Differential enhancement of dengue virus immune complex infectivity mediated by signaling-competent and signaling-incompetent human Fcgamma RIA (CD64) or FcgammaRIIA (CD32)

Fcgamma receptor (FcgammaR)-mediated entry of infectious dengue virus immune complexes into monocytes/macrophages is hypothesized to be a key event in the pathogenesis of complicated dengue fever. FcgammaRIA (CD64) and FcgammaRIIA (CD32), which predominate on the surface of such dengue virus-permissive cells, were compared for their influence on the infectivity of dengue 2 virus immune complexes formed with human dengue virus antibodies. A signaling immunoreceptor tyrosine-based activation motif (ITAM) incorporated into the accessory gamma-chain subunit that associates with FcgammaRIA and constitutively in FcgammaRIIA is required for phagocytosis mediated by these receptors. To determine whether FcgammaRIA and FcgammaRIIA activation functions are also required for internalization of infectious dengue virus immune complexes, we generated native and signaling-incompetent versions of each receptor by site-directed mutagenesis of ITAM tyrosine residues. Plasmids designed to express these receptors were transfected into COS-7 cells, and dengue virus replication was measured by plaque assay and flow cytometry. We found that both receptors mediated enhanced dengue virus immune complex infectivity but that FcgammaRIIA appeared to do so far more effectively. Abrogation of FcgammaRIA signaling competency, either by expression without gamma-chain or by coexpression with gamma-chain mutants, was associated with significant impairment of phagocytosis and of dengue virus immune complex infectivity. Abrogation of FcgammaRIIA signaling competency was also associated with equally impaired phagocytosis but had no discernible effect on dengue virus immune complex infectivity. These findings point to fundamental differences between FcgammaRIA and FcgammaRIIA with respect to their immune-enhancing capabilities and suggest that different mechanisms of dengue virus immune complex internalization may operate between these FcgammaRs.     

Tools for the SBML Community

MOTIVATION: SBML is quickly becoming the standard format to exchange biochemical models. The tools presented in this paper are loosely-coupled, and are intended to be incorporated into SBML aware applications. The rationale for this is to reduce the amount of repeated work carried out within the community and to create tools that offer a greater number of features to the end-user. AVAILABILITY: All tools described are available from http://www.basis.ncl.ac.uk/software and are licensed under GNU General Public License.     

Allele-specific differences in activity of a novel cannabinoid receptor 1 (CNR1) gene intronic enhancer in hypothalamus, dorsal root ganglia, and hippocampus

Polymorphisms within intron 2 of the CNR1 gene, which encodes cannabinoid receptor 1 (CB(1)), have been associated with addiction, obesity, and brain volume deficits. We used comparative genomics to identify a polymorphic (rs9444584-C/T) sequence (ECR1) in intron 2 of the CNR1 gene that had been conserved for 310 million years. The C-allele of ECR1 (ECR1(C)) acted as an enhancer in hypothalamic and dorsal root ganglia cells and responded to MAPK activation through the MEKK pathway but not in hippocampal cells. However, ECR1(T) was significantly more active in hypothalamic and dorsal root ganglia cells but, significantly, and in contrast to ECR1(C), was highly active in hippocampal cells where it also responded strongly to activation of MAPK. Intriguingly, rs9444584 is in strong linkage disequilibrium with two other SNPs (rs9450898 (r(2) = 0.841) and rs2023239 (r(2) = 0.920)) that have been associated with addiction, obesity (rs2023239), and reduced fronto-temporal white matter volumes in schizophrenia patients as a result of cannabis misuse (rs9450898). Considering their high linkage disequilibrium and the increased response of ECR1(T) to MAPK signaling when compared with ECR1(C), it is possible that the functional effects of the different alleles of rs9444584 may play a role in the conditions associated with rs9450898 and rs2023239. Further analysis of the different alleles of ECR1 may lead to a greater understanding of the role of CNR1 gene misregulation in these conditions as well as chronic inflammatory pain.     

Pregnancy-Specific Glycoproteins Bind Integrin αIIbβ3 and Inhibit the Platelet—Fibrinogen Interaction

Pregnancy-specific glycoproteins (PSGs) are immunoglobulin superfamily members encoded by multigene families in rodents and primates. In human pregnancy, PSGs are secreted by the syncytiotrophoblast, a fetal tissue, and reach a concentration of up to 400 ug/ml in the maternal bloodstream at term. Human and mouse PSGs induce release of anti-inflammatory cytokines such as IL-10 and TGFβ1 from monocytes, macrophages, and other cell types, suggesting an immunoregulatory function. RGD tri-peptide motifs in the majority of human PSGs suggest that they may function like snake venom disintegrins, which bind integrins and inhibit interactions with ligands. We noted that human PSG1 has a KGD, rather than an RGD motif. The presence of a KGD in barbourin, a platelet integrin αIIbβ3 antagonist found in snake venom, suggested that PSG1 may be a selective αIIbβ3 ligand. Here we show that human PSG1 binds αIIbβ3 and inhibits the platelet – fibrinogen interaction. Unexpectedly, however, the KGD is not critical as multiple PSG1 domains independently bind and inhibit αIIbβ3 function. Human PSG9 and mouse Psg23 are also inhibitory suggesting conservation of this function across primate and rodent PSG families. Our results suggest that in species with haemochorial placentation, in which maternal blood is in direct contact with fetal trophoblast, the high expression level of PSGs reflects a requirement to antagonise abundant (3 mg/ml) fibrinogen in the maternal circulation, which may be necessary to prevent platelet aggregation and thrombosis in the prothrombotic maternal environment of pregnancy.     

