Delay of phagosome maturation by a mycobacterial lipid is reversed by nitric oxide.

Mycobacterium tuberculosis is a facultative intracellular pathogen that inhibits phagosome maturation in macrophages thereby securing survival and growth. Mycobacteria reside in an early endocytic compartment of near-neutral pH where they upregulate production of complex glycolipids such as trehalose dimycolate. Here, we report that trehalose dimycolate coated onto beads increased the bead retention in early phagosomes, i.e. at a similar stage as viable mycobacteria. Thus, a single mycobacterial lipid sufficed to divert phagosome maturation and likely contributes to mycobacterial survival in macrophages. Previous studies showed that activated macrophages promote maturation of mycobacterial phagosomes and eliminate mycobacteria through bactericidal effectors including nitric oxide generated by inducible nitric-oxide synthase. We show that deceleration of bead phagosome maturation by trehalose dimycolate was abolished in immune-activated wild type, but not in activated nitric-oxide synthase-deficient macrophages, nor when hydroxyl groups of trehalose dimycolate were chemically modified by reactive nitrogen intermediates. Thus, specific host defence effectors of activated macrophages directly target a specific virulence function of mycobacteria

Impact of humoral alloreactivity early after transplantation on the long-term survival of renal allografts

Impact of humoral alloreactivity early after transplantation on the long-term survival of renal allografts.BackgroundThe contribution of humoral alloreactivity to the rejection of renal allografts is not well defined because humoral antigraft reactions are not easily detectable in transplant biopsies, and serial measurements of circulating allo-antibodies in the post-transplantation period are not routinely performed. We have developed diagnostic techniques that improve the assessment of humoral alloreactivity in vivo and in vitro.MethodsHumoral alloreactivity in transplant biopsies derived from 218 single kidney grafts was detected by assessing the deposition of complement fragment C4d in interstitial capillaries. Circulating alloantibodies were determined in corresponding serum samples by flow cytometry using lymphoblastoid cell lines of donor DR-type as target cells and by a conventional microcytotoxicity test. The impact of capillary C4d and other selected variables on renal graft survival was calculated by univariate and multivariate analysis.ResultsCapillary C4d, present in 46% of biopsies from first grafts and 72% of regrafts, is related to circulating alloantibodies. Grafts with capillary C4d have a markedly shorter survival than grafts without capillary C4d (50% graft survival, 4 vs. 8 years, P = 0.0001). Among several risk factors, capillary C4d is the strongest predictor of subsequent graft loss in a multivariate analysis (relative risk, 2.1, 95% CI, 1.4 to 3.1). Humoral alloreactivity detectable within six months after transplantation has a much stronger impact on graft survival than alloreactivity detected beyond this period.ConclusionsHumoral alloreactivity, manifested by the capillary deposition of complement C4d in about 50% of biopsied renal grafts, exerts a strong impact on graft survival when it operates within six months after transplantation

Contamination of human DNA samples with mouse DNA can lead to false detection of XMRV-like sequences

<p>Abstract</p> <p>Background</p> <p>In 2006, a novel gammaretrovirus, XMRV (xenotropic murine leukemia virus-related virus), was discovered in some prostate tumors. A more recent study indicated that this infectious retrovirus can be detected in 67% of patients suffering from chronic fatigue syndrome (CFS), but only very few healthy controls (4%). However, several groups have published to date that they could not identify XMRV RNA or DNA sequences in other cohorts of CFS patients, while another group detected murine leukemia virus (MLV)-like sequences in 87% of such patients, but only 7% of healthy controls. Since there is a high degree of similarity between XMRV and abundant endogenous MLV proviruses, it is important to distinguish contaminating mouse sequences from true infections.</p> <p>Results</p> <p>DNA from the peripheral blood of 112 CFS patients and 36 healthy controls was tested for XMRV with two different PCR assays. A TaqMan qPCR assay specific for XMRV <it>pol </it>sequences was able to detect viral DNA from 2 XMRV-infected cells (~ 10-12 pg DNA) in up to 5 μg of human genomic DNA, but yielded negative results in the test of 600 ng genomic DNA from 100,000 peripheral blood cells of all samples tested. However, positive results were obtained with some of these samples, using a less specific nested PCR assay for a different XMRV sequence. DNA sequencing of the PCR products revealed a wide variety of virus-related sequences, some identical to those found in prostate cancer and CFS patients, others more closely related to known endogenous MLVs. However, all samples that tested positive for XMRV and/or MLV DNA were also positive for the highly abundant intracisternal A-type particle (IAP) long terminal repeat and most were positive for murine mitochondrial cytochrome oxidase sequences. No contamination was observed in any of the negative control samples, containing those with no DNA template, which were included in each assay.</p> <p>Conclusions</p> <p>Mouse cells contain upwards of 100 copies each of endogenous MLV DNA. Even much less than one cell's worth of DNA can yield a detectable product using highly sensitive PCR technology. It is, therefore, vital that contamination by mouse DNA be monitored with adequately sensitive assays in all samples tested.</p

