L'impact des cellules dendritiques dans la dérégulation des cellules B dans un contexte d'infection au virus d'immunodéficience humaine

Les cellules dendritiques (DC) sont parmi les premières cellules à rencontrer le virus d’immunodéficience humaine (VIH) au niveau des muqueuses. De plus, le fait que les DC sont, de manière directe ou indirecte par le virus et ses composantes, altérées tant par leur nombre, leur phénotype et leur fonction suggère leur implication dans les dérégulations des cellules B. Selon cette hypothèse, des études longitudinales impliquant des individus infectés au VIH-1 présentant différents profils de progression clinique menées dans notre laboratoire ont démontré que les altérations des cellules B sont concomitantes à une augmentation de l’expression de BLyS/BAFF dans le sang ainsi que par les DC myéloïdes (mDC) sanguines. De plus, lors de travaux antérieurs utilisant le modèle murin VIH-transgénique, les altérations des cellules B ont démontré une implication des DC et d’un excès de BLyS/BAFF, et ce, dépendamment du facteur négatif du VIH (Nef). Dans cette optique, nous investiguons dans cette présente étude l’implication de Nef dans la modulation du phénotype des DC ainsi que dans les dérégulations des cellules B. Chez tous les patients virémiques infectés au VIH-1, nous avons détecté la présence de Nef dans le plasma ainsi qu’au niveau des mDC et de leurs précurseurs d’origine monocytaire, tout au long du suivi de la progression clinique et au-delà de la thérapie antirétrovirale (ART). La surexpression de BLyS/BAFF est associée à la présence de Nef au niveau des mDC et de leur précurseur.. Des essais in vitro ont permis de démontrer l’induction d’un phénotype proinflammatoire par des mDC dérivés de monocytes lorsqu’en présence de Nef soluble, via l’augmentation de l’expression de BLyS/BAFF et de TNF-α, et où cet effet est bloqué par l’ajout de l’acide rétinoïque. Nos résultats suggèrent donc que Nef est impliquée dans le déclenchement et la persistance des dérégulations des cellules B retrouvées chez les individus infectés au VIH-1. Basé sur nos observations, une thérapie adjointe impliquant le blocage de BLyS/BAFF et/ou Nef pourrait contribuer au contrôle de l’inflammation et des altérations des cellules B. De plus, la quantification de Nef post-ART pourrait s’avérer utile dans l’évaluation du statut des réservoirs. Précédemment, nous avons démontré que les dérégulations des cellules B sanguines de ces mêmes individus présentant un profil de progression rapide et classique sont accompagnées par l’augmentation de la fréquence d’une population partageant des caractéristiques des cellules B transitionnelles immatures (TI) et des cellules B de la zone marginale (ZM), que nous avons nommé les cellules B précurseur de la ZM. Toutefois, cette population est préservée chez les contrôleurs élites, chez qui nous avons trouvé une diminution significative de la fréquence des cellules B de la ZM présentant des marqueurs phénotypiques plus matures. Récemment, ces cellules ont été associées à un potentiel de fonction régulatrice (Breg), motivant ainsi notre poursuite, dans cette étude, de la caractérisation de ces cellules B. Comme pour les individus non infectés au VIH-1, nous avons démontré que les cellules B matures de la ZM contrôlent leur capacité de production d’IL-10 chez les contrôleurs élites, contrairement à une augmentation chez les progresseurs rapides et classiques. Aussi, les cellules B précurseur de la ZM des contrôleurs élites fournissent une expression importante de LT-α lorsque comparés aux individus non infectés au VIH-1, alors que cet apport de LT-α est attribué aux cellules B TI chez les progresseurs. Le contrôle de la progression clinique semble associé à un ratio en faveur de LT-α vs IL-10 au niveau des cellules B précurseur de la ZM. Nos résultats suggèrent qu’un maintien de l’intégrité du potentiel régulateur ainsi qu’une expression augmentée de LT-α par les cellules B de première ligne, telles les populations de la ZM, sont impliqués dans le contrôle de la progression clinique du VIH-1, possiblement par leur contribution à la modulation et l’homéostasie immunitaire. De telles populations doivent être considérées lors de l’élaboration de vaccins, ces derniers cherchant à générer une réponse protectrice de première ligne et adaptative.

