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We study the response of hot Jupiters to a static tidal perturbation using the concentric MacLaurin spheroid method. For strongly irradiated planets, we first performed radiative transfer calculations to relate the planet's equilibrium temperature, T_(eq), to its interior entropy. We then determined the gravity harmonics, shape, moment of inertia, and static Love numbers for a range of two-layer interior models that assume a rocky core plus a homogeneous and isentropic envelope composed of hydrogen, helium, and heavier elements. We identify general trends and then study HAT-P-13b, the WASP planets 4b, 12b, 18b, 103b, and 121b, and Kepler-75b and CoRot-3b. We compute the Love numbers, k_(nm), and transit radius correction, ΔR, which we compare with predictions in the literature. We find that the Love number, k₂₂, of tidally locked giant planets cannot exceed a value of 0.6, and that the high T_(eq) consistent with strongly irradiated hot Jupiters tends to further lower k₂₂. While most tidally locked planets are well described by a linear regime response of k₂₂ = 3J₂/q₀ (where q₀ is the rotation parameter of the gravitational potential), for extreme cases such as WASP-12b, WASP-103b, and WASP-121b, nonlinear effects can account for over 10% of the predicted k₂₂. The k₂₂ values larger than 0.6, as they have been reported for planets WASP-4b and HAT-P13B, cannot result from a static tidal response without extremely rapid rotation and thus are inconsistent with their expected tidally locked state.

Since its arrival at Jupiter in 2016, the Juno spacecraft has measured the planet’s gravity fields with unprecedented precision. The interpretation of these measurements has been challenging because the magnitudes of the gravity coefficients J4 and J6 were smaller than predicted by traditional interiors models that included a dense inner core composed of rock and ice. Here we instead present models with dilute cores [Geophys. Res. Lett. 44 (2017) 4649] and deep-winds that conform to theoretical predictions of hydrogen-helium phase separation in the interior layer from approximately 0.8 to 0.85 Jupiter radii. Such models match the entire set of zonal gravity measurements by the Juno spacecraft. Our work is based on the accelerated version of the Concentric Maclaurin Spheroid method [Astrophysical J. 879 (2019) 78]. We conclude by comparing with models for Saturn’s interior.

International audience; Jupiter’s atmosphere is rotating differentially, with zones and belts rotating at speeds that differ by up to 100 metres per second. Whether this is also true of the gas giant’s interior has been unknown1,2, limiting our ability to probe the structure and composition of the planet3,4. The discovery by the Juno spacecraft that Jupiter’s gravity field is north–south asymmetric5 and the determination of its non-zero odd gravitational harmonics J3, J5, J7 and J9 demonstrates that the observed zonal cloud flow must persist to a depth of about 3,000 kilometres from the cloud tops6. Here we report an analysis of Jupiter’s even gravitational harmonics J4, J6, J8 and J10 as observed by Juno5 and compared to the predictions of interior models. We find that the deep interior of the planet rotates nearly as a rigid body, with differential rotation decreasing by at least an order of magnitude compared to the atmosphere. Moreover, we find that the atmospheric zonal flow extends to more than 2,000 kilometres and to less than 3,500 kilometres, making it fully consistent with the constraints obtained independently from the odd gravitational harmonics. This depth corresponds to the point at which the electric conductivity becomes large and magnetic drag should suppress differential rotation7. Given that electric conductivity is dependent on planetary mass, we expect the outer, differentially rotating region to be at least three times deeper in Saturn and to be shallower in massive giant planets and brown dwarfs.

Israeli Ministry of Science; Minerva foundation; Federal German Ministry of Education and Research; Helen Kimmel Center for Planetary Science at the Weizmann Institute of Science; CNES; BSF; NSF; Juno project

The Cassini spacecraft's Grand Finale orbits provided a unique opportunity to probe Saturn's gravity field and interior structure. Doppler measurements yielded unexpectedly large values for the gravity harmonics J_6, J_8, and J_10 that cannot be matched with planetary interior models that assume uniform rotation. Instead we present a suite of models that assume the planet's interior rotates on cylinders, which allows us to match all the observed even gravity harmonics. For every interior model, the gravity field is calculated self-consistently with high precision using the Concentric Maclaurin Spheroid (CMS) method. We present an acceleration technique for this method, which drastically reduces the computational cost, allows us to efficiently optimize model parameters, map out allowed parameter regions with Monte Carlo sampling, and increases the precision of the calculated J_2n gravity harmonics to match the error bars of the observations, which would be difficult without acceleration. Based on our models, Saturn is predicted to have a dense central core of 15-18 Earth masses and an additional 1.5-5 Earth masses of heavy elements in the envelope. Finally, we vary the rotation period in the planet's deep interior and determine the resulting oblateness, which we compare with the value from radio occultation measurements by the Voyager spacecraft. We predict a rotation period of 10:33:34 h +- 55s, which is in agreement with recent estimates derived from ring seismology., 12 color figures, 5 tables, Astrophysical Journal, in press (2019)

This article provides an overview of how models of giant planet interiors are constructed. We review measurements from past space missions that provide constraints for the interior structure of Jupiter. We discuss typical three-layer interior models that consist of a dense central core and an inner metallic and an outer molecular hydrogen-helium layer. These models rely heavily on experiments, analytical theory, and first-principle computer simulations of hydrogen and helium to understand their behavior up to the extreme pressures ~10 Mbar and temperatures ~10,000 K. We review the various equations of state used in Jupiter models and compare them with shock wave experiments. We discuss the possibility of helium rain, core erosion and double diffusive convection may have important consequences for the structure and evolution of giant planets. In July 2016 the Juno spacecraft entered orbit around Jupiter, promising high-precision measurements of the gravitational field that will allow us to test our understanding of gas giant interiors better than ever before.

Cassini's last look at Saturn's rings During the final stages of the Cassini mission, the spacecraft flew between the planet and its rings, providing a new view on this spectacular system (see the Perspective by Ida). Setting the scene, Spilker reviews the numerous discoveries made using Cassini during the 13 years it spent orbiting Saturn. Iess et al. measured the gravitational pull on Cassini, separating the contributions from the planet and the rings. This allowed them to determine the interior structure of Saturn and the mass of its rings. Buratti et al. present observations of five small moons located in and around the rings. The moons each have distinctive shapes and compositions, owing to accretion of ring material. Tiscareno et al. observed the rings directly at close range, finding complex features sculpted by the gravitational interactions between moons and ring particles. Together, these results show that Saturn's rings are substantially younger than the planet itself and constrain models of their origin. Science , this issue p. 1046 , p. eaat2965 , p. eaat2349 , p. eaau1017 ; see also p. 1028

The depth to which Jupiter’s observed east–west jet streams extend has been a long-standing question. Resolving this puzzle has been a primary goal for the Juno spacecraft, which has been in orbit around the gas giant since July 2016. Juno’s gravitational measurements have revealed that Jupiter’s gravitational field is north–south asymmetric, which is a signature of the planet’s atmospheric and interior flows. Here we report that the measured odd gravitational harmonics J_3, J_5, J_7 and J_9 indicate that the observed jet streams, as they appear at the cloud level, extend down to depths of thousands of kilometres beneath the cloud level, probably to the region of magnetic dissipation at a depth of about 3,000 kilometres. By inverting the measured gravity values into a wind field, we calculate the most likely vertical profile of the deep atmospheric and interior flow, and the latitudinal dependence of its depth. Furthermore, the even gravity harmonics J_8 and J_(10) resulting from this flow profile also match the measurements, when taking into account the contribution of the interior structure. These results indicate that the mass of the dynamical atmosphere is about one per cent of Jupiter’s total mass.

