A truncated IL-12rβ1 receptor ameliorates chronic graft-versus-host disease-induced lupus nephritis by inhibiting Th1 and Th17 cells

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Highlights

  • We report a soluble receptor, tIL12rβ1 for the treatment of lupus nephritis.

  • We present a new therapeutic strategy targeting IL-12/IL-23 p40 for lupus nephritis.

  • Both Th1 and Th17 cells are involved in the pathogenesis of cGVHD-induced SLE.

Abstract

Th1 and Th17 cells have been strongly implicated in the pathogenesis of systemic lupus erythematosus (SLE). Interleukin (IL)-12 and IL-23 respectively drive the polarization of Th1 and Th17 cells and share a common p40 subunit. In this study, the protective and therapeutic effects of a truncated human IL-12rβ1 receptor (tIL12rβ1) targeting IL-12/IL-23 p40 were evaluated in chronic graft-versus-host disease (cGVHD)-induced SLE-like model. The results indicated that tIL12rβ1 treatment effectively delayed the proteinuria onset and induced a significant remission of proteinuria, autoantibody production, and immune complex deposition in the mouse model. Remarkably, the therapeutic effects of tIL12rβ1 were predominantly dependent on the suppression of pathogenic Th1 and Th17 cell commitment through the reduction of RORγt and T-bet expression. Collectively, this receptor molecule may offer a new treatment option for SLE.

Introduction

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder with an annual incidence of 50–70 per million [1]. SLE is characterized by the production and accumulation of abnormally high amounts of anti-nuclear antibodies (ANAs), especially anti-double-stranded (ds) DNA antibodies and immune complexes (IC), which attack the cells and tissues causing inflammation and tissue damage [2], [3].

During the past several decades, considerable efforts have demonstrated critical pathogenic roles of Th1 cells and secreted IFN-γ in human and murine lupus [4], [5]. Recently, experimental and clinical evidence has suggested that Th17 cells, a distinct CD4 + T cell subset characterized by the secretion of IL-17 [6], are also involved in SLE pathogenesis [7]. Indeed, an increased level of IL-17 or greater abundance of IL-17-producing T helper cells have been found in SLE patients [8], [9], and studies based on several lupus-like mouse models have also supported the notion that the proinflammatory cytokine, IL-17, is involved in the development of SLE [10], [11], [12]. Moreover, IL-17 could act in synergy with B cell–activating factor (BAFF) to affect B cell behaviors and the pathophysiology of SLE, which demonstrates that IL-17 abundance correlates with the disease severity of SLE [13]. Therefore, both Th1 and Th17 cytokines might be involved in the pathogenesis of active SLE, possibly via different mechanisms. CD4 + T cells can be induced to differentiate toward Th1 and Th17 phenotypes, according to the distinct cytokine milieu. In fact, the transcription factor RORγt directs the development of Th17 cells [14], while T-bet is involved in the polarization of Th1 cells [15]. Despite the developmental signals that distinguish Th1 and Th17 differentiation, there are interesting cross-talks between the two developmental programs. During the development of Th17 cells, IL-23R is induced. In the case of Th1 cell development, IL-12Rβ2 is induced to pair with constitutively expressed IL-12Rβ1 chain. Subsequently, these receptors bind a common p40 subunit of IL-23 and IL-12 heterodimers to confer responsiveness to IL-23 and IL-12, respectively [16], [17].

The multifactorial pathogenesis of SLE has resulted in a complicated situation with regard to therapeutic approaches. Currently, treatment of SLE with corticosteroids and other immunosuppressive agents may be useful in many cases; however, these drugs have shown marginal efficacy or significant safety issues [18]. Therefore, it is necessary to develop new treatment strategies. Given that the p40 subunit shared by IL-12 and IL-23 play critical roles in the development of Th1 and Th17 cells, targeting p40 subunit should be able to attenuate proinflammatory T-cell responses. Indeed, anti-p40 antibody or p40 peptide-based vaccines have been tested in several animal models of autoimmunity to show positive effects [19], [20], [21], [22]. On the basis of these facts, it is conceivable to expect that the benefits of targeting IL-12p40 will eventually address unmet medical needs for a broad spectrum of immune disorders including the refractory SLE.

