Cathepsin S inhibition combines control of systemic and peripheral pathomechanisms of autoimmune tissue injury (original) (raw)
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Cathepsin S inhibition suppresses autoimmune-triggered inflammatory responses in macrophages
Biochemical pharmacology, 2017
In several types of antigen-presenting cells (APCs), Cathepsin S (CatS) plays a crucial role in the regulation of MHC class II surface expression and consequently influences antigen (Ag) presentation of APCs to CD4(+) T cells. During the assembly of MHC class II-Ag peptide complexes, CatS cleaves the invariant chain p10 (Lip10) - a fragment of the MHC class II-associated invariant chain peptide. In this report, we used a selective, high-affinity CatS inhibitor to suppress the proteolytic activity of CatS in lymphoid and myeloid cells. CatS inhibition resulted in a concentration-dependent Lip10 accumulation in B cells from both healthy donors and patients with systemic lupus erythematosus (SLE). Furthermore, CatS inhibition led to a decreased MHC class II expression on B cells, monocytes, and proinflammatory macrophages. In SLE patient-derived peripheral blood mononuclear cells, CatS inhibition led to a suppressed secretion of IL-6, TNFα, and IL-10. In a second step, we tested the ef...
Annals of the Rheumatic Diseases, 2013
Objectives Major histocompatibility complex (MHC) class II-mediated priming of T and B lymphocytes is a central element of autoimmunity in systemic lupus erythematosus (SLE) and lupus nephritis. The cysteine protease cathepsin S degrades the invariant peptide chain during MHC II assembly with antigenic peptide in antigen-presenting cells; therefore, we hypothesised that cathepsin S inhibition would be therapeutic in SLE. Methods We developed a highly specific small molecule, orally available, cathepsin S antagonist, RO5461111, with suitable pharmacodynamic and pharmacokinetic properties that efficiently suppressed antigen-specific T cell and B cell priming in vitro and in vivo. Results When given to MRL-Fas(lpr) mice with SLE and lupus nephritis, RO5461111 significantly reduced the activation of spleen dendritic cells and the subsequent expansion and activation of CD4 T cells and CD4/CD8 double-negative T cells. Cathepsin S inhibition impaired the spatial organisation of germinal centres, suppressed follicular B cell maturation to plasma cells and Ig class switch. This reversed hypergammaglobulinemia and significantly suppressed the plasma levels of numerous IgG (but not IgM) autoantibodies below baseline, including anti-dsDNA. This effect was associated with less glomerular IgG deposits, which protected kidneys from lupus nephritis. Conclusions Together, cathepsin S promotes SLE by driving MHC class II-mediated T and B cell priming, germinal centre formation and B cell maturation towards plasma cells. These afferent immune pathways can be specifically reversed with the cathepsin S antagonist RO5461111, which prevents lupus nephritis progression even when given after disease onset. This novel therapeutic strategy could correct a common pathomechanism of SLE and other immune complexrelated autoimmune diseases.
Cathepsin S activity regulates antigen presentation and immunity
Journal of Clinical Investigation, 1998
MHC class II molecules display antigenic peptides on cell surfaces for recognition by CD4 ϩ T cells. Proteolysis is required in this process both for degradation of invariant chain (Ii) from class II-Ii complexes to allow subsequent binding of peptides, and for generation of the antigenic peptides. The cysteine endoprotease, cathepsin S, mediates Ii degradation in human and mouse antigen-presenting cells.