The gravity harmonics of a fluid, rotating planet can be decomposed into static components arising from solid-body rotation and dynamic components arising from flows. In the absence of internal dynamics, the gravity field is axially and hemispherically symmetric and is dominated by even zonal gravity harmonics J_(2n) that are approximately proportional to q^n, where q is the ratio between centrifugal acceleration and gravity at the planet’s equator. Any asymmetry in the gravity field is attributed to differential rotation and deep atmospheric flows. The odd harmonics, J_3, J_5, J_7, J_9 and higher, are a measure of the depth of the winds in the different zones of the atmosphere. Here we report measurements of Jupiter’s gravity harmonics (both even and odd) through precise Doppler tracking of the Juno spacecraft in its polar orbit around Jupiter. We find a north–south asymmetry, which is a signature of atmospheric and interior flows. Analysis of the harmonics, described in two accompanying papers, provides the vertical profile of the winds and precise constraints for the depth of Jupiter’s dynamical atmosphere.

The Juno spacecraft has measured Jupiter's low-order, even gravitational moments, $J_2$--$J_8$, to an unprecedented precision, providing important constraints on the density profile and core mass of the planet. Here we report on a selection of interior models based on ab initio computer simulations of hydrogen-helium mixtures. We demonstrate that a dilute core, expanded to a significant fraction of the planet's radius, is helpful in reconciling the calculated $J_n$ with Juno's observations. Although model predictions are strongly affected by the chosen equation of state, the prediction of an enrichment of $Z$ in the deep, metallic envelope over that in the shallow, molecular envelope holds. We estimate Jupiter's core to contain an 7--25 Earth mass of heavy elements. We discuss the current difficulties in reconciling measured $J_n$ with the equations of state, and with theory for formation and evolution of the planet., 21 Pages, 3 figures, plus Supplementary Information

We investigated the role of secretory leukocyte protease inhibitor (SLPI) in ischemia/reperfusion injury in cardiac transplantation. SLPI-/- mouse hearts and wild-type (WT) controls were transplanted immediately or after 10 h of cold ischemia (CI). Recombinant SLPI (rSLPI) was added to the preservation solution or given systemically. After evaluation of myocardial performance, grafts were investigated for histology, SLPI, TNF-alpha, TGF-beta, NF-kappaB and protease expression at indicated time points. Early myocardial contraction was profoundly impaired in SLPI-/- hearts exposed to CI and associated with high intra-graft protease expression. Systemic administration of rSLPI had no effect, however, when SLPI was added to the preservation solution, myocardial contraction was restored to normal. At 10 days, inflammation, myocyte vacuolization and necrosis were significantly more severe in SLPI-/- hearts. SLPI gene expression was detected in WT mice at 12 and 24 h and was significantly higher after CI. SLPI protein was observed at 24 h and 10 days. High intra-graft concentrations of SLPI after administration of rSLPI were inversely correlated with protease levels early and TGF-beta expression late after reperfusion. SLPI plays a crucial role in early myocardial performance and postischemic inflammation after cardiac transplantation. A dual inhibitory effect on protease and TGF-beta expression might be the underlying mechanism.

In anticipation of improved observational data for Jupiter's gravitational field from the Juno spacecraft, we predict the static tidal response for a variety of Jupiter interior models based on ab initio computer simulations of hydrogen-helium mixtures. We calculate hydrostatic-equilibrium gravity terms using the non-perturbative concentric Maclaurin Spheroid (CMS) method that eliminates lengthy expansions used in the theory of figures. Our method captures terms arising from the coupled tidal and rotational perturbations, which we find to be important for a rapidly-rotating planet like Jupiter. Our predicted static tidal Love number $k_2 = 0.5900$ is $\sim$10\% larger than previous estimates. The value is, as expected, highly correlated with the zonal harmonic coefficient $J_2$, and is thus nearly constant when plausible changes are made to interior structure while holding $J_2$ fixed at the observed value. We note that the predicted static $k_2$ might change due to Jupiter's dynamical response to the Galilean moons, and find reasons to argue that the change may be detectable, although we do not present here a theory of dynamical tides for highly oblate Jovian planets. An accurate model of Jupiter's tidal response will be essential for interpreting Juno observations and identifying tidal signals from effects of other interior dynamics in Jupiter's gravitational field., Comment: 10 Pages, 6 figures, 4 tables

The accretion of a terrestrial body and differentiation of its silicate/oxide mantle from iron core provide abundant energy for heating its interior to temperatures much higher than the present day Earth. The consequences of differentiation on the structure and composition of planets are typically addressed considering only the interaction of molten iron with an immiscible `rocky' phase. We demonstrate that mixing in a representative system of liquid or solid MgO and liquid iron to a single homogeneous liquid occurs at sufficiently low temperature to be present in the aftermath of a giant impact. Applying the thermodynamic integration technique to density functional theory molecular dynamics simulations, we determine the solvus closure temperature for the Fe-MgO system for pressures up to 400 GPa. Solvus closure occurs at $\sim$4000 K at low pressure, and has a weak positive pressure dependence, such that its gradient with respect to depth is less steep than an adiabatic temperature profile. This predicts a new mode of core-mantle differentiation following the most energetic giant impacts, with exsolution of iron from the mixture beginning in the outer layers of the planet. We demonstrate that high-temperature equilibration results in delivery of nominally insoluble Mg-rich material to the early core. Since MgO is the least soluble major mantle component in iron at low temperatures, these results may represent an upper bound on temperature for mixing in terrestrial planets., Comment: 10 pages, 9 figures, supplementary material

We extend to three dimensions the Concentric Maclaurin Spheroid method for obtaining the self-consistent shape and gravitational field of a rotating liquid planet, to include a tidal potential from a satellite. We exhibit, for the first time, the important effect of the planetary rotation rate on tidal response of gas giants. Simulations of planets with fast rotation rates like those of Jupiter and Saturn, exhibit significant changes in calculated tidal love numbers $k_{nm}$ when compared with non-rotating bodies. A test model of Saturn fitted to observed zonal gravitational multipole harmonics yields $k_2=0.413$, consistent with a recent observational determination from Cassini astrometry data (Lainey et al., 2016). The calculated love number is robust under reasonable assumptions of interior rotation rate, satellite parameters, and details of Saturn's interior structure. The method is benchmarked against several published test cases., Comment: 40 pages, 8 figures, 3 tables

Transforming growth factor beta is a potent immunomodulator with both pro- and antiinflammatory activities. Based on its immunosuppressive actions, exogenous TGF-beta has been shown to inhibit autoimmune and chronic inflammatory diseases. To further explore the potential therapeutic role of TGF-beta, we administered a plasmid DNA encoding human TGF-beta1 intramuscularly to rats with streptococcal cell wall-induced arthritis. A single dose of 300 microg plasmid DNA encoding TGF-beta1, but not vector DNA, administered at the peak of the acute phase profoundly suppressed the subsequent evolution of chronic erosive disease typified by disabling joint swelling and deformity (articular index = 8.17+/-0. 17 vs. 1.25+/-0.76, n = 6, day 26, P < 0.01). Moreover, delivery of the TGF-beta1 DNA even as the chronic phase commenced virtually eliminated subsequent inflammation and arthritis. Both radiologic and histopathologic as well as molecular evidence supported the marked inhibitory effect of TGF-beta1 DNA on synovial pathology, with decreases in the inflammatory cell infiltration, pannus formation, cartilage and bone destruction, and the expression of proinflammatory cytokines that characterize this model. Increases in TGF-beta1 protein were detected in the circulation of TGF-beta1 DNA-treated animals, consistent with the observed therapeutic effects being TGF-beta1 dependent. These observations provide the first evidence that gene transfer of plasmid DNA encoding TGF-beta1 provides a mechanism to deliver this potent cytokine that effectively suppresses ongoing inflammatory pathology in arthritis.

