Les complications des lymphadénectomies coelioscopiques pelviennes et lombo-aortiques en oncologie gynécologique

De décembre 1988 à mars 2004, 915 patientes ont fait l'objet d'une lymphadénectomie pelvienne et/ou aortique sous contrôle coelioscopique. Les interventions ont été réalisées dans le cadre d'une (re)stadification pour 98 cancers débutants de l'ovaire, 237 cancers du col aux stades précoces et 216 cancers avancés du col. Elles ont été associées à un traitement chirurgical radical ou conservateur (trachélectomie), par coelioscopie ou voie vaginale, dans 161 cas de cancer de l'endomètre et 203 cancers du col utérin. Un total de 1102 lymphadénectomies pelviennes ou aortiques a été pratiqué : 714 pelviennes (694 transpéritonéales, 20 extra péritonéales) et 388 aortiques (154 transpéritonéales, 234 extra péritonéales. 17 laparotomies (1,85%) ont été nécessaires pour impossibilité technique ou complications. La technique peut être considérée comme sûre, avec un taux de complications non supérieur à celui de la laparotomie, et une mortalité jusqu'ici nulle.TOULOUSE3-BU Santé-Centrale (315552105) / SudocTOULOUSE3-BU Santé-Allées (315552109) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Human Cytomegalovirus Infection Elicits New Decidual Natural Killer Cell Effector Functions

International audienceDuring the first trimester of pregnancy the uterus is massively infiltrated by decidual natural killer cells (dNK). These cells are not killers, but they rather provide a microenvironment that is propitious to healthy placentation. Human cytomegalovirus (HCMV) is the most common cause of intrauterine viral infections and a known cause of severe birth defects or fetal death. The rate of HCMV congenital infection is often low in the first trimester of pregnancy. The mechanisms controlling HCMV spreading during pregnancy are not yet fully revealed, but evidence indicating that the innate immune system plays a role in controlling HCMV infection in healthy adults exists. In this study, we investigated whether dNK cells could be involved in controlling viral spreading and in protecting the fetus against congenital HCMV infection. We found that freshly isolated dNK cells acquire major functional and phenotypic changes when they are exposed to HCMV-infected decidual autologous fibroblasts. Functional studies revealed that dNK cells, which are mainly cytokines and chemokines producers during normal pregnancy, become cytotoxic effectors upon their exposure to HCMV-infected autologous decidual fibroblasts. Both the NKG2D and the CD94/NKG2C or 2E activating receptors are involved in the acquired cytotoxic function. Moreover, we demonstrate that CD56 pos dNK cells are able to infiltrate HCMV-infected trophoblast organ culture ex-vivo and to co-localize with infected cells in situ in HCMV-infected placenta. Taken together, our results present the first evidence suggesting the involvement of dNK cells in controlling HCMV intrauterine infection and provide insights into the mechanisms through which these cells may operate to limit the spreading of viral infection to fetal tissues

Natural cytotoxicity receptor splice variants orchestrate the distinct functions of human natural killer cell subtypes

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Usutu Virus Infects Human Placental Explants and Induces Congenital Defects in Mice

International audienceUsutu virus (USUV) is a neurotropic mosquito-borne flavivirus that has dispersed quickly in Europe these past years. This arbovirus mainly follows an enzootic cycle involving mosquitoes and birds, but can also infect other mammals, causing notably sporadic cases in humans. Although it is mainly asymptomatic or responsible for mild clinical symptoms, USUV has been associated with neurological disorders, such as encephalitis and meningoencephalitis, highlighting the potential health threat of this virus. Among the different transmission routes described for other flaviviruses, the capacity for some of them to be transmitted vertically has been demonstrated, notably for Zika virus or West Nile virus, which are closely related to USUV. To evaluate the ability of USUV to replicate in the placenta and gain access to the fetus, we combined the use of several trophoblast model cell lines, ex vivo human placental explant cultures from first and third trimester of pregnancy, and in vivo USUV-infected pregnant mice. Our data demonstrate that human placental cells and tissues are permissive to USUV replication, and suggest that viral transmission can occur in mice during gestation. Hence, our observations suggest that USUV could be efficiently transmitted by the vertical route

Metabolic reprogramming by Zika virus provokes inflammation in human placenta

International audienceThe recent outbreak of Zika virus (ZIKV) was associated with birth defects and pregnancy loss when maternal infection occurs in early pregnancy, but specific mechanisms driving placental insufficiency and subsequent ZIKV-mediated pathogenesis remain unclear. Here we show, using large scale metabolomics, that ZIKV infection reprograms placental lipidome by impairing the lipogenesis pathways. ZIKV-induced metabolic alterations provide building blocks for lipid droplet biogenesis and intracellular membrane rearrangements to support viral replication. Furthermore, lipidome reprogramming by ZIKV is paralleled by the mitochondrial dysfunction and inflammatory immune imbalance, which contribute to placental damage. In addition, we demonstrate the efficacy of a commercially available inhibitor in limiting ZIKV infection, provides a proof-of-concept for blocking congenital infection by targeting metabolic pathways. Collectively, our study provides mechanistic insights on how ZIKV targets essential hubs of the lipid metabolism that may lead to placental dysfunction and loss of barrier function

Genotype specific pathogenicity of hepatitis E virus at the human maternal-fetal interface

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dNK cells are cytotoxic against HCMV-infected autologous fibroblasts.