The effects of infection by Tetracapsuloides bryosalmonae (Myxozoa) and temperature on Fredericella sultana (Bryozoa)

The myxozoan, Tetracapsuloides bryosalmonae, exploits freshwater bryozoans as definitive hosts, occurring as cryptic stages in bryozoan colonies during covert infections and as spore-forming sacs during overt infections. Spores released from sacs are infective to salmonid fish, causing the devastating Proliferative Kidney Disease (PKD). We undertook laboratory studies using mesocosm systems running at 10, 14 and 20 degrees C to determine how infection by T bryosalmonae and water temperature influence fitness of one of its most important bryozoan hosts, Fredericella sultana, over a period of 4 weeks. The effects of infection were context-dependent and often undetectable. Covert infections appear to pose very low energetic costs. Thus, we found that growth of covertly infected F. sultana colonies was similar to that of uninfected colonies regardless of temperature, as was the propensity to produce dormant resting stages (statoblasts). Production of statoblasts, however, was associated with decreased growth. Overt infections imposed greater effects on correlates of host fitness by: (i) reducing growth rates at the two higher temperatures: (ii) increasing mortality rates at the highest temperature: (iii) inhibiting statoblast production. Our results indicate that parasitism should have a relatively small effect on host fitness in the field as the negative effects of infection were mainly expressed in environmentally extreme conditions (20 degrees C for 4 weeks). The generally low virulence of T. bryosalmonae is similar to that recently demonstrated for another myxozoan endoparasite of freshwater bryozoans. The unique opportunity for extensive vertical transmission in these colonial invertebrate hosts couples the reproductive interests of host and parasite and may well give rise to the low virulence that characterises these systems. Our study implies that climate change can be expected to exacerbate PKD outbreaks and increase the geographic range of PKD as a result of the combined responses of T. bryosalmonae and its bryozoan hosts to higher temperatures. Crown Copyright (C) 2009 Published by Elsevier Ltd. All rights reserved.

The influence of powdery mildew infection on photosynthesis, chlorophyll fluorescence, leaf chlorophyll and carotenoid content of three woody plant species

The effect of powdery mildew development on photosynthesis, chlorophyll fluorescence, leaf chlorophyll and carotenoid concentrations on three woody plants frequently planted in urban environments was studied. Rates of photosynthetic CO2 fixation were rapidly reduced in two of the three genotypes tested prior to visible signs of infection. Effects on chlorophyll fluorescence (Fo, Fv/Fo, Fv/Fm), leaf chlorophyll and carotenoid content were not manifest until >25 per cent of the leaf area was observed to be covered by mycelial growth indicating reduced photo-synthetic rates during the early stages of infection were not due to degradation of the leaf chloroplast structure. Observation of the fluorescence transient (OJIP curves) showed powdery mildew infection impairs photosynthetic electron transport system by reducing the size but not heterogeneity of the plastoquninone pool, effecting both the acceptor and donor side of photosystem II. Impairment of the photosynthetic electron transport system was reflected by reduced values of a performance index used in this investigation as a measure of photochemical events within photosystem II electron transport. In addition interpretation of the fluorescence data indicated powdery mildew infection may impair the photo-protective process that facilitates the dissipation of excess energy within leaf tissue.

