Takayoshi Okada

Production of interleukin-6 by normal human trophoblast.

The placenta plays a number of important roles during pregnancy, some of which might be mediated by cytokines with multiple activities such as IL-6. Using an IL-6-dependent cell line, MH6o.BSF2, we showed that the placenta released IL-6 into the culture supernatant. Analysis of single-cell suspensions of placental cells determined the major source of IL-6 to be trophoblast. Using a mouse monoclonal antibody specific for IL-6 (alpha BSF2-I66), immuno-histochemical analysis of placental specimens demonstrated the localization of IL-6 only in the trophoblast layer. Additional immunocytochemical studies with single-cell suspensions of trophoblasts demonstrated the preferential presence of IL-6 molecules in syncytiotrophoblasts rather than cytotrophoblasts. The evidence that a high titer of IL-6 is produced spontaneously by syncytiotrophoblasts indicates that IL-6 may play immunological roles in fetomaternal interactions by means of IL-6-driven multiple immunoregulatory activities.

Placental interleukin-6 production is enhanced in intrauterine infection but not in labor

OBJECTIVE Because interleukin-6 is an important mediator in the host defense mechanism against infection and tissue damage, we studied the capacity of placentas with or without either labor or chorioamnionitis in the third trimester to produce interleukin-6. STUDY DESIGN The placental blocks were cultured, and their interleukin-6 titers were measured by a bioassay. RESULTS Placentas with labor produced a similar amount of interleukin-6 to placentas without labor. In contrast, placentas with chorioamnionitis produced much more interleukin-6 than the placentas with or without labor (p < 0.0001). CONCLUSION Placental interleukin-6 is thus surmised to participate in potentiation of the placental and fetomaternal defense mechanisms together with placental interleukin-1 during chorioamnionitis.

Fetal mononuclear cells show a comparable capacity with maternal mononuclear cells to produce IL-8 in response to lipopolysaccharide in chorioamnionitis

IL-8 is a chemotactic and activating cytokine for neutrophils which eliminate invading bacteria by releasing bactericidal metabolites. Cord blood mononuclear cells (CBMCs) obtained from neonates born to mothers with chorioamnionitis actively produced a significantly higher amount of IL-8 than those of neonates without chorioamnionitis, suggesting that the mononuclear cells of fetuses with chorioamnionitis had been activated in utero. As lipopolysaccharide (LPS) can often be detected in the uteroplacental space in chorioamnionitis, the LPS-mediated activation mechanism of neonatal mononuclear cells was analyzed in vitro to produce IL-8. Neonatal mononuclear cells stimulated with LPS increased IL-8 production in a time- and dose-dependent manner. The ability of term or preterm neonatal mononuclear cells to produce IL-8 was comparable with that of adult (maternal) mononuclear cells, suggesting functional maturity of the neonatal or fetal mononuclear cells to produce IL-8. However, IL-8 production by neonatal CBMCs was down-regulated by dexamethasone, a glucocorticoid which is clinically administered to mothers to promote fetal lung maturity in preterm delivery. Our present study revealed a regulatory mechanism of fetal IL-8 production, suggesting that functionally mature fetal mononuclear cells produce IL-8 in response to LPS in chorioamnionitis and activate the fetal defense mechanism against infection.

Detection of monocyte chemotactic and activating factor (MCAF) and interleukin (IL)-6 in human seminal plasma and effect of leukospermia on these cytokine levels

PROBLEM: To demonstrate whether monocyte chemotactic and activating factor (MCAF) and interleukin‐6 (IL‐6) are present in the seminal plasma, and whether these presence is modulated by leukospermia.

