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Imlifidase (IdeS).

Security, immunogenicity, pharmacokinetics and effectiveness of destruction of anti-HLA antibodies by IdeS (imlifidase) in chronic kidney illness individuals.Lorant et al., Am J Transplant. 2018 Mar 21. doi: 10.1111/ ajt.14733. [Epub ahead of print] Abstract.

Safety, immunogenicity, pharmacokinetics, and efficacy of the IgG-degrading enzyme of Streptococcus pyogenes (IdeS [imlifidase] were analyzed in a single-center, open-label ascending-dose research in very sensitized patients with chronic kidney disease. Eight individuals with cytotoxic PRAs (average cytotoxic PRAs of 64%) at registration got 1 or 2 intravenous mixtures of IdeS on successive days (0.12 mg/kg body weight × 2 [n = 3]; 0.25 mg/kg × 1 [n = 3], or 0.25 mg/kg × 2 [n = 2]. IgG degradation was observed in all topics after IdeS therapy, with <1% plasma IgG remaining within 48 hours and remaining low up to 7 days. Mean fluorescence intensity values of HLA class I and II reactivity were substantially reduced in all patients, and C1q binding to anti-HLA was abolished. IdeS also cleaved the IgG-type B cell receptor on CD19+ memory B cells. Anti-IdeS antibodies developed 1 week after treatment, peaking at 2 weeks. A few hours after the second IdeS infusion, 1 patient received a deceased donor kidney offer. At enrollment, the patient had a positive serum crossmatch (HLA-B7), detected by complement-dependent cytotoxicity, flow cytometry, and multiplex bead assays. After IdeS infusion (0.12 mg/kg ×2) and when the HLA-incompatible donor (HLA-B7+ ) kidney was offered, the HLA antibody profile was negative. The kidney was transplanted successfully.

  • IgG Endopeptidase in Highly Sensitized Patients Undergoing Transplantation.
  • Jordan et al, N Eng J Med 2017;377: 442-53.

ABSTRACT BACKGROUND

Donor-specific antibodies create an immunologic barrier to transplantation. Current therapies to modify donor-specific antibodies are limited and ineffective in the most highly HLA-sensitized patients. The IgG-degrading enzyme derived from Streptococcus pyogenes(IdeS), an endopeptidase, cleaves human IgG into F(ab′)2 and Fc fragments inhibiting complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity, which suggests that IdeS might be useful for desensitization. We report on the combined experience of two independently performed open-label, phase 1–2 trials (conducted in Sweden and the United States) that assessed the efficacy of IdeS with regard to desensitization and transplantation of a kidney from an HLA-incompatible donor.

METHODS.

We administered IdeS to 25 highly HLA-sensitized patients (11 patients in Uppsala or Stockholm, Sweden, and 14 in Los Angeles) before the transplantation of a kidney from an HLA-incompatible donor. Frequent monitoring for adverse events, outcomes, donor-specific antibodies, and renal function was performed, as were renal biopsies. Immunosuppression after transplantation consisted of tacrolimus, mycophenolate mofetil, and glucocorticoids. Patients in the U.S. study also received intravenous immune globulin and rituximab after transplantation to prevent antibody rebound.

RESULTS.

Recipients in the U.S. study had a significantly longer cold ischemia time (the time elapsed between procurement of the organ and transplantation), a significantly higher rate of delayed graft function, and significantly higher levels of class I donor-specific antibodies than those in the Swedish study. A total of 38 serious adverse events occurred in 15 patients (5 events were adjudicated as being possibly related to IdeS). At transplantation, total IgG and HLA antibodies were eliminated. A total of 24 of 25 patients had perfusion of allografts after transplantation. Antibody-mediated rejection occurred in 10 patients (7 patients in the U.S. study and 3 in the Swedish study) at 2 weeks to 5 months after transplantation; all these patients had a response to treatment. One graft loss, mediated by non-HLA IgM and IgA antibodies, occurred.

CONCLUSIONS.

IdeS reduced or eliminated donor-specific antibodies and permitted HLA-incompatible transplantation in 24 of 25 patients. (Funded by Hansa Medical; ClinicalTrials.gov numbers, NCT02224820, NCT02426684, and NCT02475551.).

Read more (external link) ›.

