7 research outputs found
HIV-1 replication in colonic tissue obtained through surgical resection and flexible sigmoidoscopy.
<p>Polarized cultures were set-up in duplicate. Titrations of HIV-1<sub>BaL</sub> and HIV-1<sub>JR-CSF</sub> were applied to the apical surface and allowed to culture overnight. The following day, the tissues were thoroughly washed and followed through day 21 of culture. HIV-1 replication (p24 ELISA) was monitored in the basolateral supernatant collected every 3 to 4 days of culture. The data presented are the median ±95% confidence interval of 7 tissues for HIV-1<sub>BaL</sub> and 5 tissues for HIV-1<sub>JR-CSF</sub> for surgical resections; and 12 tissues for HIV-1<sub>BaL</sub> and 9 tissues for HIV-1<sub>JR-CSF</sub> for flexible sigmoidoscopy.</p
Rectal-specific microbicide (RM) products protect surgically resected colonic tissue from HIV-1 infection and are safe.
<p>Colonic tissue obtained through surgical resections was used to create polarized explant cultures. The explants were set-up in duplicated. (A) Efficacy: Tenofovir (TFV) aqueous-based RMs were diluted 1∶5 in medium with HIV-1<sub>BaL</sub> and applied to the apical surface. The UC781 lipid-based RMs were used neat and mixed with HIV-1<sub>BaL</sub>, and then applied to the apical surface. The corresponding placebo products were handled in a similar fashion. After an overnight culture, the tissues were thoroughly washed and followed through day 21 of culture. HIV-1 replication (p24 ELISA) was monitored in the basolateral supernatant collected every 3 to 4 days of culture. The data presented are the median ±95% confidence interval of 5 independent tissues for each treatment. (B) Tissue viability: After an overnight culture with the indicated RM product, medium alone (untreated control) or medium containing a 1∶5 dilution of a 3% nonoxynol-9 (N9)-containing gel, tissues were washed with one piece further cultured in medium containing 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan for the MTT assay or the other piece fixed in paraformaldehyde for hematoxylin and eosin staining. The MTT assay results are presented as scatter plots with the horizontal lines denoting the mean ± standard deviation of 5 independent tissues. The histology is representative of one of those tissues.</p
Physiochemical characterization of the rectal-specific microbicide formulations.
<p>*not required.</p>a<p>Data are presented as the average ± standard deviation of three independent tests.</p
Rectal-specific microbicide (RM) products protect colonic tissue obtained through flexible sigmoidoscopy from HIV-1 infection and are safe.
<p>Colonic tissue obtained through flexible sigmoidoscopy was used to create polarized explant cultures. The explants were set-up in duplicate. (A) Efficacy: Tenofovir (TFV) aqueous-based RMs were diluted 1∶5 in medium with HIV-1<sub>BaL</sub> and applied to the apical surface. The UC781 lipid-based RMs were used neat and mixed with HIV-1<sub>BaL</sub>, and then applied to the apical surface. The corresponding placebo products were handled in a similar fashion. After an overnight culture, the tissues were thoroughly washed and followed through day 21 of culture. HIV-1 replication (p24 ELISA) was monitored in the basolateral supernatant collected every 3 to 4 days of culture. The data presented are the median ±95% confidence interval of 3 to 5 independent donors for each treatment. (B) Tissue viability: After an overnight culture with the indicated RM product, medium alone (untreated control) or medium containing a 1∶5 dilution of a 3% nonoxynol-9 (N9)-containing gel, tissues were washed with one piece further cultured in medium containing 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan for the MTT assay or the other piece fixed in paraformaldehyde for hematoxylin and eosin staining. The MTT assay results are presented as scatter plots with the horizontal lines denoting the mean ± standard deviation of 4 independent donors. The histology is representative of one of those donors.</p
HIV-1 replication across four major areas of the colon.
<p>Tissues from the ascending, transverse, descending, and sigmoid colon were evaluated for the ability of HIV-1<sub>BaL</sub> to infect and replicate. Tissues were set-up in polarized conditions and HIV-1 was applied to the apical, luminal surface. After overnight culture, the tissues were washed and followed through day 21 of culture. HIV-1 replication (p24 ELISA) was monitored in the basolateral supernatant collected every 3 to 4 days of culture. The data presented are a median of 7, 6 (ascending and sigmoid), and 4 (transverse and descending) tissues ±95% confidence interval.</p
The impact of semen on HIV-1 replication by colonic tissue.
<p>Tissues were set-up in duplicate in polarized cultures to evaluate the effects of semen on HIV-1 infection (A) or on tissue viability/architecture (B). (A) HIV-1<sub>BaL</sub> or HIV-1<sub>JR-CSF</sub> was mixed with 50% whole semen and applied to the apical surface and cultured overnight. The following day, the tissues were thoroughly washed and followed through day 21 of culture. HIV-1 replication (p24 ELISA) was monitored in the basolateral supernatant collected every 3 to 4 days of culture. The data presented are the median ±95% confidence interval of 5 tissues for HIV-1<sub>BaL</sub> and HIV-1<sub>JR-CSF</sub> each for surgical resections; and 7 tissues for HIV-1<sub>BaL</sub> and 5 tissues for HIV-1<sub>JR-CSF</sub> for flexible sigmoidoscopy. (B) 50% semen was added to the apical surface of surgical resections (SR) or flexible sigmoidoscopy (FS). As controls, medium alone (untreated control) or medium containing a 1∶5 dilution of a 3% nonoxynol-9 (N9)-containing gel were used. After an overnight culture, tissues were washed with one piece further cultured in medium containing 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan for the MTT assay or the other piece fixed in paraformaldehyde for hematoxylin and eosin staining. The MTT assay results are presented as scatter plots with the horizontal lines denoting the mean ± standard deviation of 5 independent tissues. The histology is representative of one of those tissues.</p