Tumor-derived macrophage migration inhibitory factor (MIF) inhibits T lymphocyte activation
Introduction
Macrophage migration inhibitory factor (MIF) was first described as an activator of macrophage function that inhibits random macrophage migration. MIF is a 12.5 kDa protein secreted by macrophages, T cells, and other tissues, notably the anterior pituitary, that induces both gram-negative septic shock and delayed-type hypersensitivity reactions [1], [2], [3], [4]. MIF is unique in that the protein sequence and structural studies have not identified any cytokine family within which MIF can be placed. MIF is the only cytokine that is upregulated by glucocorticoids, and thus MIF plays a role in regulating host global responses to infection as glucocorticoid hormones are released from the hypothalamus-pituitary-adrenal-axis [3], [5]. Although a naturally occurring substrate has not been identified, MIF also has enzymatic properties and exhibits tautomerase and redox activity. This structural aspect of MIF is somewhat reminiscent of cyclophilin, which functions intracellularly as a peptidy-prolyl cis-trans isomerase; and can be secreted in response to proinflammatory stimuli [6], [7]. Studies have shown that MIF can override glucocorticoid inhibition of T-cell activation [8]. Furthermore, studies with a neutralizing anti-MIF antibody have shown that depletion of MIF from activated T-cell cultures inhibits T-cell proliferation in vivo and can inhibit the development of DTH lesions [8], [9].
Malignant tissues are able to blunt or alter anti-tumor immune responses by the production of soluble factors that act directly on lymphocytes [10], [11]. In our studies of the immune response to neuroblastoma, we found that both murine neuroblastoma cell lines and low passage clinical tumor isolates from either surgical resection of primary disease or bone marrow metastases produce MIF [12]. The list of other cytokines produced by neuroblastoma that could potentially influence the anti-tumor immune response also includes tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, and IL-15 [13], [14], [15], [16]. The prevalence of each of these factors remains to be established, as in our limited analysis of clinical isolates only 1 in 5 produced IL-6, while 4 of 5 produced MIF. MIF has pleiotropic effects on both tumor cells and lymphocytes. MIF derived from neuroblastoma displays the classical functional activity of decreasing macrophage migration from glass capillary tubes, while also increasing transwell migration of tumor [12]. Studies with either tumor-derived, a parasite-encoded homolog, or recombinantly expressed MIF demonstrated that MIF not only inhibits macrophage random migration, but that it also plays a role in directional movement of tumor cells and macrophages [12], [17], [18].
The production of MIF by tumors appears contrary to its reported immuno-stimulatory activity. However, MIF plays a broad role in growth regulation in a number of systems. MIF is able to antagonize p53-mediated gene activation and apoptosis, and benzo[a]pyrene-induced fibrosarcomas in MIF knockout mice were smaller in size and had a lower mitotic index [19]. The MIF receptor has not been identified and is still under active investigation. CD74, the cell surface form of major histocompatibility class II-associated invariant chain, has been demonstrated to bind MIF with high affinity and is required for MIF-mediated extracellular signal-regulated kinase (ERK)-1/2 phosphorylation and PGE2 production in defined systems [20]. MIF has also been shown to have intracellular function in the regulation of cell growth via Jab1, a co-activator of AP-1 transcription [21].
With regard to the immune system, MIF is considered a broad-spectrum proinflammatory cytokine. MIF has been shown to stimulate TNF-α synthesis, is detected in DTH-reactive cells, and has been found to stimulate cognate T-cell activation [3], [22]. T cells activated by specific antigen, mitogen, or anti-CD3 antibody increase MIF mRNA expression and secretion of protein. Anti-MIF antibodies inhibit T-cell proliferation and B cell antibody production in vivo [8]. However, some studies have demonstrated negative effects of MIF on T-cell immunity. EG7 tumor-bearing mice treated with anti-MIF antibody show an increase in CD4 and CD8 T-cell accumulation at the tumor site and an increase in T-cell accumulation of adoptively transferred T cells [23]. Thus, decreasing MIF augmented the immune response to EG7. In studies described here, we sought to determine if tumor-encoded MIF has the potential to impact anti-tumor immunity. We found that high levels of tumor-derived MIF inhibited T-cell activation by preventing cell cycle progression and augmenting apoptosis through a specific signaling pathway that involved T lymphocyte-secreted interferon-gamma (IFN-γ). We speculate that the high levels of MIF produced by tumor cells results in activation-induced cell death of tumor-infiltrating T lymphocytes, thereby suppressing anti-tumor immunity.
Section snippets
Mice, tumor cells and reagents
A/J, C57BL/6 (termed B6) mice and IFN-γ knockout mice (B6.129S7-Ifngtm1Ts/J) were purchased from The Jackson Laboratory (Bar Harbor, MA) and used at 4–6 weeks of age. Mice were housed in the Biomedical Research Center at the Medical College of Wisconsin according to institutional guidelines. AGN2a is an aggressive subclone of the mouse Neuro-2a neuroblastoma cell line generated by serial in vivo passage [24]. Permanent cell lines transfected to express CD80 and CD137L (AGN2a/CD80-137L) were
In vitro demonstration of MIF knockdown
In previous work we described the production of soluble MIF in low passage cultures of patient-derived tumors and in a mouse neuroblastoma cell line, AGN2a [12]. To determine the functional consequences of inhibiting MIF production by AGN2a, endogenous MIF RNA was blocked with d-siRNA (dicer-generated small interfering RNA). d-siRNA was transfected into wild-type AGN2a, or an AGN2a-derived tumor vaccine cell line, AGN2a/CD80-137L (AGN2a permanently transfected with CMV-based mammalian
Discussion
During our study of the in vitro immune response to neuroblastoma we found that murine neuroblastoma cell lines inhibited rather than stimulated T lymphocyte activation in vitro. Unexpectedly, we determined that MIF is in large part the responsible agent. The MIF produced by neuroblastoma is secreted at super-physiological levels, exceeding 100 ng/ml in 3-day cultures of AGN2a. To date, MIF was thought to promote T-cell immunity as antibody depletion studies showed a critical requirement for MIF
Acknowledgments
We would like to thank Natalia Natalia for technical support, and acknowledge support from the Midwest Athletes Against Childhood Cancer (MACC Fund, Inc.) and from National Institutes of Health, NCI grant CA100030.