1996
is the proteolytic destruction of Ii, as intact ␣Ii trimers and Harold A. Chapman* themselves are unable to bind peptides (Roche and *Department of Medicine Cresswell, 1990). In vivo, inhibition of all cysteine class Brigham and Women's Hospital proteases impairs Ii breakdown and induces accumulaand Harvard Medical School tion of class II-associated Ii fragments in B lymphoblas-Boston, Massachusetts 02115 toid cells (Blum and Cresswell, 1988; Nguyen et al., 1988; † Center for Cancer Research Newcomb and Cresswell, 1993). Consequently, acquisi-Department of Biology tion of antigenic peptide by class II ␣ dimers is pre-Massachusetts Institute of Technology vented (Neefjes and Ploegh, 1992), expression of class Cambridge, Massachusetts 02139 II molecules at the cell surface is decreased (Neefjes ‡ Arris Pharmaceuticals, Incorporated and Ploegh, 1992; Bé naroch et al., 1995), and antigen-385 Oyster Point Boulevard stimulated T cell proliferation is attenuated (Buus and South San Francisco, California 94080 Werdelin, 1986; Diment, 1990). Despite the absolute requirement for Ii destruction to render class II molecules capable of binding peptide, the primary proteases that perform this task remain unidentified. Previous attempts Summary at identifying these proteases are difficult to interpret, owing to the rather nonspecific action of the inhibitors Destruction of Ii by proteolysis is required for MHC used and possible contamination of commercially availclass II molecules to bind antigenic peptides, and for able protease preparations with other proteases. For transport of the resulting complexes to the cell surexample, leupeptin, a commonly utilized protease inhibiface. The cysteine protease cathepsin S is highly tor (Nguyen et al., 1988), acts on many proteases of the expressed in spleen, lymphocytes, monocytes, and cysteine and serine class. Lysosomotropic agents, such other class II-positive cells, and is inducible with as quinidine (Humbert et al., 1993) and concanamycin interferon-␥. Specific inhibition of cathepsin S in B B (Bé naroch et al., 1995), neutralize acidic compartlymphoblastoid cells prevented complete proteolysis ments in a nonspecific manner. Also, analysis of some of Ii, resulting in accumulation of a class II-associated commercially available cathepsin B preparations in our 13 kDa Ii fragment in vivo. Consequently, the formation laboratory have demonstrated the presence of addiof SDS-stable complexes was markedly reduced. Puritional cysteine proteases (H. A. C., unpublished data). fied cathepsin S, but not cathepsin B, H, or D, specifi-Cathepsin S, a cysteine protease originally cloned cally digested Ii from ␣Ii trimers, generating ␣-CLIP from human alveolar macrophages, is highly expressed complexes capable of binding exogenously added in the spleen and professional antigen-presenting cells, peptide in vitro. Thus, cathepsin S is essential in B including B lymphocytes, macrophages, and other class cells for effective Ii proteolysis necessary to render II-positive cells (Shi et al., 1992, 1994; Morton et al., class II molecules competent for binding peptides. 1995). Moreover, it is inducible with interferon-␥ (IFN␥), it is a potent endoprotease, and it has a broad pH activity profile (Shi et al.
Differential Regulation of Cathepsin S and Cathepsin L in Interferon γ–treated Macrophages
Journal of Experimental Medicine, 2003
Cathepsin S (catS) and cathepsin L (catL) mediate late stages of invariant chain (Ii) degradation in discrete antigen-presenting cell types. Macrophages (Mϕs) are unique in that they express both proteases and here we sought to determine the relative contribution of each enzyme. We observe that catL plays no significant role in Ii cleavage in interferon (IFN)-γ–stimulated Mϕs. In addition, our studies show that the level of catL activity is significantly decreased in Mϕs cultured in the presence of IFN-γ whereas catS activity increases. The decrease in catL activity upon cytokine treatment occurs despite the persistence of high levels of mature catL protein, suggesting that a specific inhibitor of the enzyme is up-regulated in IFN-γ–stimulated peritoneal Mϕs. Similar inhibition of activity is observed in dendritic cells engineered to overexpress catL. Such enzymatic inhibition in Mϕs exhibits only partial dependence upon Ii and therefore, other mechanisms of catL inhibition are regu...
The Journal of Experimental Medicine, 2000
The major histocompatibility complex (MHC) class II–associated invariant chain (Ii) regulates intracellular trafficking and peptide loading of MHC class II molecules. Such loading occurs after endosomal degradation of the invariant chain to a ∼3-kD peptide termed CLIP (class II–associated invariant chain peptide). Cathepsins L and S have both been implicated in degradation of Ii to CLIP in thymus and peripheral lymphoid organs, respectively. However, macrophages from mice deficient in both cathepsins S and L can process Ii and load peptides onto MHC class II dimers normally. Both processes are blocked by a cysteine protease inhibitor, indicating the involvement of an additional Ii-processing enzyme(s). Comparison of cysteine proteases expressed by macrophages with those found in splenocytes and dendritic cells revealed two enzymes expressed exclusively in macrophages, cathepsins Z and F. Recombinant cathepsin Z did not generate CLIP from Ii–MHC class II complexes, whereas cathepsin ...
Cathepsin S inhibitors as novel immunomodulators
Current opinion in investigational drugs (London, England : 2000), 2005
Cathepsin S is one of the major cysteine proteases, and is expressed in the lysosome of antigen presenting cells; primarily dendritic cells, B-cells and macrophages. Cathepsin S is most well known for its critical function in the proteolytic digestion of the invariant chain chaperone molecules, thus controlling antigen presentation to CD4+ T-cells by major histocompatibility complex (MHC) class II molecules or to NK1.1+ T-cells via CD1 molecules. Cathepsin S also appears to participate in direct processing of exogenous antigens for presentation by MHC class II to CD4+ T-cells, or in cross-presentation by MHC class I molecules to CD8+ T-cells. In addition, although direct evidence is still lacking, in its secreted form cathepsin S is implicated in degradation of the extracellular matrix, which may contribute to the pathology of a number of diseases, including arthritis, atherosclerosis and chronic obstructive pulmonary disease. Therefore, inhibition of cathepsin S is a promising targ...