  • Enzymatic inactivation of endogenous IgG by IdeS enhances therapeutic antibody efficacy.
  • Järnum et al., Mol Cancer Ther. 2017 May 22. pii: molcanther.0108.2017.

Abstract.

Endogenous plasma IgG sets an immunological threshold that dictates the activity of tumor-directed therapeutic antibodies. Saturation of cellular antibody receptors by endogenous antibody limits antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP). Here we show how enzymatic cleavage of IgG using the bacterial enzyme IdeS can be utilized to empty both high and low affinity Fcγ-receptors and clear the entire endogenous antibody pool. Using in vitro models, tumor animal models as well as ex vivo analysis of sera collected during a previous clinical trial with IdeS, we show how clearing of competing plasma antibody levels with IdeS unblocks cellular antibody receptors. We show that therapeutic antibodies against breast cancer (trastuzumab), colon cancer (cetuximab) and lymphomas (rituximab and alemtuzumab) can be potentiated when endogenous IgG is removed. Overall, IdeS is shown to be a potent tool to reboot the human antibody repertoire and to generate a window to preferentially load therapeutic antibodies onto effector cells and thereby create an armada of dedicated tumor seeking immune cells.

  • IgG-degrading enzyme of Streptococcus pyogenes (IdeS) prevents disease progression and facilitates improvement in a rabbit model of Guillain-Barré syndrome.
  • Wang et al., Exp Neurol. 2017 May;291:134-140.

Abstract.

Autoantibodies binding to peripheral nerves followed by complement deposition and membrane attack complex formation results in nerve damage in Guillain-Barré syndrome (GBS). Strategies to remove the pathogenic autoantibodies or block the complement deposition benefit most patients with GBS. Immunoglobulin G-degrading enzyme of Streptococcus pyogenes (IdeS) is a cysteine protease which cleaves IgG antibodies into F(ab’)2 and Fc fragments. In this study, using a rabbit model of axonal GBS, acute motor axonal neuropathy (AMAN), we demonstrated that IdeS treatment significantly reduced the disruption of Nav channels as well as activated C3 deposition at the anterior spinal root nodes of Ranvier in AMAN rabbits. IdeS significantly promoted the clinical recovery of AMAN rabbits and there were significant lower frequencies of axonal degeneration in anterior spinal roots of AMAN rabbits with IdeS treatment compared to the saline controls. Our data support that IdeS treatment is a promising therapeutic strategy for GBS.

  • The Bacterial Enzyme IdeS Cleaves the IgG-Type of B Cell Receptor (BCR), Abolishes BCR-Mediated Cell Signaling and Inhibits Memory B Cell Activation.
  • Jarnum et al, J Immunol. 2015 Nov 9. pii: 1501929.

Abstract.

Ag binding to the BCR is a critical step in B cell development and activation, initiating a cascade of signaling events ultimately leading to proliferation, differentiation, or cell death. A bacterial enzyme, IgG-degrading enzyme of Streptococcus pyogenes (IdeS), was shown to specifically cleave IgG molecules below the hinge region of soluble IgG and when IgG is bound to Ag, resulting in one F(ab’)2 molecule and one homodimeric Fc fragment. Whether IdeS could also cleave the IgG molecule when it is present in the BCR attached to the B cell membrane in a complex with CD79a and CD79b is unknown. In this article, we present human in vitro and ex vivo data showing that IdeS cleaves the IgG present in the BCR complex and very efficiently blocks Ag binding to the BCR. As a consequence of IdeS cleaving the BCR, signaling cascades downstream of the BCR are blocked, and memory B cells are temporarily silenced, preventing them from responding to antigenic stimulation and their transition into Ab-producing cells.

  • Streptococcal IdeS: therapeutic potential for Guillain-Barré syndrome.
  • Takahashi et al., Sci Rep. 2015 Jul 21;5:10809.

Abstract.

Plasma exchange and intravenous immunoglobulin are effective in treating Guillain–Barré syndrome (GBS) probably because the former removes IgG autoantibodies and complement and the latter inhibits complement activation subsequent to the autoantibody binding to peripheral nerve antigens. IgG degrading enzyme of Streptococcus pyogenes (IdeS) can cleave the pathogenic autoantibodies into F(ab’)2 and Fc. The purpose of this study is to show whether IdeS has novel therapeutic potential for GBS. Sera with anti-ganglioside IgG antibodies from 15 patients with GBS or Miller Fisher syndrome were used. We tested whether IdeS cleaved the anti-ganglioside IgG antibodies and inhibited deposition of activated complement component on ELISA plates. IdeS efficiently cleaved IgG and blocked complement activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies. IdeS has therapeutic potential for GBS and related conditions.

