Abnormal lymphatic development in trisomy 16 mouse embryos precedes nuchal edema.

Ultrasound measurement of increased nuchal translucency is a method of risk assessment for heart malformations and trisomy 21 in human pregnancy. The developmental background of this nuchal edema is still not sufficiently understood. We have studied the process in trisomy 16 mice that show nuchal edema and heart malformations. We used trisomy 16 and wild-type (WT) embryos from embryonic day (E) 12.5 to E18.5. In WT embryos at E13, bilateral jugular lymphatic sacs are visible that share a lymphatic-venous membrane with the jugular vein. We could not in any case discern a valve between these vessels. At E14 in the TS16 embryos, the lymphatic sacs become enlarged showing abnormally thickened endothelium, specifically at the site of the membrane. In these embryos, severe edema develops in the nuchal region. There is a very close colocalisation of the nerves with the vascular structures. The start of reorganization of the jugular lymphatic sac to a lymph node is observed in both wild-type and TS16 but is diminished in the latter. In conclusion, abnormal size and structure of the jugular lymphatic sacs coincides with the development of nuchal edema. A disturbance of lymphangiogenesis might be the basis for increased nuchal translucency that is often observed in diseased human fetuses.     

Altered interleukin-1 production in mice exposed to rotation stress.

This study examined the effects of rotation stress (78 circles per 1 min for 1 h: 10 min rotation + 5 min interval) on peritoneal macrophage release of lymphocyte-activating factors (LAF) and interleukin-1 alpha (IL-1 alpha) concentration in mice (CBA x C57BL6)F1. Rotation stress induced the macrophages' release of LAF with peak reactions in 0-1 h after termination of stress, followed by elevation of plasma IL-1 alpha. Induction of LAF release was accompanied by a higher concentration of corticosterone (Cs) in the blood, most prominent at 0 and 0.5 h after termination of the stressful procedure, and recovering to normal values 1 h later. It is suggested that the stimulating effect of stress on LAF and IL-1 production possibly involves glucocorticoid hormones, as well as other stress-induced neurohumoral shifts accompanying the reaction of glucocorticoids. It is also possible that the activated production of LAF and IL-1 by macrophages has a stress-limiting role in cases of mild stress.     

Inhibitory effects of indomethacin on interleukin-1 and nitric oxide production in rat microglia in vitro.

Indomethacin, as a nonsteroidal antiinflammatory drug, is reported to be effective in some degree in the prevention and treatment of Alzheimer's disease (AD). Effects of indomethacin on proinflammatory cytokines interleukin-1 (IL-1), tumor necrosis factor alpha and nitric oxide (NO) on rat microglia in vitro were investigated. The biological activities of IL-1 were assayed by thymocyte proliferation assay and the activities of TNG-alpha, by L929 cytotoxicity assay. NO concentration was represented by nitrite and determined by Griess reaction. Indomethacin inhibited IL-1 and NO production by rat microglia stimulated at the concentration of 0.1-10 micromol/l. However, it did not show any inhibitory effect on TNF-alpha production by resting and LPS-stimulated rat microglia. The results suggest that the mechanism by which indomethacin might be beneficial in treatment of AD might be due to the inhibition IL-1 and NO production from microglia.     

Abnormal production of T helper 2 cytokines interleukin-4 and interleukin-5 by T cells from newborns with atopic parents

T cell clones were generated from umbelical cord blood lymphocytes (UCBL) of nine newborns with atopic or nonatopic parents and their cytokine secretion profile was assessed. Both phytohemagglutinin-induced and Dermatophagoides pteronyssinus-spetific T cell clones from newborns with atopic parents exhibited an enhanced ability to produce the Th2 cytokines interleukin (IL)-4 and IL-5, compared to T cell clones from newborns with nonatopic parents. In contrast, the ability to produce interferon-γ by UCBL from the two groups of newborns was not different. Of the five children who could be followed up to 3 years after birth, four with atopic parents developed clinical and/or biological atopic manifestations, whereas one without atopic parents did not. Thus, the pronounced production of IL-4 and IL-5 by UCBL not only appears to be related to the atopic status of parents, but also associates with the subsequent development of atopy in childhood.     

Chlamydia trachomatis-induced production of interleukin-1 by human monocytes.

Human diseases caused by the intracellular bacterium Chlamydia trachomatis include genital tract infections and blinding trachoma. Chlamydial infections are characterized by chronic inflammation and scarring, and development of such complications is thought to be immunologically mediated. In this study, we show that coculture of C. trachomatis (serovar L2) with human blood monocytes induced the production of interleukin-1 (IL-1), an important mediator of inflammation, tissue remodeling, and scarring. IL-1 was produced in response to UV-inactivated elementary bodies containing from 0.1 to 50 micrograms of protein per ml, with a maximal response at 5 to 10 micrograms/ml. IL-1 activity was detected by 6 h of incubation and was maximal by 24 h. Peak levels were maintained throughout 96 h of incubation. Rabbit antibody to human IL-1(alpha + beta) effectively neutralized the thymocyte-stimulating activity of the supernatants. The apparent molecular weight of chlamydia-induced IL-1 was 16,000, as determined by gel filtration on a Bio-Gel P-60 column. Isoelectric focusing yielded two peaks of activity, with pIs of 5.5 and 6.9. Neutralization studies with antisera against human IL-1 alpha and IL-1 beta showed that the acidic and neutral peaks corresponded to IL-1 alpha and IL-1 beta, respectively, with IL-1 beta predominating. Heat-killed chlamydiae, which are not internalized by monocytes, were effective IL-1 inducers, indicating that phagocytosis was not required for IL-1 induction. Purified C. trachomatis lipopolysaccharide was also an effective IL-1 inducer, suggesting that the response to intact organisms may be largely a response to chlamydial lipopolysaccharide. Finally, purified chlamydial major outer membrane protein induced low but detectable IL-1 activity.     

