solvothermal synthesis by adjusting the stirring time of the precursor solution. Ethanol-based solutions of the WCl6 precursor with various colors (dark green, yellow, white, blue, and blue-black) were prepared, and this triggered a significant change in the growth behavior during the evolution of WO3 nanostructures. Controlling the initial state of the precursors in solution enabled sequential nucleation and growth which resulted in the production of zero-to-three-dimensional nanostructures including nanoparticles, a mixture of nanosheets and nanoparticles, jointed-nanosheets, and three-dimensionally clustered jointed-nanosheets. The crystallographic characteristics (preferred orientation along the (002) plane) and the concentration of surface oxygen vacancies were also controllable, suggesting the formation of nanostructures with tuneable surface reactivity. Differing NO2 sensing performances were observed because of the variation in configurations of the WO3 nanostructures.

Herein, we report a simple and scalable synthesis route to prepare Ag-functionalized porous ZnO sheets and their enhanced NO2 gas sensing properties. Porous ZnO sheets functionalized with well-dispersed submicron Ag particles were prepared by using a hydrothermal method-based one-pot synthesis route from Zn and Ag precursors. NO2 gas sensing performance (response, selectivity, response time, and recovery time) was optimized at 200°C in the gas sensor fabricated with 3 at% Ag-functionalized porous ZnO sheets. We demonstrated a response (Rg/Ra) of 17.18 to 10 ppm NO2 gas and also obtained a high response of 14.05 even at 60% relative humidity due to the synergetic effect of improved NO2 gas adsorption in the presence of Ag particles and increased resistance by the formation of Schottky barrier at Ag-ZnO heterojunctions.

Since the increase in the emission of various Volatiles Organic Compounds, gas and formaldehyde gas have had a harmful effect on the human body, and gas sensors that can measure those gases were fabricated in this study. After Pt coating was performed on the alumina substrate, Zn seed layers were fabricated. Nanostructures were formed through sonochemical synthesis by varying the ratio of ZnO and TiO2. Thereafter, the reactivity and recovery properties were compared and evaluated according to the concentrations of formaldehyde and toluene gas. The ZnO(99%)-TiO2(1%) gas sensor showed meaningful selectivity of about 40% or more at a concentration ranging from 5 to 20 ppm (high concentration) of formaldehyde and toluene gas, and showed a low selectivity of about 5% or more for a concentration ranging from 0.1 to 1 ppm (low concentration) of formaldehyde and toluene gas. This sensor can be optimized to have a meaningful selectivity of formaldehyde gas compared to other Volatiles Organic Compounds gases by optimizing the ZnO-TiO2 nanostructure.

Herein, we present the effect of electron-beam irradiation (EBI) on the gas-sensing properties of Pd-functionalized reduced graphene oxide (RGO). Scanning electron microscopy, transmission electron microscopy, and X-ray photoelectron spectroscopy were used to characterize the synthesized products. The samples were irradiated using electron beams at doses of 0 (Pd-RGO-0), 100 (Pd-RGO-100), and 500 kGy (Pd-RGO-500), and the NO2 gas-sensing properties were investigated. It was found that irradiation by electron beams has a critical effect on the gas-sensing properties of the samples, and the Pd-RGO-500 sensor showed the highest response to NO2 gas. In particular, the response of the unirradiated sensor and the sensor irradiated at doses of 100 and 500 kGy to 10 ppm NO2 were 1.027, 1.045, and 1.047, respectively. The response times of Pd-RGO-0, Pd-RGO-100, and Pd-RGO-500 to 10 ppm NO2 gas were 389, 335, and 345 s, respectively. The corresponding recovery times for these sensors were 808, 766, and 816 s, respectively. The increased numbers of oxygen functional groups and high-energy defects were the main reasons for the increased gas response. This study will eventually lead to increased performance levels of RGO-based sensors that use EBI, as this technology can introduce changes into materials in a non-contact, clean, and powerful manner.

This technical note provides some considerations for the metallographic observation of intergranular attack (IGA) in Alloy 600 steam generator tubes. The IGA region was crazed along the grain boundaries through a deformation by an applied stress. The direction and extent of the crazing depended on those of the applied stress. It was found that an IGA defect can be misevaluated as a stress corrosion crack. Therefore, special caution should be taken during the destructive examination of the pulled-out tubes from operating steam generators.