Drinking water application of Denargard Tiamulin for control of Brachyspira pilosicoli infection of laying poultry

Avian intestinal spirochaetosis (AIS) caused by Brachyspira spp., and notably Brachyspira pilosicoli, is common in layer flocks and reportedly of increasing incidence in broilers and broiler breeders. Disease manifests as diar- rhoea, increased feed consumption, reduced growth rates and occasional mortality in broilers and these signs are shown in layers also associated with a delayed onset of lay, reduced egg weights, faecal staining of eggshells and non-productive ovaries. Treatment with Denagard® Tiamulin has been used to protect against B. pilosicoli colonisation, persistence and clinical presentation of AIS in commercial layers, but to date there has been no de- finitive study validating efficacy. Here, we used a poultry model of B. pilosicoli infection of layers to compare the impact of three doses of Denagard® Tiamulin. Four groups of thirty 17 week old commercial pre-lay birds were all challenged with B. pilosicoli strain B2904 with three oral doses two days apart. All birds were colonised within 2 days after the final oral challenge and mild onset of clinical signs were observed thereafter. A fifth group that was unchallenged and untreated was also included for comparison as healthy birds. Five days after the final oral Brachypira challenge three groups were given Denagard® Tiamulin in drinking water made up following the manufacturer's recommendations with doses verified as 58.7 ppm, 113 ppm and 225 ppm. Weight gain body condition and the level of diarrhoea of birds infected with B. pilosicoli were improved and shedding of the organism reduced significantly (p = 0.001) following treatment with Denagard® Tiamulin irrespective of dose given. The level and duration of colonisation of organs of birds infected with B. pilosicoli was also reduced. Confirming previous findings we showed that the ileum, caeca, colon, and both liver and spleen were colonised and here we demonstrated that treatment with Denagard® Tiamulin resulted in significant reduction in the numbers of Brachyspira found in each of these sites and dramatic reduction in faecal shedding (p b 0.001) to ap- proaching zero as assessed by culture of cloacal swabs. Although the number of eggs produced per bird and the level of eggshell staining appeared unaffected, egg weights of treated birds were greater than those of untreated birds for a period of approximately two weeks following treatment. These data conclusively demonstrate the ef- fectiveness of Denagard® Tiamulin in reducing B. pilosicoli infection in laying hens.

Drinking water application of Denagard® Tiamulin for control of Brachyspira pilosicoli infection of laying poultry

Avian intestinal spirochaetosis (AIS) caused by Brachyspira spp., and notably Brachyspira pilosicoli, is common in layer flocks and reportedly of increasing incidence in broilers and broiler breeders. Disease manifests as diarrhoea,increased feed consumption, reduced growth rates and occasional mortality in broilers and these signs are shown in layers also associated with a delayed onset of lay, reduced egg weights, faecal staining of eggshells and non-productive ovaries. Treatment with Denagard® Tiamulin has been used to protect against B. pilosicoli colonisation, persistence and clinical presentation of AIS in commercial layers, but to date there has been no definitive study validating efficacy. Here, we used a poultry model of B. pilosicoli infection of layers to compare the impact of three doses of Denagard® Tiamulin. Four groups of thirty 17 week old commercial pre-lay birds were all challengedwith B. pilosicoli strain B2904with three oral doses two days apart. All birdswere colonised within 2 days after the final oral challenge and mild onset of clinical signs were observed thereafter. A fifth group that was unchallenged and untreated was also included for comparison as healthy birds. Five days after the final oral Brachypira challenge three groups were given Denagard® Tiamulin in drinking water made up following the manufacturer's recommendations with doses verified as 58.7 ppm, 113 ppm and 225 ppm. Weight gain body condition and the level of diarrhoea of birds infected with B. pilosicoli were improved and shedding of the organism reduced significantly (p = 0.001) following treatment with Denagard® Tiamulin irrespective of dose given. The level and duration of colonisation of organs of birds infected with B. pilosicoli was also reduced. Confirming previous findings we showed that the ileum, caeca, colon, and both liver and spleen were colonised and here we demonstrated that treatment with Denagard® Tiamulin resulted in significant reduction in the numbers of Brachyspira found in each of these sites and dramatic reduction in faecal shedding (p b 0.001) to approaching zero as assessed by culture of cloacal swabs. Although the number of eggs produced per bird and the level of eggshell staining appeared unaffected, egg weights of treated birds were greater than those of untreated birds for a period of approximately two weeks following treatment. These data conclusively demonstrate the effectiveness of Denagard® Tiamulin in reducing B. pilosicoli infection in laying hens.