Lessons from interleukin-deficient mice: the interleukin-1 system.

The study of cytokine-deficient mice has provided important information for a better understanding of inflammatory processes. In this report, the characterization of mice deficient for various components of the interleukin (IL)-1 system is reviewed. Results obtained by studying mice deficient for IL-1alpha, IL-1beta, IL-1 receptor antagonist, IL-1 receptor type I, IL-1 receptor accessory protein, IL-1 receptor-associated kinase, and the IL-1beta-converting enzyme caspase-1 are summarized. Because some of the components of the IL-1 system are shared with IL-18, similarities between IL-1beta and IL-18 are also discussed.     

Influence of age on the production and regulation of interleukin-1 in mice.

The decrease in T-cell proliferation with age is due, in part, to the decline in the production of IL-2. Since IL-1 is needed to trigger IL-2 production, we determined the IL-1 producing capacity of peritoneal macrophages of young (2-4 months) and old (24-26 months) BALB/c and C57BL/6 mice. Mice were stimulated with LPS, and their peritoneal macrophages were obtained 3 days later, purified, and assessed for IL-1 production by coculturing them with splenic T cells at a ratio of 1:5 in the presence of LPS. Supernatants were obtained 4 days later when the PGE2 and IL-2 activities were minimal and IL-1 activity maximal. IL-1 activity was assessed for their ability to augment the proliferative activity of indicator thymocytes in their response to PHA stimulation. The results revealed that (i) IL-1 production by cells of old BALB/c and C57BL/6 mice is reduced to about 40% and 30% that of young mice, respectively; (ii) indomethacin enhances IL-1 production by cells of both young and old mice to the same extent; and (iii) reduction in the IL-1 producing capacity by cells of old mice results from altered activities of both the IL-1 producing peritoneal macrophages and the augmenting T cells.     

Interleukin-6 production by thyroid epithelial cells. Enhancement by interleukin-1.

Interleukin-1 is a potent inhibitor of thyroglobulin and cAMP production in human thyroid cells and the inhibitory effect is enhanced by tumor necrosis factor-alpha and interferon-gamma. In the present study secondary cultures of human thyroid cells produced interleukin-6 and the production was significantly increased after exposure of the cells to recombinant interleukin-1 alpha and -1 beta. This increase was dose-dependent and concomitant of the IL-1 induced decrease in cAMP and thyroglobulin production. Both tumor necrosis factor-alpha and -beta also augmented interleukin-6 production, but less potently than interleukin-1. Interferon-gamma did not affect the production of interleukin-6. The rat thyroid cell line FRTL-5 produced interleukin-6 spontaneously, and the production was enhanced after addition of recombinant interleukin-1 beta. A pathogenetic role of interleukin-6 in autoimmune thyroid disease is suggested.     

Inhibitory effect of interleukin-16 on interleukin-2 production by CD4+ T cells

Signalling through CD4 by human immunodeficiency virus (HIV)-1 envelope glycoprotein (gpl20) and/or anti-CD4 antibodies can promote T-cell activation and anergy. Interleukin (IL)-16 is a competence growth factor for CD4+ T cells that can induce a G0 to G1 cell cycle transition but cannot induce cell division. The receptor of this cytokine is thought to be the CD4 molecule, although the binding epitope of IL-16 differs from that of HIV. We have demonstrated that both HIV-1/gp120 and IL-16 induced CD4+ T-cell dysfunction, as indicated by suppression of mitogen-induced IL-2 production. Two anti-CD4 antibodies with different binding sites on CD4 also showed an inhibitory effect on IL-2 production. These results indicate that promotion of CD4+ T-cell anergy via the CD4 molecule does not depend on the binding sites of the CD4 ligands.     

Hypotelorism in trisomy 1-producing mice

Trisomy 1 embryos in mice are smaller in all dimensions, showing a developmental retardation as compared with euploid mice. Very rarely the trisomic embryos develop a typical hypotelorism with holoprosencephalon and missing olfactory nerves. Corneal distances, angles between the optic nerves and further microscopic examination showed no intermediate forms between the trisomy 1 embryos and the rare trisomy 1 embryo with hypotelorism. There seems to be a threshold, beyond which the major developmental derangement occurred. This is an experimental model showing parallels to the occasional varying phenotypes in human trisomies.     

