Highly Frequent Mutations in Negative Regulators of Multiple Virulence Genes in Group A Streptococcal Toxic Shock Syndrome Isolates

Streptococcal toxic shock syndrome (STSS) is a severe invasive infection characterized by the sudden onset of shock and multiorgan failure; it has a high mortality rate. Although a number of studies have attempted to determine the crucial factors behind the onset of STSS, the responsible genes in group A Streptococcus have not been clarified. We previously reported that mutations of csrS/csrR genes, a two-component negative regulator system for multiple virulence genes of Streptococcus pyogenes, are found among the isolates from STSS patients. In the present study, mutations of another negative regulator, rgg, were also found in clinical isolates of STSS patients. The rgg mutants from STSS clinical isolates enhanced lethality and impaired various organs in the mouse models, similar to the csrS mutants, and precluded their being killed by human neutrophils, mainly due to an overproduction of SLO. When we assessed the mutation frequency of csrS, csrR, and rgg genes among S. pyogenes isolates from STSS (164 isolates) and non-invasive infections (59 isolates), 57.3% of the STSS isolates had mutations of one or more genes among three genes, while isolates from patients with non-invasive disease had significantly fewer mutations in these genes (1.7%). The results of the present study suggest that mutations in the negative regulators csrS/csrR and rgg of S. pyogenes are crucial factors in the pathogenesis of STSS, as they lead to the overproduction of multiple virulence factors

Discrimination of water quality monitoring sites in River Vouga using a mixed-effect state space model

The surface water quality monitoring is an important concern of public organizations due to its relevance to the public health. Statistical methods are taken as consistent and essential tools in the monitoring procedures in order to prevent and identify environmental problems. This work presents the study case of the hydrological basin of the river Vouga, in Portugal. The main goal is discriminate the water monitoring sites using the monthly dissolved oxygen concentration dataset between January 2002 and May 2013. This is achieved through the extraction of trend and seasonal components in a linear mixed-effect state space model. The parameters estimation is performed with both maximum likelihood method and distribution-free estimators in a two-step procedure. The application of the Kalman smoother algorithm allows to obtain predictions of the structural components as trend and seasonality. The water monitoring sites are discriminated through the structural components by a hierarchical agglomerative clustering procedure. This procedure identified different homogenous groups relatively to the trend and seasonality components and some characteristics of the hydrological basin are presented in order to support the results

VCAM-1 and VLA-4 Modulate Dendritic Cell IL-12p40 Production in Experimental Visceral Leishmaniasis

Vascular cell adhesion molecule-1 (VCAM-1) interacts with its major ligand very late antigen-4 (VLA-4) to mediate cell adhesion and transendothelial migration of leukocytes. We report an important role for VCAM-1/VLA-4 interactions in the generation of immune responses during experimental visceral leishmaniasis caused by Leishmania donovani. Our studies demonstrate that these molecules play no direct role in the recruitment of leukocytes to the infected liver, but instead contribute to IL-12p40-production by splenic CD8+ dendritic cells (DC). Blockade of VCAM-1/VLA-4 interactions using whole antibody or anti-VCAM-1 Fab′ fragments reduced IL-12p40 mRNA accumulation by splenic DC 5 hours after L. donovani infection. This was associated with reduced anti-parasitic CD4+ T cell activation in the spleen and lowered hepatic IFNγ, TNF and nitric oxide production by 14 days post infection. Importantly, these effects were associated with enhanced parasite growth in the liver in studies with either anti-VCAM-1 or anti-VLA-4 antibodies. These data indicate a role for VCAM-1 and VLA-4 in DC activation during infectious disease

Incompetence of Neutrophils to Invasive Group A streptococcus Is Attributed to Induction of Plural Virulence Factors by Dysfunction of a Regulator

Group A streptococcus (GAS) causes variety of diseases ranging from common pharyngitis to life-threatening severe invasive diseases, including necrotizing fasciitis and streptococcal toxic shock-like syndrome. The characteristic of invasive GAS infections has been thought to attribute to genetic changes in bacteria, however, no clear evidence has shown due to lack of an intriguingly study using serotype-matched isolates from clinical severe invasive GAS infections. In addition, rare outbreaks of invasive infections and their distinctive pathology in which infectious foci without neutrophil infiltration hypothesized us invasive GAS could evade host defense, especially neutrophil functions. Herein we report that a panel of serotype-matched GAS, which were clinically isolated from severe invasive but not from non-invaive infections, could abrogate functions of human polymorphnuclear neutrophils (PMN) in at least two independent ways; due to inducing necrosis to PMN by enhanced production of a pore-forming toxin streptolysin O (SLO) and due to impairment of PMN migration via digesting interleukin-8, a PMN attracting chemokine, by increased production of a serine protease ScpC. Expression of genes was upregulated by a loss of repressive function with the mutation of csrS gene in the all emm49 severe invasive GAS isolates. The csrS mutants from clinical severe invasive GAS isolates exhibited high mortality and disseminated infection with paucity of neutrophils, a characteristic pathology seen in human invasive GAS infection, in a mouse model. However, GAS which lack either SLO or ScpC exhibit much less mortality than the csrS-mutated parent invasive GAS isolate to the infected mice. These results suggest that the abilities of GAS to abrogate PMN functions can determine the onset and severity of invasive GAS infection

PGL-III, a rare intermediate of Mycobacterium leprae phenolic glycolipid biosynthesis, is a potent Mincle ligand