Toll-Like Receptor 4 Signaling Leads to Severe Fungal Infection Associated with Enhanced Proinflammatory Immunity and Impaired Expansion of Regulatory T Cells

Toll-like receptors (TLRs) present in innate immune cells recognize pathogen molecular patterns and influence immunity to control the host-parasite interaction. The objective of this study was to characterize the involvement of TLR4 in the innate and adaptive immunity to Paracoccidioides brasiliensis, the most important primary fungal pathogen of Latin America. We compared the responses of C3H/HeJ mice, which are naturally defective in TLR4 signaling, with those of C3H/HePas mice, which express functional receptors, after in vitro and in vivo infection with P. brasiliensis. Unexpectedly, we verified that TLR4-defective macrophages infected in vitro with P. brasiliensis presented decreased fungal loads associated with impaired synthesis of nitric oxide, interleukin-12 (IL-12), and macrophage chemotactic protein 1 (MCP-1). After intratracheal infection with 1 million yeasts, TLR4-defective mice developed reduced fungal burdens and decreased levels of pulmonary nitric oxide, proinflammatory cytokines, and antibodies. TLR4-competent mice produced elevated levels of IL-12 and tumor necrosis factor alpha (TNF-alpha), besides cytokines of the Th17 pattern, indicating a proinflammatory role for TLR4 signaling. The more severe infection of TLR4-normal mice resulted in increased influx of activated macrophages and T cells to the lungs and progressive control of fungal burdens but impaired expansion of regulatory T cells (Treg cells). In contrast, TLR4-defective mice were not able to clear their diminished fungal burdens totally, a defect associated with deficient activation of T-cell immunity and enhanced development of Treg cells. These divergent patterns of immunity, however, resulted in equivalent mortality rates, indicating that control of elevated fungal growth mediated by vigorous inflammatory reactions is as deleterious to the hosts as low fungal loads inefficiently controlled by limited inflammatory reactions.

Rates of common communicable illnesses in non-anaemic 12-24 month old South Island, New Zealand children

Aims : To describe the incidence of parentally reported illness in otherwise healthy South Island toddlers; characterise the predictors of illness; and determine whether there was a relationship between teething and illness in this population.

Methods : A 20-week randomised controlled trial was conducted on 1-year-old children (n=225) from Otago and Southland between February 2004 and December 2005. Information on symptoms of morbidity, occurrence of teething, and childcare attendance were recorded daily throughout the intervention period. Morbidity symptoms were categorised into respiratory illness (RI), gastrointestinal illness (GII), ear infection, and total illness, and the number and duration of events were determined.

Results : The mean (SD) number of total illnesses was 3.4 (2.3) per 20 weeks, with an average duration of 4.5 days. Episodes of RI were most common (50% of total illness events), and tended to be the longest in duration (mean of 3.7 days). Having siblings aged less than 5 years (23% increase, 95%CI 6%–42%, p=0.007) and attending childcare (72% increase, 95%CI 38%–113%, p<0.001)), were positively associated with the number of total illness events but not duration. In addition, teething was positively associated with total events (OR 1.94, 95%CI 1.45–2.60, p<0.001), RI events (OR 2.03, 95%CI 1.41–2.93, p<0.001) and GII events (OR 1.90, 95%CI 1.36–2.67, p<0.001).

Conclusion : This study has shown that illness (particularly RI) is common in the second year of life. It has also confirmed that attending childcare and having siblings aged under 5 years increases the number of illness events. An association between teething and the occurrence of illness was also seen but the exact nature of this relationship requires verification.