Leukocyte integrins are fundamentally important in modulating adhesion to extracellular matrix components and to other cells. This integrin-mediated adhesion controls leukocyte arrest and extravasation during the onset of inflammatory responses. Moreover, integrin-ligand interactions trigger signaling pathways that may influence leukocyte phenotype and function at sites of inflammation. In the current studies, we evaluated the combinatorial effects of monocyte adhesion and IFN-gamma on intracellular signaling pathways. IFN-gamma triggers a well-defined signal transduction pathway, which although not directly stimulated by monocyte adherence to fibronectin or arginine-glycine-aspartate (RGD)-coated substrata, was enhanced significantly in these matrix-adherent cells. Compared with monocytes in suspension or adherent on plastic surfaces, monocytes adherent to fibronectin or RGD exhibited a greater than threefold increase in steady state levels of IFN-gamma-induced mRNA for the high affinity Fc gammaRI receptor. By electrophoretic mobility shift assays, this increase in mRNA was associated with a 5- to 10-fold increase in the STAT1-containing DNA-binding complex that binds to Fc gammaRI promoter elements. Furthermore, the tyrosine phosphorylation of STAT1 and the tyrosine kinases JAK1 and JAK2 was enhanced significantly in RGD-adherent monocytes compared with control cells. These results suggest a novel mechanism by which integrin-mediated cell adhesion can modulate the magnitude of cytokine-induced signal transduction pathways, thereby amplifying cellular events leading to monocyte activation and inflammation.

Cytokine-induced neutrophil chemoattractant (CINC), a member of the IL-8 superfamily of chemokines, is one rat homologue of the three human GRO proteins. Neutralizing Abs against CINC have been shown previously to be efficacious in several models of inflammation, indicating that CINC is an important proinflammatory mediator in vivo. By introducing the N-terminal mutation delta1-5,ELR>AAR into CINC, we have developed a rat alpha-chemokine receptor antagonist (ra) analogous to a previously described mutant of human IL-8. Bacterially expressed CINCra had no chemotactic activity itself, but completely blocked the activity of physiologic concentrations of CINC when used as an antagonist in vitro. Inhibition by CINCra was specific, since it had approximately 10-fold less antagonist activity on the related, but distinct rat alpha-chemokine macrophage-inflammatory protein 2. When coinjected with zymosan into the peritoneal cavities of Lewis rats, 100 microg of CINCra inhibited neutrophil influx by approximately 40%, which was comparable with inhibition caused by polyclonal anti-CINC Abs.

The formation of the giant planets in our solar system, and likely a majority of giant exoplanets, is commonly explained by the accretion of nebular hydrogen and helium onto a large core of terrestrial-like composition. The fate of this core has important consequences for the evolution of the interior structure of the planet. It has recently been shown that H2O, MgO and SiO2 dissolve in liquid metallic hydrogen at high temperature and pressure. In this study, we perform ab initio calculations to study the solubility of an innermost metallic core. We find dissolution of iron to be strongly favored above 2000 K over the entire pressure range (0.4-4 TPa) considered. We compare with and summarize the results for solubilities on other probable core constituents. The calculations imply that giant planet cores are in thermodynamic disequilibrium with surrounding layers, promoting erosion and redistribution of heavy elements. Differences in solubility behavior between iron and rock may influence evolution of interiors, particularly for Saturn-mass planets. Understanding the distribution of iron and other heavy elements in gas giants may be relevant in understanding mass-radius relationships, as well as deviations in transport properties from pure hydrogen-helium mixtures., 8 pages, 5 figures, 2 tables. Submitted to the Astrophysical Journal on March 26, 2013

The targeted disruption of the TGF-beta1 gene in mice (TGF-beta1 -/-) leads to extensive inflammation in vital organs, cachexia, and death within 3 to 4 wk. Significant inflammatory lesions develop initially in the periductal regions of the salivary glands and escalate as the animals become symptomatic. These inflammatory sites, characterized by lymphocytic infiltration and increased proliferation, cytokine mRNA expression, and IgG-positive cells, resemble lesions of Sjögren's syndrome. Moreover, the inflammatory pathology, enhanced MHC expression, and Ab production are consistent with an autoimmune-like etiology. Glandular atrophy and loss of acini with reduced saliva production appear to contribute to the wasting syndrome characteristic of the TGF-beta1 -/- mice. To determine whether the structural and functional defects were developmental due to the absence of TGF-beta1 or secondary to the inflammation, TGF-beta1 -/- mice were treated with synthetic fibronectin peptides, which block leukocyte infiltration. Daily systemic injections of RGD, CS-1, and/or peptides derived from the heparin-binding region of the A chain not only prevented leukocyte infiltration in the salivary glands of the TGF-beta1 -/- mice, but also reversed the acinar and ductal derangements. These data suggested that salivary gland development is not jeopardized in the absence of TGF-beta1, but that the extensive infiltration of inflammatory cells compromises glandular structure and function. The essential nature of TGF-beta1 in controlling inflammatory and immune processes is confirmed by these studies. Moreover, these TGF-beta1 -/- mice provide an important model of autoimmune disease that can be used in the design of therapeutic interventions.

Monocyte apoptosis has emerged as a central regulatory event in hemopoiesis and inflammation. Inflammatory cytokines can either promote or prevent monocyte apoptosis. To study the possible role of Fas Ag, a member of the TNF/nerve growth factor receptor family, in monocyte apoptosis, human peripheral blood monocytes activated by IL-1 beta or TNF-alpha were exposed to anti-Fas mAb. Engagement of the Fas Ag resulted in apoptosis of monocytes, as monitored by propidium iodide uptake, decrease in cell size, DNA fragmentation, and characteristic ultrastructural changes. The apoptotic action of Fas was abolished completely by antioxidants such as N-acetylcysteine and glutathione, suggesting a role for reactive oxygen intermediates (ROI) in the death process. Consistent with this observation, Fas stimulation enhanced the fluorescence associated with oxidation of 2',7'-dichlorofluorescein, indicating increased levels of intracellular ROI. Moreover, the exogenous addition of hydrogen peroxide or menadione, an intracellular generator of superoxide anion, was sufficient for the induction of monocyte apoptosis. These data indicate that ROI are key mediators of Fas-induced apoptosis. In contrast to IL-1 beta and TNF-alpha, LPS-treated monocytes were resistant to the apoptotic action of Fas. Under these conditions, LPS did not down-regulate Fas, but inhibited the Fas-dependent elevation of ROI. Therefore, monocytes appear to have a protective mechanism that can interfere directly with the Fas-induced pathway of cell suicide, thereby controlling their destiny.