Trophoblast-Derived Immunoregulatory Factor: Demonstration of the Biological Function and the Physicochemical Characteristics of the Factor Derived From Choriocarcinoma Cell Lines

ABSTRACT: An immunosuppressive factor released by choriocarcinoma cell lines was analyzed in the present study. It inhibited the proliferative responses of human T cells stimulated by lectins or alloantigens. It also blocked the generation of alloreactive cytotoxic T cells. The suppressive activity of the factor was detected in the responses of the T cells costimulated with 1 nM 12‐0‐tetradecanoyl phorbol 13‐acetate and 1 μM A23187, suggesting the possibility that the factor acted on the intracellular signal transduction in T cells rather than interfering with early events such as T cell receptor signal transduction through cell membranes. Moreover, the factor acted directly on T cell proliferation pathways without activation of suppressor cells but did not act on T cell activation pathways. Taken together, all these findings expanded our previous reports on a factor released by normal trophoblasts, indicating the possible identity of the two factors. The physico‐chemical properties of the choriocarcinoma‐derived factor were examined, and the biological significance of the factor during pregnancy was discussed in this paper.

Analysis of site of action of a choriocarcinoma-derived immunoregulatory factor on IL-2-mediated T cell responses

We have investigated the functional ability of a choriocarcinoma-cell-derived factor to block human T cell responses and the factors immunoregulatory site of action on the T cell signal transduction pathway. The factor completely suppressed human T cell responses activated by phorbol ester and calcium ionophore, reagents which strongly stimulate IL-2-mediated T cell responses. It failed to inhibit CD 25 expression and IL-2 production by T cell blasts in the T cell activation phase, but completely blocked recombinant IL-2-induced proliferation of T cell blasts in the T cell proliferation phase. Absorption experiments with the factor and Con A-induced T cell blasts as well as [125I]IL-2 binding experiments with T cell blasts revealed that the factor acted on the physiological events occurring after IL-2-mediated stimulation of IL-2 receptor complexes, demonstrating no interaction of the factor with either IL-2 molecules or IL-2 receptor complexes. Moreover, it suppressed murine IL-2 dependent T cell line proliferation, suggesting the presence of common pathways in human and murine T cell proliferation. The biological and immunological significance of the factor during pregnancy and in the immunosuppressed tumor-bearing hosts are discussed.

Demonstration of functional immaturity of signal transduction pathways in human cord T cells

We examined the nature of cytoplasmic signal transduction pathways in cord blood T cells by stimulating them with tumor promoter (TPA) and calcium ionophore (A23187). Costimulation of T cells with TPA and A23187 induced optimal proliferative responses on Day 2 in cord T cells but on Day 4 in adult T cells. The maximal responses observed in cord T cells were much less than those of adult T cells, whereas the Con A-induced proliferative responses of these cells showed no significant differences. The reduced responses of cord T cells were due to their lower efficiency in activating the cellular events in T cell activation and proliferation phase, because cord T cells have significantly less ability than adult T cells to express IL-2 receptor as well as HLA-DR and produce IL-2 molecules, thereby inducing proliferation. These data show immature characteristics of intracellular signal transduction pathways in cord T cells, which are directly related with the functional immaturity of cord T cells.

Analysis of immunoregulatory activity of a choriocarcinoma-derived factor: specific suppression of proliferative process of cell-mediated immune responses including LAK cell generation

The immunosuppressive activity of a JEG-3 choriocarcinoma-derived factor in human IL-2-dependent T cell responses has been studied, together with its effect on IL-2-independent T cell responses induced by 10 nM TPA. The factor completely suppressed the IL-2-independent proliferative responses of T cells but failed to suppress antigenic expression of activation-associated CD 25 molecules. Further studies examined the effect of the factor on LAK cell generation induced by rIL-2. Recombinant IL-2-induced LAK cell proliferation was observed on Day 4 and Day 5, but not on Day 3. As the factor suppressed the responses of LAK cell proliferation, we tested whether it blocked the generation of Day 3, Day 4 and Day 5 LAK cells. The addition of the factor failed to suppress the generation of Day 3 LAK cells, while it partially suppressed the lytic activity of Day 4 LAK cells and completely suppressed that of Day 5 LAK cells. The data suggest the presence of a heterogeneous pattern for LAK cell generation; one without proliferation, but the other requiring proliferation, to acquire killer activity. Taken together with the evidence that the factor failed to suppress NK activity, the choriocarcinoma-derived factor suppressed only the proliferative events of immunocompetent cells, but inhibited neither their activation nor the differentiation events. This immunosuppressive factor might be involved in the prevention of host-mediated rejection of choriocarcinoma cells or maternal rejection of the fetus.