Inhibition of interferon γ induced interleukin 12 production: A potential mechanism for the anti-inflammatory activities of tumor necrosis factor

J Hodge-Dufour

M Marino

M Horton

A Jungbluth

^{Immunology Graduate Group, University of Pennsylvania, Philadelphia, PA 19104;}^{Ludwig Institute for Cancer Research, New York, NY 10105;}^{Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205;}^{Department of Medicine, University of Michigan Medical School, Ann Arbor, MI 48109;}^{Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520 and The VA Connecticut Healthcare System, West Haven, CT 06516;}^{Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA 19104; and}^{The Wistar Institute, Philadelphia, PA 19104}

^{To whom reprint requests should be addressed at: The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104-4268. e-mail:}ude.nnepu.ratsiw.atsiw@erup.

Edited by Anthony Cerami, The Kenneth S. Warren Laboratories, Tarrytown, NY, and approved September 23, 1998

Received 1998 Apr 23

Abstract

Inflammation is associated with production of cytokines and chemokines that recruit and activate inflammatory cells. Interleukin (IL) 12 produced by macrophages in response to various stimuli is a potent inducer of interferon (IFN) γ production. IFN-γ, in turn, markedly enhances IL-12 production. Although the immune response is typically self-limiting, the mechanisms involved are unclear. We demonstrate that IFN-γ inhibits production of chemokines (macrophage inflammatory proteins MIP-1α and MIP-1β). Furthermore, pre-exposure to tumor necrosis factor (TNF) inhibited IFN-γ priming for production of high levels of IL-12 by macrophages in vitro. Inhibition of IL-12 by TNF can be mediated by both IL-10-dependent and IL-10-independent mechanisms. To determine whether TNF inhibition of IFN-γ-induced IL-12 production contributed to the resolution of an inflammatory response in vivo, the response of TNF^{and TNF}^{mice injected with}Corynebacterium parvum were compared. TNF^{mice developed a delayed, but vigorous, inflammatory response leading to death, whereas TNF}^{mice exhibited a prompt response that resolved. Serum IL-12 levels were elevated 3-fold in}C. parvum-treated TNF^{mice compared with TNF}^{mice. Treatment with a neutralizing anti-IL-12 antibody led to resolution of the response to}C. parvum in TNF^{mice. We conclude that the role of TNF in limiting the extent and duration of inflammatory responses}in vivo involves its capacity to regulate macrophage IL-12 production. IFN-γ inhibition of chemokine production and inhibition of IFN-γ-induced IL-12 production by TNF provide potential mechanisms by which these cytokines can exert anti-inflammatory/repair function(s).

Abstract

Inflammation is normally a localized, protective response to tissue injury. The accumulation and activation of leukocytes at sites of inflammation occurs through a tightly regulated program involving cell adhesion receptors, chemoattractants, and proinflammatory cytokines. Cytokines such as tumor necrosis factor (TNF) and interleukin (IL) 1β are released early and alter blood flow, increase vascular permeability, augment leukocyte adhesion, promote migration into tissue space, and stimulate leukocytes to destroy inciting agents.

Components of the extracellular matrix (ECM), in combination with adhesion receptors, provide cells with the necessary traffic signals to migrate to an inflammatory site (1). The ECM can be modified by an evolving inflammatory process by binding and anchoring proinflammatory cytokines and chemokines and by being processed into biologically active products or fragments. Such modifications can confer proinflammatory activities on matrix components.

Infiltrating leukocytes produce cytokines that amplify the ongoing response. One such cytokine is IL-12, a potent proinflammatory cytokine produced mainly by phagocytic cells in response to bacteria and parasites or, as has been recently demonstrated, by low molecular weight forms of the ECM component hyaluronan (LMW-HA) (2, 3). IL-12 plays a critical role in bridging the innate and adaptive immune responses by inducing interferon (IFN) γ production by T and NK cells and thereby a TH1 type immune response (2). In turn, IFN-γ markedly augments IL-12 production, thus providing an important amplifying mechanism in inflammation (2).

The inflammatory response typically is self-limiting, but the regulatory mechanisms remain unclear. States of chronic inflammation, such as those seen in rheumatoid arthritis, involve the unremitting recruitment and activation of monocytes/macrophages, neutrophils, and T lymphocytes, resulting in excessive cytokine production and ECM turnover and tissue damage. Ultimately, this chronic inflammation can lead to scar tissue formation and end organ dysfunction. However, ECM components and proinflammatory cytokines, although required for an inflammatory response, under appropriate conditions also may play a negative regulatory role.

In this study we investigated the regulation of LMW-HA- and lipopolysaccharide (LPS)-induced chemokine/cytokine production by IFN-γ and TNF. We demonstrate that although IFN-γ enhanced LMW-HA-induced macrophage IL-12 production, it inhibited the production of macrophage inflammatory proteins MIP-1α and MIP-1β in response to LMW-HA, thereby having the potential to promote leukocyte activation at an inflammatory site while limiting further recruitment. Additionally, while concomitant treatment with TNF, IFN-γ, and LMW-HA, or LPS led to increased IL-12 production, pre-exposure to TNF markedly inhibited IFN-γ-enhanced IL-12 production. This activity was specific to TNF, was mediated through the p55 subunit of the TNF receptor (TNFR), and can occur by IL-10-dependent and IL-10-independent mechanisms. Further, TNF inhibits IL-12 production in part by inhibiting the accumulation of IL-12 p40 mRNA. To determine whether TNF inhibition of IL-12 plays a role in the recently reported homeostatic function of TNF in limiting the inflammatory process in vivo, we compared the response of TNF^{and TNF}^{mice to heat-killed}Corynebacterium parvum. Our results provide further evidence that TNF and IFN-γ are not only critical for the initiation and progression of an inflammatory response, but also for the down-regulation and resolution of the response both in vitro and in vivo.

Acknowledgments

We thank Drs. Lloyd J. Old, Aili L. Lazaar, and Giorgio Trinchieri for many helpful discussions and critical review of this manuscript. This work was supported by National Institutes of Health Grants HL50057 (to R.M.S), HL60539 (to P.W.N), and AI42334 (to C.A.H) and a grant from the Arthritis Foundation (to E.P.). J.H.D. is supported by National Institutes of Health Training Grant CA 09171.

Acknowledgments

ABBREVIATIONS

ECM	extracellular matrix
TNF	tumor necrosis factor
IL	interleukin
IFN	interferon
HA	hyaluronan
LMW-HA	low molecular weight hyaluronan
LPS	lipopolysaccharide
MIP	macrophage inflammatory protein
TNFR	tumor necrosis factor receptor

ABBREVIATIONS

Footnotes

This paper was submitted directly (Track II) to the Proceedings Office.

Footnotes

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