Human choriocarcinoma JAR cells constitutively express pro-interleukin-1beta that can be released with Fc(gamma) receptor engagement.

Some authors have suggested that fetally derived syncytiotrophoblasts, which form the barrier between mother and the fetus, are an integral part of a complex macrophage-cytokine network involving maternal leukocytes, decidual cells, placental tissues, and even the fetus itself. We report here that syncytiotrophoblast-like JAR cells, a human choriocarcinoma cell line, share another feature common to cells of the monocyte-macrophage lineage, the ability to secrete IL-1beta when stimulated through their Fc(gamma) receptors. We incubated JAR cells with physiologically relevant concentrations of model BSA-rabbit IgG-anti-BSA immune complexes or monomeric rabbit IgG for periods of up to 72 h. Both monomeric IgG and immune complexes induced IL-1beta from JAR cells, although levels produced by immune complexes were approximately twice those induced by monomeric IgG. IL-1beta secretion was not inhibited by cycloheximide, and Western blots of JAR cell lysates using pro-IL-1beta MAb revealed constitutive expression of a 31-kD protein, whose levels declined within 2 h of stimulation by either IgG or immune complexes, but returned to baseline within 18 h.

In vitro induction of proinflammatory cytokine secretion by juvenile rheumatoid arthritis synovial fluid immune complexes.

OBJECTIVE: To characterize juvenile rheumatoid arthritis synovial fluid (SF) immune complexes and to examine their interaction with leukocytes. METHODS: SF immunoglobulin-containing fractions were prepared by sequential chromatography on protein A and Sephacryl 300. Fractions were subdivided according to molecular weight, characterized for immunoglobulin and complement content, and incubated with either promonocytic U937 cells or normal human peripheral blood mononuclear cells (PBMC). RESULTS: High molecular weight SF immunoglobulin-containing fractions stimulated the release of interleukin-1beta (IL-1beta) from U937 cells. These same complexes stimulated tumor necrosis factor alpha (TNFalpha), IL-1beta, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) from PBMC. Lower molecular weight material was less efficient in inducing any of the cytokines. TNFalpha and IL-1beta were the earliest of the messenger RNAs examined to be induced by the high molecular weight complexes. However, the secretion of IL-6, IL-8, and GM-CSF stimulated by the complexes was not completely dependent upon the secretion of IL-1beta. Addition of IL-1 receptor antagonist to the cell cultures reduced GM-CSF and IL-6 production by 40% and IL-8 production by 25% in PBMC. CONCLUSION: SF immunoglobulin fractions contain immune complexes that vary in size, composition, and phlogistic potential. High molecular weight complexes are capable of inducing a spectrum of proinflammatory cytokines, all of which have been implicated in the pathogenesis of rheumatic disease.

Constitutive expression c-fos, c-jun, and NF kappa B mRNA is in nucleated fetal blood cells and up-regulation of c-fos and c-jun with anti-CD3 stimulation.

Fetal and neonatal lymphocytes are relatively resistant to activation and cytokine production when stimulated either via their T-cell antigen receptors or lectins. The molecular mechanism(s) responsible for this phenomenon have not been clearly elucidated. We have hypothesized that such defects in fetal/neonatal T-cell activation may be due to lack of expression of the transcriptional regulatory elements required for T-cell activation. We used reverse transcriptase-polymerase chain reaction to examine both fetal and term neonatal cord bloods for mRNA expression of three transcription factors implicated in T-cell activation: c-jun, c-fos, and NF kappa B (p50 subunit). We demonstrate that mRNAs for all three of these regulatory factors are expressed in fetal blood cells by the 27th week of gestation and in term cord bloods. Activation of term infant cord blood mononuclear cells with anti-CD3 monoclonal antibodies resulted in up-regulation of both c-jun and c-fos mRNAs within 15 min of stimulation. However, secretion of IL-2 by anti-CD3-stimulated cord blood mononuclear cells was still blunted compared with control cells from adults. We conclude that fetal nucleated blood cells constitutively express important genes for cytokine regulation and are able to increase intracellular accumulation of the mRNAs for these factors in response to anti-CD3 stimulation. Thus, qualitative differences in the capacity to regulate these factors could not be shown in fetal blood cells. Quantitative experiments comparing binding of these transcription factors to the IL-2 promoter are currently under investigation.

Regulation of cytokine mRNA expression in activated lymphocytes by human choriocarcinoma JAR cells.

Because the fetus is semiallogenic to the mother, considerable modulation of the maternal immune response must occur in order for pregnancy to be successfully carried to term. Some authors have hypothesized that the immunomodulation of pregnancy includes an adjustment of cytokine responses away from the Th1 paradigm and toward the Th2 pattern. In vivo data from murine pregnancy support this hypothesis. However, in humans, the Th1/Th2 model appears to be more complex than that in mice, and cytokine expression of mRNA in human decidual tissue does not reflect a clear-cut Th2 bias. The experiments described here were undertaken to determine whether and how trophoblastic cells modulate cytokine expression in activated lymphocytes, and whether there is a trend toward the use of the Th2 pattern in an experimental model of the maternal-fetal interface. We used reverse transcriptase polymerase chain reaction (rtPCR) to detect cytokine mRNA expression in human peripheral blood mononuclear cells cocultivated with human choriocarcinoma JAR cells. We found that although IL-2 (a paradigmatic Th1 cytokine) was significantly down-regulated by JAR cells at the mRNA level, similar decreases were also seen in IL-10, which participates in the Th2 paradigm. We were unable to detect changes in either interferon-gamma (IFN-gamma, a Th1 cytokine) or IL-4 (a Th2 cytokine) mRNA's or in IL-2R expression by fluorescence-activated cell sorting. These studies indicate that human choriocarcinoma JAR cells are capable of modifying cytokines in activated lymphocytes other than those involved in the Th1 paradigm. While it may be useful to view human responses against the background of these patterns established from murine systems, it is reasonable to conclude that human pregnancy may not involve regulation of Th1 immune responses exclusively.

Expression of complement regulatory proteins on fetal blood cells in utero.

Since the fetus is semiallogenic to the mother, mechanisms have evolved to protect fetal tissue from the maternal immune response. Among these mechanisms is the expression of cell-surface complement regulatory proteins at the maternal-fetal interface. However, beginning in the third trimester, fetal blood cells are exposed to actively-transported IgG antibody. Thus, we speculated that fetal blood cells would require expression of one or more complement regulators by the early third trimester. Using flow cytometry and Western blots, we have demonstrated the presence of three important complement regulatory proteins in the circulating blood cells of human fetuses. These findings are consistent with the putative biological role of the cell-surface complement regulatory proteins.

Composition and biological behaviour of immune complexes isolated from synovial fluid of patients with juvenile rheumatoid arthritis (JRA).

Data published from in vitro studies have shown that IgM-rheumatoid factor (RF)-bearing immune complexes possess several biological features that may contribute to their pathogenicity. However, no studies have demonstrated that such complexes exist at sites of inflammation in children with rheumatoid disease. We used two methods of sequential column chromatography to purify immune complexes from synovial fluids of children with JRA. We demonstrate that high molecular weight complexes contain IgM-RF, have not bound C4 in vivo, but activate the classical pathway in vitro. In contrast, complexes which have bound C3 in vivo do not contain IgM-RF and are weak complement activators in vitro.