This work presents the influence of noise originating from the tube itself on the detectability and sizing accuracy for laboratory-induced outer diameter axial cracks in nuclear steam generator tubes. The variations of signal amplitude and phase angle of the same cracks were analyzed when increasing the signal-to-noise ratio of the tube itself from 9 to 18. It was experimentally verified that the detectability for small cracks was enhanced by increasing the signal-to-noise ratio. The phase angle also rotated to a value representing the actual position and depth of a crack when increasing the signal-to-noise ratio.

We prepared a highly sensitive and selective NO2 sensor, based on the TeO2 branched SnO2 nanowires (NWs), in terms of vapor-liquid-solid method, with subsequent growing of branches on the stems of SnO2 NWs. Fabricated sensors showed a high response higher than 10 to 10 ppm of NO2 gas at 100 °C. We investigated the associated sensing mechanisms, with respect to the enhancement of sensing behaviors by the addition of TeO2 branches. Based on the results obtained in this work, we believe that the present sensor with an efficient fabrication technique, and high sensitivity and selectivity can be used for detection of NO2 gas in real applications.

The unexpectedly large outbreak of Middle East respiratory syndrome in South Korea in 2015 was initiated by an infected traveler and amplified by several “superspreading” events. Previously, we reported the emergence and spread of mutant Middle East respiratory syndrome coronavirus bearing spike mutations (I529T or D510G) with reduced affinity to human receptor CD26 during the outbreak. To assess the potential association of spike mutations with superspreading events, we collected virus genetic information reported during the outbreak and systemically analyzed the relationship of spike sequences and epidemiology. We found sequential emergence of the spike mutations in 2 superspreaders. In vivo virulence of the mutant viruses seems to decline in human patients, as assessed by fever duration in affected persons. In addition, neutralizing activity against these 2 mutant viruses in serum samples from mice immunized with wild-type spike antigen were gradually reduced, suggesting emergence and wide spread of neutralization escapers during the outbreak.

ABSTRACTScrub typhus is an acute febrile disease caused by Orientia tsutsugamushi infection. Despite the wide range of approaches explored during the last seventy years, an effective prophylactic vaccine is not yet available. Here, we developed a novel recombinant antigen derived from conserved regions of 56 kDa type-specific antigen (TSA56), a major outer membrane protein responsible for genetic heterogeneity and antigenicity, and evaluated it as a protective vaccine antigen. Our findings demonstrate that immunization with conserved blocks of TSA56 (cTSA56) not only provides protective immunity against lethal challenges with the homologous genotype, but also confers significantly better protection against heterologous genotypes than TSA56. Adoptive transfer of CD4+ or CD8+ T cells from immunized mice provided significantly enhanced protection against lethal challenge, whereas immune B cells failed to do so, indicating that cellular immunity against the conserved epitopes plays a protective role. Moreover, immunization with a 10-mer peptide mixture, screened from CD8+ T cell epitopes within the conserved region of TSA56, provided enhanced protection against lethal challenge with O. tsutsugamushi. Therefore, this novel recombinant antigen is a promising candidate for scrub typhus vaccine against a wide range of O. tsutsugamushi genotypes.