We have previously eucaryotically expressed a novel human truncated IL12rβ1-Fc fusion protein (tIL12rβ1/Fc) binding the p40 subunit and found it efficacious in the amelioration of MOG35–55-induced EAE [23].In the present work, we investigated the efficacy of a truncated human IL-12rβ1 receptor (tIL12rβ1) previously constructed in our laboratory in cGVHD-induced lupus nephritis (CLN) and its mechanism of action on the lupus immune system. We found that tIL12rβ1 treatment could effectively ameliorate autoimmune syndromes in the cGVHD-induced SLE-like model. Moreover, the decreased production of Th1- and Th17- polarized pro-inflammatory cytokines was resulted from the significant reduction of RORγt and T-bet expression. This treatment approach may offer a new therapeutic option for SLE patients.

Section snippets

Preparation of tIL12rβ1

Based on the amino acid sequence of human IL12rβ1 receptor (NP_005526) and the codon preference in E. coli, the tIL12rβ1 gene (639 bp) was obtained by RT-PCR from human spleen cDNA library (Biomics, China) using the following primers: 5′-CGGGATCCTGCAGAACCAGTGAGTGCT-3′ and 5′-CCGCTCGAGTTAGGGGTTTTCAGGGGGA-3′ and then cloned into the expression vector PGEX-4T-2. Correctly constructed plasmids were transformed into E. coli BL21 (DE3) (Novagen, Germany) and the expression of the protein was achieved

Preparation of tIL12rβ1

In the present study, we chose p40 subunit as the target molecule. For the construction of expression plasmid, the tIL12rβ1 gene was cloned into pGEX-4T-2, where it was fused with a GST-tag and under the control of tac promoter (Fig. 1A) and the GST-tIL12rβ1 fusion protein was successfully induced by IPTG in E. coli BL21 (DE3) (Fig. 1B). GST-tag can facilitate soluble expression of tIL12rβ1 in high yield and simplify the downstream purification. The bacterial lysates containing GST-tIL12rβ1

Discussion

IL-12 and IL-23 respectively drive polarization of Th1 and Th17 cells and share the p40 subunit. Consequently, any intervention targeting IL-12/23p40 and thus down-regulating both Th1 and Th17 responses is potentially beneficial to the treatment of Th1/Th17-related diseases. The extracellular domain of the cytokine receptor is exactly the region at which the receptor strongly binds the ligand for the activation of downstream biological events. Therefore, utilizing extracellular binding domains

Acknowledgments

This work was supported by a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), National Natural Science Foundation of China (81202450, 81430082 and 81573444), and the Fundamental Research Funds for the Central Universities (JKPZ2013013 and YD2014SK0002).

References (33)

  • W. Ouyang et al.

    The biological functions of T helper 17 cell effector cytokines in inflammation

    Immunity

    (2008)
  • K. Masutani et al.

    Predominance of Th1 immune response in diffuse proliferative lupus nephritis

    Arthritis and Rheumatism

    (2001)
  • D. Balomenos et al.

    Interferon-gamma is required for lupus-like disease and lymphoaccumulation in MRL-lpr mice

    Journal of Clinical Investigation

    (1998)
  • L. Steinman

    A brief history of TH17, the first major revision in the TH1/TH2 hypothesis of T cell-mediated tissue damage

    Nature Medicine

    (2007)
  • Martin JC, Baeten DL, Josien R. Emerging role of IL-17 and Th17 cells in systemic lupus erythematosus. Clinical...
  • J.C. Crispin et al.

    Expanded double negative T cells in patients with systemic lupus erythematosus produce IL-17 and infiltrate the kidneys

    Journal of Immunology

    (2008)
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