  • Complete Removal of Extracellular IgG Antibodies in a Randomized Dose-Escalation Phase I Study with the Bacterial Enzyme IdeS – A Novel Therapeutic Opportunity.
  • Winstedt et al. PLOS ONE, July 15, 2015.

Abstract.

IdeS is a streptococcal protease that cleaves IgG antibodies into F(ab’)2 and Fc fragments with a unique degree of specificity, thereby providing a novel treatment opportunity of IgG-driven autoimmune conditions and antibody mediated transplant rejection. Here we report the results from a first in man, double blinded and randomized study with single ascending doses of IdeS in healthy, male subjects. Twenty healthy subjects were given intravenous single ascending doses of IdeS. With impressive efficacy IdeS cleaved the entire plasma IgG-pool only minutes after dosing. IgG reached nadir 6-24 hours after dosing and then slowly recovered. The half-life of IdeS was 4.9 (±2.8) hours at 0.24 mg/kg with the main fraction eliminated during 24 hours. Already two hours after IdeS-dosing, the phagocytic capacity of IgG/IgG-fragments was reduced to background levels. Importantly, IdeS has the capacity to inactivate Fc-mediated effector function in vivo, was considered safe with no serious adverse events, and without dose limiting toxicity in this study. The complete, rapid, but temporary removal of IgG provides a new potent therapeutic opportunity in IgG-mediated pathogenic conditions.

  • Successful treatment of experimental glomerulonephritis with IdeS and EndoS, IgG-degrading streptococcal enzymes.
  • Yang et al. NEPHROLOGY DIALYSIS TRANSPLANTATION, Vol. 25, Issue 8, 2479-2486, March 2010.

Abstract.

BACKGROUND: Anti-glomerular basement membrane (anti-GBM) disease often results in end-stage renal failure despite therapy with plasma exchange and immunosuppressive drugs. The newly discovered streptococcal enzymes IgG-degrading enzyme of S.pyogenes (IdeS) and endoglycosidase S (EndoS) act with remarkable specificity on circulating IgG. In this study, we investigate their ability in vivo to prevent damage mediated by kidney-bound antibodies in a mouse model of anti-GBM disease.

METHODS: Anti-GBM disease was induced in mice by injection of subnephritogenic doses of rabbit anti-mouse GBM, followed a week later by injection of monoclonal mouse anti-rabbit IgG antibodies. By administrating IdeS or EndoS as fusion partners with GST between these antibody injections, we tested their ability to prevent damage by acting on kidney-bound rabbit anti-GBM. Control animals received placebo injections.

RESULTS: All animals in the positive control groups developed severe albuminuria immediately after the second antibody injection (mean, 2.51 mg/24 h; range, 0.13-8.20). This was significantly diminished by EndoS (1.3 +/- 1.3 mg/24 h) and completely prevented by IdeS (0.017 +/- 0.014 mg/24 h). Immunofluorescence studies showed that IdeS treatment effectively removed the Fc fragments of the rabbit IgG. This was accompanied by a significant reduction of the deposition of the complement components C3 and C1q, and this diminished the recruitment of leukocytes to the glomeruli.

CONCLUSION: IdeS degrades IgG bound to the GBM in vivo, thereby preventing renal damage in this animal model. Most likely, IdeS would degrade both circulating and kidney-bound anti-GBM in patients with Goodpasture’s disease. Whether this would lead to a halt in disease progression and a better prognosis remains to be determined.

  • Therapeutic cleavage of IgG: new avenues for treating inflammation.
  • Nandakumar et al. TRENDS IN IMMUNOLOGY, Vol. 29, Issue 4, Apr 2008.

Abstract.