Intracellular dynamics of HIV infection

Early studies of HIV infection dynamics suggested that virus-producing HIV-infected cells had an average half-life of approximately 1 day. However, whether this average behavior is reflective of the dynamics of individual infected cells is unclear. Here, we use HIV-enhanced green fluorescent protein (EGFP) constructs and flow cytometry sorting to explore the dynamics of cell infection, viral protein production, and cell death in vitro. By following the numbers of productively infected cells expressing EGFP over time, we show that infected cell death slows down over time. Although infected cell death in vivo could be very different, our results suggest that the constant decay of cell numbers observed in vivo during antiretroviral treatment could reflect a balance of cell death and delayed viral protein production. We observe no correlation between viral protein production and death rate of productively infected cells, showing that viral protein production is not likely to be the sole determinant of the death of HIV-infected cells. Finally, we show that all observed features can be reproduced by a simple model in which infected cells have broad distributions of productive life spans, times to start viral protein production, and viral protein production rates. This broad spectrum of the level and timing of viral protein production provides new insights into the behavior and characteristics of HIV-infected cells.

Community-onset Escherichia coli infection resistant to expanded- spectrum cephalosporins in low-prevalence countries

Background
By global standards the prevalence of community onset expanded-spectrum cephalosporin resistant Escherichia coli (ESC-R-EC) remains low in Australia and New Zealand. Of concern, our countries are in a unique position with high extramural resistance pressure from close population and trade links to Asia-Pacific neighbours with high ESC-R-EC rates. We aim to characterize the risks and dynamics of community onset ESC-R-EC in our low-prevalence region.

Methods
A case-control methodology was used. Patients with ESC-R-EC or susceptible E. coli isolated from blood or urine were recruited at six geographically dispersed tertiary hospitals in Australia and New Zealand. Epidemiological data was prospectively collected and bacteria were retained for analysis.

Results
In total, 182 patients (91 cases and 91 controls) were recruited. Multivariate logistic regression identified risk factors for ESC-R amongst E. coli including birth on the Indian subcontinent (OR=11.13, 2.17-56.98, p=0.003), urinary tract infection in the past year (per infection OR=1.430, 1.13-1.82, p=0.003), travel to South East Asia, China, Indian subcontinent, Africa and the Middle East (OR=3.089, 1.29-7.38, p=0.011), prior exposure to trimethoprim+/-sulfamethoxazole &/or an expanded-spectrum cephalosporin (OR=3.665, 1.30-10.35, p=0.014) and healthcare exposure in the previous six months (OR=3.16, 1.54-6.46, p=0.02).

Amongst our ESC-R-EC the blaCTX-M ESBLs was dominant (83% of ESC-R-EC), and the worldwide pandemic clone ST-131 was frequent (45% of ESC-R-EC).

Conclusion
In our low prevalence setting, ESC-R amongst community onset E. coli may be associated with both ‘export’ from healthcare facilities into the community and direct ‘import’ into the community from high-prevalence regions.

Surgical Site Infection in a University Hospital in Northeast Brazil

We examined prevention of surgical site infection (SSI) in a tertiary teaching hospital in northeast Brazil, from January 1994 to December 2003. The survey included 5,742 patients subjected to thoracic, urologic, vascular and general surgery. The criteria for diagnosing SSI were those of the Centers for Disease Control, USA, and the variables of the National Nosocomial Infection Surveillance risk index were used. Data analysis revealed that anesthetic risk scores, wound class and duration of surgery were significantly associated with SSI. A total of 296 SSIs were detected among the 5,742 patients (5.1%). The overall incidence of SSI was 8.8% in 1994; it decreased to 3.3% in 2003. In conclusion, the use of educational strategies, based on guidelines for SSI prevention reduced SSI incidence. Appropriate management of preoperative, intraoperative, and postoperative incision care, and a surveillance system based on international criteria, were useful in reducing SSI rates in our hospital

Effect of Fusarium graminearum and infection index on germination and vigor of maize seeds