Immunity, 1999
Cathepsins have been implicated in the degradation Rodriguez and Diment, 1995; Riese et al., 1996). Howof proteins destined for the MHC class II processing ever, in sharp contrast with many biochemical in vitro pathway and in the proteolytic removal of invariant experiments, genetic analyses using cathepsin B-and chain (Ii), a critical regulator of MHC class II function. D-deficient mice revealed that these two major lyso-Mice lacking the lysosomal cysteine proteinase casomal proteinases are not necessary for Ii degradation thepsin S (catS) demonstrated a profound inhibition and processing of a number of exogenous and endogeof Ii degradation in professional APC in vivo. A marked nous protein antigens (Villadangos et al., 1997; Deussing variation in the generation of MHC class II-bound Ii et al., 1998; T. N. et al., unpublished data). This sugfragments and presentation of exogenous proteins gested that the complexity and redundancy of lysowas observed between B cells, dendritic cells, and somal proteinases may impede the elucidation of the macrophages lacking catS. CatS-deficient mice showed function of individual enzymes. However, our recent diminished susceptibility to collagen-induced arthritis, analysis of cathepsin L (catL)-deficient mice revealed suggesting a potential therapeutic target for regulaa profound defect in Ii degradation in thymic cortical tion of immune responsiveness. epithelial cells (cTEC) but not in bone marrow-derived APCs (B cells, DC, and Mph). This tissue-specific defect correlates with the restricted pattern of expression of later. Macrophages were cultured in teflon plates in the presence of 50 U/ml recombinant IFN␥ for 48 hr. Nonadherent cells were We thank M. Appleby, F. Ramsdell, and D. Walker (Chiroscience, washed with warm medium, and macrophages were harvested by Bothell, WA) for generous assistance with cell sorting; Immunex trypsinization. The class II expression on activated Mph was as-Corporation for providing recombinant human Flt3 ligand; D. Wilson sessed by staining in the presence of Fc receptor blocking MAb and M. Gomez for technical assistance; H. Caldwell (NIAID Rocky 24G.2 (ATCC 197) with F4/80-FITC MAb (Pharmingen) and biotinyl-Mountain Laboratory) for C. trachomatis and 116.3 T hybrids; S. ated anti-class II MAbs followed by streptavidin-Tricolor. Purity of Kahn (University of Washington) for recombinant SA85 T. cruzi prothe macrophage population was over 95% as determined by two tein and 71.5 T hybrids; N. Shastri (UC Berkeley) for BO4 T hybrids; color flow cytometry. M. Bevan, P. Fink, and P. Wong for reading of the manuscript; and members of our laboratories for helpful comments and discussion. . Kovats et al., 1998) were cultured in duplicates for 20 hr with variable numbers of control or mutant SC. In parallel, 1 ϫ 10 5 SC were References incubated with I-A b -restricted T hybrids (Nakagawa et al., 1998) specific for HEL (BO4), KLH (2BH11. A1), myoglobin (1BE6A1), T. Avva, R.R., and Cresswell, P. (1994). In vivo and in vitro formation cruzi SA85 (71.5), and C. trachomatis (116.3) and I-A q -restricted T and dissociation of HLA-DR complexes with invariant chain-derived cell hybrids specific for HEL (qLy30.0), OVA (qO/H166.0), and bovine peptides. Immunity 1, 763-774. collagen type II (qCII85.33) (Rosloniec et al., 1996) with variable amounts of exogenous antigens. IL-2 production was assessed us-Barrett, A.J. (1987). The cystatins: a new class of peptidase inhibiing HT-2 proliferation as determined by the Alamar Blue colorimetric tors. Trends Biochem. Sci. 12, 193-196. assay. The results are expressed as arbitrary OD units (A570-A600). Baumgart, M., Moos, V., Schuhbauer, D., and Muller, B. (1998). Differential expression of major histocompatibility complex class II Induction and Scoring of Collagen-Induced Arthritis genes on murine macrophages associated with T cell cytokine pro-CIA was induced and scored in male catS Ϫ/Ϫ or wt mice (9-13 weeks file and protective/suppressive effects. Proc. Natl. Acad. Sci., USA old; 10-12 animals per group) immunized at the base of the tail on 95, 6936-6940. day 0 and 21 with chick type II collagen in CFA as described (Griffiths Bennett, K., Levine, T., Ellis, J.S., Peanasky, R.J., Samloff, I.M., Kay, et al., 1995). Severity of the disease was evaluated by inspection of J., and Chain, B.M. (1992). Antigen processing for presentation by the paws and joints and a score was assigned (0, normal paw; 1, class II major histocompatibility complex requires cleavage by caswelling and/or redness of one toe or finger joint; 2, two or more thepsin E. Eur. J. Immunol. 