IL-4 has diverse effects on hematopoietic cells, including the ability to suppress certain mononuclear cell functions. To evaluate the effect of IL-4 on the evolution of acute and chronic arthritis, murine recombinant IL-4 was administered systemically to animals receiving an arthropathic dose of group A streptococcal cell wall fragments. Daily treatment with IL-4 had a minimal effect on the acute phase, but significantly suppressed the chronic, destructive phase. By 4 wk after initiation of disease, the articular index of IL-4-treated animals was reduced > 60% (articular index = 4 +/- 0.9) compared with the untreated rats (11.5 +/- 0.48, p < 0.001). A substantial decrease in the influx of inflammatory cells and virtual elimination of pannus and erosions occurred after IL-4 therapy. Associated with the reduced accumulation of mononuclear leukocytes was a decrease in their proinflammatory functions including cytokine production and reactive oxygen intermediate metabolism. These observations are consistent with the selective effects of IL-4 on phagocytic cell function demonstrated in vitro. Furthermore, IL-4 induced gene expression for IL-1ra, a protein that antagonizes the action of IL-1 by binding to the IL-1 receptor without agonist activity. Through an expanding spectrum of effects on monocyte-macrophage phenotypic and functional parameters, IL-4 is emerging as an important inhibitor of cell-mediated immune responses and pathogenic processes.

Transforming growth factor-beta (TGF-beta) is a potent immunomodulatory molecule that promotes inflammation through recruitment of monocytes and induction of IL-1 and other cytokines. These proinflammatory processes may be modulated by the ability of TGF-beta to induce de novo synthesis and secretion of an IL-1 receptor antagonist (IL-1ra) that binds to and blocks IL-1 receptors. In this study, we show that the addition of TGF-beta to human peripheral blood monocytes induced the sequential transcription of the 1.8-kb mRNA for IL-1 beta and for IL-1ra. The expression of detectable mRNA and synthesis of IL-1 beta peptide routinely preceded that for IL-1ra, suggesting possible dependency of IL-1ra generation on IL-1 beta. Antibody to IL-1 blocked TGF-beta induction of IL-1ra mRNA expression demonstrating an unique IL-1-dependent induction of its own antagonist. Confirmatory evidence that IL-1 participates in the generation of IL-1ra was obtained when exogenously added IL-1 induced and, IL-1 receptor antagonist blocked, IL-1ra transcription. Thus, these data suggest that TGF-beta, after release at a site of inflammation, induces synthesis of IL-1 that participates in the initial cytokine cascade central to an inflammatory response, and then triggers the generation of its own natural inhibitor by autocrine or paracrine pathways. This TGF-beta-induced IL-1-dependent induction of IL-1ra may provide a negative feedback loop, thereby promoting the resolution of an inflammatory response.

As a multifunctional cytokine that can augment certain T cell responses, IL-4 is being evaluated currently as a possible therapeutic agent in the treatment of cancer patients. In this report, PBMC were isolated from such patients before and after IL-4 therapy and were analyzed for phenotypic and functional alterations. Differential blood counts showed that the relative percentages of lymphocytes and, to a lesser degree, monocytes decreased after treatment with IL-4. However, when phenotypically analyzed by FACS, monocyte numbers generally increased, whereas there was a decrease in lymphocyte numbers. Monocytes taken from patients before therapy and cultured with and without LPS exhibited normal patterns of monokine-specific (IL-1 beta and TNF-alpha) mRNA expression, as well as secretion of peptide. The addition of rIL-4 to these cultures, however, resulted in the down-regulation of both gene expression and peptide release. Although monocytes taken from post-therapy patients also displayed normal patterns of monokine gene expression and cytokine release after stimulation with LPS, they were no longer inhibited by the addition of exogenous rIL-4 in vitro. These data suggest that monocytes become refractory to the inhibitory effects of IL-4 after in vivo exposure to this cytokine. However, when these monocytes were examined for expression of CD14, CD32, and CD64, exogenous IL-4 was observed to decrease markedly the appearance of these markers, regardless of whether the cells were obtained before or after therapy. Furthermore, monocytes from post-therapy patients exhibited reduced production of PGE2 and superoxide anion, compared with cells obtained before therapy. This effect persisted in culture independent of the further addition of exogenous IL-4. These data suggest a dichotomy between the IL-4-dependent mechanisms that regulate monokine production and those that regulate certain other monocyte functions. The exact mechanisms that govern these two pathways are not known. These results have important clinical implications for the use of IL-4 as an immunotherapeutic agent, as well as providing insight into the physiologic role of IL-4.

Because IL-4 down-regulates several proinflammatory functions associated with human monocytes/macrophages, we explored the possibility that IL-4 also decreases monocyte survival. IL-4 caused a concentration-dependent decrease in viability of IL-1 or LPS stimulated, but not unstimulated, monocytes. Nonviable cells demonstrated classic features of programmed cell death or apoptosis, in that they were condensed and contained oligonucleosome-sized (200 bp) DNA fragments. When compared with several other cytokines commonly associated with inflammatory lesions, IL-4 was uniquely effective in enhancing cell death. We found that IL-4 enhanced death more quickly in IL-1-stimulated cells than in LPS-stimulated cells, that stimulated monocytes did not become resistant to the effects of IL-4 during culture, and that the effects of IL-4 on viability were antagonized by IFN-gamma. Enhanced cell death was stimulus-specific in that monocyte viability maintained by certain activating agents, such as Con A or CSF, was unaffected by IL-4. These findings represent the first evidence of cytokine-enhanced programmed cell death in monocytes and suggest that the antiinflammatory effects of IL-4 are mediated in part by reducing survival of stimulated monocytes in chronic lesions.

Mononuclear phagocytes in the synovium of patients with arthritis, in contrast to blood monocytes, were found to express a third receptor for the constant region of Ig (Fc gamma RIII), in addition to Fc gamma RI and Fc gamma RII. Previously identified on mature mononuclear phagocytes or phagocytes exposed to transforming growth factor-beta (TGF-beta) in vitro, this study documents the presence of Fc gamma RIII (CD16) expressing cells at an inflammatory site. Furthermore, the presence of CD16 on the majority of the LeuM3 (CD14) positive synovial monocytic cells could be mimicked by exposing blood monocytes to synovial fluids from patients with rheumatoid arthritis (17 of 19) and synovial fluids from patients with osteoarthritis (4 of 4). In additional studies, the soluble factor in inflammatory synovial fluids responsible for regulating CD16 expression was found to be consistent with the presence of TGF-beta. Inhibition of the activity in synovial fluids with a neutralizing antibody to TGF-beta confirmed a role for this peptide in synovial phagocytic cell CD16 expression. Moreover, signal transduction through CD16 on synovial phagocytes resulted in augmented extracellular release of superoxide anion that may contribute to tissue damage and other inflammatory sequelae. Identification of TGF-beta and its association with upregulation of CD16 at sites of chronic inflammation may provide insight into the destructive lesions associated with inflammatory arthropathies.

In the absence of appropriate stimuli, monocytes undergo programmed cell death (PCD) or apoptosis. IL-1 beta and TNF-alpha prevent monocyte PCD, which suggests that viability may be regulated by biologically active peptides released during inflammation. To explore this possibility, we evaluated several chemotactic factors and pro-inflammatory cytokines for their ability to regulate PCD. The recruitment factors, FMLP, C fragment C5a, monocyte chemotactic protein-1, or transforming growth factor-beta 1, were incapable of rescuing monocytes from PCD nor did they enhance PCD, whereas several inflammatory cytokines in addition to IL-1 beta and TNF-alpha, including granulocyte-monocyte-CSF and IFN-gamma, prevented monocyte PCD provided that sufficient levels of these cytokines were continuously maintained in the cultures. Cytokine-mediated inhibition of PCD could be blocked by specific antisera, ruling out potential effects caused by LPS contamination. When tested at equivalent concentrations, IL-2, IL-4, and IL-6 had no effect on PCD indicating selectivity in cytokine modulation of monocyte PCD. Because monocytes produce IL-1 beta, TNF-alpha, and granulocyte-monocyte CSF when activated, the data suggest autocrine as well as paracrine control of cell survival and accumulation. The results also suggest that monocytes recruited to a site of inflammation will undergo PCD in the absence of specific cytokines and/or other stimuli that block this process.

