Corticosteroids Inhibit Il-12 Production In Human Monocytes And Enhance Their Capacity To Induce Il-4 Synthesis In Cd4+ Lymphocytes

We examined the effects of corticosteroids on IL-12 production by human monocytes and on cytokine synthesis in T cells. To distinguish the effects of corticosteroids on the APC used to activate the T cell from direct effects of corticosteroids on the T cell, experiments were performed by exposing the APC and not the T cell to corticosteroids. We found that corticosteroids significantly inhibited the production in monocytes of IL-12, a cytokine that is extremely potent in enhancing IFN-γ and inhibiting IL-4 synthesis in T cells. We demonstrated that reduced production of IL-12 in corticosteroid-treated monocytes resulted in a decreased capacity of the monocytes to induce IFN-γ and an increased ability to induce IL-4 in T cells. These results suggest that although corticosteroids may be beneficial for the treatment of asthma or allergic disease due to direct inhibitory effects of corticosteroids on cytokine synthesis in T cells, chronic corticosteroid therapy may indirectly exacerbate the long-term course of allergic disease. This deleterious effect of corticosteroids would result from a limitation in IL-12 production in tissue monocytes and macrophages, which would enhance production of Th2 cytokines (which augment allergic disease), and would reduce production of Th1 cytokines (which attenuate allergic disease) in T cells that subsequently infiltrate the tissues.1581255895595Goulding, N.J., Guyre, P.M., Glucocorticoids, lipocortins and the immune response (1993) Curr. Opin. Immunol., 5, p. 108Gillis, S., Crabtree, G.R., Smith, K.A., Glucocorticoid-induced inhibition of T cell growth factor production. I. The effect on mitogen-induced lymphocyte proliferation (1979) J. Immunol., 123, p. 1624Arya, S.K., Wong-Staal, F., Gallo, R.C., Dexamethasone-mediated inhibition of human T cell growth factor and γ-interferon messenger RNA (1984) J. Immunol., 133, p. 273Schleimer, R.P., Bochner, B.S., The effects of glucocorticoids on human eosinophils (1994) J. Allergy Clin. Immunol., 94, p. 1202(1992) International Consensus Report on Diagnosis and Management of Asthma, , U.S. Public Health Service, Washington, DCRobinson, D.S., Hamid, Q., Ying, S., Tsicopoulos, A., Barkans, J., Bentley, A.M., Corrigan, C., Kay, A.B., Predominant Th2-like bronchoalveolar T-lymphocyte population in atopic asthma (1992) N. Engl. J. Med., 326, p. 298Busse, W.W., Coffman, R.L., Gelfand, E.W., Kay, A.B., Rosenwasser, L.J., Mechanisms of persistent airway inflammation in asthma: A role for T cells and T-cell products (1995) Am. J. Respir. Crit. Care Med., 152, p. 388Umetsu, D.T., Bocian, R.C., DeKruyff, R.H., Restricted lymphokine synthesis in CD4+ helper T cells: New insights into the regulation of IgE synthesis and into the pathogenesis of allergic disease (1992) Pediatr. Asthma Allergy Immunol., 6, p. 1Romagnani, S., Lymphokine production by human T cells in disease states (1994) Annu. Rev. Immunol., 12, p. 227Bousquet, J., Chanez, P., Lacoste, J.Y., Barneon, G., Ghavanian, N., Enander, I., Venge, P., Godard, P., Eosinophilic inflammation in asthma (1990) N. Engl. J. Med., 323, p. 1033Gundel, R.H., Letts, L.G., Gleich, G.J., Human eosinophil major basic-protein induces airway constriction and airway hyperresponsiveness in primates (1991) J. Clin. Invest., 87, p. 1470Scheinman, R.I., Cogswell, P.C., Lofquist, A.K., Baldwin Jr., A.S., Role of transcriptional activation of IκB alpha in mediation of immunosuppression by glucocorticoids (1995) Science, 270, p. 283Auphan, N., Didonato, J.A., Rosette, C., Helmberg, A., Karin, M., Immunosuppression by glucocorticoids: Inhibition of NF-κB activity through induction of I kappa B synthesis (1995) Science, 270, p. 286Barnes, P.J., Molecular mechanisms of steroid action in asthma (1996) J. Allergy Clin. Immunol., 97, p. 159Wu, C.Y., Fargeas, C., Nakajima, T., Delespesse, G., Glucocorticoids supress the production of interleukin 4 by human lymphocytes (1991) Eur. J. Immunol., 21, p. 2645Leung, D.Y., Martin, R.J., Szefler, S.J., Sher, E.R., Ying, S., Kay, A.B., Hamid, Q., Dysregulation of interleukin 4, interleukin 5, and interferon γ gene expression in steroid-resistant asthma (1995) J. Exp. Med., 181, p. 33Ramirez, F., Fowell, D.J., Puklavec, M., Simmonds, S., Mason, D., Glucocorticoids promote a Th2 cytokine response by CD4+ T cells in vitro (1996) J. Immunol., 156, p. 2406Daynes, R.A., Araneo, B.A., Contrasting effects of glucocorticoids on the capacity of T cells to produce the growth factors interleukin 2 and interleukin 4 (1989) Eur. J. Immunol., 19, p. 2319Zieg, G., Lack, G., Harbeck, R.J., Gelfand, E.W., Leung, D.Y., In vivo effects of glucocorticoids on IgE production (1994) J. Allergy Clin. Immunol., 94, p. 222Trinchieri, G., InterIeukin-12: A proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity (1995) Annu. Rev. Immunol., 13, p. 251Marshall, J.D., Secrist, H., DeKruyff, R.H., Wolf, S.F., Umetsu, D.T., IL-12 inhibits the production of IL-4 and IL-10 in allergen-specific human CD4+ T lymphocytes (1995) J. Immunol., 155, p. 111Secrist, H., DeKruyff, R.H., Umetsu, D.T., Interleukin 4 production by CD4+ cells from allergic