Treatment of lupus patients with a tolerogenic peptide, hCDR1 (Edratide): Immunomodulation of gene expression
Introduction
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies and by impaired functions of B- and T-cells, accompanied by systemic clinical manifestations [1], [2]. Previous studies demonstrated the role of cytokines [3], [4], apoptosis [5], [6], and dysfunction of regulatory T-cells [7], [8] in the pathogenesis of murine and human SLE. For a specific treatment of SLE, our laboratory designed a peptide, designated hCDR1 [9], which is based on the sequence of the complementarity-determining region (CDR)1 of a human anti-DNA monoclonal antibody that bears the major idiotype designated 16/6Id [10], [11]. hCDR1 was shown to ameliorate the serological and clinical manifestations of induced or spontaneously developed lupus in mice [12]. The beneficial effects of hCDR1 were associated with the down-regulation of the pathogenic cytokines IL-1β, IFN-γ, and IL-10 [12], and the up-regulation of the immunosuppressive cytokine TGF-β [13]. Treatment with hCDR1 also diminished significantly the production of the B-cell stimulator (BLyS, BAFF) in lupus-prone mice [14]. Furthermore, treatment of SLE-afflicted mice with hCDR1 resulted in reduced apoptosis rates of T-cells [15], [16], [17], and in specific induction of CD4+CD25+Foxp3+ regulatory cells [13], [18].
We have previously shown that hCDR1 down-regulated, in vitro, autoreactive T-cell responses of peripheral blood mononuclear cells (PBMC) of SLE patients in association with an increased production of TGF-β [9]. Moreover, we established a human model of SLE by transferring PBMC of lupus patients into severe combined immunodeficient mice [19]. Treatment with hCDR1 ameliorated the serological (human anti-DNA antibodies) and clinical lupus-related manifestations in the latter model of human SLE [20]. Furthermore, we have recently shown that in vitro incubation of PBMC of lupus patients with hCDR1, but not with a control peptide, down-regulated gene expression of IL-1β, TNF-α, IFN-γ, IL-10, and caspase-3 and up-regulated the expression of TGF-β, FoxP3, the anti-apoptotic molecule Bcl-xL and of the negative regulators Foxj1 and Foxo3a [21]. In addition, the in vitro incubation with hCDR1 resulted in an increase of CD4+CD25+FoxP3+ functional regulatory T-cells [21].
It has been of great interest to determine whether hCDR1 functions via similar mechanisms of actions when administered to SLE patients. The present study was therefore aimed at determining the in vivo effects of hCDR1 on human SLE. To this end, blood samples were taken from 9 lupus patients that were treated with hCDR1 or with the vehicle for a period of 26 weeks. Gene expression of the pathogenic cytokines (IL-1β, TNF-α, IFN-γ, and IL-10), pro-apoptotic molecules (caspase-3, caspase-8), BLyS, and immunoregulatory TGF-β and FoxP3 molecules was determined by real-time RT-PCR. The results indicate that hCDR1 immunomodulated in vivo the expression of the above genes in SLE patients in a manner similar to that demonstrated for SLE-afflicted mice.
Section snippets
Patients
The data presented here are of nine SLE patients from two Israeli Medical Centers (Kaplan, Rehovot; Rabin, Petah-Tikva). These patients participated in a large double-blinded clinical trial. Included are all patients from the two medical centers who completed the study and from whom blood samples were taken at least twice (before treatment initiation and at week 24) for mRNA preparation. We present here the effect of treatment with hCDR1 on cytokine, apoptosis, BLyS, and regulatory gene
Patients
The mean (±SD) age of our nine patients (one male, eight females) at the time of the study was 46.4 ± 10.5 (range 29–61) years. All had antinuclear antibodies and anti-dsDNA autoantibodies in their sera at study entry. Hypocomplementemia (either C3 or C4) was observed in five patients. The main SLE-related clinical manifestations at study entry were skin involvement (6/9), arthritis (5/9), alopecia (4/9), thrombocytopenia and lymphopenia (2/9), mucosal ulcers (2/9), and pleuritis (1/9). The mean
Discussion
The main finding of the present study is that the tolerogenic peptide hCDR1 is capable of suppressing, in vivo, gene expression of pro-inflammatory cytokines, apoptosis, and of B-cell stimulation. Furthermore, in vivo treatment with hCDR1 up-regulated immunosuppressive and regulatory molecules in PBMC of SLE patients. To the best of our knowledge, this is the first report demonstrating in vivo immunomodulation of gene expression in SLE patients by a specific peptide previously shown to
Acknowledgement
The clinical trial was conducted by Teva Pharmaceutical Industries, Ltd., and the blood samples used in our study were of patients that participated in this trial. However, the study reported here was performed independently at the Weizmann Institute of Science.
References (38)
- et al.
Amelioration of SLE-like manifestations in (NZBxNZW)F1 mice following treatment with a peptide based on the complementarity determining region 1 of an autoantibody is associated with a down-regulation of apoptosis and of the pro-apoptotic factor JNK kinase
Clin Immunol
(2005) - et al.
Amelioration of murine lupus by a peptide, based on the complementarity determining region-1 of an autoantibody as compared to dexamethasone: different effects on cytokines and apoptosis
Clin Immunol
(2006) - et al.
The tolerogenic peptide hCDR1 downregulates pathogenic cytokines and apoptosis and upregulates immunosuppressive molecules and regulatory T cells in peripheral blood mononuclear cells of lupus patients
Hum Immunol
(2009) - et al.
Immunization with peptides derived from the idiotypic region of lupus-associated autoantibodies delays the development of lupus nephritis in the (SWR x NZB)F1 murine model
J Autoimmun
(2007) An overview of the pathogenesis of systemic lupus erythematosus
- et al.
T- and B-cell abnormalities in systemic lupus erythematosus
Crit Rev Immunol
(2005) - et al.
The cytokine network in the pathogenesis of systemic lupus erythematosus and possible therapeutic implications
Springer Semin Immumopathol
(1994) - et al.
Kinetics of cytokine production in experimental systemic lupus erythematosus: involvement of T helper cell 1/T helper cell 2-type cytokines in disease
J Immunol
(1997) - et al.
B cell apoptosis accelerates the onset of murine lupus
Eur J Immunol
(2003) - et al.
Accelerated in vitro apoptosis of lymphocytes from patients with systemic lupus erythematosus
J Immunol
(1994)