Autoantibodies developing in humans contribute to the pathogenesis of several diseases, and injected therapeutic antibodies can also trigger adverse side effects. An efficient and rapid elimination of these antibodies are therefore critically needed. Antibody removal by plasmapheresis and immunoadsorption are commonly used methods but have their own limitations. Bacterial enzymes that can cleave IgG molecules or remove carbohydrate moieties to ameliorate their immunogenicity or effector functions in vivo offer new avenues for drug development. Recent discoveries highlight the possibility of cleaving or modifying IgG in vivo by injection of enzymes. Such an approach opens up new therapeutic possibilities not only for the control of pathogenic antibody-mediated inflammatory diseases but also allograft rejection or the treatment of side-effects of ‘biologicals’ such as monoclonal antibodies.

  • Cleaving antibodies in vivo with a bacterial enzyme – a novel therapeutic concept.
  • Johansson et al. PLOS ONE, Feb 2008, Vol 3, Issue 2.

Abstract.

BACKGROUND: IdeS, a proteinase from Streptococcus pyogenes, cleaves immunoglobulin (Ig)G antibodies with a unique degree of specificity. Pathogenic IgG antibodies constitute an important clinical problem contributing to the pathogenesis of a number of autoimmune conditions and acute transplant rejection. To be able to effectively remove such antibodies is therefore an important clinical challenge.

METHODOLOGY/PRINCIPAL FINDINGS: IdeS was found to specifically and efficiently cleave IgG in human blood in vitro (20 µg of IdeS caused a complete degradation of IgG in one ml of human whole blood in 15 minutes) and to clear IgG from the blood stream of rabbits in vivo (no IgG was detected six hours following an intravenous injection of 5 mg of IdeS) without any side effects. In a mouse model of immune thrombocytopenic purpura (ITP), polyclonal IgG antibodies against platelet surface antigens were used to induce a lethal disease. These profoundly thrombocytopenic animals were treated and cured by a single injection of IdeS.

CONCLUSIONS/SIGNIFICANCE: Novel information is provided concerning the IgG-cleaving activity of IdeS in vitro and in vivo. The highly specific and rapid elimination of IgG in vivo, the dramatic effect in a mouse model of ITP, and the lack of side effects in the treated animals, indicate that IdeS could also be used to treat IgG-driven diseases in humans.

  • Blocking of experimental arthritis by cleavage of IgG antibodies in vivo.
  • Nandakumar et al. ARTHRITIS & RHEUMATISM, Vol. 56, Issue 10, pp. 3253-3260, Oct. 2007.

Abstract.

OBJECTIVE: To investigate whether IgG-degrading enzyme of Streptococcus pyogenes (IdeS), a bacterial cysteine endopeptidase that cleaves human IgG in the hinge region, can be used for blocking the development of arthritis.

METHODS: Recombinant IdeS was purified and tested for specificity against mouse IgG. IdeS was injected intravenously into mice with collagen antibody-induced arthritis (CAIA), collagen-induced arthritis (CIA), or relapsing CIA, and its effects on arthritis development and severity were assessed.

RESULTS: IdeS efficiently cleaved mouse IgG2a/c and IgG3 in vitro. Even at low dosage (10 microg), IdeS specifically cleaved IgG2a in vivo without any apparent side effects. IdeS treatment efficiently blocked CAIA induced by IgG2a antibodies. No effect was observed when arthritis was induced with IgG2b anti-type II collagen antibodies; since IdeS does not cleave IgG2b, this indicated that IgG cleavage was the mechanism of action. IdeS treatment reduced the severity of arthritis if administered within 24 hours after the onset of clinical arthritis, but did not block ongoing severe arthritis. IdeS treatment also significantly prevented an antibody-induced relapse in mice that had chronic arthritis, and delayed the onset and reduced the severity of arthritis in classic CIA.

CONCLUSION: IdeS has therapeutic potential in IgG antibody-mediated autoimmune arthritis, representing a new and unique means of blocking pathogenic antibodies.

  • Enzymatic characterization of the streptococcal endopeptidase, IdeS, reveals that it is a cysteine protease with strict specificity for IgG cleavage due to exosite binding.
  • Vincents et al. BIOCHEMISTRY 2004, 43, 15540-15549.

Abstract.