Peritoneal and peripheral blood monocyte-macrophages from inbred Lewis (LEW) rats generate higher levels of reactive oxygen intermediates (ROI) in response to group A streptococcal cell walls (SCW) than do similar populations of cells from histocompatible Fischer rats. This differential sensitivity of the phagocytes to SCW is reflected in differences in susceptibility of the two strains to the development of arthritis in response to SCW. After systemic administration of the SCW, LEW rats develop acute and chronic erosive polyarthritis, whereas the Fischer rats are arthritis resistant. Inasmuch as these data suggested that the SCW-induced release of inflammatory cell products such as ROI might be an important contributory factor in the pathogenesis of arthritis in the LEW rats, the animals were injected with SCW and treated with ROI inhibitors. A single intraarticular injection of superoxide dismutase or catalase significantly reduced the SCW-induced inflammatory response and evolution of erosive arthritis in the treated animals (articular index 3.6 +/- 0.36 for SCW only vs 1.4 +/- 0.3 for SCW + SOD; p less than 0.001; n = 6). These data indicate that ROI play a pivotal role in synovitis and, furthermore, that suppression of these inflammatory mediators modulates both acute and chronic SCW-induced inflammation of the joint.

To evaluate the integrity of humoral immunologic memory among persons with HIV infection, we measured the levels, specificity, and functional affinity of circulating antibodies to vaccine-related recall Ag, tetanus (TT) and diphtheria toxoids (DT), and to naturally acquired measles virus, in sera from 17 HIV-seronegative control subjects, 17 asymptomatic HIV-seropositive patients, and 10 patients with AIDS. Preimmunization levels of TT- and measles-specific IgG were similar in all groups, although DT-specific IgG was lower in AIDS patients. Four wk after immunization with TT3 and DT, all groups showed significantly increased specific antibody levels (p less than 0.02). The asymptomatic HIV+ patients and control subjects achieved similar peak serum levels of TT-specific IgG (102 +/- 32 and 169 +/- 36 micrograms/ml, respectively). In contrast, the AIDS patients had lower peak values of both TT- and DT- specific IgG (p less than 0.05). Peak levels correlated directly with the number of CD4+ T cells (p less than 0.05). However, 80 to 100% of all participants tested, independent of HIV status, showed higher levels of TT- and DT-specific IgG 6 mo after immunization compared with preimmunization levels. The antitoxoid antibodies were specific as they did not cross-react with other Ag in competitive inhibition experiments. In addition, all groups exhibited antibodies to TT and DT both pre- and postimmunization of equivalent functional affinity (avidity) (Kd = 10(-10)-10(-11) mol/liter). We conclude that, in contrast to the profoundly depressed humoral responses to new Ag, persons with asymptomatic HIV infection retain humoral immunity to certain recall Ag. These levels of specific IgG to three recall Ag are not proportional to elevated levels of total serum IgG in HIV-infected patients. In addition, many patients with HIV respond to challenge with recall Ag by producing significant amounts of high affinity IgG that may persist over time.

The multifunctional cytokines, transforming growth factor (TGF)-beta 1 and beta 2, have the capability of inducing human peripheral blood monocytes to express the type III receptor for the Fc portion of IgG (Fc gamma RIII/CD16). In this study we show that the T cell-derived cytokine, IL-4, antagonizes the ability of TGF-beta to induce the expression of CD16. Furthermore, this ability to down-regulate expression of CD16 is completely abrogated after treatment with polyclonal anti-IL-4, suggesting that IL-4 is solely responsible for the observed inhibition. The mechanism for negating the effect of TGF-beta is not due to decreased expression of surface receptors for TGF-beta, but appears to occur at the mRNA level. Nuclear run-off assays indicate that regulation occurs predominantly through a posttranscriptional mechanism(s), although a transcriptional process cannot be ruled out. Normally, CD16 appears on only a small population of circulating monocytes, however, expression is apparent on the majority of mature tissue and inflammatory macrophages likely due to the release of TGF-beta in these sites. Inasmuch as this receptor binds immune complexes and opsonized particles, it is associated with enhanced immunophagocytosis. Suppression of CD16 expression and its ability to suppress a number of other monocyte functions suggests that IL-4 may play an important role in the resolution of inflammatory and tissue repair responses.

Participation of human polymorphonuclear neutrophils in the inflammatory response is mediated, in part, by soluble factors such as chemotactic peptides and cytokines. Although the cytokine, transforming growth factor beta (TGF-beta), has been shown to recruit monocytes and promote the inflammatory process, its effects on neutrophils are unknown. In this investigation, [125I]TGF-beta 1 affinity binding studies were employed to show that neutrophils express TGF-beta receptors (350 +/- 20 receptors/cell), which exhibit high affinity for the ligand (dissociation constant, 50 pM). Affinity cross-linking studies identified the receptors to be primarily of the type I class. In contrast to the receptors on monocytes, neutrophil TGF-beta receptors were not down-regulated by exposure to specific inflammatory mediators. Additional studies examined whether exposure of neutrophils to TGF-beta could enhance specific functions, as occurs with monocytes. TGF-beta was shown to cause directed migration of neutrophils at femtomolar concentrations, thus it is the most potent neutrophil chemotactic factor yet identified. Neutrophil production of reactive oxygen intermediates was not stimulated by TGF-beta, nor did TGF-beta enhance or depress subsequent PMA- or FMLP-stimulated superoxide production. However, the stable expression of neutrophil TGF-beta receptors, and the capacity of this cytokine to stimulate neutrophil chemotaxis, suggest that the pro-inflammatory effects of TGF-beta are mediated by neutrophils in addition to monocytes.

Human peripheral blood monocytes progressively lose viability when cultured in the absence of serum, cytokines, or other stimuli. In this study, we investigated whether monocyte death results from membrane damage (i.e., necrosis) or internally regulated processes [i.e., programmed cell death (PCD) or apoptosis]. Our results clearly indicated that monocytes die by PCD when cultured without stimulation. Death was associated with fragmentation of DNA into integer multiples of approximately 200 bp, a decrease in cell size, condensation of the nucleus and cytoplasmic organelles, and membrane blebbing, all of which are cardinal features of PCD. Monocytes exposed to nonphysiologic conditions such as acidic media (pH 4.2), 56 degrees C for 30 min, or freezing and thawing were killed without concomitant DNA fragmentation, indicating that DNA fragmentation was not a result of cell death per se. Addition of Escherichia coli LPS, a potent monocyte activating agent, in concentrations as low as 0.1 ng/ml caused a marked increase in monocyte survival and prevented DNA fragmentation. Moreover, exogenous human rTNF-alpha or IL-1 beta also prevented PCD, suggesting that PCD is regulated by certain cytokines released from LPS-stimulated monocytes. The results indicate that in the absence of appropriate stimulation, monocytes are programmed to undergo a sequence of molecular events leading to cell death. Regulation of PCD may be an important homeostatic mechanism for controlling the number of monocytes available to respond to infection, wound healing, and tumor growth.