Scrub typhus, caused by Orientia tsutsugamushi infection, has been a serious public health issue in the Asia-Pacific region, with rising incidence and sporadic outbreaks. However, human protective immunity against specific antigens has been poorly characterized for this bacterium. In addition, immunity produced in early vaccine trials or even after natural infections, did not last long and had poor cross-reactivity among various genotypes. Here, we systematically investigated the kinetics and magnitude of specific adaptive immunity against two membrane antigens, 56 kDa type-specific antigen (TSA56) and surface cell antigen A (ScaA), that are involved in bacterial adhesion and invasion of the host in 64 recovered scrub typhus patients. Antibody responses to the bacterial antigens in patients were generally short-lived and waned to baseline levels 2 years after recovery. The anti-TSA56 IgG responses were predominantly composed of the IgG1 and IgG3 subclasses and persisted for up to 1 year after recovery, whereas IgG specific to ScaA primarily consisted of more transient IgG1, with limited responses by other subclasses. Cellular immunity, including CD4 and CD8 T-cells specific to membrane antigens, also rapidly declined from 1 year after infection, as measured by enzyme-linked immunospot (ELISPOT) assays and flow cytometry. The short longevity of antigen-specific adaptive immunity might be attributable to limited memory responses, as observed in earlier vaccine studies using whole bacterial antigens. Finally, we identified HLA-A*0201-restricted and highly conserved CD8 T-cell epitopes in the TSA56 antigen, which may be valuable tools for assessing cellular immunity against O. tsutsugamushi and developing an effective scrub typhus vaccine.Emerging Microbes & Infections (2017) 6, e116 doi:10.1038/emi.2017.106; published online 20 December 2017

Abstract Background Zinc oxide nanoparticle (ZNP) has been applied in various biomedical fields. Here, we investigated the usage of ZNP as an antigen carrier for vaccine development by combining a high affinity peptide to ZNP. Results A novel zinc oxide-binding peptide (ZBP), FPYPGGDA, with high affinity to ZNP (K a = 2.26 × 106 M−1) was isolated from a random peptide library and fused with a bacterial antigen, ScaA of Orientia tsutsugamushi, the causative agent of scrub typhus. The ZNP/ZBP-ScaA complex was efficiently phagocytosed by a dendritic cell line, DC2.4, in vitro and significantly enhanced anti-ScaA antibody responses in vivo compared to control groups. In addition, immunization with the ZNP/ZBP-ScaA complex promoted the generation of IFN-γ-secreting T cells in an antigen-dependent manner. Finally, we observed that ZNP/ZBP-ScaA immunization provided protective immunity against lethal challenge of O. tsutsugamushi, indicating that ZNP can be used as a potent adjuvant when complexed with ZBP-conjugated antigen. Conclusions ZNPs possess good adjuvant potential as a vaccine carrier when combined with an antigen having a high affinity to ZNP. When complexed with ZBP-ScaA antigen, ZNPs could induce strong antibody responses as well as protective immunity against lethal challenges of O. tsutsugamushi. Therefore, application of ZNPs combined with a specific soluble antigen could be a promising strategy as a novel vaccine carrier system.

Despite the various roles of type I interferon (type I IFN) responses during bacterial infection, its specific effects in vivo have been poorly characterized in scrub typhus caused by Orientia tsutsugamushi infection. Here, we show that type I IFNs are primarily induced via intracellular nucleic acids sensors, including RIG-I/MAVS and cGAS/STING pathways, during O. tsutsugamushi invasion. However, type I IFN signaling did not significantly affect pathogenesis, mortality, or bacterial burden during primary infection in vivo, when assessed in a mice model lacking a receptor for type I IFNs (IFNAR KO). Rather, it significantly impaired the induction of antigen-specific T cells and reduced memory T cell responses. IFNAR KO mice that recovered from primary infection showed stronger antigen-specific T cell responses, especially Th1, and more efficiently controlled bacteremia during secondary infection than wild type mice. Enhanced IL-10 expression by macrophages in the presence of type I IFN signaling might play a significant role in the suppression of antigen-specific T cell responses as neutralization or knock-out (KO) of IL-10 increased T cell responses in vitro. Therefore, induction of the type I IFN/IL-10 axis by O. tsutsugamushi infection might play a significant role in the suppression of T cell responses and contribute to the short longevity of cell-mediated immunity, often observed in scrub typhus patients.

