Monocyte-derived macrophages (A), monocyte-derived dendritic cells (B) and T cells (C) were incubated with R5 or X4 viruses in the presence of increasing concentrations of C14 or an unique dose of T 20 (5 μM) for 3 hours at 37°C

Cells were then washed and cultured in fresh medium for 3 days. DNA was extracted and the viral DNA was quantified by real time PCR. Means are representative of 3 independent experiments and assays were performed in triplicates. *, ≤ 0.05; **, ≤ 0.01 between untreated and treated cells.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical development as microbicide of zinc tetra-ascorbo-camphorate, a novel terpenoid derivative: Potent in vitro inhibitory activity against both R5- and X4-tropic HIV-1 strains without significant in vivo mucosal toxicity"</p><p>http://www.aidsrestherapy.com/content/5/1/10</p><p>AIDS Research and Therapy 2008;5():10-10.</p><p>Published online 3 Jun 2008</p><p>PMCID:PMC2426711.</p><p></p

Monocyte-derived macrophages, monocyte-derived dendritic cells and T cells were incubated with R5 viruses in the presence of increasing concentrations of C14 for 3 hours at 37°C

Cells were then washed and incubated with polyclonal human anti-gp160 antibodies. The staining was revealed with FITC-conjugated mouse anti-human IgG mAbs. Cells were then analysed by confocal microscopy. The experiment was performed 3 times with cells from three different donors. 30 cells were at least analyzed for each donor.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical development as microbicide of zinc tetra-ascorbo-camphorate, a novel terpenoid derivative: Potent in vitro inhibitory activity against both R5- and X4-tropic HIV-1 strains without significant in vivo mucosal toxicity"</p><p>http://www.aidsrestherapy.com/content/5/1/10</p><p>AIDS Research and Therapy 2008;5():10-10.</p><p>Published online 3 Jun 2008</p><p>PMCID:PMC2426711.</p><p></p

HIV-1 was adsorbed on poly-L-lysin pre-coated wells (Greiner Bio-One) at 4°C overnight and further incubated with C14 for 1 h

In positive and negative control wells, 1% Triton X-100 and medium were added, respectively. After four washes, activated peripheral blood lymphocytes were incubated with C14 or triton-treated or untreated HIV-1 particles. After 6 days, viral production was assessed by p24 Ag capture ELISA. Means are representative of 3 independent experiments were performed in triplicates. **, ≤ 0.01.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical development as microbicide of zinc tetra-ascorbo-camphorate, a novel terpenoid derivative: Potent in vitro inhibitory activity against both R5- and X4-tropic HIV-1 strains without significant in vivo mucosal toxicity"</p><p>http://www.aidsrestherapy.com/content/5/1/10</p><p>AIDS Research and Therapy 2008;5():10-10.</p><p>Published online 3 Jun 2008</p><p>PMCID:PMC2426711.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-1

domain of gp41) was derived from clonal viruses isolated from plasma of patients chronically infected with HIV-1. The ability of these viruses, which express luciferase in the place of Nef, to infect U373 cells stably expressing CD4 and either CCR5 (panel A) or CXCR4 (panel B) co-receptors was measured by evaluating luciferase expression in target cells 44 hours after infection. For patient 2, clonal viruses were obtained from plasma samples obtained 8 months apart (26 and 34 months after initial diagnosis). The ability of all recombinant viruses to infect the two cell types was evaluated, but results are shown only for viruses that induced significant luciferase activity in the indicated target cell type [>2 (RLU/sec)/(ng p24/ml)]. Viruses could be classified as having strict R5 tropism (open symbols) strict X4 tropism (solid black symbols) and dual tropism (sold gray symbols). Results for viruses expressing sequences amplified by RT-PCR from patient plasma is also shown (stars). Each symbol is the mean of at least 3 independent experiments; the mean coefficient of variation for these results is as follows: U373-R5 cells, 42% (range 3 – 83%); U373-X4 cells, 50% (range 10 – 78%). For each patient, significant differences were found comparing the viruses with the highest and lowest infectivity (p < 0.05 – 0.001 by t-test).<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-0

F encompassing C1 to V2 (corresponding to nucleotides 6440 – 6809 of HXB2) of the 70 clonal viruses evaluated in the study (open circles, strict R5 tropism, solid black circles, strict X4 tropism; solid gray circles, dual tropism), the consensus sequence of the same region for viral RNA amplified by RT-PCR from an aliquot of the plasma sample used to generate the clonal viruses (stars), and the sequence of the laboratory strain pNL4-3. Bootstrap values are also indicated. For patient 2, the consensus sequence of plasma viruses grouped with clonal viruses with X4 tropism at M26, and with clonal viruses with R5 tropism at M34.<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-5