References (56)
- et al.
Angiogenesis and neuroblastomas: interleukin-8 and interleukin-8 receptor expression in human neuroblastoma
J Urol
(2000) - et al.
Interleukin-15, a T-cell growth factor, is expressed in human neural cell lines and tissues
J Neurol Sci
(1998) - et al.
Neurotrophic role of interleukin-6 and soluble interleukin-6 receptros in N1E-115 neuroblastoma cells
J Neuroimmunol
(2000) - et al.
Dual expression of CD80 and CD86 produces a tumor vaccine superior to single expression of either molecule
Cell Immunol
(2003) - et al.
High expression of macrophage migration inhibitory factor in human melanoma cells and its role in tumor cell growth and angiogenesis
Biochem Biophys Res Commun
(1999) - et al.
Enhanced expression of macrophage migration inhibitory factor in prostatic adenocarcinoma metastases
Urology
(1996) - et al.
Loss of STAT1 expression confers resistance to IFN-γ-induced apoptosis in ME180 cells
FEBS Lett
(1999) - et al.
STAT-1 interacts with p53 to enhance DNA damage-induced apoptosis
J Biol Chem
(2004) - et al.
STAT1-induced apoptosis is mediated by caspases 2, 3, and 7
J Biol Chem
(2004) - et al.
Macrophage migration inhibitory factor (MIF) seems crucially involved in Guillain-Barre syndrome and experimental allergic neuritis
J. Neuroimmunol
(2005)
Delayed hypersensitivity in vitro: its mediation by cell-free substances formed by lymphoid cell-antigen interaction
Proc Natl Acad Sci USA
Reactions in vivo and in vitro produced by a soluble substance associated with delayed-type hypersensitivity
Proc Natl Acad Sci USA
Regulation of the immune response by macrophage migration inhibitory factor: biological and structural features
J Mol Med
Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor
Proc Natl Acad Sci USA
MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia
Nature
Dissection of the enzymatic and immunologic functions of macrophage migration inhibitory factor
Eur J Biochem
Identification of cyclophilin as a proinflammatory secretory product of lipopolysaccharide-activated macrophages
Proc Natl Acad Sci USA
An essential regulatory role for macrophage migration inhibitory factor in T-cell activation
Proc Natl Acad Sci USA
An essential role for macrophage migration inhibitory factor in tuberculin delayed-type hypersensitivity reaction
J Exp Med
Regulation of interleukin-12 gene expression and its anti-tumor activities by prostaglandin E2 derived from mammary carcinomas
J Leukoc Biol
Production of macrophage migration inhibitory factor by human and murine neuroblastoma
Tumor Biol
Relative contribution of different receptor subtypes in the response of neuroblastoma cells to tumor necrosis factor-alpha
J Neurochem
Filarial nematode parasites secrete a homologue of the human cytokine macrophage migration inhibitory factor
Infect Immun
Enzymatically inactive macrophage migration inhibitory factor inhibits monocyte chemotaxis and random migration
Biochemistry
The p53-dependent effects of macrophage migration inhibitory factor revealed by gene targeting
Proc Natl Acad Sci USA
MIF signal transduction initiated by binding to CD74
J Exp Med
Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1
Nature
Cited by (44)
The immune landscape of neuroblastoma: Challenges and opportunities for novel therapeutic strategies in pediatric oncology
2021, European Journal of CancerCitation Excerpt :In experimental neuroblastoma models, NK cell function was directly modulated by TGFβ [107], and tumoural galectin-1 or MIF knockdown resulted in increased T-cell-mediated cytotoxicity, IFNγ secretion, CD4+ and CD8+ T cell recruitment, and more efficient tumour rejection [104,108,109]. Soluble galectin-1, as well as MIF overexpression, induced T cell apoptosis, inhibited T cell proliferation and inhibited DC maturation [104,110,111]. In patients, high tumoural MIF was associated with lower abundance of CTLs, NKT cells, B cells and DC, and a poor prognosis in stage 4 tumours independent of MYCN [112].
Tumoral Immune Cell Exploitation in Colorectal Cancer Metastases Can Be Targeted Effectively by Anti-CCR5 Therapy in Cancer Patients
2016, Cancer CellCitation Excerpt :Potentially, with the influx of new effector cells the deleterious effects of CCL5 could be shifted toward the beneficial effects of IFN-α2 and IFN-γ (Hervas-Stubbs et al., 2011). Reduction of immunosuppression via MIF, angiogenesis via VEGF, and chemotherapy resistance via CXCL8 and increased IFN levels could possibly contribute to this anti-tumoral effect and also add to the momentum of CCR5 inhibition combined with chemotherapy (Hwang et al., 2011; Sistigu et al., 2014; Yan et al., 2006). How do these observations fit into previous findings that a high T cell density at the tumor site is associated with a better clinical outcome (Fridman et al., 2012; Galon et al., 2006)?
MIF contribution to progressive brain diseases
2024, Journal of Neuroinflammation