individuals is modulated by allergen concentration and antigen-presenting cell type (1995) J. Exp. Med., 181, p. 1081Marshall, J., Wen, Y., Abrams, J.S., Umetsu, D.T., In vitro synthesis of IL-4 by human CD4+ T cells requires repeated antigenic stimulation (1993) Cell. Immunol., 152, p. 18Li, Y., Ito, N., Suzuki, T., Stechschulte, D.J., Dileepan, K.N., Dexamethasone inhibits nitric oxide-mediated cytotoxicity via effects on both macrophages and target cells (1995) Immunopharmacology, 30, p. 177Constant, S., Pfeiffer, C., Woodard, A., Pasqualini, T., Bottomly, K., Extent of T cell receptor ligation can determine the functional differentiation of naive CD4+ T cells (1995) J. Exp. Med., 182, p. 1591Viola, A., Lanzavecchia, A., T cell activation determined by T cell receptor number and tunable thresholds (1996) Science, 273, p. 104Snijdewint, F.G., Kapsenberg, M.L., Wauben-Penris, P.J., Bos, J.D., Corticosteroids class-dependently inhibit in vitro Th1- And Th2-type cytokine production (1995) Immunopharmacology, 29, p. 93Masuyama, K., Jacobson, M.R., Rak, S., Meng, Q., Sudderick, R.M., Kay, A., Lowhagen, O., Topical glucocorticosteroid (fluticasone propionate) inhibits cells expressing cytokine mRNA for inlerleukin-4 in the nasal mucosa in allergen-induced rhinitis (1994) Immunology, 82, p. 192Culpepper, J.A., Lee, F., Regulation of IL-3 expression by glucocorticoids in cloned murine T lymphocytes (1985) J. Immunol., 135, p. 3191Beutler, B., Krochin, N., Milsark, I.W., Luedke, C., Cerami, A., Control of cachectin synthesis: Mechanisms of endotoxin resistance (1986) Science, 232, p. 977Truss, M., Beato, M., Steroid hormone receptors: Interaction with deoxyribonucleic acid and transcription factors (1993) Endocrinol. Rev., 14, p. 459Yang-Yen, H.F., Chambard, J.C., Sun, Y.S., Transcriptional interference between c-Jun and the glucocorticoid receptor: Mutual inhibition of DNA binding due to direct protein-protein interaction (1990) Cell, 62, p. 1205Adcock, I.M., Brown, C.R., Gelder, C.M., Shirasaki, H., Peters, M.J., Barnes, P.J., The effects of glucocorticoids on transcription factor activation in human peripheral blood mononuclear cells (1995) Am. J. Physiol., 37, pp. C331Ray, A., Prefontaine, K.E., Physical association and functional antagonism between the p65 subunit of transcription factor NF-κB and the glucocorticoid receptor (1994) Proc. Natl. Acad. Sci. USA, 91, p. 752Scheinman, R.I., Cogswell, P.C., Lofquist, A.K., Baldwin Jr., A.S., Role of transcriptional activation of IκBα in mediation of immunosuppression by glucocorticoids (1995) Science, 270, p. 283Auphan, N., DiDonato, J.A., Rosette, C., Helmberg, A., Karin, M., Immunosuppression by glucocorticoids: Inhibition of NF-κB activity through induction of IκB synthesis (1995) Science, 270, p. 286Zembowicz, A., Vane, J.R., Induction of nitric oxide synthase activity by toxic shock syndrome toxin 1 in a macrophage-monocyte cell line (1992) Proc. Natl. Acad. Sci. USA, 89, p. 2051Breuninger, L.M., Dempsey, W.L., Uhl, J., Murasko, D.M., Hydrocortisone regulation of interleukin-6 protein production by a purified population of human peripheral blood monocytes (1993) Clin. Immunol. Immunopathol., 69, p. 205Heidenreich, S., Kubis, T., Schmidt, M., Fegeler, W., Glucocorticoid-induced alterations of monocyte defense mechanisms against Candida albicans (1994) Cell. Immunol., 157, p. 320Murphy, T.L., Cleveland, M.G., Kulesza, P., Magram, J., Murphy, K.M., Regulation of interleukin 12 p40 expression through an NF-κB half-site (1995) Mol. Cell. Biol., 15, p. 5258Manetti, R., Gerosa, F., Giudizi, M.G., Biagiotti, R., Parronchi, P., Piccinni, M., Sampognaro, S., Trinchieri, G., Interleukin 12 induces stable priming for interferon γ (IFN-γ) production during differentiation of human T helper (Th) cells and transient IFN-γ production in established Th2 cell clones (1994) J. Exp. Med., 179, p. 1273DeKruyff, R.H., Fang, Y., Wolf, S.F., Umetsu, D.T., IL-12 inhibits IL-4 synthesis in keyhole limpet hemocyanin-primed CD4+ T cells through an effect on antigen-presenting cells (1995) J. Immunol., 154, p. 2578Murphy, E., Shibuya, K., Hosken, N., Openshaw, P., Maino, V., Davis, K., Murphy, K., O'Garra, A., Reversibility of T helper 1 and 2 populations is lost after long-term stimulation (1996) J. Exp. Med., 183, p. 901Jeannin, P., Delneste, Y., Seveso, M., Life, P., Bonnefoy, J.Y., IL-12 synergizes with IL-2 and other stimuli in inducing IL-10 production by human T cells (1996) J. Immunol., 156, p. 3159Windhagen, A., Anderson, D.E., Carrizosa, A., Williams, R.E., Hafler, D.A., IL-12 induces human T cells secreting IL-10 with IFN-γ (1996) J. Immunol., 157, p. 1127Magram, J., Connaughton, S.E., Warrier, R.R., Carvajal, D.M., Wu, C.Y., Ferrante, J., Stewart, C., Gately, M.K., IL-12-deficient mice are defective in IFN-γ production and type 1 cytokine responses (1996) Immunity, 4, p. 471Snyder, D.S., Unanue, E.R., Corticosteroids inhibit murine macrophage Ia expression and interleukin 1 production (1982) J. Immunol., 129, p. 1803Moser, M., De Smedt, T., Somasse, T., Tielemans, F., Chentoun, A.A., Muraille, E., Van Mechelen, M., Leo, O., Glucocorticoids down-regulate dendritic cell function in vitro and in vivo (1995) Eur. J. Immunol., 25, p. 281

