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pplied by Flexercell tension system for SMCs and DCs. IL-8nd GRO� mRNA and protein expressions were measuredy RT-PCR and ELISA. Neutrophil chemotactic activity inonditioned media was evaluated by migration assay. Pro-MP1 was measured by ELISA. To evaluate the effect of

rogesterone, SMCs and DCs were pretreated with proges-erone for 24 h. The data showed the following: (1) In SMCs,yclic stretch significantly increased IL-8 and GRO� mRNAxpression (7.11 and 3.92 times, P < 0.05 for both) andheir protein production (5.54 and 3.62 times, P < 0.0001nd P < 0.01, respectively). In DCs, cyclic stretch signifi-antly increased IL-8 and GRO� mRNA expression (3.61 and.22 times, P < 0.05 for both) and their protein production4.65 and 3.08 times, P < 0.05 and P < 0.01, respectively). (2)upernatants from stretched cells had higher neutrophilhemotactic activity compared with control (2.53 and 6.88imes, in SMCs and DCs respectively) and this increase wasanceled by anti-IL8 and GRO� antibody. (3) Cyclic stretchignificantly increased pro-MMP1 production both in SMCs3.65 times) and DCs (5.26 times) (P < 0.05 for both). (4)hese effects of stretch were reduced by progesterone.hese results suggest that during the labor, cyclic stretchnduces chemokines in SMCs and DCs, which results ineutrophil infiltrations in the myometrium-decidual layer.tretch also activates MMPs, which may cause cervicalipening and rupture of membrane. These indicate thatterine contraction per se contributes to the augmentationf parturition. The inhibitory effects of progesterone mayxplain the therapeutic property of progestin for pretermabor.

oi:10.1016/j.jri.2010.06.060

14

iral infection sensitizes pregnancy to LPS leading toreterm labor

. Cardenas a,∗, P. Aldo a, R. Means b, S. Lang b, P. Stabach b,. Mor a

OBGYN, Yale University, New Haven, Connecticut, USAPathology, Yale University, New Haven, Connecticut, USA

Among pregnant women, an acquired viral infectionith a concurrent bacterial infection is a detrimental fac-

or associated to poor prognosis and death. Here, we usedmurine model to evaluate the effect of a viral infection

hat does not lead to preterm labor on the response to lowoses of LPS. Our objectives were: (1) to characterize theffect of a viral infection with a bacterial infection on preg-ancy outcome, (2) to characterize the placental and fetal

mmune responses to the viral sensitization to LPS, and (3)o characterize the mechanisms of viral sensitization to LPSuring pregnancy. C57B/6wt mice were injected intraperi-oneally with murine gammaherpesvirus 68 (MHV68) at8.5. Either PBS or LPS was injected i.p. at E15.5. Preg-ancy outcome, fetal weight, viral load (plaques forming

nits/ml of tissue) at the placenta, were determined at17.5. Placental morphology and immune cells infiltrationere evaluated by immunohistochemistry. Cytokine pro-le and TLR expression were determined by Multiplex andRT-PCR respectively. LPS treatment of MHV-68 infected

tive Immunology 86 (2010) 11–34 33

animals was found to induce preterm delivery and fetaldeath in 100% of the mice in less than 24 h comparedto 29% in the LPS treated mice without a previous viralinfection. Low doses of LPS (0.8 �g/kg) are able to inducea significant increase on inflammatory cytokines only inthe viral infected mice. Similarly, morphological evalua-tion of the placenta from MHV68 and LPS treated micerevealed necrosis and polymorphonuclear infiltration. Theobserved changes in LPS response correlated with viralinduced upregulation of TLR2, TLR4 and TLR6. We describefor the first time that a silent viral infection in pregnantmice sensitizes to a bacterial infection leading to pretermdelivery and IUGR. This effect is mediated by a specific reg-ulation of TLRs in the trophoblast and increased expressionof pro-inflammatory cytokines. Our data suggest that anongoing viral infection may increase the risk for pregnancycomplications due to subsequent bacterial infection.

doi:10.1016/j.jri.2010.06.061

O15

Development of a vaginally applied, non-hormonal con-traceptive: the contraceptive efficacy and impact onbone turnover of PEGLA, a long-acting LIF antagonist

E. Menkhorst a,∗, J.G. Zhang b, P.O. Morgan b, I.J. Poulton c,D. Metcalf b, L.A. Salamonsen a, N.A. Sims c, N.A. Nicola b, E.Dimitriadis a

a Prince Henry’s Institute, Clayton, VIC, Australiab Walter and Eliza Hall Institute, Parkville, VIC, Australiac St Vincent’s Institute of Medical Research, Melbourne, VIC,Australia

The WHO has called for the urgent development of phar-macological, non-hormonal contraceptives. Leukaemiainhibitory factor (LIF) is obligatory for embryo implan-tation in mice and associated with infertility in women.Injection of a long-acting LIF antagonist (PEGLA) blocksuterine LIF preventing implantation in mice, making PEGLAa promising non-hormonal contraceptive. LIF and LIFRnull mice show decreased bone volume associated withincreased osteoclast number and size, suggesting PEGLAmay target bone. Vaginally administered PEGLA could bea ‘dual-role’ contraceptive: carried in a microbicide thatblocks the vaginal transmission of sexually transmittedinfections. We aimed to establish the contraceptive efficacyof vaginally administered PEGLA and identify non-uterinetargets of PEGLA in mice. PEGLA was administered to matedfemale mice by intraperitoneal (IP) injection or vaginally(n = 4/group) during the peri-implantation period to deter-mine its effect on implantation and bone turnover. Thetissue and blood accumulation of 125I-PEGLA or controlwas identified at various time-points following IP injection(≤120 h) or vaginal administration (≤24 h) (n = 3/group).PEGLA administered via vaginal gel blocked implanta-tion (0.0 + 0.0 vs 8.5 + 0.5) at a lower dose (500 �g) than

IP injection (1500 �g). PEGLA administered by IP injec-tion resulted in fewer (4.0 + 0.3% vs 7.7 + 1.5%; p < 0.05)but larger (20.9 + 0.9 �m vs 18.1 + 0.5 �m; p < 0.05) osteo-clasts and increased trabecular bone volume (6.8 + 0.9% vs3.1 + 1.1%; p < 0.05) but vaginally administered PEGLA had