Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue

Human cytomegalovirus (HCMV) can be acquired sexually and is shed from the genital tract. Cross-sectional studies in women show that changes in genital tract microbial flora affect HCMV infection and/or shedding. Since genital microbial flora may affect HCMV infection or replication by stimulating cells through Toll-like receptors (TLR), we assessed the effects of defined TLR-ligands on HCMV replication in foreskin fibroblasts and ectocervical tissue. Poly I:C (a TLR3-ligand) and lipopolysaccharide (LPS, a TLR4-ligand) inhibited HCMV and induced secretion of IL-8 and Interferon-beta (IFNβ) in both foreskin fibroblasts and ectocervical tissue. The anti-HCMV effect was reversed by antibody to IFNβ. CpG (TLR9 ligand) and lipoteichoic acid (LTA, TLR2 ligand) also inhibited HCMV infection in ectocervical tissue and this anti-HCMV effect was also reversed by anti-IFNβ antibody. In contrast, LTA and CpG did not inhibit HCMV infection in foreskin fibroblasts. This study shows that TLR ligands induce an HCMV-antiviral effect that is mediated by IFNβ suggesting that changes in genital tract flora may affect HCMV infection or shedding by stimulating TLR. This study also contrasts the utility of two models that can be used for assessing the interaction of microbial flora with HCMV in the genital tract. Clear differences in the response to different TLR ligands suggests the explant model more closely reflects in vivo responses to genital infections

Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue-7

<p><b>Copyright information:</b></p><p>Taken from "Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue"</p><p>http://www.virologyj.com/content/4/1/133</p><p>Virology Journal 2007;4():133-133.</p><p>Published online 5 Dec 2007</p><p>PMCID:PMC2222636.</p><p></p>ere then collected and assayed for IL-8 by ELISA. IL-8 secretion from one experiment representative of three. Bars represent mean ± SD of triplicate cultures. B and C. After treatment of HFF with medium alone, LTA, Poly I:C, LPS, or CpG 2395 for 24 hours, cells were washed and CMVPT30-gfp was added. After four hours, the virus innoculum was removed and replaced with fresh culture medium. HCMV infection was quantified on day 10 post-infection by counting fluorescent (GFP expressing) cells in each well. B. Shown is a representative culture well from cells treated with medium alone. C. Percent inhibition compared to medium control. Results of one experiment, representative of 3 independent experiments, is shown. Bars represent mean ± SD of triplicate cultures. * indicates P ≤ 0.05 compared to control. ** indicates P ≤ 0.01 compared to control. *** indicates P ≤ 0.001 compared to control

Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue-0

<p><b>Copyright information:</b></p><p>Taken from "Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue"</p><p>http://www.virologyj.com/content/4/1/133</p><p>Virology Journal 2007;4():133-133.</p><p>Published online 5 Dec 2007</p><p>PMCID:PMC2222636.</p><p></p>ere then collected and assayed for IL-8 by ELISA. IL-8 secretion from one experiment representative of three. Bars represent mean ± SD of triplicate cultures. B and C. After treatment of HFF with medium alone, LTA, Poly I:C, LPS, or CpG 2395 for 24 hours, cells were washed and CMVPT30-gfp was added. After four hours, the virus innoculum was removed and replaced with fresh culture medium. HCMV infection was quantified on day 10 post-infection by counting fluorescent (GFP expressing) cells in each well. B. Shown is a representative culture well from cells treated with medium alone. C. Percent inhibition compared to medium control. Results of one experiment, representative of 3 independent experiments, is shown. Bars represent mean ± SD of triplicate cultures. * indicates P ≤ 0.05 compared to control. ** indicates P ≤ 0.01 compared to control. *** indicates P ≤ 0.001 compared to control

Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue-3

<p><b>Copyright information:</b></p><p>Taken from "Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue"</p><p>http://www.virologyj.com/content/4/1/133</p><p>Virology Journal 2007;4():133-133.</p><p>Published online 5 Dec 2007</p><p>PMCID:PMC2222636.</p><p></p>itional 24 period in one ml of fresh medium. These conditioned supernatants were collected and incubated in the presence of either medium as a control, rabbit polyclonal anti-IFNβ antibody, or normal rabbit serum for 1 hour at 37°C. Recombinant IFNβ (IFN) was also incubated in the presence of either medium as a control, rabbit polyclonal anti-IFNβ antibody, or normal rabbit serum for 1 hour at 37°C. The treated supernatants were then transferred to wells of confluent HFF fibroblasts and cultured for 24 hours. The conditioned medium was removed and the fresh HFF were challenged with CMVPT30-gfp. Fluorescent cells were then counted on day 10 post-infection (PI). The data shown is representative of 3 independent experiments. *** indicates P ≤ 0.001 compared to control

Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue-2

<p><b>Copyright information:</b></p><p>Taken from "Differential inhibition of human cytomegalovirus (HCMV) by toll-like receptor ligands mediated by interferon-beta in human foreskin fibroblasts and cervical tissue"</p><p>http://www.virologyj.com/content/4/1/133</p><p>Virology Journal 2007;4():133-133.</p><p>Published online 5 Dec 2007</p><p>PMCID:PMC2222636.</p><p></p>ected and tested for IFNβ by ELISA. The limit of detection of this assay was 1000 pg/ml. * indicates P ≤ 0.05 compared to control. ** indicates P ≤ 0.01 compared to control. *** indicates P ≤ 0.001 compared to control