<p>Decidual fibroblasts were kept uninfected or infected for 48 h with HCMV. dNK cell cytotoxicity was determined by ⁵¹Cr-release assay after 18 hours of contact at different E/T ratios. (A) Fibroblasts were infected with VHLE clinical isolate (n = 3), (B) cells infected with AD169 laboratory strain of HCMV (n = 5). (C) Analysis of dNK cell cytotoxicity from a cohort of 10 <i>decidua</i> samples at the 50 to 1 ratio. (D) dNK cells were pre-incubated with anti-FasL or (E) anti-TRAIL blocking mAbs at the final concentration of 10 µg/ml for 20 min and cytotoxicity was monitored after 18 h. Control (CTRL), lysis performed in the presence of IgG control. Each data point is calculated as the mean lysis ± S.D. from at least five independent experiments done in replicate tissue culture wells. Statistical comparisons of mean lysis of uninfected versus HCMV-infected were performed using two-way ANOVA test. ***, <i>p</i><0.001; **, <i>p</i><0.01; ns, not significant, <i>p</i>>0.05.</p

dNK cells infiltrate infected placental tissues.

<p>(A) Two-color 3D-images of chorionic <i>villi</i> explants organ cultures established from first trimester trophoblast either uninfected (AD169⁻) or HCMV-infected (AD169⁺). Nuclei were stained with dapi (cyan). Infiltrating dNK cells (red). Lower and side panels show orthogonal XZ and YZ slices, respectively. Images are from two-photon Z-stack (total of 200 µm). Scale bars = 100 µm. (B) Two-color IHC of 6-µm-thick sections from paraffin-embedded whole placental biopsies of HCMV⁺ pregnancy termination. HCMV⁺ and HCMV⁻ tissue sections are presented. Representative immunostaining of HCMV-IE (blue, alkaline phosphatase staining) and CD56⁺ NK cells (brown, peroxidase staining) (n = 2).</p

Polarization of the MTOC and lytic granules to the immune synapse formed with HCMV-infected fibroblasts.

<p>Uninfected (AD169⁻) or HCMV-infected (AD169⁺) decidual fibroblasts (F) plated on glass coverslips were incubated with autologous dNK cells (dNK) for 20 min at 37°C. (A) Representative images of maximum intensity projection. Microtubules (α-tubulin in green), lytic granules containing Perforin (red), HCMV-IE antigen (blue). Arrowhead points to the MTOC polarization (aster). Bar represent 20 µm. (B) Left cartoon shows schematic representation of the immunological synapse (IS), D (distance in µm). The center of IS was defined as the center of the contact zone between dNK and target cell (see cartoon, red line). Zero on the Y axis (µm) represents synaptic area; blue dot represents the microtubule organizing center (MTOC) and microtubules are in green. The MTOC polarization (Right graph) defined by the distance between the MTOC and the center of IS formed with uninfected (AD169⁻) and HCMV-infected (AD169⁺) fibroblasts. Distances were calculated for 50 conjugates from five independent experiments. Statistical analysis was performed using unpaired Student's <i>t</i>-test. ***, <i>p</i><0.001. (C) Percentage of conjugates showing polarized perforin containing granules to the NKIS. Results from 5 independent conjugations were averaged, values represent means and S.D.s. At least 300 conjugates were analyzed. Statistical analysis was performed using unpaired Student's <i>t</i>-test. *** <i>p</i> = 0.0002. (D) Kinetic of CD107a cell surface expression was analyzed by flow cytometry on dNK cells that were in contact with uninfected or AD169-infected autologous fibroblasts. Values presented in the bar graphs are mean values calculated from three independent experiments done in triplicates at the ratio 1 to 1. Error bars are SEM. Statistical comparisons were performed using unpaired Student's <i>t</i>-test, ** <i>p</i><0.01.</p

HCMV infection modulates dNK cells cytokine/chemokine production.

<p>dNK cells were stimulated with uninfected (AD169⁻, gray) or AD169-infected (AD169⁺, black) autologous decidual fibroblasts for 24 h. Cytokines were quantified in the supernatants using a 42-multi-plexed cytokine assay. Representative histograms from selected cytokines-chemokines that showed significant differences are presented. (A & B) Soluble factors that are produced at high levels by dNK cells. (C) Soluble factors that are produced at low levels by dNK cells. Concentrations are given as differences between secretions of dNK cell in presence of uninfected or infected fibroblasts and the corresponding uninfected or infected fibroblasts. Normalized data points are given as mean ± S.D. calculated as from four independent experiments. Statistical comparisons were performed using Mann & Whitney test. **, <i>p</i><0.01; *, <i>p</i><0.05.</p