Increased CCL2 Expression and Macrophage/Monocyte Migration During Microbicide-Induced Vaginal Irritation

Abstract

Despite availability of successful prevention strategies, HIV continues to spread at alarming rates, especially among women in developing countries. Vaginal microbicides offer a promising approach for blocking transmission of HIV when condom use cannot be negotiated with male partners. A major problem in the development of vaginal microbicides is chemically induced vaginal irritation, which can enhance the risk of HIV transmission. Evaluation of vaginal irritation prior to clinical trials typically uses an expensive and animal-intensive rabbit vaginal irritation model, which could be supplemented by measuring additional inflammatory biomarkers. We studied several immunological parameters as potential biomarkers of vaginal irritation, using the spermicides nonoxynol-9 and benzalkonium chloride as test microbicides. We measured amounts of cytokines, as well as inflammatory cells, in vaginal tissue lysates and on the vaginal surface. We observed that treatment with the selected microbicides increases quantities of the inflammatory cytokines interleukin-1β, CXCL8, and CCL2 in the vaginal tissue parenchyma, and of CCL2 on the vaginal surface. This observation was correlated with increases in macrophages in the vaginal parenchyma. We suggest that measurements of CCL2 and macrophages can serve as new inflammatory biomarkers to evaluate the safety of promising novel microbicides for prevention of HIV.

Amyloid-binding Small Molecules Efficiently Block SEVI (Semen-derived Enhancer of Virus Infection)- and Semen-mediated Enhancement of HIV-1 Infection

“…5 C ), IL-1β, and TNF-α (data not shown). These cytokines and chemokines were selected because they have been shown to be up-regulated by other candidate microbicides and because they may play a role in microbicide-mediated enhancement of HIV-1 infection (16–18). BTA-EG 6 did not lead to the release of any of these cytokines or chemokines, even at the highest doses tested.…”

Effects of Benzalkonium Chloride on THP-1 Differentiated Macrophages In Vitro

“…Although, we similarly observed that a low concentration of BAK (33 nM or 10 −5 %) did not induce CCL4 production by THP-1 cells, this concentration of BAK could induce cytokines production by macrophages (THP-1 differentiated cells). It has been previously shown that BAK induced the release of inflammatory cytokines (IL-1β, CXCL1, CCL3, CCL5) and IL-1β, CXCL8/IL-8 and CCL2/MCP-1 by vaginal parenchyma [34], [35]. On ear tissues, application of BAK has been shown to release inflammatory cytokines such as TNF-α, IL-1β and MIP-1α and MIP-1β [36].…”

L-Selectin and P-Selectin Are Novel Biomarkers of Cervicovaginal Inflammation for Preclinical Mucosal Safety Assessment of Anti-HIV-1 Microbicide

“…While N9, BZK, and SDS were selected as positive controls in models of vaginal irritation and inflammation, as reported in previous works (2,9,13,15,16), carboxymethyl cellulose (CMC; which is widely used as a microbicide excipient and control) and TFV (a microbicide candidate which is proven to be efficacious and safe [1]) were chosen as negative controls. N9 and TFV were purchased from LKT Laboratories, Inc., and Molekula Limited (United Kingdom), respectively.…”

“…Most interestingly, we also found that three adhesion molecules, L-selectin, P-selectin, and VCAM-1, exhibited significant increases after N9 treatment. In previous studies of soluble biomarkers for microbicidal safety, most attention was focused on proinflammatory cytokines and chemokines, while little attention was given to soluble adhesion molecules (2,9,13,15,16,39). Actually, it is well documented that adhesion molecules play a pivotal role in the onset and progress of inflammation and inflammatory diseases (5,23,25).…”

“…In fact, a number of animal and human studies have produced evidence that the levels of proinflammatory cytokine interleukin 1 (IL-1), IL-6, and chemokines IL-8, monocyte chemotactic protein 1 (MCP-1), and macrophage inflammatory protein 3␣ (MIP-3␣) in cervicovaginal lavage (CVL) fluid positively correlate with histopathological and clinical signs of vaginal inflammation (2,9,(13)(14)(15)(16). Accordingly, these cytokines/chemokines have been defined as biomarkers of cervicovaginal inflammation.…”