Patógenos em sementes de milho (Zea mays) causam sérios problemas, como a perda de sua capacidade germinativa. O objetivo do trabalho foi determinar qual o melhor tempo para infecção das sementes de milho com Fusarium graminearum, para posterior avaliação dos danos causados pelo fungo na germinação e vigor das mesmas. As sementes foram colocadas sobre meio de BDA contendo o patógeno e incubadas por 4, 8, 16 e 32 h. Após os respectivos períodos de incubação, estas foram submetidas ao teste de sanidade (papel de filtro), com duas variações, sem e com assepsia superficial, usando hipoclorito de sódio a 1% de cloro ativo, por 3 min. Determinado o melhor tempo para infecção, outras sementes foram infetadas com o patógeno, para realização dos testes de germinação e vigor (envelhecimento acelerado e teste de frio) com uma mistura de sementes sadias (colocadas sobre o meio BDA) e sementes inoculadas, resultando em 0, 20, 40, 60, 80 e 100% de sementes infetadas com o fungo em estudo. Os resultados obtidos mostraram que o período de incubação de 32 h foi suficiente para se obter sementes infetadas. Com relação à germinação, não houve diferenças significativas entre os diferentes níveis de infecção, provavelmente devido ao alto vigor das sementes de milho testadas. Quanto aos testes de vigor, os níveis de infecção diferiram significativamente da testemunha, apesar de não terem diferido entre si.

Neospora caninum infection in birds: experimental infections in chicken and embryonated eggs

Neospora caninum causes economical impact in cattle-raising farms since it is implicated as the major cause of bovine abortions. Although infection by the parasite has been widely described in mammals, the role of birds in its life-cycle is still obscure. Therefore, this work aimed to evaluate the infection by N. caninum in different chicken models. Experimental infections were conducted in 7-day-old chicks, laying hens and embryonated eggs, where samples were analysed for parasite burden, IgG antibodies and lesions promoted. Chickens demonstrated an asymptomatic infection, although with seroconversion and systemic replication of the parasite. In laying hens, no signs of vertical transmission were observed. However, embryonated eggs inoculated by the allantoic cavity route demonstrated susceptibility to infection, with mortality rates around 50% independent of the inoculum dose. Additionally, dogs became infected after ingestion of different amounts of inoculated eggs, producing either oocysts or specific IgG antibodies. The results herein presented demonstrate that chickens may be intermediate hosts of N. caninum and that embryonated eggs could be a useful model to study the parasite's biology.

Organic acids and/or compound with defined microorganisms to control Salmonella enterica serovar Enteritidis experimental infection in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Organ tropism during the acute and chronic phases of Trypanosoma cruzi infection in BALB/c mice

The aim of the present study was to investigate the presence of Trypanosoma cruzi in the heart, liver, lung, and kidneys, using hemoculture and PCR analysis, of mice infected with different parasite strains during the acute and chronic phases of infection. Parasitemia curves revealed strain-specific biological behaviors. For the Y and JLP strains, the acute phase of infection started at days six and ten post-infection, parasitemia peaked at days seven and 15 post-infection, the chronic phase started at days nine and 28 post-infection, and animals started dying at days 19 and 120 post-infection, respectively. When the two strains were compared, the JLP strain exhibited reduced and slower replication rates associated with a delayed peak of parasitism and reduced parasite burdens. However, parasites were detected in all studied organs using PCR analysis. The capacity of both strains to infect different organs likely influences disease pathogenesis.

Factors contributing to the differences in peritonitis rates between centers and regions

Despite improvements in connectology, peritoneal dialysis (PD) - associated peritonitis contributes significantly to morbidity and modality failure in patients maintained on PD therapy. A broad spectrum of organisms -gram-positive, gram-negative, fungal, anaerobic - are involved in this complication. In addition, a significant percentage of episodes involve polymicrobial and culture-negative infection. Technological advances are being developed to minimize the incidence of access-related complications such as peritonitis. Many traditional factors such as exit-site infection and poor technique have been already identified. In the present review, we discuss the geographic, patient selection, and clinical issues that can affect peritonitis rates in different areas of the world and in different centers in the same area.