Ted monoclonal antibodies or isotype-matched control antibodies conjugated with the same flurochrome. (A) CD4 expression on U373-R5 cells (thin solid line) and MT4-R5 cells (heavy solid line) detected using FITC-labelled mouse anti-human CD4 monoclonal antibody (clone RPA-T4). Binding of an isotype-matched control antibody conjugated with FITC is shown in the corresponding dashed lines. (B) CCR5 expression on U373-R5 cells (thin solid line) and MT4-R5 cells (heavy solid line) detected using phycoerythrin-conjugated mouse anti-human CCR5 monoclonal antibody (clone 2D7). Binding of an isotype-matched control antibody conjugated with phycoerythrin to these cells is shown in the corresponding dashed lines.<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-6

(panel A) and/or MT4-R5 cells (panel B) were selected from among those obtained from patients 1–4. For each of these 20 viruses, the IC50s were determined on U373-R5 cells for soluble CD4 (C), enfuvirtide (E), TAK-799 (G), and neutralizing monoclonal antibodies 2F5 (D), 2G12 (F), and 48d (H). Each symbol represents the mean of three independent determinations. For all inhibitors except TAK-799, significant patient-specific differences in IC50 were observed using the Kruskal-Wallis test. Brackets indicate significant pairwise differences in post-test comparisons performed using Dunn's multiple comparison test (* p < 0.05; ** p < 0.01). For patient 2, clonal viruses were obtained from plasma samples obtained at both 26 months (solid symbols) and 34 months (open symbols) after initial diagnosis.<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-3

Ty of these viruses to infect MT4-R5 cells was measured by evaluating luciferase expression in the target cells 44 hours after infection. For patient 2, clonal viruses were obtained from plasma samples obtained 8 months apart (26 and 34 months after initial diagnosis). The tropism of the recombinant viruses, as defined by their ability to infect U373-R5 and U373-X4 cells is shown: strict R5, open symbols; dual, solid gray symbols; strict X4, solid black symbols. Results for viruses expressing sequences amplified by RT-PCR from patient plasma is also shown (stars). Each symbol is the mean of at least 3 independent experiments; the mean coefficient of variation for these results is 37% (range 1 – 78%). For each patient, significant differences were found comparing the viruses with the highest and lowest infectivity (p < 0.05 – 0.005 by t-test). (B) For each recombinant virus, the infectivity [(RLU/sec)/(ng p24/ml)] observed using U373 target cells and MT4-R5 target cells is expressed as a ratio.<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-7

Ed by the 17 clonal viruses from patient 2 with strict R5 tropism are shown. The consensus amino sequence is shown on the top line, and only amio acids differing from the consensus sequence are shown for each clone. For each variable region, sequences that are identical or that differ by a single amino acid substitution not identified in another sequence are highlighted by the same color, and these haplotypes are also identifed by numbers adjacent to the brackets. The red arrow indicates the envelope expressing the V2 region haplotype 4 associated with the V3 region haplotype 1 (see text). (B) The infectivity of recombinant viruses expressing these Env proteins using U373-R5 cells is shown. For each of the variable regions, the color of the symbols corresponds to that of the V region haplotype expressed by that virus. The red arrow indicates the infectivity of the clone expressing the V2 region haplotype 4 associated with the V3 region haplotype 1.<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p

Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses-4

F these viruses to infect MT4-R5 cells and U373-R5 cells was measured by evaluating luciferase expression in the target cells 44 hours after infection. For each of the 53 viruses with strict R5 tropism, the infectivity ratio (infectivity for U373-R5 cells/infectivity for MT4-R5 cells) is expressed as a function of the infectivity for MT4-R5 cells [(RLU/sec)/(ng p24/ml)]. A significant inverse correlation between these parameters was observed (Spearman r = -0.64, p < 0.0001).<p><b>Copyright information:</b></p><p>Taken from "Functional diversity of HIV-1 envelope proteins expressed by contemporaneous plasma viruses"</p><p>http://www.retrovirology.com/content/5/1/23</p><p>Retrovirology 2008;5():23-23.</p><p>Published online 29 Feb 2008</p><p>PMCID:PMC2270869.</p><p></p