Interactions between lithology and biology drive the long-term response of stream chemistry to major hurricanes in a tropical landscape

Humid tropical forests play a dominant role in many global biogeochemical cycles, yet long-term records of tropical stream chemistry and its response to disturbance events such as severe storms and droughts are rare. Here we document the long-term variability in chemistry of two streams in the Luquillo Mountains, Puerto Rico over a period of 27 years. Our two focal study watersheds, the Río Icacos and Quebrada Sonadora, both drain several hundred hectares of tropical wet forests, and each received direct hits from Hurricanes Hugo (1989) and Georges (1998). They differ primarily in lithology (granitic vs. volcaniclastic) and elevation. Changes in major cations, anions, silica, and dissolved organic carbon were minimal over the study period, but the concentrations of nitrate show a strong response to hurricane disturbance and the longest time to recovery. Potassium also showed a large, although less consistent, response to disturbance. In the granitic terrain, nitrate concentrations exceeded long-term pre-hurricane background levels for over a decade, but were elevated in the volcaniclastic terrain for only 1–2 years. Lithology appears to be the primary driver explaining the different response trajectories of the two watersheds. In the granitic terrain, which showed slow recovery to pre-hurricane conditions, the quartz diorite bedrock weathers to produce coarser soils, deeper groundwater flow paths, and riparian zones with sharp spatial variation in redox conditions and very high nitrogen levels immediately adjacent to the stream. Groundwater flow paths are shallow and the levels of N in streamside groundwater are much lower in the volcaniclastic terrain. The recovery of vegetation following hurricane disturbance appears similar in the two watersheds, suggesting that the extent of structural damage to canopy trees determines the magnitude of NO₃ increases, but that the duration of elevated concentrations in stream water is a function of lithology.     

Comparisons of watershed sulfur budgets in southeast Canada and northeast US: new approaches and implications

Most of eastern North America receives elevated levels of atmospheric deposition of sulfur (S) that result from anthropogenic SO₂ emissions from fossil fuel combustion. Atmospheric S deposition has acidified sensitive terrestrial and aquatic ecosystems in this region; however, deposition has been declining since the 1970s, resulting in some recovery in previously acidified aquatic ecosystems. Accurate watershed S mass balances help to evaluate the extent to which atmospheric S deposition is retained within ecosystems, and whether internal cycling sources and biogeochemical processes may be affecting the rate of recovery from decreasing S atmospheric loads. This study evaluated S mass balances for 15 sites with watersheds in southeastern Canada and northeastern US for the period 1985 to 2002. These 15 sites included nine in Canada (Turkey Lakes, ON; Harp Lake, ON; Plastic Lake, ON; Hermine, QC; Lake Laflamme, QC; Lake Clair, QC; Lake Tirasse, QC; Mersey, NS; Moosepit, NS) and six in the US (Arbutus Lake, NY; Biscuit Brook, NY; Sleepers River, VT; Hubbard Brook Experimental Forest, NH; Cone Pond, NH; Bear Brook Watershed, ME). Annual S wet deposition inputs were derived from measured bulk or wet-only deposition and stream export was obtained by combining drainage water fluxes with SO₄ ^2? concentrations. Dry deposition has the greatest uncertainty of any of the mass flux calculations necessary to develop accurate watershed balances, and here we developed a new method to calculate this quantity. We utilized historical information from both the US National Emissions Inventory and the US (CASTNET) and the Canadian (CAPMoN) dry deposition networks to develop a formulation that predicted SO₂ concentrations as a function of SO₂ emissions, latitude and longitude. The SO₂ concentrations were used to predict dry deposition using relationships between concentrations and deposition flux derived from the CASTNET or CAPMoN networks. For the year 2002, we compared the SO₂ concentrations and deposition predictions with the predictions of two continental-scale air quality models, the Community Multiscale Air Quality (CMAQ) model and A Unified Regional Air-quality Modeling System (AURAMS) that utilize complete inventories of emissions and chemical budgets. The results of this comparison indicated that the predictive relationship provides an accurate representation of SO₂ concentrations and S deposition for the region that is generally consistent with these models, and thus provides confidence that our approach could be used to develop accurate watershed S budgets for these 15 sites. Most watersheds showed large net losses of SO₄ ^2? on an annual basis, and the watershed mass balances were grouped into five categories based on the relative value of mean annual net losses or net gains. The net annual fluxes of SO₄ ^2? showed a strong relationship with hydrology; the largest net annual negative fluxes were associated with years of greatest precipitation amount and highest discharge. The important role of catchment hydrology on S budgets suggests implications for future predicted climate change as it affects patterns of precipitation and drought. The sensitivity of S budgets is likely to be greatest in watersheds with the greatest wetland area, which are particularly sensitive to drying and wetting cycles. A small number of the watersheds in this analysis were shown to have substantial S sources from mineral weathering, but most showed evidence of an internal source of SO₄ ^2?, which is likely from the mineralization of organic S stored from decades of increased S deposition. Mobilization of this internal S appears to contribute about 1?C6 kg S ha^?1 year^?1 to stream fluxes at these sites and is affecting the rate and extent of recovery from acidification as S deposition rates have declined in recent years. This internal S source should be considered when developing critical deposition loads that will promote ecosystem recovery from acidification and the depletion of nutrient cations in the northeastern US and southeastern Canada.