Régénération assistée du karité (Vitellaria paradoxa C. F. Gaertn.) dans les parcs agroforestiers au Burkina Faso

Le karité (Vitellaria paradoxa C. F. Gaertn.) présente une importance sociale, culturelle, économique et alimentaire pour le Burkina Faso où il occupe la quatrième place dans les produits d’exportation. La pérennité de cette espèce dont les populations rurales sont tributaires, est compromise par de multiples phénomènes dont le manque de régénération, les pratiques culturales et la coupe abusive du bois pour satisfaire les besoins énergétiques. L’objectif de cette étude est de proposer des méthodes adéquates pour rajeunir les parcs à karité au Burkina Faso. Cinq sites de recherche ont été choisis suivant un gradient phytogéographique : Sobaka, Noumoudara et Kakoumana (secteur sud soudanien), Gonsé (secteur nord soudanien) et Bouria (secteur sub sahélien). Les essais effectués dans chaque parcelle choisie, ont comporté trois répétitions et 7 traitements, représentés par les différentes techniques de régénération (plantation, transplantation, semis direct libre, semis dans les buissons, régénération naturelle assistéeRNA-, induction de drageon, induction de pousse adventive). Les résultats indiquent que le taux de survie de la régénération naturelle assistée est audessus de 70% après deux années de suivi. Les plantations et les semis directs dans les buissons sont aussi des techniques efficaces pour la régénération/restauration des parcs à karité, avec respectivement des taux de survie de 13,33% et 6,67% dans le secteur sub sahélien, 12,22% et 6% dans le nord soudanien, et 55,56% et 25,33% dans le sud soudanien après la troisième année de suivi. En conclusion, la régénération assistée est la technique la plus efficiente pour restaurer les parcs à karité. The Shea (Vitellaria paradoxa C. F. Gaertn.) is of enormous social, cultural, economic and nutritional importance for Burkina Faso where it ranks fourth in exports. The sustainability of this species on which rural populations depend is compromised by multiple phenomena, including the lack of regeneration of shea tree and the abusive cutting of wood to meet energy needs. This work aimed to propose adequate methods for rejuvenating shea parkland in Burkina Faso. Five sites distributed along a phytogeographic gradient were selected for tests : Sobaka, Noumoudara and Kakoumana (South-Sudanian phytogeographic zone), Gonsé (NorthSudanian phytogeographic zone) and Bouria (sub-Sahelian phytogeographic zone). The field trials included three repetitions by regeneration technique (planting, transplanting, direct sowing free, sowing in bushes and assisted natural regeneration -ANR-, sucker induction, induction of adventitious growth). The results indicate that the survival rate of assisted natural regeneration is above 70% after two years of follow-up. Planting and direct seeding in bushes are also effective techniques for the regeneration / restoration of shea tree parks with respectively survival rates of 13.33% and 6.67% in the sub sahelian sector, 12.22% and 6% in north sudanian, and 55.56% and 25.33% in south sudanian, after the third year of monitoring. In conclusion, in order to restore the shea parks, all seedlings regardless of their origins, will need to be assisted i

A Mouse Model for Chikungunya: Young Age and Inefficient Type-I Interferon Signaling Are Risk Factors for Severe Disease

Chikungunya virus (CHIKV) is a re-emerging arbovirus responsible for a massive outbreak currently afflicting the Indian Ocean region and India. Infection from CHIKV typically induces a mild disease in humans, characterized by fever, myalgia, arthralgia, and rash. Cases of severe CHIKV infection involving the central nervous system (CNS) have recently been described in neonates as well as in adults with underlying conditions. The pathophysiology of CHIKV infection and the basis for disease severity are unknown. To address these critical issues, we have developed an animal model of CHIKV infection. We show here that whereas wild type (WT) adult mice are resistant to CHIKV infection, WT mouse neonates are susceptible and neonatal disease severity is age-dependent. Adult mice with a partially (IFN-α/βR+/−) or totally (IFN-α/βR−/−) abrogated type-I IFN pathway develop a mild or severe infection, respectively. In mice with a mild infection, after a burst of viral replication in the liver, CHIKV primarily targets muscle, joint, and skin fibroblasts, a cell and tissue tropism similar to that observed in biopsy samples of CHIKV-infected humans. In case of severe infections, CHIKV also disseminates to other tissues including the CNS, where it specifically targets the choroid plexuses and the leptomeninges. Together, these data indicate that CHIKV-associated symptoms match viral tissue and cell tropisms, and demonstrate that the fibroblast is a predominant target cell of CHIKV. These data also identify the neonatal phase and inefficient type-I IFN signaling as risk factors for severe CHIKV-associated disease. The development of a permissive small animal model will expedite the testing of future vaccines and therapeutic candidates