22, 1519-1524. joints involved; 3, severe arthritis in the entire paw; maximum score Bevec, T., Stoka, V., Pungercic, G., Dolenc, I., and Turk, V. (1996). for each animal-12). Collagen-specific antibodies were measured Major histocompatibility complex class II-associated p41 invariant in the serum of mice at the termination of the experiment (day 58) by ELISA. Immune and control sera were titrated on Nunc-Immuno chain fragment is a strong inhibitor of lysosomal cathepsin L. J. Maxisorp plates coated with chick collagen (Marie Griffith, University Exp. Med. 183, 1331-1338. of Utah). The presence of collagen-specific antibodies was deter-Blum, J.S., and Cresswell, P. (1988). Role of intracellular proteases mined with goat anti-mouse IgG1, IgG2a, IgG2b, and IgG3 HRPin the processing and transport of class II HLA antigens. Proc. Natl. conjugated antibodies (Southern Biotech). Total collagen-specific Acad. Sci., USA 85, 3975-3980. IgG and IgM was measured with analogous HRP-conjugated goat Brachet, V., Raposo, G., Amigorena, S., and Mellman, I. (1997). Ii antibodies (Jackson ImmunoResearch Laboratory, Inc.). chain controls the transport of major histocompatibility complex class II molecules to and from lysosomes. . (1994). Characterization of the T cell Proteinase Active Site Labeling determinants in the induction of autoimmune arthritis by bovine Splenocytes, DC, B cells, and Mph were preincubated for 1 hr at 37ЊC in methionine/cysteine-free RPMI 1640 supplemented with 200 ␣1(II)-CB11 in H-2 q Mice. J. Immunol. 152, 3088-3097.
Journal of Leukocyte Biology, 2003
Endocytic proteolysis represents a major functional component of the major histocompatibility complex class II antigen-presentation machinery. Although transport and assembly of class II molecules in the endocytic compartment are well characterized, we lack information about the pattern of endocytic protease activity along this pathway. Here, we used chemical tools that visualize endocytic proteases in an activity-dependent manner in combination with subcellular fractionation to dissect the subcellular distribution of the major cathepsins (Cat) CatS, CatB, CatH, CatD, CatC, and CatZ as well as the asparagine-specific endoprotease (AEP) in human B-lymphoblastoid cells (BLC). Endocytic proteases were distributed in two distinct patterns: CatB and CatZ were most prominent in early and late endosomes but absent from lysosomes, and CatH, CatS, CatD, CatC, and AEP distributed between late endosomes and lysosomes, suggesting that CatB and CatZ might be involved in the initial proteolytic attack on a given antigen. The entire spectrum of protease activity colocalized with human leukocyte antigen-DM and the C-terminal and N-terminal processing of invariant chain (Ii) in late endosomes. CatS was active in all endocytic compartments. Surprisingly and in contrast with results from dendritic cells, inhibition of CatS activity by leucine-homophenylalanine-vinylsulfone-phenol prevented N-terminal processing of Ii but did not alter the subcellular trafficking or surface delivery of class II complexes, as deferred from pulse-chase analysis in combination with subcellular fractionation and biotinylation of cell-surface protein. Thus, BLC contain distinct activity patterns of proteases in endocytic compartments and regulate the intracellular transport and surface-delivery of class II in a CatS-independent manner. J. Leukoc. Biol. 75: 000 -000; 2004.
Scientific Reports, 2021
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that affects multiple organs. Recent studies suggest relevance between cysteine protease cathepsin S (CTSS) expression and SLE. To investigate the mechanism of CTSS in SLE, CTSS-overexpressing transgenic (TG) mice were generated, and induced lupus-like symptoms. Eight months later, the TG mice spontaneously developed typical SLE symptoms regardless of the inducement. Furthermore, we observed increased toll-like receptor 7 (TLR7) expression with increased monocyte and neutrophil populations in the TG mice. In conclusion, overexpression of CTSS in mice influences TLR7 expression, autoantibodies and IFN-α, which leads to an autoimmune reaction and exacerbates lupus-like symptoms.