Intercellular adhesion molecule (ICAM)-1-mediated cell-cell adhesion is essential for various immunological functions, including natural killer (NK) cell-mediated cytotoxicity against endometrium. The present study was designed to establish whether shedding of ICAM-1 from cultured endometrial stromal cells occurred and to characterize its potential functional significance in endometrial physiology and pathology. The shed sICAM-1 molecule was detected and quantified in supernatants from endometrial stromal cultures and in peritoneal fluid by a specific enzyme-linked immunosorbent assay. The results of this study indicate that cultured endometrial stromal cells constitutively shed ICAM-1 from their surface. This ability is regulated during the menstrual cycle, as it appears to be higher in the proliferative than in the secretory phase of the cycle (16.93 +/- 2.2 and 7.7 +/- 1.76 ng/ml respectively). In order to evaluate whether the release of sICAM-1 could interfere with cell-mediated lysis of endometrium we compared the determinations of sICAM-1 in endometrial supernatants with the ability of such supernatants to suppress NK cell-mediated cytotoxicity toward endometrial targets. A significant correlation (r = 0.6, P < 0.05) was found between the sICAM-1 concentration in endometrial supernatants and the percentage of inhibition of NK cell-mediated lysis exerted by the same supernatant samples. Finally, endometrial stromal shedding of sICAM-1 appears to be related to endometriosis since endometrial stromal cultures obtained from patients with advanced stages of the disease released significantly higher amounts of the soluble protein compared to the control group (P < 0.05). sICAM-1 is a soluble molecule which can interfere with immunological functions, and its shedding may be one of the mechanisms by which refluxed endometrial cells escape immunosurveillance.
Nineteen pregnant women with uncomplicated pregnancies were studied during the first, second, and third trimesters. We measured the following hemostatic parameters: prothrombin time, activated partial thromboplastin time, fibrinogen, antithrombin III, protein C, protein S, platelet number and volume. Platelet function was examined by a cytofluorimetric method, using an anti-GPM-140 antibody which is directed against a platelet alpha granule membrane protein. Activated platelets were expressed as a percentage of the GMP-140-positive platelets over total platelets. Fibrinogen levels showed a steady increase during pregnancy; conversely prothrombin time, activated partial thromboplastin time, protein C, and antithrombin III showed no significant modifications and remained within the reference range. There was a decrease of protein S activity throughout pregnancy, although protein S antigen did not follow this trend. The decrease occurred early in pregnancy and persisted during the second and third trimesters, reaching a stable plateau. We observed no platelet volume change or activation: the percentage of activated platelets was within the normal reference range, even in late pregnancy.
Platelet function was investigated in full-term infants on the first, the fourth and the tenth days of life and compared to normal adult controls. Platelet function was analyzed through a new cytofluorimetric technique with two murine monoclonal antibodies, PAC-1 and anti-GMP-140, directed against two membrane proteins expressed on the activated platelets’ surface. The percentage of activated platelets detected with PAC-1 and anti-GMP-140 was evaluated at basal condition and after in vitro stimulation with a weak agonist (ADP) and a strong Txa2 analogue inducer (U 46619). At day 1 platelet activation at basal condition was negligible and similar to adult controls both with PAC-1 (1.2 vs. 1.1%) and anti-GMP-140 (2.6 vs. 3.3%). On the contrary, after ADP stimulation the percentage of PAC-1-positive activated platelets was significantly reduced in neonates compared to adults (22 vs. 66%; p < 0.001) and even more after U 46619 (11 vs. 72%; p < 0.001). The percentage of anti-GMP-140-positive activated platelets behaved similarly after adding both ADP (26 vs. 46%; p < 0.01) and U 46619 (37 vs. 67%; p < 0.001). The reduced platelet activation after ADP and U 46619 persisted at day 4 both with PAC-1 and with anti-GMP-140. On the contrary, at day 10 newborn platelets analyzed with anti-GMP-140 behaved similarly to the adult ones both at basal condition and after stimulation with ADP or U 46619 (6 vs. 3% at basal state, 42 vs. 46% after ADP addition, and 55 vs. 67% after U 46619). These data demonstrate that the reduced platelet activation present in newborns is restored by the tenth day after birth.
Peripheral blood lymphocytes from fetal, term and adult subjects were analyzed with a panel of B- and T-lymphoid differentiation markers. CDlc +ve and CD5 +ve B cell (IgM +ve) percentage in fetal, cord and adult blood decreased significantly. Fetal and cord blood contained less CD2+ and CD 3+ T lymphocytes and a higher percentage of CD38 +ve cells than adult blood. The phenotypic changes occurring during perinatal age in humans relate to the changes in the functional activities of the B and T lymphoid subsets from fetal to postnatal age.
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