Blockade of Tim-1 and Tim-4 Enhances Atherosclerosis in Low-Density Lipoprotein Receptor-Deficient Mice

Objective - T cell immunoglobulin and mucin domain (Tim) proteins are expressed by numerous immune cells, recognize phosphatidylserine on apoptotic cells, and function as costimulators or coinhibitors. Tim-1 is expressed by activated T cells but is also found on dendritic cells and B cells. Tim-4, present on macrophages and dendritic cells, plays a critical role in apoptotic cell clearance, regulates the number of phosphatidylserine-expressing activated T cells, and is genetically associated with low low-density lipoprotein and triglyceride levels. Because these functions of Tim-1 and Tim-4 could affect atherosclerosis, their modulation has potential therapeutic value in cardiovascular disease. Approach and Results - ldlr-/- mice were fed a high-fat diet for 4 weeks while being treated with control (rat immunoglobulin G1) or anti-Tim-1 (3D10) or -Tim-4 (21H12) monoclonal antibodies that block phosphatidylserine recognition and phagocytosis. Both anti-Tim-1 and anti-Tim-4 treatments enhanced atherosclerosis by 45% compared with controls by impairment of efferocytosis and increasing aortic CD4+T cells. Consistently, anti-Tim-4-treated mice showed increased percentages of activated T cells and late apoptotic cells in the circulation. Moreover, in vitro blockade of Tim-4 inhibited efferocytosis of oxidized low-density lipoprotein-induced apoptotic macrophages. Although anti-Tim-4 treatment increased T helper cell (Th)1 and Th2 responses, anti-Tim-1 induced Th2 responses but dramatically reduced the percentage of regulatory T cells. Finally, combined blockade of Tim-1 and Tim-4 increased atherosclerotic lesion size by 59%. Conclusions - Blockade of Tim-4 aggravates atherosclerosis likely by prevention of phagocytosis of phosphatidylserine-expressing apoptotic cells and activated T cells by Tim-4-expressing cells, whereas Tim-1-associated effects on atherosclerosis are related to changes in Th1/Th2 balance and reduced circulating regulatory T cells

