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In vitro secretion profiles of interleukin (IL)-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes.

Zaga-Clavellina V, Garcia-Lopez G, Flores-Herrera H, Espejel-Nuñez A, Flores-Pliego A, Soriano-Becerril D, Maida-Claros R, Merchant-Larios H, Vadillo-Ortega F - Reprod. Biol. Endocrinol. (2007)

Bottom Line: When the amnion was stimulated directly, the level of IL-1beta (13-fold) rose significantly; however, the increase in IL-8 secretion was larger (20-fold), regardless of the primary site of infection.This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending intrauterine infection, being the main barrier to progression of the infection into the amniotic cavity.Therefore, the tissue-specific capacities for the secretion of these immune modulators could be a determining factor for the degree of severity of the inflammation process in fetal membranes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Biomedical Research Branch, Instituto Nacional de Perinatologia Isidro Espinosa de los Reyes, México City, México. zaga@universo.com

ABSTRACT

Background: Chorioamniotic membranes infection is a pathologic condition in which an abnormal secretion of proinflammatory cytokines halts fetal immune tolerance. The aim of the present study was to evaluate the functional response of human chorioamniotic membranes, as well as the individual contribution of the amnion and choriodecidua after stimulation with Escherichia coli, a pathogen associated with preterm labor.

Methods: Explants of chorioamniotic membranes from 10 women (37-40 weeks of gestation) were mounted and cultured in a Transwell system, which allowed us to test the amnion and choriodecidua compartments independently. Escherichia coli (1 x 10 6 CFU/mL) was added to either the amniotic or the choriodecidual regions or both; after a 24-h incubation, the secretion of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10 in both compartments was measured using a specific ELISA. Data were analyzed by Kruskal-Wallis one-way analysis of variance.

Results: After stimulation with Escherichia coli, the choriodecidua compartment showed an increase in the secretion of IL-1beta (21-fold), IL-6 (2-fold), IL-8 (6-fold), and IL-10 (37-fold), regardless of which side of the membrane was stimulated; TNFalpha secretion augmented (22-fold) also but only when the stimulus was applied simultaneously to both sides. When the amnion was stimulated directly, the level of IL-1beta (13-fold) rose significantly; however, the increase in IL-8 secretion was larger (20-fold), regardless of the primary site of infection. TNFalpha secretion in the amnion compartment rose markedly only when Escherichia coli was simultaneously applied to both sides.

Conclusion: Selective stimulation of fetal membranes with Escherichia coli results in a differential production of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10. These tissues were less responsive when the amnion side was stimulated. This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending intrauterine infection, being the main barrier to progression of the infection into the amniotic cavity. Therefore, the tissue-specific capacities for the secretion of these immune modulators could be a determining factor for the degree of severity of the inflammation process in fetal membranes.

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Experimental model. The fetal membranes are held together with silicone rubber rings in the upper chamber of a Transwell® device. In this system, the choriodecidua region delimits the upper chamber and the amnion region, the lower chamber.
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Figure 1: Experimental model. The fetal membranes are held together with silicone rubber rings in the upper chamber of a Transwell® device. In this system, the choriodecidua region delimits the upper chamber and the amnion region, the lower chamber.

Mentions: The chorioamniotic membranes were cut at a distance of 5–6 cm from the placental disc, transported to the laboratory in sterile Dulbecco Modified Eagle Medium (DMEM; Gibco BRL, Bethesda, MD), and rinsed in sterile saline solution to remove adherent blood clots. Segments representing all zones of membranes were manually cut into 18 mm diameter discs and held together with silicone rubber rings to be placed on the upper chamber of a Transwell® system (CORNING, New York, NY) from which the original polycarbonate membrane had been previously removed. In this model the upper chamber of the Transwell® system is delimited by choriodecidual tissue and the lower chamber by amniotic tissue, which allowed testing the two compartments independently (Figure 1). A detailed description and validation of this model has been published previously [13]. One milliliter of DMEM (Gibco BRL), supplemented with 10% fetal calf serum (FCS), 1 mM sodium pyruvate, and 1× antibiotic-antimycotic solution (penicillin 100 U/mL, streptomycin 100 μg/mL) (DMEM-FCS) was added to each compartment. The mounted explants were placed in a 12-well tissue culture plate (CORNING, New York, NY) and incubated in 5% CO2 at 37°C for 24 h.