BACKGROUND:Scrub typhus is a mite-borne febrile disease caused by O. tsutsugamushi infection. Recently, emergence of scrub typhus has attracted considerable attention in several endemic countries in Asia and the western Pacific. In addition, the antigenic diversity of the intracellular pathogen has been a serious obstacle for developing effective diagnostics and vaccine. METHODOLOGY/PRINCIPAL FINDINGS:To understand the evolutionary pathway of genotypic diversification of O. tsutsugamushi and the environmental factors associated with the epidemiological features of scrub typhus, we analyzed sequence data, including spatiotemporal information, of the tsa56 gene encoding a major outer membrane protein responsible for antigenic variation. A total of 324 tsa56 sequences covering more than 85% of its open reading frame were analyzed and classified into 17 genotypes based on phylogenetic relationship. Extensive sequence analysis of tsa56 genes using diverse informatics tools revealed multiple intragenic recombination events, as well as a substantially higher mutation rate than other house-keeping genes. This suggests that genetic diversification occurred via frequent point mutations and subsequent genetic recombination. Interestingly, more diverse bacterial genotypes and dominant vector species prevail in Taiwan compared to other endemic regions. Furthermore, the co-presence of identical and sub-identical clones of tsa56 gene in geographically distant areas implies potential spread of O. tsutsugamushi genotypes. CONCLUSIONS/SIGNIFICANCE:Fluctuation and diversification of vector species harboring O. tsutsugamushi in local endemic areas may facilitate genetic recombination among diverse genotypes. Therefore, careful monitoring of dominant vector species, as well as the prevalence of O. tsutsugamushi genotypes may be advisable to enable proper anticipation of epidemiological changes of scrub typhus.

Type I interferons (IFNs) play an important role in antiviral immunity as well as immunopathogenesis of diverse chronic viral infections. However, the precise mechanisms regulating the multifaceted effects of type I IFNs on the immune system and pathological inflammation still remain unclear. In order to assess the immunological dynamics associated with rapid viral clearance in chronic hepatitis C patients during the acute phase of type I IFN therapy, we analyzed multiple parameters of virological and immunological responses in a cohort of 59 Korean hepatitis C patients who received pegylated IFN-α and ribavirin (IFN/RBV). Most of the Korean patients had favorable alleles in the IFN-λ loci for responsiveness to IFN/RBV (i.e., C/C in rs12979860, T/T in rs8099917, and TT/TT in rs368234815). Rapid virological response (RVR) was determined mainly by the hepatitis C virus genotype. Among the cytokines analyzed, higher plasma levels of IL-17A and FGF were observed in non-RVR patients infected with viral genotype 1 and IP-10 was consistently elevated in RVR group infected with genotype 2 during the early phase of antiviral therapy. In addition, these three cytokines were correlated each other, suggesting a functional linkage of the cytokines in antiviral responses during IFN/RBV therapy. A low baseline frequencies of regulatory T cells and γδ T cells, but high level of group 2 innate lymphoid cells, in peripheral bloods were also significantly associated with the RVR group, implicating a potential role of the cellular immunity during the early phase of IFN/RBV therapy. Therefore, the immunological programs established by chronic hepatitis C and rapid disruption of the delicate balance by exogenous type I IFN might be associated with the subsequent virological outcomes in chronic hepatitis C patients.

ABSTRACT The newly emerging Middle East respiratory syndrome coronavirus (MERS-CoV) causes a severe respiratory infection with a high mortality rate (~35%). MERS-CoV has been a global threat due to continuous outbreaks in the Arabian peninsula and international spread by infected travelers since 2012. From May to July 2015, a large outbreak initiated by an infected traveler from the Arabian peninsula swept South Korea and resulted in 186 confirmed cases with 38 deaths (case fatality rate, 20.4%). Here, we show the rapid emergence and spread of a mutant MERS-CoV with reduced affinity to the human CD26 receptor during the South Korean outbreak. We isolated 13 new viral genomes from 14 infected patients treated at a hospital and found that 12 of these genomes possess a point mutation in the receptor-binding domain (RBD) of viral spike (S) protein. Specifically, 11 of these genomes have an I529T mutation in RBD, and 1 has a D510G mutation. Strikingly, both mutations result in reduced affinity of RBD to human CD26 compared to wild-type RBD, as measured by surface plasmon resonance analysis and cellular binding assay. Additionally, pseudotyped virus bearing an I529T mutation in S protein showed reduced entry into host cells compared to virus with wild-type S protein. These unexpected findings suggest that MERS-CoV adaptation during human-to-human spread may be driven by host immunological pressure such as neutralizing antibodies, resulting in reduced affinity to host receptor, and thereby impairs viral fitness and virulence, rather than positive selection for a better affinity to CD26. IMPORTANCE Recently, a large outbreak initiated by an MERS-CoV-infected traveler from the Middle East swept South Korea and resulted in 186 confirmed cases with 38 deaths. This is the largest outbreak outside the Middle East, and it raised strong concerns about the possible emergence of MERS-CoV mutations. Here, we isolated 13 new viral genomes and found that 12 of them possess a point mutation in the receptor-binding domain of viral spike protein, resulting in reduced affinity to the human cognate receptor, CD26, compared to the wild-type virus. These unexpected findings suggest that MERS-CoV adaptation in humans may be driven by host immunological pressure.