Enhancement of interleukin-1 and interleukin-2 production by soluble glucan

Soluble glucan, a beta-1,3-linked polyglucose, is a biologic response modifier effective in the therapy of experimental neoplasia, infectious diseases and immunosuppression. Interleukin-1 (IL-1) and interleukin-2 (IL-2) are endogenous immunomodulators which are essential for effective immune responsiveness. In view of its broad spectrum of immunobiological activity, the ability of glucan to enhance the production of IL-1 and IL-2 was evaluated. Splenic IL-1 and IL-2 secretion as well as plasma IL-1 and IL-2 levels were determined in Sprague-Dawley rats receiving glucan (100 mg/kg, i.p.) at intervals ranging from 12 days to 1 h prior to collection of splenocytes and plasma. Glucan (100 mg/kg) was also injected either s.c., i.p. or i.v. on days -4, -3 and -2 prior to harvesting splenocytes on day 0. Splenic macrophage IL-1 production was initially elevated 12 h following glucan injection and was maintained for a 5 day period. IL-2 secretion by splenic lymphocytes was enhanced 6 h post-glucan and remained elevated for an additional 9 days. Plasma IL-1 activity was elevated 12 h post-injection, while IL-2 activity in plasma was enhanced at 1 h post-glucan. Peak IL-1 and IL-2 activity in plasma occurred 9 and 12 days, respectively, following glucan administration. With regard to route of administration, IV glucan was most effective in inducing lymphokine production. This study demonstrates that: (1) glucan will enhance IL-1 and IL-2 production and (2) elevations in lymphokine production can be maintained up to 12 days post-glucan.     

Suppressor cells induced by influenza virus inhibit interleukin-2 production in mice.

PR8 virus depressed interleukin-2 (IL-2) and natural killer (NK) cell activity in BALB/c infected mice. IL-2 production was not dependent on (i) a decreased number of T cells or (ii) a primary defect in IL-1 production, but on a T-suppressor cell subpopulation. In fact, when T suppressor cells were removed from infected spleen cells, we observed normal levels of IL-2 activity.     

Decrease in neutrophil migration induced by endotoxin and suppression of interleukin-1 production by macrophages in lactic dehydrogenase virus-infected mice.

Neutrophil (PMN) migration into the peritoneal cavity after intraperitoneal injection of lipopolysaccharide (LPS), chemotactic activity of PMN, interleukin-1 (IL-1) production by macrophages (M phi) and its ability to attract PMN in mice chronically infected with lactic dehydrogenase virus (LDV) were compared with those in uninfected control mice. PMN migration into the peritoneal cavity decreased in infected mice when LPS was injected intraperitoneally. PMN chemotactic activity did not show any difference following infection. To assess the mechanism of this decreased PMN migration, IL-1 production, which is responsible for PMN attraction, was studied in LDV-infected mice. IL-1 production by M phi derived from infected mice decreased and its ability to attract PMN was weak. IL-1 production by M phi from control and infected mice increased after treatment by indomethacin and LPS. PMN migration into the peritoneal cavity increased after treatment with indomethacin and LPS in both control and infected mice. However, the rate of increase of IL-1 production and PMN migration was greater in infected mice. These results suggest that the excess activation of cyclo-oxygenase-derived products (prostaglandins) in infected mice might be responsible for the suppression of IL-1 production by M phi, resulting in decreased PMN migration induced by endotoxin.     

Studies of depressed interleukin-2 production by spleen cells from mice following infection with cytomegalovirus

Summary Spleen cells from mice infected with virulent and avirulent strains of murine cytomegalovirus showed depressed interleukin-2 production after ConA stimulation. Cell-mixing experiments indicated that this was due to a primary defect in non adherent cells. Spleen cells infected in vitro were also studied.     

Molecular studies of non-disjunction in trisomy 16.

The origin of the additional chromosome in 26 trisomy 16 spontaneous abortions was studied using DNA probes for chromosome 16, including a probe for centromeric alpha sequences. We were able to determine the parent and meiotic stage of origin of trisomy in 22 cases, with all being attributable to maternal meiosis I non-disjunction. Furthermore, in each of the remaining four cases the results were compatible with this origin. Thus, it is likely that the high incidence of trisomy 16 results from an abnormal process acting at maternal meiosis I which more frequently involves chromosome 16 than other similar sized chromosomes. In studies of recombination, we found little evidence for an association between reduced or absent recombination and chromosome 16 non-disjunction; however, we were unable to rule out an effect of hyperrecombination.     

Induction of human interleukin-1 production by polymyxin B

Lipopolysaccharide (LPS) is a well known interleukin-1 inducer. Polymyxin B sulphate (PMB) is a cyclic antibiotic which neutralizes different biological properties of LPS. Under the conditions used in our laboratory, we were able to show that PMB alone could stimulate human monocytes to produce and release interleukin-1 activity, and that PMB was unable to inhibit the production of interleukin-1 by human monocytes stimulated with LPS. On the contrary, a synergistic effect was obtained which was not observed with another stimulant such as muramyl dipeptide.