Monocyte recruitment and accumulation in the synovial tissue is pivotal in the evolution of rheumatoid arthritis (RA). In the present study we examined the chemotactic potential of monocytes obtained from synovial fluid (SF) of patients with RA. Functionally, SF monocytes exhibited greatly diminished chemotactic activity to C5a compared with monocytes from the peripheral blood. In contrast, their chemotactic responsiveness to the synthetic peptide, FMLP, was nearly normal. To define a mechanism for this differential chemotactic dysfunction, cell-surface receptors for C5a (C5aR) and FMLP (FMLP-R) were evaluated. Whereas FMLP-R expression was similar on both blood and inflammatory monocytes, C5aR expression was markedly reduced on SF cells. Because decreased C5a binding in certain RA SF samples could not be attributed to free C5a, known or suspected components of inflammatory SF were evaluated for their ability to modulate chemotactic ligand receptors. Bacterial products including LPS and streptococcal cell walls, which are potent monocyte activators, down-regulated C5aR without affecting FMLP-R. Moreover, the cytokines IFN-gamma and granulocyte-macrophage-CSF selectively decreased C5aR in parallel with decreased in vitro chemotactic activity to C5a. Thus, these data indicate that 1) synovial effusions may contain C5a and/or inflammatory mediators that modulate phenotypic and functional changes in monocytes, 2) chemotactic ligand receptors are independently regulated in inflammatory lesions, and 3) decreased C5aR expression and chemotactic potential likely provide a mechanism whereby monocyte-macrophages persist within the inflamed synovium.

Localization of streptococcal cell wall Ag (SCW) in the synovial tissue of treated rats induces an influx of leukocytes and a cell-mediated immune response leading to arthritis and joint destruction. Systemic administration of the T cell product, IFN-gamma (10(6) U/kg/day), suppresses the recruitment of leukocytes into the synovium and effectively inhibits the inflammation and pathology characteristic of SCW-induced arthritis (articular index 10.4 +/- 0.6 for SCW vs 2.0 +/- 0.7 for SCW with IFN-gamma, p less than 0.005). Monocyte-macrophages from animals treated with IFN-gamma exhibited defective chemotactic responses when tested in vitro and furthermore, monocytes cultured with IFN-gamma (25 to 500 U/ml) in vitro had significantly suppressed chemotactic responses to the complement fragment C5a (p less than 0.005). The decreased ability to migrate to C5a was associated with decreased binding of fluorochrome-conjugated C5a indicative of reduced expression of C5a receptors. Based on these data, IFN-gamma that induces monocyte maturation as reflected by increased Ia expression conversely inhibits C5a receptor expression. Although locally elevated IFN-gamma levels may serve to inhibit recruitment away from an inflammatory site, systemic exposure to IFN-gamma appears to inhibit leukocyte recruitment to the inflammatory site by its ability to induce premature maturation and concomitant inability to respond to certain chemotactic ligands. Inasmuch as monocyte recruitment to the synovium is pivotal in the development of SCW-induced polyarthritis, the ability of IFN-gamma to inhibit this event effectively inhibits the synovial pathology.

Transforming growth factor (TGF-beta) is an immunoregulatory cytokine produced by many different cell types, including lymphocytes and monocytes-macrophages. Two assays are described for the quantitation of TGF-beta activity in supernatant fluids. The bioassay for TGF-beta is based on the potent ability of TGF-beta to suppress lymphocyte proliferation. The radioreceptor assay is based on the ability of TGF-beta-containing supernatant fluids to inhibit the binding of radiolabeled TGF-beta to its cellular receptor. Two support protocols describe acid-activation and neutralization of TGF-beta.

We investigated potential mechanisms by which lymphocytes infiltrating rheumatoid synovium become immunosuppressed. In 20 of 22 synovial fluids and 12 of 13 synovial tissue culture supernatants, no IL-1 bioactivity could be detected in the thymocyte proliferation assay. These same preparations could, however, support proliferation of fibroblast monolayers, consistent with the presence of IL-1 and/or other fibroblast growth factors. Addition of either rheumatoid synovial fluids or synovial culture supernatants to exogenous IL-1 in the IL-1 bioassay caused marked inhibition of the assay indicative of an IL-1 inhibitor. This inhibition of IL-1 could be reversed by treating the effusions or supernatants with a neutralizing antibody to transforming growth factor-beta (TGF-beta). Furthermore, monocyte-macrophages isolated from rheumatoid synovial fluid constitutively released both latent and active TGF-beta in culture at levels sufficient to completely block the biologic activity of 100 U/ml IL-1. The production of substantial levels of TGF-beta by synovial macrophages, as well as the apparent ability of these inflammatory macrophages to activate latent TGF-beta, implicates TGF-beta not only as an important inhibitor of IL-1-induced lymphocyte proliferation, but also as a key cytokine in promoting synovial fibroblast hyperplasia and pathology.

We investigated the effect of recombinant human granulocyte-macrophage CSF (rhGM-CSF) on the accessory cell function of purified human monocytes. Compared with untreated monocytes, rhGM-CSF-treated monocytes promoted enhanced mitogen- and Ag-stimulated lymphocyte proliferation. This enhancement was significantly inhibited by mAb to rhGM-CSF. In experiments designed to define the mechanism of rhGM-CSF augmentation of accessory cell function, rhGM-CSF was shown to cause a dose-dependent increase in monocyte expression of surface HLA-DR molecules and stimulated secretion of IL-1, both important in monocyte T cell interactions. Further studies demonstrated that levels of HLA-DR and IL-1 mRNA were increased by rhGM-CSF, indicating transcriptional regulation of gene expression for HLA-DR and IL-1. Thus, rhGM-CSF augments accessory cell function by human monocytes, and this augmentation correlates with rhGM-CSF-induced increases in transcription of the HLA-DR and IL-1 genes leading to increased expression of surface HLA-DR and secretion of IL-1.

Transforming growth factor-beta (TGF-beta) is a polypeptide that participates in morphogenesis, inflammation, and would repair. Emerging as an important immunomodulatory molecule, TGF-beta may also regulate the phenotype and function of mononuclear cells within an inflammatory site. In this study TGF-beta is shown to induce expression of the gp50-65 receptor for Fc gamma RIII on otherwise negative peripheral blood monocytes. Circulating monocytes express Fc gamma RI and Fc gamma RII, but not Fc gamma RIII, and no inducing agent for Fc gamma RIII has previously been identified. Within hours after exposure of freshly isolated peripheral blood monocytes to picomolar concentrations of TGF-beta 1 or TGF-beta 2, mRNA specific for Fc gamma RIII is detectable followed by expression of the receptors on the cell surface. Neutralizing antibodies to TGF-beta completely inhibit the TGF-beta-induced up-regulation of Fc gamma RIII. In contrast to TGF-beta, other cytokines involved in the inflammatory response are ineffective in regulating Fc gamma RIII expression on human monocytes. Thus, the early release of TGF-beta by platelets in an inflammatory site may foster enhanced immunophagocytosis and contribute to host defense by inducing monocyte Fc gamma RIII expression.

The oral epithelium is the site of first exposure of HIV-1 to host tissues during oral sex with an infected partner or through breast-feeding by an infected mother. Although the oral epithelium is distinguishable by its apparent resistance, the mucosal surfaces represent a primary target of HIV-1. After oral exposure and swallowing, infection is detected prominently in the gastrointestinal tract, which becomes depleted of CD4+ T-cells. The oral cavity and palatine tonsils appear to resist infection and transfer to susceptible lymphoid cells in the lamina propria by local anti-HIV-1 mechanisms. In some cases, expression of these antiviral mechanisms increases after exposure to HIV-1. During primary exposure and before seroconversion, based on limited in vitro and primate data, a window of opportunity for capture of HIV-1 by the oral epithelium may exist. After seroconversion, the risk of infectious HIV-1 appearing in saliva is negligible. This report considers evidence that oral epithelium has the potential both to enable and to resist infection by HIV-1.