In vitro secretion profiles of interleukin (IL)-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes.

Zaga-Clavellina V, Garcia-Lopez G, Flores-Herrera H, Espejel-Nuñez A, Flores-Pliego A, Soriano-Becerril D, Maida-Claros R, Merchant-Larios H, Vadillo-Ortega F - Reprod. Biol. Endocrinol. (2007)

Experimental model. The fetal membranes are held together with silicone rubber rings in the upper chamber of a Transwell® device. In this system, the choriodecidua region delimits the upper chamber and the amnion region, the lower chamber.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2175507&req=5

Figure 1: Experimental model. The fetal membranes are held together with silicone rubber rings in the upper chamber of a Transwell® device. In this system, the choriodecidua region delimits the upper chamber and the amnion region, the lower chamber.
Mentions: The chorioamniotic membranes were cut at a distance of 5–6 cm from the placental disc, transported to the laboratory in sterile Dulbecco Modified Eagle Medium (DMEM; Gibco BRL, Bethesda, MD), and rinsed in sterile saline solution to remove adherent blood clots. Segments representing all zones of membranes were manually cut into 18 mm diameter discs and held together with silicone rubber rings to be placed on the upper chamber of a Transwell® system (CORNING, New York, NY) from which the original polycarbonate membrane had been previously removed. In this model the upper chamber of the Transwell® system is delimited by choriodecidual tissue and the lower chamber by amniotic tissue, which allowed testing the two compartments independently (Figure 1). A detailed description and validation of this model has been published previously [13]. One milliliter of DMEM (Gibco BRL), supplemented with 10% fetal calf serum (FCS), 1 mM sodium pyruvate, and 1× antibiotic-antimycotic solution (penicillin 100 U/mL, streptomycin 100 μg/mL) (DMEM-FCS) was added to each compartment. The mounted explants were placed in a 12-well tissue culture plate (CORNING, New York, NY) and incubated in 5% CO2 at 37°C for 24 h.

Bottom Line: When the amnion was stimulated directly, the level of IL-1beta (13-fold) rose significantly; however, the increase in IL-8 secretion was larger (20-fold), regardless of the primary site of infection.This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending intrauterine infection, being the main barrier to progression of the infection into the amniotic cavity.Therefore, the tissue-specific capacities for the secretion of these immune modulators could be a determining factor for the degree of severity of the inflammation process in fetal membranes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Biomedical Research Branch, Instituto Nacional de Perinatologia Isidro Espinosa de los Reyes, México City, México. zaga@universo.com

ABSTRACT

Background: Chorioamniotic membranes infection is a pathologic condition in which an abnormal secretion of proinflammatory cytokines halts fetal immune tolerance. The aim of the present study was to evaluate the functional response of human chorioamniotic membranes, as well as the individual contribution of the amnion and choriodecidua after stimulation with Escherichia coli, a pathogen associated with preterm labor.

Methods: Explants of chorioamniotic membranes from 10 women (37-40 weeks of gestation) were mounted and cultured in a Transwell system, which allowed us to test the amnion and choriodecidua compartments independently. Escherichia coli (1 x 10 6 CFU/mL) was added to either the amniotic or the choriodecidual regions or both; after a 24-h incubation, the secretion of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10 in both compartments was measured using a specific ELISA. Data were analyzed by Kruskal-Wallis one-way analysis of variance.

Results: After stimulation with Escherichia coli, the choriodecidua compartment showed an increase in the secretion of IL-1beta (21-fold), IL-6 (2-fold), IL-8 (6-fold), and IL-10 (37-fold), regardless of which side of the membrane was stimulated; TNFalpha secretion augmented (22-fold) also but only when the stimulus was applied simultaneously to both sides. When the amnion was stimulated directly, the level of IL-1beta (13-fold) rose significantly; however, the increase in IL-8 secretion was larger (20-fold), regardless of the primary site of infection. TNFalpha secretion in the amnion compartment rose markedly only when Escherichia coli was simultaneously applied to both sides.

Conclusion: Selective stimulation of fetal membranes with Escherichia coli results in a differential production of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10. These tissues were less responsive when the amnion side was stimulated. This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending intrauterine infection, being the main barrier to progression of the infection into the amniotic cavity. Therefore, the tissue-specific capacities for the secretion of these immune modulators could be a determining factor for the degree of severity of the inflammation process in fetal membranes.

Show MeSH
Related in: MedlinePlus