Year-wise trends of VL cases reported in Syria and Saudi Arabia.

<p>Data is based on WHO estimates.</p

Relative distribution of VL cases reported in the year 2008.

<p>Relative distribution of VL cases reported in the year 2008.</p

Year-wise trend of VL cases reported in Iraq.

<p>Data is based on WHO estimates.</p

Year-wise trend of CL cases reported in Middle East.

<p>Data is based on WHO reports for each country <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003208#pntd.0003208-World6" target="_blank">[48]</a>.</p

DCIR modelling.

<p><b>A) Partial sequence alignment</b> between CLEC4M (GenBank™ #AAI10615) and DCIR (GenBank™ # NP_057268) indicating completely conserved regions and residues. <b>B)</b><b>Three-dimensional model</b> showing positions of the selected docking sites on the dendritic cell immunoreceptor (DCIR) model (103-233) for virtual screening runs according to the examples. Residues forming site A are represented by the <i>blue</i> portions while <i>orange</i> indicates those forming site B.</p

ADMET.

<p>Modelled ADMET (Absorption, Distribution, Metabolism, Elimination and Toxicity) properties of our first four lead compounds using ADMET predictor. These results shown that toxicity of our molecules is minimal compare to protease inhibitor or Maraviroc, both molecules currently used in clinic.</p

Three-dimensional schematic representation of the docking pose of selected active molecules.

<p><b>Panel A</b>: Compound A1 (1,5-diphenyl-2,4-pentadien-1-one) in site A. <b>Panel B</b>: Compound A4∶3,6-di(2 pyridyl) pyridazine in site A. <b>Panel C</b>: Compound B1 (1-benzofuran-2-yl-phenylmethanone in site B. <b>Panel D</b>: Compound B2 (1-methyl-4-[(4-methylphenyl)-NNO-azoxy]benzene) in site B.</p

Impact of DCIR inhibitors on HIV-1 transmission by apoptotic CD4⁺ T cells.

<p>Target CD4⁺ T cells (1×10⁶) were treated for 16 h with H₂O₂ (30 µM) to induce apoptosis and the surface expression of DCIR. Cells (± H₂O₂ treatment) were incubated for 10 min with a site A inhibitor (1 and 4) or a site B inhibitor (1 and 2) or with 10 mM DMSO. A) Cells were next exposed to NL4-3 for 1 h at 37°C, washed thoroughly to remove un-adsorbed virions before assessing p24 content. B) Cells were first incubated with NL4-3 for 2 h at 37°C, washed thoroughly to remove un-adsorbed virions and cultured with autologous PHA-L/IL-2-activated CD4TL in complete RPMI-1640 supplemented with rhIL-2. Cell-free supernatants were collected on day 3 and assayed for p24 content. Data correspond to mean ± SEM of 4 independent experiments performed in triplicate for panel A and mean ± SEM of 4 independent experiments for panel B. Asterisks denote statistically significant values (*, <i>P</i><0.05; **, <i>P</i><0.01).</p

Selected compounds screened and tested <i>in vitro</i>.

<p><b>A)</b> Chemical structure of potential inhibitors of HIV-1 attachment to DCIR, selected by virtual screening, Compounds selected from CRD site are depicted as A1 to A4. Those selected from EPS site are named as B1 to B3 Virtual interacting compounds with both site of DCIR modeling have anti-HIV activity. <b>B)</b> Inhibitors decrease HIV-1 attachment to DCIR: Raji-CD4-DCIR cells were treated with one of four site-A inhibitors or one of three site-B inhibitors (inhibitor concentration = 10 µM) or with solvent (10 mM DMSO) only for 10 min at 37°C. Cells were thereafter exposed to NL4-3 virus for 60 min. After three washes with PBS to remove un-adsorbed virus, the abundance of cell-associated virions was quantified by measuring p24 content. Data correspond to mean ± SEM of three independent experiments performed with triplicate samples. Asterisks (*) denote statistically significant values (*, <i>P</i><0.05, ** P<0.01, *** P<0.001).</p

HIV-1 replication is diminished in DCIR-expressing cells by inhibitors specific for the site A and site B.

<p>Raji-CD4 and Raji-CD4-DCIR were treated with the inhibitors selected in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067873#pone-0067873-g002" target="_blank">figure 2</a>, or DMSO. Cells were then exposed to NL4-3 for 2 h, rinsed and maintained in culture for 3d. Cell-free culture supernatants were collected and assayed for p24 content. Data shown correspond to the means ± SEM from 3 independent experiments performed with triplicate samples. Asterisks denote statistically significant values (*, <i>P</i><0.05, ** P<0.01, *** P<0.001).</p