BACKGROUND:Scrub typhus is an acute febrile disease caused by Orientia tsutsugamushi infection. Recently, the rapid increase of scrub typhus incidence in several countries within the endemic region has become a serious public health issue. Despite the wide range of preventative approaches that have been attempted in the past 70 years, all have failed to develop an effective prophylactic vaccine. Currently, the selection of the proper antigens is one of the critical barriers to generating cross-protective immunity against antigenically-variable strains of O. tsutsugamushi. METHODOLOGY/PRINCIPAL FINDINGS:We examined the potential role of ScaA protein, an autotransporter protein of O. tsutsugamushi, in bacterial pathogenesis and evaluated the protective attributes of ScaA immunization in lethal O. tsutsugamushi infection in mice. Our findings demonstrate that ScaA functions as a bacterial adhesion factor, and anti-ScaA antibody significantly neutralizes bacterial infection of host cells. In addition, immunization with ScaA not only provides protective immunity against lethal challenges with the homologous strain, but also confers significant protection against heterologous strains when combined with TSA56, a major outer membrane protein of O. tsutsugamushi. CONCLUSIONS/SIGNIFICANCE:Immunization of ScaA proteins provides protective immunity in mice when challenged with the homologous strain and significantly enhanced protective immunity against infection with heterologous strains. To our knowledge, this is the most promising result of scrub typhus vaccination trials against infection of heterologous strains in mouse models thus far.

The presence of the nucleic acid of the spotted fever group (SPG) and typhus group (TG) rickettsiae was investigated in 200 serum specimens seropositive for SFG rickettsiae by multiplex-nested polymerase chain reaction with primers derived from the rickettsial outer membrane protein B gene. The DNA of SFG, TG, or both rickettsiae was amplified in the 24 serum specimens, and sequence analysis showed Rickettsia conorii, R. japonica, and R. felis in the specimens. R. conorii and R. typhi were found in 7 serum specimens, which indicated the possibility of dual infection in these patients. These findings suggest that several kinds of rickettsial diseases, including boutonneuse fever, rickettsialpox, R. felis infection, and Japanese spotted fever, as well as scrub typhus and murine typhus, are occurring in Korea.

Orientia tsutsugamushi, the causative agent of scrub typhus, is an obligate intracellular bacterium. Previously, a large number of genes that encode proteins containing eukaryotic protein-protein interaction motifs such as ankyrin-repeat (Ank) domains were identified in the O. tsutsugamushi genome. However, little is known about the Ank protein function in O. tsutsugamushi.To characterize the function of Ank proteins, we investigated a group of Ank proteins containing an F-box-like domain in the C-terminus in addition to the Ank domains. All nine selected ank genes were expressed at the transcriptional level in host cells infected with O. tsutsugamushi, and specific antibody responses against three Ank proteins were detected in the serum from human patients, indicating an active expression of the bacterial Ank proteins post infection. When ectopically expressed in HeLa cells, the Ank proteins of O. tsutsugamushi were consistently found in the nucleus and/or cytoplasm. In GST pull-down assays, multiple Ank proteins specifically interacted with Cullin1 and Skp1, core components of the SCF1 ubiquitin ligase complex, as well as the eukaryotic elongation factor 1 α (EF1α). Moreover, one Ank protein co-localized with the identified host targets and induced downregulation of EF1α potentially via enhanced ubiquitination. The downregulation of EF1α was observed consistently in diverse host cell types infected with O. tsutsugamushi.These results suggest that conserved targeting and subsequent degradation of EF1α by multiple O. tsutsugamushi Ank proteins could be a novel bacterial strategy for replication and/or pathogenesis during mammalian host infection.