Streptococcus pyogenes, an important pathogen in humans, secretes an IgG specific endopeptidase named IdeS. To elucidate the mechanism that is responsible for this specificity, we have here characterized the activity of IdeS in detail. Both gamma chains of human IgG or its Fc fragment were cleaved in the hinge region after Gly236 by IdeS, but other proteins or synthetic peptides containing sequences such as the P( 4 )-P( 1 ) segment in the IgG cleavage site, or long peptides resembling the IgG hinge, were not hydrolyzed at all. This is likely due to a second binding site interacting with the Fc part of IgG. The lack of IdeS activity on peptide substrates necessitated the development of an assay with IgG as the substrate for kinetic studies. IdeS showed a sigmoidal velocity curve at physiological IgG concentrations, and a declining enzyme rate at higher IgG concentrations. This atypical velocity curve suggests product inhibition and/or allosteric control, which again indicates the presence of an exosite involved in substrate binding. The pseudoequilibrium constant for IdeS hydrolysis of IgG was 90 microM. The enzyme exhibited activity in the pH range of 5.1-7.6, with an optimum at pH 6.6. IdeS was stable above pH 10 but not at acidic pH. It exhibited an activity maximum around 37 degrees C and a decreased thermal stability at 42 degrees C. Iodoacetate and iodoacetamide inhibited IdeS, as expected for a cysteine protease, and biochemical evidence verified this classification. E-64 and chicken cystatin, specific inhibitors of family C1 and C13 cysteine proteases, were without effect on enzyme activity, as were class specific serine, aspartic, and metallo protease inhibitors. No significant similarities were found in protein sequence comparisons with known enzyme families, suggesting that IdeS represents a novel family of cysteine proteases.

  • Structure of the streptococcal endopeptidase IdeS, a cysteine proteinase with strict specificity for IgG.
  • Wenig et al. PNAS Dec. 2004, Vol. 101, No. 50, pp. 17371-17376.

Abstract.

Pathogenic bacteria have developed complex and diverse virulence mechanisms that weaken or disable the host immune defense system. IdeS (IgG-degrading enzyme of Streptococcus pyogenes) is a secreted cysteine endopeptidase from the human pathogen S. pyogenes with an extraordinarily high degree of substrate specificity, catalyzing a single proteolytic cleavage at the lower hinge of human IgG. This proteolytic degradation promotes inhibition of opsonophagocytosis and interferes with the killing of group A Streptococcus. We have determined the crystal structure of the catalytically inactive mutant IdeS-C94S by x-ray crystallography at 1.9-A resolution. Despite negligible sequence homology to known proteinases, the core of the structure resembles the canonical papain fold although with major insertions and a distinct substrate-binding site. Therefore IdeS belongs to a unique family within the CA clan of cysteine proteinases. Based on analogy with inhibitor complexes of papain-like proteinases, we propose a model for substrate binding by IdeS.

  • IdeS, a novel streptococcal cysteine protease with unique specificity for immunoglobulin G.
  • Von Pawel Rammingen et al. THE EMBO JOURNAL Vol. 21, No. 7 pp. 1607-1615, 2002.

Abstract.

Recent work from several laboratories has demonstrated that proteolytic mechanisms significantly contribute to the molecular interplay between Streptococcus pyogenes, an important human pathogen, and its host. Here we describe the identification, purification and characterization of a novel extracellular cysteine proteinase produced by S.pyogenes. This enzyme, designated IdeS for Immunoglobulin G-degrading enzyme of S.pyogenes, is distinct from the well-characterized streptococcal cysteine proteinase, SpeB, and cleaves human IgG in the hinge region with a high degree of specificity. Thus, other human proteins, including immunoglobulins M, A, D and E, are not degraded by IdeS. The enzyme efficiently cleaves IgG antibodies bound to streptococcal surface structures, thereby inhibiting the killing of S.pyogenes by phagocytic cells. This and additional observations on the distribution and expression of the ideS gene indicate that IdeS represents a novel and significant bacterial virulence determinant, and a potential therapeutic target.

  • HBP-assay.
  • Heparin-Binding Protein Measurement Improves the Prediction of Severe Infection With Organ Dysfunction in the Emergency Department.
  • Crit Care Med. 2015 Nov;43( 11 ):2378-86.

Abstract.

Objectives: Early identification of patients with infection and at risk of developing severe disease with organ dysfunction remains a difficult challenge. We aimed to evaluate and validate the heparin-binding protein, a neutrophil-derived mediator of vascular leakage, as a prognostic biomarker for risk of progression to severe sepsis with circulatory failure in a multicenter setting.