The host response to infection or injury initiates a cascade of events involving recruitment of leukocytes and the release of multiple inflammatory mediators. One of these mediators, nitric oxide (NO), not only represents an important microbicidal agent in host defense, but also functions as a biological signaling and effector molecule in inflammation and immunity. However, overproduction of NO can be autotoxic and contribute to tissue damage and has been implicated in pathogenesis of tumors, and infectious, autoimmune and chronic degenerative diseases. NO is generated via constitutive and inducible nitric oxide synthases (iNOS) which catalyze the oxidation of a guanidino nitrogen associated with L-arginine. Whereas endothelial NOS (eNOS) and neuronal NOS (nNOS) are constitutively expressed, iNOS is transcriptionally induced by bacterial constituents and inflammatory mediators, including TNF alpha and IL-1. In an experimental model of bacterial component-induced joint inflammation and tissue degradation, functionally distinct roles of the constitutive NOS and iNOS were demonstrated. Following systemic delivery of an arthritogenic dose of streptococcal cell walls (SCW), these bacterial peptidoglycan-polysaccharide complexes disseminate and target the peripheral joints, liver and spleen of the treated animals. Following deposition of the SCW in the peripheral joints, an initial innate inflammatory response to the bacterial components progresses into an adaptive immune response with the recruitment and activation of mononuclear phagocytes and T lymphocytes. With the release of cytokines and inflammatory mediators, there is an upregulation of gene expression for iNOS, but not the constitutive nNOS or eNOS. Nonetheless, the constitutive NOS isoforms, regulated by calcium fluxes and interaction with calmodulin, may also enhance NO production. Increased release of NO was detected not only in the synovium, but also in the circulation, and plasma levels of nitrate plus nitrite, the stable products of NO reactions, correlated with disease progression. Following inhibition of NO production with nonspecific NOS inhibitors, such as N(G)-monomethyl-L-arginine, which target all three isoforms, there is a striking therapeutic benefit with reduced signs and symptoms of erosive arthritis. In contrast, selective targeting of iNOS with N-iminoethyl-L-lysine resulted in exacerbation of the synovial inflammation and degradation of joint structures. Based on these data, it appears that the constitutive isoforms of NOS contribute to the pathophysiology of the arthropathy, and that induced NOS and NO may function, in part, in a protective pathway. Moreover, the suppression of NO following treatment with TNF alpha antagonists results in reduced inflammation and the associated synovial pathology. Collectively, these data implicate discrete roles for the NOS isoforms in the emergence of local tissue pathology and underscore the need to define the specific pathways that are being targeted for interventional strategies.

Transforming growth factor-beta 1 (TGF-beta1) is a cytokine member of the TGF-beta superfamily involved in the control of proliferation and differentiation of various cell types. TGF-beta1 plays an important role in bone formation and resorption. To determine the effect of TGF-beta1 deficiency on bone mineral and matrix, tibias from mice in which TGF-beta1 expression had been ablated (TGF-beta1 null) were analyzed and compared with background- and age-matched wild-type (WT) control animals by Fourier transform-infrared imaging (FTIRI) and histochemistry. FTIRI allows the characterization of nondemineralized thin tissue sections at the ultrastructural level with a spatial resolution of approximately 7 microm. The spectroscopic parameters calculated were: mineral-to-matrix ratio (previously shown to correspond to ash weight); mineral crystallinity (related to the crystallographically determined crystallite size and perfection in the apatite c-axis direction); and collagen maturity (related to the ratio of pyridinoline:deH-DHLNL collagen cross-links). Several fields were selected to represent different stages of bone development within the same specimen from the secondary ossification center to the distal diaphysis. Anatomically equivalent areas were compared as a function of age and genotype. The spectroscopic results were expressed both as color-coded images and as pixel population distributions for each of the three parameters monitored. Based on comparisons of histochemistry and FTIRI, there were distinctive age and genotype variations. At all ages examined, in the TGF-beta1 null mice growth plates, alkaline phosphatase (ALP) activity and collagen maturity were reduced, but no effect on mineral content or crystallinity was noted. In the TGF-beta1 null mice metaphyses, there was a persistence of trabeculae, but no significant alterations in mineral content or crystallinity. In contrast, mineral content, mineral crystallinity, and collagen maturity were reduced in the secondary ossification center and cortical bone of the TGF-beta1 null mice. These results, consistent with a mechanism of impaired bone maturation in the TGF-beta1 null mice, may be directly related to TGF-beta1 deficiency and indirectly to increased expression of inflammatory cytokines in the TGFbeta1 null mice.

To determine whether human chorionic gonadotropin (HCG) contributes to pregnancy-associated immunosuppression, as observed clinically by an amelioration of symptoms in human autoimmune diseases, including rheumatoid arthritis, during pregnancy.Administration of HCG was initiated 2 days prior to an arthritogenic dose of streptococcal cell wall (SCW) in nonpregnant female rats, and the development and severity of SCW-induced arthritis was monitored. Inflammatory mediators, including plasma nitrite/nitrate and cytokine levels, were measured. Inducible nitric oxide synthase (iNOS) protein and cytokine messenger RNA expression in joint tissue were compared between treated and untreated arthritic animals.Systemic administration of HCG resulted in a dose-dependent reduction in the clinical arthritis index. Consistent with the amelioration of clinical symptoms, HCG significantly reduced the inflammatory cell infiltration, pannus formation, and bone and cartilage degradation. Mechanistically, HCG therapy was associated with suppression of the overzealous production of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-1beta, which contribute to synovial pathology in animals with SCW-induced arthritis. Circulating nitric oxide and the amount of iNOS protein were also reduced. Furthermore, circulating transforming growth factor beta levels were elevated by the HCG, all of which suggest monocytes/macrophages as a potential target.These findings indicate that HCG exerts a protective effect in this experimental arthritis model, through modulation of inflammatory mediators.

Immunodeficiency, the consequence of HIV-1 infection, predisposes the host to opportunistic infections. In turn, opportunistic pathogens influence target cell susceptibility to HIV-1 infection and replication. Although the advent of highly active antiretroviral therapy (HAART) has altered these sequelae, co-infections may prevail in some parts of the world and in failed HAART regimens. Moreover, immune activation as occurs in tonsil and non-infectious mucosal inflammatory lesions may also be associated with proximal sites of viral replication. These connections between enhancement of HIV-1 infection and activation/inflammation warrant further elucidation of the factors promoting permissiveness to HIV-1 infection. Using the opportunistic pathogen Mycobacterium avium as an in vitro model, we demonstrated that co-infection facilitated HIV-1 infection of monocyte-macrophages by multiple pathways. M. avium activated NF-kappaB, the downstream consequences of which included augmented expression of tumor necrosis factor alpha and CCR5 receptors, both permissive for sustaining HIV-1 infection. Pronounced viral replication in lymph nodes co-infected with M. avium and HIV-1 paralleled these in vitro findings. Furthermore, reduction in viral burden is associated with treatment of infected or inflamed tissues, underscoring the link between immune activation and viral replication.

