Lipopolysaccharide induces normal human term placentae to secrete high levels of TNFa and IL-6

Lipopolysaccharide induces normal human term placentae to secrete high levels of TNFa and IL-6

Holcberg, M. Huleihe, O. Sapir, S. Levi, M. Katz, L. Myatt,

M. Mazor department of Obstetrics and Gynecology, Soroka University Medical Center; department of Microbiology & Immunology, Faculty of Health Sciences, Ben Gurion University of the Negev, Israel; 3University of Cincinnati, College of Medicine, Cincinnati, OH, USA

Objective. To examine the stimulatory effect of lipopolysaccha- ride (LPS) on IL-6 and TNFa secretion in the isolated cotyledons of human term placentas.

Study design. Isolated placental cotyledons were dually perfused in 5 normal term placentas obtained after birth. LPS (100 ng/kg) was perfused into the maternal site during 10 hours. Perfusate samples from fetal and maternal sites were collected every 30 min. The perfusates were examined by ELISA for IL-6 and TNFa levels. Statistical significance was determined by paired t test and ANOVA.

Results. A significant increase in IL-6 and TNFa concentrations was observed within 8—10 hours of perfusion in maternal and fetal circulation in term placentas perfused with either or without LPS. The increase of TNFa and IL-6 in perfusate was time course dependent. The TNFa and IL-6 levels were significantly higher in maternal compartment after exposure to LPS as compared to control placentas (from 421 ± 217 pg/ml to 560 ± 202 pg/ml, p < 0,05 and from 971 ± 390 pg/ml to 2136 ± 688 pg/ml respectively, p < 0,05). The same pattern of LPS stimulation of TNFa and IL-6 was shown in the fetal compartment of the term placentas (from 20 ± 9 pg/ml to 53 ± 15 pg/ml, p < 0,05 and from 90 ± 39 pg/ml to 388 ± 154 pg/ml, p < 0,05 respectively).

Conclusions. In normal human placentas the elevation of TNFa and IL-6 is time course dependent and increases after 10 hours of perfusion. LPS had significant increased the capacity of term placentas to secrete TNFa and IL-6. Thus, LPS may affect parturition by mechanisms that involve the regulation of cytokine production.

Elevation of TNFa and IL-6 levels in preterm human placentae compared to term placentae

M. Huleihel, G. Holcberg, O. Sapir, S. Levi, M. Katz, L. Myatt,

M. Mazor department of Microbiology and Immunology; department of Obstetrics and Gynecology, Soroka University Medical Center, Faculty of Health Sciences, Ben- Gurion University of the Negev, Beer-Sheva, Israel; 3University of Cincinnati, College of Medicine, Cincinnati, OH, USA

Aim of the study. To determine the concentrations of TNFa and IL6 in the perfusate of isolated human placental cotyledon from term and preterm deliveries.

Study design. Isolated placental cotyledons were dually perfused in 5 term placentas and 4 placentas obtained after preterm birth (28-34 W of gestation). Perfusate samples from fetal and maternal sites were collected every 30 min (for 10 hours) and were examined by ELISA for IL-6 and TNFa levels. Statistical analysis was performed by ANOVA and paired t test.

Results. A significant increase of TNFa and IL-6 concentrations was observed in perfusate of maternal and fetal circulation in term and preterm placentas during 10 hours of perfusion. The increase of these cytokines in perfusate was time course dependent. The TNFa levels were significantly higher in maternal compartment in both groups (421 ± 217 and 522 ± 179 pg/ml respectively) as compared to fetal compartment (20 ± 9 and 65 ± 25 pg/ml respectively, p < 0,05). Moreover the TNFa levels were significantly higher in perfusate of preterm placentas in maternal and fetal compartments (522 ±179 and 65 ± 25 pg/ml respectively) as compared to term placentas (20 ± 9 and 421 ±217 pg/ml respectively, p < 0,05) after 10 hours of perfusion. The IL-6 levels were significantly higher in preterm placentae as compared to term placentae in maternal and fetal compartments (1945 ± 688 pg/ml and 971 ± 390 pg/ml in maternal compartment respectively, and 419 ±154 pg/ml and 90 ± 39 pg/ml in fetal compartment respectively p < 0,05).

Conclusions. Perfusate of maternal sites from term and preterm placentas show higher levels of TNFa and IL-6 than of the fetal sites. Perfusates of preterm placentas shows higher levels of TNFa and IL-6 than term placentas. Our results may indicate the involvement of these cytokines in the process of the preterm parturition.

Distinct expression of pro-inflammatory cytokines and their modulators (IL-1Ra and IL-10) in testicular tissue cells of fertile and infertile men

M. Huleihel1, E. Lunenfeld2, V. Dyomin3, N. Vardi3, I. Ynai-Inbar3, L. Harosh1,

Potashnik2

department of Microbiology and Immunology; department of Obstetrics and

Gynecology; 3Pathology Institute, Soroka University Medical Center, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel

Introduction. Cytokines are produced and involved in the regulation of testicular cell functions.

Our aim: to evaluate the levels of inflammatory cytokines (IL- 1a, IL-10, IL-6 and TNFa) and their down-regulators (IL-1Ra and IL-10) in testicular biopsies of fertile and infertile men.

Methods. Biopsies from 6 fertile men, 6 infertile men with incomplete maturation arrest and 6 infertile men with Sertoli only syndrom men were examined by immunohistochemical staining for the expression levels and cellular localization of IL-1a, IL-10, IL-6, TNFa, IL-1Ra and IL-10.

Results see in table.

Conclusions. This is the first report showing expression of im- muno-regulatory cytokines in human testicular tissues. IL-1a, IL- 1Ra, IL-6 and TNFa are differently expressed in testicular cells of pathological conditions as compared to normal. The distinct expression of the cytokines in the different testicular cells may indicate their involvement in the function of these cells. The expression of high levels of IL-10 in testicular tissues may suggest its involvement in a general suppression mechanism against autoimmune response against sperm cells in the testicular compartment. The mechanism by which cytokines involved in male fertility is

Table 1

L — Leydig cells; S — Sertoli cells; G — Germ cells; (+) intensity of staining.

not yet clear, however our results may indicate that some cytokines may be involved in the regulation of testicular functions, under physiological and pathological conditions, and thus could be involved in male fertility.