BACKGROUND: Dendritic cells (DCs) are the most potent antigen-presenting cells that link innate and adaptive immune responses, playing a pivotal role in triggering antigen-specific immunity. Antigen uptake by DCs induces maturational changes that include increased surface expression of major histocompatibility complex (MHC) and costimulatory molecules. In addition, DCs actively migrate to regional lymph nodes and activate antigen-specific naive T cells after capturing antigens. We characterize the functional changes of DCs infected with Orientia tsutsugamushi, the causative agent of scrub typhus, since there is limited knowledge of the role played by DCs in O. tsutsugamushi infection. METHODOLOGY/PRINCIPAL FINDING: O. tsutsugamushi efficiently infected bone marrow-derived DCs and induced surface expression of MHC II and costimulatory molecules. In addition, O. tsutsugamushi induced autophagy activation, but actively escaped from this innate defense system. Infected DCs also secreted cytokines and chemokines such as IL-6, IL-12, MCP5, MIP-1α, and RANTES. Furthermore, in vitro migration of DCs in the presence of a CCL19 gradient within a 3D collagen matrix was drastically impaired when infected with O. tsutsugamushi. The infected cells migrated much less efficiently into lymphatic vessels of ear dermis ex vivo when compared to LPS-stimulated DCs. In vivo migration of O. tsutsugamushi-infected DCs to regional lymph nodes was significantly impaired and similar to that of immature DCs. Finally, we found that MAP kinases involved in chemotactic signaling were differentially activated in O. tsutsugamushi-infected DCs. CONCLUSION/SIGNIFICANCE: These results suggest that O. tsutsugamushi can target DCs to exploit these sentinel cells as replication reservoirs and delay or impair the functional maturation of DCs during the bacterial infection in mammals.

BACKGROUND: Scrub typhus, caused by Orientia tsutsugamushi infection, is one of the main causes of febrile illness in the Asia-Pacific region. Although cell-mediated immunity plays an important role in protection, little is known about the phenotypic changes and dynamics of leukocytes in scrub typhus patients. METHODOLOGY/PRINCIPAL FINDINGS: To reveal the underlying mechanisms of immunological pathogenesis, we extensively analyzed peripheral blood leukocytes, especially T cells, during acute and convalescent phases of infection in human patients and compared with healthy volunteers. We observed neutrophilia and CD4(+) T lymphopenia in the acute phase of infection, followed by proliferation of CD8(+) T cells during the convalescent phase. Massive T cell apoptosis was detected in the acute phase and preferential increase of CD8(+) T cells with activated phenotypes was observed in both acute and convalescent phases, which might be associated or correlated with elevated serum IL-7 and IL-15. Interestingly, peripheral Treg cells were significantly down-regulated throughout the disease course. CONCLUSIONS/SIGNIFICANCE: The remarkable decrease of CD4(+) T cells, including Treg cells, during the acute phase of infection may contribute to the loss of immunological memory that are often observed in vaccine studies and recurrent human infection.

Lipid rafts are membrane microdomains that function as platforms for signal transduction and membrane trafficking. Tyrosine kinase interacting protein (Tip) of T lymphotropic Herpesvirus saimiri (HVS) is targeted to lipid rafts in T cells and downregulates TCR and CD4 surface expression. Here, we report that the membrane-proximal amphipathic helix preceding Tip's transmembrane (TM) domain mediates lipid raft localization and membrane deformation. In turn, this motif directs Tip's lysosomal trafficking and selective TCR downregulation. The amphipathic helix binds to the negatively charged lipids and induces liposome tubulation, the TM domain mediates oligomerization, and cooperation of the membrane-proximal helix with the TM domain is sufficient for localization to lipid rafts and lysosomal compartments, especially the mutivesicular bodies. These findings suggest that the membrane-proximal amphipathic helix and TM domain provide HVS Tip with the unique ability to deform the cellular membranes in lipid rafts and to downregulate TCRs potentially through MVB formation.