CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates-0

<p><b>Copyright information:</b></p><p>Taken from "CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates"</p><p>http://www.retrovirology.com/content/4/1/50</p><p>Retrovirology 2007;4():50-50.</p><p>Published online 23 Jul 2007</p><p>PMCID:PMC1950888.</p><p></p>RT/well. The day after infection cultures were washed extensively and fresh medium was added. Infected NP-2/CCR5 cells were followed for syncytia induction up to seven days after infection. RT was analyzed in supernatants from NP-2 cells at day 1 after wash and before start of cocultivation. Cocultivation of NP-2/CCR5 cells with hPBMC was started seven days after infection and virus production was measured after additional 6 days. CD4-independent-HIGH, virus production and/or syncytia induction could be detected directly in NP-2/CCR5 cells (dark grey). CD4-independent-LOW, productive infection in NP-2/CCR5 cells revealed only after cocultivation of infected NP-2/CCR5 cells with hPBMC (light grey). RT was analyzed with undiluted supernatants and therefore values above 1000 pg RT/ml cannot be separated. Detection limit for RT was 50 pg/ml. Values are means of duplicate infections

CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates-2

<p><b>Copyright information:</b></p><p>Taken from "CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates"</p><p>http://www.retrovirology.com/content/4/1/50</p><p>Retrovirology 2007;4():50-50.</p><p>Published online 23 Jul 2007</p><p>PMCID:PMC1950888.</p><p></p>he CCR5 part is represented as grey and the CXCR4 part as black. B. Mode of CCR5-use was analyzed by infection of U87.CD4 cells expressing CCR5 or chimeric receptors. Length of bars indicates degree of infection and syncytia induction as observed in a light microscope 5 and 7 days after infection. Degree of infection follows a scale from 0 to 5 where 0 is no syncytia and RT negative; 1 was 90% of the wells. RT production was positive in grades ranging from 2 to 5 and in concordance with syncytia induction

CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates-4

<p><b>Copyright information:</b></p><p>Taken from "CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates"</p><p>http://www.retrovirology.com/content/4/1/50</p><p>Retrovirology 2007;4():50-50.</p><p>Published online 23 Jul 2007</p><p>PMCID:PMC1950888.</p><p></p>time in 13 macaques. Neutralization sensitivity of three isolates (A, 2-week isolates, B, 3 or 4-month isolates and C, late isolates) from each macaque was tested with 1:20 dilution of serum [30]. Neutralization was also performed with autologous serum which gave similar results (data not shown). Neutralization sensitivity was measured using the GHOST(3) cell plaque reduction assay which has a cut-off for neutralization at 30% (marked with a line), that is results below 30% are negative [67]. The majority of newly infected macaques harbored virus populations with a CD4-independent-HIGH (dark grey bars) and neutralization sensitive phenotype. This phenotype gradually changed to become a CD4-independent-LOW (light grey bars) and neutralization resistant (below 30%) virus population. CD4-dependent isolates (white bars) were seen in both neutralization sensitive and neutralization resistant populations. Values are mean neutralization (+/- SD) of two independent assays performed in triplicates

CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates-1

<p><b>Copyright information:</b></p><p>Taken from "CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates"</p><p>http://www.retrovirology.com/content/4/1/50</p><p>Retrovirology 2007;4():50-50.</p><p>Published online 23 Jul 2007</p><p>PMCID:PMC1950888.</p><p></p>ysed by 0.001% Triton X-100 followed by one cycle of freeze-thawing step. Lysates were titrated at five-fold dilution steps on hPBMC. Supernatant culture fluids from hPBMC infections were collected at day 7 and production of RT was analyzed with undiluted supernatants. The RT cut-off detection level was 50 pg/ml and values above 1000 pg/ml could not be separated. Dark grey bars represent mean virus production in NP-2/CD4/CCR5 cells and. light grey bars represent virus production in NP-2/CCR5 cells. White bars represent virus production measured by RT in PBMC infected with cell lysates diluted 1:5 from infected NP-2/CCR5 cells. Positive syncytia induction (SI) are indicated with +. Means of RT production in duplicates of infection are indicated

CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates-3

<p><b>Copyright information:</b></p><p>Taken from "CD4-independent use of the CCR5 receptor by sequential primary SIVsm isolates"</p><p>http://www.retrovirology.com/content/4/1/50</p><p>Retrovirology 2007;4():50-50.</p><p>Published online 23 Jul 2007</p><p>PMCID:PMC1950888.</p><p></p>ell (in 88% of the cultures) and RT production was measured in MDM 15 days after infection. Values are means from at least two experiments with MDM from different donors