Inability of interleukin-12 to modulate T-helper 0 effectors to T-helper 1 effectors: a possible distinct subset of T cells

Interleukin-12 (IL-12) strongly favours the development of T-helper 1 (Th1)-type cells through its ability to induce interferon-γ (IFN-γ) production by natural killer cells and T cells. In the present work we analysed the effects of IL-12 on the synthesis and secretion of IFN-γ and IL-4 by human T-cell clones. Several previously described human T-cell clones exhibiting Th1, Th2 or Th0 phenotypes were used for these analyses. We demonstrated, by enzyme-linked immunosorbent assay (ELISA) and intracytoplasmic staining, that, in Th0 clones, IL-12 up-regulated the production of both IFN-γ and IL-4 and was unable to modulate these cells to Th1-type. The up-regulation of cytokine gene expression was transcriptionally regulated and was not due to differences in mRNA stability. In Th1 cells, IL-12 up-regulated only IFN-γ and not IL-4. However, in Th2 cells, both IFN-γ and IL-4 were up-regulated by IL-12. This suggests that Th2 cells may be less stable than Th1 cells. We also observed that human Th2 cells expressed the IL-12β2 receptor, in contrast to murine Th2, which lacks this receptor. The observed differences in the effects of IL-12 on the three T-cell subsets may have important ramifications for IL-12-based therapies

Sulfasalazine prevents T-helper 1 immune response by suppressing interleukin-12 production in macrophages

Interleukin-12 (IL-12) plays a pivotal role in the development of T-helper 1 (Th1) immune response, which may be involved in the pathogenesis of chronic inflammatory autoimmune disorders. In this study we investigated the effects of sulfasalazine, a drug for treating inflammatory bowel disease and rheumatoid arthritis, on the production of IL-12 from mouse macrophages stimulated with lipopolysaccharide (LPS). Sulfasalazine potently inhibited the production of IL-12 in a dose-dependent manner, in part through the down-regulation of nuclear factor κB (NFκB) activation in IL-12 p40 gene. Activation of macrophages by LPS resulted in markedly enhanced binding activities to the κB site, which significantly decreased upon addition of sulfasalazine as demonstrated by an electrophoretic gel shift assay. Importantly, macrophages pretreated with sulfasalazine either in vitro or in vivo reduced their ability to induce interferon-γ (IFN-γ) and increased the ability to induce IL-4 in antigen-primed CD4+ T cells. From these results, sulfasalazine may induce the Th2 cytokine profile in CD4+ T cells by suppressing IL-12 production in macrophages, and sulfasalazine-induced inhibition of IL-12 production in macrophages may explain some of the known biological effects of sulfasalazine

The radiation-attenuated schistosome vaccine induces high levels of protective immunity in the absence of B cells

Radiation-attenuated cercariae of Schistosoma mansoni elicit consistently high levels of protective immunity in mice. The cell-mediated pulmonary effector mechanisms have been well characterized but the role of B cells and antibodies remains ill defined. We have compared the immune responses of B-cell-deficient (μMT) mice and their wild-type (WT) counterparts following exposure to the attenuated vaccine. Both groups mounted a T helper type 1 (Th1)-biased response in the skin-draining lymph nodes after vaccination. Interferon-γ was the dominant cytokine secreted by airway leucocytes after challenge in both μMT and WT mice, but there was a somewhat greater Th2 component in the former animals. The cellular infiltrates observed in the airways, and the pulmonary effector foci, were of similar composition in the two groups although some large foci were present in the μMT mice. There was a marked dichotomy in the protection induced in μMT animals by a single vaccination, with two-thirds showing levels similar to their WT counterparts, demonstrating that cell-mediated mechanisms alone can provide adequate protection. The remaining μMT mice had a mean worm burden identical to that of their challenge controls. A possible explanation is that a proportion of the μMT animals have a genetic defect closely associated with the μ-heavy-chain locus on chromosome 12, which affects their ability to mount a protective cell-mediated response. Three vaccinations enhanced the immunity of WT animals, most likely by augmenting antibody-mediated mechanisms. In contrast, no enhancement was seen in μMT mice, suggesting that the cell-mediated response is not boosted by multiple exposures to attenuated larvae