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Toll-like receptors and cytokines as surrogate biomarkers for evaluating vaginal immune response following microbicide administration.

Gupta SM, Aranha CC, Mohanty MC, Reddy KV - Mediators Inflamm. (2008)

Bottom Line: The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels.However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade.Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides.

View Article: PubMed Central - PubMed

Affiliation: Immunology Division, National Institute for Research in Reproductive Health, Mumbai, India.

ABSTRACT
Topical microbicides are intended for frequent use by women in reproductive age. Hence, it is essential to evaluate their impact on mucosal immune function in the vagina. In the present study, we evaluated nisin, a naturally occurring antimicrobial peptide (AMP), for its efficacy as an intravaginal microbicide. Its effect on the vaginal immune function was determined by localizing Toll-like receptors (TLRs-3, 9) and cytokines (IL-4, 6 , 10 and TNF-alpha) in the rabbit cervicovaginal epithelium following intravaginal administration of high dose of nisin gel for 14 consecutive days. The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels. However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade. Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides.

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Flow cytometric analysis of TLR expression in rabbit CVE cells after intravaginal administration of placebo, nisin (15.15 mM in 2 mL of 1% polycarbophil gel) SDS (56 μM in 2 mL of 1% polycarbophil gel) and 2 mL of 1% polycarbophil gel. Mean fluorescence intensity for TLR-3 and 9 were significantly increased in SDS gel-treated animals compared to their respective placebo and nisin gel-treated groups. The mean and error bars indicate standard deviation of triplicate measurements obtained from three separate experiments.
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fig2: Flow cytometric analysis of TLR expression in rabbit CVE cells after intravaginal administration of placebo, nisin (15.15 mM in 2 mL of 1% polycarbophil gel) SDS (56 μM in 2 mL of 1% polycarbophil gel) and 2 mL of 1% polycarbophil gel. Mean fluorescence intensity for TLR-3 and 9 were significantly increased in SDS gel-treated animals compared to their respective placebo and nisin gel-treated groups. The mean and error bars indicate standard deviation of triplicate measurements obtained from three separate experiments.

Mentions: Theinflammatory potential of nisin gel was further evaluated by flow cytometry.The results indicated no significant difference in the number of cells positivefor TLR-3 and 9 between placebo and nisin gel-treated groups (Figure 2).However, in the SDS gel-treated group, mean fluorescence intensity for TLR-3and 9 increased significantly. The experiment was repeated twice in duplicatesand similar results were obtained.


Toll-like receptors and cytokines as surrogate biomarkers for evaluating vaginal immune response following microbicide administration.

Gupta SM, Aranha CC, Mohanty MC, Reddy KV - Mediators Inflamm. (2008)

Flow cytometric analysis of TLR expression in rabbit CVE cells after intravaginal administration of placebo, nisin (15.15 mM in 2 mL of 1% polycarbophil gel) SDS (56 μM in 2 mL of 1% polycarbophil gel) and 2 mL of 1% polycarbophil gel. Mean fluorescence intensity for TLR-3 and 9 were significantly increased in SDS gel-treated animals compared to their respective placebo and nisin gel-treated groups. The mean and error bars indicate standard deviation of triplicate measurements obtained from three separate experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2607029&req=5

fig2: Flow cytometric analysis of TLR expression in rabbit CVE cells after intravaginal administration of placebo, nisin (15.15 mM in 2 mL of 1% polycarbophil gel) SDS (56 μM in 2 mL of 1% polycarbophil gel) and 2 mL of 1% polycarbophil gel. Mean fluorescence intensity for TLR-3 and 9 were significantly increased in SDS gel-treated animals compared to their respective placebo and nisin gel-treated groups. The mean and error bars indicate standard deviation of triplicate measurements obtained from three separate experiments.
Mentions: Theinflammatory potential of nisin gel was further evaluated by flow cytometry.The results indicated no significant difference in the number of cells positivefor TLR-3 and 9 between placebo and nisin gel-treated groups (Figure 2).However, in the SDS gel-treated group, mean fluorescence intensity for TLR-3and 9 increased significantly. The experiment was repeated twice in duplicatesand similar results were obtained.

Bottom Line: The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels.However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade.Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides.

View Article: PubMed Central - PubMed

Affiliation: Immunology Division, National Institute for Research in Reproductive Health, Mumbai, India.

ABSTRACT
Topical microbicides are intended for frequent use by women in reproductive age. Hence, it is essential to evaluate their impact on mucosal immune function in the vagina. In the present study, we evaluated nisin, a naturally occurring antimicrobial peptide (AMP), for its efficacy as an intravaginal microbicide. Its effect on the vaginal immune function was determined by localizing Toll-like receptors (TLRs-3, 9) and cytokines (IL-4, 6 , 10 and TNF-alpha) in the rabbit cervicovaginal epithelium following intravaginal administration of high dose of nisin gel for 14 consecutive days. The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels. However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade. Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides.

Show MeSH