Although leprosy(Hansen's disease) is one ofthe oldestknown diseases, the pathogenicity of Mycobacterium leprae (M. leprae) remains enigmatic. Indeed, the cellwall components responsible for the immune response against M. leprae are as yet largely unidentified. We reveal herephenolic glycolipid-III (PGL-III) as an M. leprae-specific ligand for the immune receptor Mincle. PGL-III is a scarcelypresent trisaccharide intermediate in the biosynthetic pathway toPGL-I, an abundant and characteristic M. leprae glycolipid.Using activity-based purification, we identified PGL-III as a Mincleligand that is more potent than the well-known M. tuberculosis trehalose dimycolate. The cocrystal structure of Mincle and a syntheticPGL-III analogue revealed a unique recognition mode, implying thatit can engage multiple Mincle molecules. In Mincle-deficient miceinfected with M. leprae, increased bacterial burdenwith gross pathologies were observed. These results show that PGL-IIIis a noncanonical ligand recognized by Mincle, triggering protectiveimmunity.PGL-III, a potent immunostimulatory glycolipid,is limitedin M. leprae by the quick addition of a single methylgroup to convert it into immunosuppressive PGL-I, which confers immuneescape.Bio-organic Synthesi

Discrepancy between magnetic resonance imaging and cranial nerve neuropathies associated with the involvement of diffuse large B-cell lymphoma（DLBL）.

An 83-year-old female developed diffuse large B-cell lymphoma（DLBL） of the left nasal cavity. Complete remission was achieved after two courses of Rituximab and CHOP（R-CHOP） . During the fourth course of R-CHOP, sensory disturbance and palsy of the left face developed. Left trigeminal nerve swelling was observed in magnetic resonance imaging（MRI） followed by double vision in the left eye, and MRI revealed swelling of both trigeminal nerves but not of the abducens nerve. Although the swelling of the trigeminal nerves and the double vision subside after administration of prednisolone, the palsy of the left face persisted. Two months after the fourth course of R-CHOP, symptoms of the palsy of the left face progressed and palsy of the right face, double vision, and palsy of the left facialis nerve developed. Then,blepharoptosis of the right eye developed and palsy of the right oculomotorius nerve was observed. MRI showed the presence of trigeminal nerve and oculomotorius nerve swelling but no swelling of the other cranial nerves. Furthermore, skin eruption developed around the left eye.Cytology of this lesion revealed the invasion of lymphoma cells

Higher Expression of CCL2, CCL4, CCL5, CCL21, and CXCL8 Chemokines in the Skin Associated with Parasite Density in Canine Visceral Leishmaniasis

Several previous studies correlated immunopathological aspects of canine visceral leishmaniasis (CVL) with tissue parasite load and/or the clinical status of the disease. Recently, different aspects of the immune response in Leishmania-infected dogs have been studied, particularly the profile of cytokines in distinct compartments. However, the role of chemokines in disease progression or parasite burdens of the visceralising species represents an important approach for understanding immunopathology in CVL. We found an increase in inflammatory infiltrate, which was mainly composed of mononuclear cells, in the skin of animals presenting severe forms of CVL and high parasite density. Our data also demonstrated that enhanced parasite density is positively correlated with the expression of CCL2, CCL4, CCL5, CCL21, and CXCL8. In contrast, there was a negative correlation between parasite density and CCL24 expression. These findings represent an advance in the knowledge of the involvement of skin inflammatory infiltrates in CVL and the systemic consequences and may contribute to developing a rational strategy for the design of new and more efficient prophylactic tools and immunological therapies against CVL

Interferon-γ-producing immature myeloid cells confer protection against severe invasive group A Streptococcus infections

Cytokine-activated neutrophils are known to be essential for protection against group A Streptococcus infections. However, during severe invasive group A Streptococcus infections that are accompanied by neutropenia, it remains unclear which factors are protective against such infections, and which cell population is the source of them. Here we show that mice infected with severe invasive group A Streptococcus isolates, but not with non-invasive group A Streptococcus isolates, exhibit high concentrations of plasma interferon-γ during the early stage of infection. Interferon-γ is necessary to protect mice, and is produced by a novel population of granulocyte–macrophage colony-stimulating factor-dependent immature myeloid cells with ring-shaped nuclei. These interferon-γ-producing immature myeloid cells express monocyte and granulocyte markers, and also produce nitric oxide. The adoptive transfer of interferon-γ-producing immature myeloid cells ameliorates infection in wild-type and interferon-γ-deficient mice. Our results indicate that interferon-γ-producing immature myeloid cells have a protective role during the early stage of severe invasive group A Streptococcus infections

Innate Killing of Leishmania donovani by Macrophages of the Splenic Marginal Zone Requires IRF-7

Highly phagocytic macrophages line the marginal zone (MZ) of the spleen and the lymph node subcapsular sinus. Although these macrophages have been attributed with a variety of functions, including the uptake and clearance of blood and lymph-borne pathogens, little is known about the effector mechanisms they employ after pathogen uptake. Here, we have combined gene expression profiling and RNAi using a stromal macrophage cell line with in situ analysis of the leishmanicidal activity of marginal zone macrophages (MZM) and marginal metallophilic macrophages (MMM) in wild type and gene targeted mice. Our data demonstrate a critical role for interferon regulatory factor-7 (IRF-7) in regulating the killing of intracellular Leishmania donovani by these specialised splenic macrophage sub-populations. This study, therefore, identifies a new role for IRF-7 as a regulator of innate microbicidal activity against this, and perhaps other, non-viral intracellular pathogens. This study also highlights the importance of selecting appropriate macrophage populations when studying pathogen interactions with this functionally diverse lineage of cells