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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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In vitro secretion of IL-6 in amnion and choriodecidua regions after selective infection with Escherichia coli. IL-6 secreted to the culture medium after 24 h of infection with 1 × 106CFU of E. coli. Data were normalized in function of protein concentration (pg/μg protein) and each bar represents the 95% confidence intervals and the median (solid line) of 10 different experiments. Significant difference between basal and stimulated values is indicated (*P < 0.05) C. Choriodecidua; A. Amnion.
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Figure 3: In vitro secretion of IL-6 in amnion and choriodecidua regions after selective infection with Escherichia coli. IL-6 secreted to the culture medium after 24 h of infection with 1 × 106CFU of E. coli. Data were normalized in function of protein concentration (pg/μg protein) and each bar represents the 95% confidence intervals and the median (solid line) of 10 different experiments. Significant difference between basal and stimulated values is indicated (*P < 0.05) C. Choriodecidua; A. Amnion.

Mentions: IL-6 (Figure 3) and IL-10 (Figure 4) showed a similar secretion profile, with the choriodecidua as the most active tissue after Escherichia coli stimulation whether applied directly or indirectly to this membrane (P < 0.05). In both cases, the largest response was observed when the stimulus was applied directly in the choriodecidual zone (13.45 [9.39–18.3] and 32.38 [23.6–44.2] pg/μg protein, respectively). Again, the amnion showed only a mild but significant increase in the secretion of IL-6 and IL-10 (10.8 [8.3–16.75] and 15.97 [13.02–21.49] pg/μg protein, respectively).


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)

In vitro secretion of IL-6 in amnion and choriodecidua regions after selective infection with Escherichia coli. IL-6 secreted to the culture medium after 24 h of infection with 1 × 106CFU of E. coli. Data were normalized in function of protein concentration (pg/μg protein) and each bar represents the 95% confidence intervals and the median (solid line) of 10 different experiments. Significant difference between basal and stimulated values is indicated (*P < 0.05) C. Choriodecidua; A. Amnion.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: In vitro secretion of IL-6 in amnion and choriodecidua regions after selective infection with Escherichia coli. IL-6 secreted to the culture medium after 24 h of infection with 1 × 106CFU of E. coli. Data were normalized in function of protein concentration (pg/μg protein) and each bar represents the 95% confidence intervals and the median (solid line) of 10 different experiments. Significant difference between basal and stimulated values is indicated (*P < 0.05) C. Choriodecidua; A. Amnion.
Mentions: IL-6 (Figure 3) and IL-10 (Figure 4) showed a similar secretion profile, with the choriodecidua as the most active tissue after Escherichia coli stimulation whether applied directly or indirectly to this membrane (P < 0.05). In both cases, the largest response was observed when the stimulus was applied directly in the choriodecidual zone (13.45 [9.39–18.3] and 32.38 [23.6–44.2] pg/μg protein, respectively). Again, the amnion showed only a mild but significant increase in the secretion of IL-6 and IL-10 (10.8 [8.3–16.75] and 15.97 [13.02–21.49] pg/μg protein, respectively).

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