The stoichiometry of trimeric SIV glycoprotein interaction with CD4 differs from that of anti-envelope antibody Fab fragments

Human and simian immunodeficiency viruses infect host lymphoid cells by binding CD4 molecules via their gp160 envelope glycoproteins. Biochemical studies on recombinant SIVmac32H (pJ5) envelope ectodomain gp140 precursor protein show that the envelope is a trimer. Using size exclusion chromatography, quantitative amino acid analysis, analytical ultracentrifugation, and CD4-based competition assay, we demonstrate that the stoichiometry of CD4 receptor-oligomeric envelope interaction is 1:1. By contrast, Fab fragments of both neutralizing and non-neutralizing monoclonal antibodies bind at a 3:1 ratio. Thus, despite displaying equivalent CD4 binding sites on each of the three gp140 protomers within an uncleaved trimer, only one site binds the soluble 4-domain human CD4 extracellular segment. The anti-cooperativity and the faster k(off) of gp140 trimer:CD4 versus gp120 monomer:CD4 interaction suggest that CD4-induced conformational change is impeded in the intact envelope. The implications of these findings for immunity against human immunodeficiency virus and simian immunodeficiency virus are discussed

The demography of space occupancy: measuring plant colonization and survival probabilities using repeated pin-point measurements

International audience1. The study of plant demography has long been an important component of plant ecological studies. However, in plant communities, e.g. grasslands, where individual plants are not easily distinguished and often vary in size, a convenient method of describing the demography and its ecological consequences has been lacking. 2. The aim of this study was to discuss the potential for using the change in the probability of space occupancy as a measure of ecological success in plant population biology. This will be done by demonstrating how the change in the probability of space occupancy depends on the processes of coloni-zation and survival, and demonstrating how colonization and survival probabilities may be estimated from repeated pinpoint recordings at the same pin-position. Furthermore, it will be demonstrated how to calculate the sensitivity and elasticity of the change in the probability of space occupancy to colonization and survival probabilities. 3. The method is applied to a case of repeated pinpoint data of the perennial grass Brachypodium phoenicoı¨es that underwent different grazing regimes from 2001 to 2008 on xero-halophytic grasslands. 4. It was demonstrated that grazing affected both plant survival and colonization probabilities so that the estimated colonization and survival probabilities of B. phoenicoı¨es were highest in the non-grazed regime. Furthermore, using the calculated elasticity of the change in space occupancy, we found that survival was more important than colonization events in determining the ecological success of B. phoenicoı¨es in the non-grazed regime, whereas colonization events were more important than survival in the grazed regime. 5. Synthesis and applications. We expect that the relatively simple method may be widely used on existing and future repeated pinpoint recordings from the same pin-position and, consequently, that plant demographic questions may be addressed with larger precision in plant communities where individual plants are not easily distinguished

Thigh-length compression stockings and DVT after stroke

Controversy exists as to whether neoadjuvant chemotherapy improves survival in patients with invasive bladder cancer, despite randomised controlled trials of more than 3000 patients. We undertook a systematic review and meta-analysis to assess the effect of such treatment on survival in patients with this disease

Evidence against Extracellular Exposure of a Highly Immunogenic Region in the C-Terminal Domain of the Simian Immunodeficiency Virus gp41 Transmembrane Protein

The generally accepted model for human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein topology includes a single membrane-spanning domain. An alternate model has been proposed which features multiple membrane-spanning domains. Consistent with the alternate model, a high percentage of HIV-1-infected individuals produce unusually robust antibody responses to a region of envelope, the so-called “Kennedy epitope,” that in the conventional model should be in the cytoplasm. Here we show analogous, robust antibody responses in simian immunodeficiency virus SIVmac239-infected rhesus macaques to a region of SIVmac239 envelope located in the C-terminal domain, which in the conventional model should be inside the cell. Sera from SIV-infected rhesus macaques consistently reacted with overlapping oligopeptides corresponding to a region located within the cytoplasmic domain of gp41 by the generally accepted model, at intensities comparable to those observed for immunodominant areas of the surface component gp120. Rabbit serum raised against this highly immunogenic region (HIR) reacted with SIV envelope in cell surface-staining experiments, as did monoclonal anti-HIR antibodies isolated from an SIVmac239-infected rhesus macaque. However, control experiments demonstrated that this surface staining could be explained in whole or in part by the release of envelope protein from expressing cells into the supernatant and the subsequent attachment to the surfaces of cells in the culture. Serum and monoclonal antibodies directed against the HIR failed to neutralize even the highly neutralization-sensitive strain SIVmac316. Furthermore, a potential N-linked glycosylation site located close to the HIR and postulated to be outside the cell in the alternate model was not glycosylated. An artificially introduced glycosylation site within the HIR was also not utilized for glycosylation. Together, these data support the conventional model of SIV envelope as a type Ia transmembrane protein with a single membrane-spanning domain and without any extracellular loops