Oxygen reactive intermediates released from phagocytic cells are important for microbicidal activity, but they may also be harmful to surrounding cells and matrix components at the inflammation site. In different forms of inflammatory periodontal disease, peripheral and crevicular polymorphonuclear leukocytes, as well as mononuclear phagocytes and gingival fibroblasts, are exposed to bacterial cell wall components and cytokines. The aim of this study was to evaluate if some bacterial components and cytokines induce superoxide release and superoxide dismutase (SOD) expression in gingival fibroblasts. Lipopolysaccharide (LPS), streptococcal cell walls (SCW), and formyl-methionyl-leucyl-phenylalanine were found to stimulate O2- release from gingival fibroblasts, which increased when Ca2+ was added. Phorbol myristate acetate, a potent activator of respiratory burst in phagocytes, was found to be a weak stimulator of O2- release in gingival fibroblasts. Of the cytokines tested, tumor necrosis factor (TNF)-alpha was found to activate superoxide release in gingival fibroblasts. Gene expression for manganese superoxide dismutase (MnSOD), but not for copper/zinc superoxide dismutase (CuZnSOD), was demonstrated in fibroblasts exposed to LPS, SCW and TNF-alpha using Northern blot analysis. The production of MnSOD may be protective for these cells. We conclude that bacterial cell wall components and cytokines modulate O2- release by gingival fibroblasts which may contribute to periodontal pathology.

Intraperitoneal injection of streptococcal cell walls (SCW) into Lewis rats results in dissemination of SCW to the liver, spleen, bone marrow, and peripheral joints. The uptake of SCW by Kupffer cells in the liver initiates a chain of events largely mediated by T lymphocytes and macrophages. Local synthesis and secretion of cytokines and growth factors in response to the persistent SCW lead to the evolution and maintenance of a chronic T cell-dependent granulomatous response and result in granuloma formation and irreversible hepatic fibrosis. In an attempt to impede the development of the chronic granulomatous lesions in the liver, we injected a plasmid DNA encoding TGF-beta 1 i.m. to the SCW animals to determine the effect of TGF-beta 1 gene transfer on the course of liver inflammation and fibrosis. A single injection of plasmid DNA encoding TGF-beta 1 resulted in virtual abolition of the development of the SCW-induced hepatic granuloma formation and matrix expansion. TGF-beta 1 DNA not only reduced key proinflammatory cytokines including TNF-alpha, IL-1 beta, IFN-gamma, and IL-18, but also inhibited both CXC and CC chemokine production, thereby blocking inflammatory cell recruitment and accumulation in the liver. Moreover, TGF-beta 1 gene delivery inhibited its own expression in the liver tissue, which is otherwise up-regulated in SCW-injected animals. Our study suggests that TGF-beta 1 gene transfer suppresses hepatic granuloma formation by blocking the recruitment of inflammatory cells to the liver, and thus may provide a new approach to the control of hepatic granulomatous and fibrotic diseases.

Though it has been shown that TGF-beta 1 directs B cells to switch to IgA in vitro, no studies have assessed TGF-beta 1 effects on mucosal vs systemic immunity in vivo. When the B cell functions of TGF-beta 1 gene-disrupted (TGF-beta 1-/-) mice were analyzed, significantly decreased IgA levels and increased IgG and IgM levels in serum and external secretions were observed. Further, analysis of Ab forming cells (AFC) isolated from both mucosal and systemic lymphoid tissue showed elevated IgM, IgG, and IgE, with decreased IgA AFC. A lack of IgA-committed B cells was seen in TGF-beta 1-/- mice, especially in the gastrointestinal (GI) tract. Splenic T cells triggered via the TCR expressed elevated Th2-type cytokines and, consistent with this observation, a 31-fold increase in serum IgE was seen in TGF-beta 1-/- mice. Thus, uncontrolled B cell responses, which include elevated IgE levels, a lack of antiinflammatory IgA, and an excess of complement-binding IgG and IgM Abs, will promote inflammation at mucosal surfaces in TGF-beta 1-/- mice and likely contribute to pulmonary and GI tract lesions, ultimately leading to the early death of these mice.

To investigate the ability of hemoglobin (Hgb), a nitric oxide (NO) scavenger, to deplete excess NO and reduce inflammation and injury in synovial tissue from joints with inflammatory arthritis.The severity of streptococcal cell wall-induced arthritis was monitored following administration of Hgb. Plasma nitrite and nitrate levels were measured, and inducible NO synthase (iNOS) and cytokine messenger RNA (mRNA) expression in peripheral blood mononuclear cells (PBMC) and joint tissue were evaluated.Following systemic administration of Hgb to arthritic rats, plasma levels of nitrite and nitrate as well as iNOS mRNA expression in the joints and PBMC were significantly reduced. Moreover, inflammatory cell accumulation and disease pathology in the joint tissue were dramatically attenuated without obvious side effects. Consistent with this reduction in the inflammatory response, cytokine gene expression was decreased in the synovium of Hgb-treated rats.Modulation of NO levels through the use of a NO scavenger, Hgb, influences the development and severity of arthritis. These findings suggest that depletion of excess NO by NO scavengers provides a prototype for further exploration of potential treatments for chronic arthritis.

To determine CD4+ T-cell count and circulating and tissue levels of HIV before and after surgery in a patient with recent-onset ulcerative colitis.CD4 lymphocytes and circulating and tissue HIV RNA levels were measured in an HIV-infected patient with ulcerative colitis before and after proctocolectomy.Approximately 3 weeks prior to surgery for ulcerative colitis that was unresponsive to corticosteroids, the patient's CD4 count was 930 cells/mm3 and fell to 313 cells/mm3 within 10 days; the viral burden was approximately 80,000 RNA copies/mL. Tissue macrophages and lymphocytes in biopsy and resection specimens were shown to express high levels of HIV RNA by in situ hybridization. Five days postoperatively, the patient became asymptomatic and was discharged on tapering prednisone without antiretroviral agents. After surgery, the patient's CD4 count progressively rose, while viral RNA levels precipitously dropped. At 3, 6, and 15 weeks postoperatively, CD4 and viral RNA counts were 622 cells/mm3 and 31,300 RNA copies/mL, 843 cells/mm3 and 11,400 RNA copies/mL, and 747 cells/mm3 and 1500 RNA copies/mL, respectively.Circulating levels of HIV and CD4+ cells, as well as tissue expression of HIV, apparently can be influenced by localized inflammatory processes such as those occurring in inflammatory bowel disease.

Oral administration of autoantigens can influence the outcome of experimental autoimmune diseases, yet little is known about nonself Ag-induced tolerance. In this study, we administered group A streptococcal cell wall (SCW) peptidoglycan-polysaccharide complexes orally and monitored the impact on SCW-induced erosive polyarthritis. Oral administration of low dose SCW (3 microg/day), initiated 7 days before an arthritogenic dose of systemic SCW, virtually eliminated the joint swelling and destruction typically observed during both the acute and chronic phases of the arthritis. High (300 microg), but not intermediate (30 microg), dose regimens also profoundly inhibited the disease. Most previous studies have demonstrated that prior feeding is required for efficacy, yet oral feeding of low dose SCW suppressed the evolution of arthritis even when administration was begun 10-15 days after induction of the arthritis. While the synovial inflammatory cell infiltration and expression of proinflammatory cytokines were markedly suppressed, no local enhancement of the regulatory cytokines IL-4, IL-10, and TGF-beta was detected. Oral administration of low dose SCW, however, up-regulated circulating levels of TGF-beta, concomitant with decreased circulating TNF-alpha and suppression of chronic arthritis. Moreover, IL-10 was increased in tolerized spleen lymphocytes, and unexpectedly, this SCW-specific IL-10 production was TGF-beta dependent. These data support a pivotal role for TGF-beta, although not necessarily in the joint, in the regulation of specific immune tolerance responsible for suppressed synovial inflammation and matrix destruction. The distant induction and up-regulation of regulatory cytokines and/or cells may contribute to the inhibition of the immune response through blunted infiltration of inflammatory cells to the joint.