  • Design: A prospective international multicenter cohort study.
  • Setting: Seven different emergency departments in Sweden, Canada, and the United States.
  • Patients: Adult patients with a suspected infection and at least one of three clinical systemic inflammatory response syndrome criteria (excluding leukocyte count).
  • Intervention: None.

Measurements and Main Results: Plasma levels of heparin-binding protein, procalcitonin, C-reactive protein, lactate, and leukocyte count were determined at admission and 12-24 hours after admission in 759 emergency department patients with suspected infection. Patients were defined depending on the presence of infection and organ dysfunction. Plasma samples from 104 emergency department patients with suspected sepsis collected at an independent center were used to validate the results. Of the 674 patients diagnosed with an infection, 487 did not have organ dysfunction at enrollment. Of these 487 patients, 141 (29%) developed organ dysfunction within the 72-hour study period; 78.0% of the latter patients had an elevated plasma heparin-binding protein level (> < 1% plasma IgG staying within 48 hours as well as continuing to be reduced as much as 7 days. Mean fluorescence intensity values of HLA class I and II reactivity were substantially decreased in all clients, as well as C1q binding to anti-HLA was eliminated. IdeS additionally cleaved the IgG-type B cell receptor on CD19 + memory B cells. Anti-IdeS antibodies created 1 week after treatment, peaking at 2 weeks. A couple of hrs after the second IdeS mixture, 1 person got a departed donor kidney deal. At registration, the client had a positive serum crossmatch (HLA-B7), spotted by complement-dependent cytotoxicity, circulation cytometry, and multiplex bead assays. After IdeS infusion( 0.12 mg/kg × 2) as well as when the HLA-incompatible donor( HLA-B7 +) kidney was provided, the HLA antibody profile was adverse. The kidney was transplanted successfully. IgG Endopeptidase in Extremely Animated Clients Undergoing Transplant. Jordan et alia, N Eng J Med 2017; 377: 442-53. ABSTRACT. BACKGROUND. Donor-specific antibodies create an immunologic obstacle to transplantation. Current therapies to modify donor-specific antibodies are restricted and also inefficient in the most very HLA-sensitized clients. The IgG-degrading enzyme stemmed from Streptococcus pyogenes( IdeS), an endopeptidase, cleaves human IgG right into

F( abdominal ′) 2 as well as Fc fragments hindering complement-dependent cytotoxicity and antibody-dependent mobile cytotoxicity, which suggests that IdeS could be useful for desensitization. We report on the consolidated experience of two independently done open-label, phase 1– 2 tests( carried out in Sweden and the United States) that analyzed the effectiveness of IdeS with regard to desensitization and transplantation of a kidney from an HLA-incompatible contributor. APPROACHES. We administered IdeS to 25 highly HLA-sensitized patients( 11 clients in Uppsala or Stockholm, Sweden, and also 14 in Los Angeles) prior to the transplantation of a kidney from an HLA-incompatible donor. Regular surveillance for adverse events, outcomes, donor-specific antibodies, and renal function was performed, as were kidney biopsies. Immunosuppression after hair transplant included tacrolimus, mycophenolate mofetil, and also glucocorticoids.

Individuals in the united state study also got intravenous immune globulin and rituximab after transplant to avoid antibody rebound. OUTCOMES. Receivers in the U.S. research had a dramatically much longer cold anemia time( the moment expired in between procurement of the organ and also hair transplant), a considerably higher rate of postponed graft function, and also significantly greater degrees of class I

donor-specific antibodies than those in the Swedish study. A total of 38 significant damaging occasions occurred in 15 people (5 occasions were settled as being perhaps pertaining to IdeS ). At transplantation, overall IgG and HLA antibodies were gotten rid of. A total of 24 of 25 people had perfusion of allografts after hair transplant. Antibody-mediated denial happened in 10 clients( 7 individuals in the U.S. research study and 3 in the Swedish research )at 2 weeks to 5 months after transplantation; all these individuals had a feedback to therapy. One graft loss, mediated by non-HLA IgM and also IgA antibodies, took place. FINAL THOUGHTS. IdeS reduced or got rid of donor-specific antibodies and permitted HLA-incompatible hair transplant in 24 of 25 patients.( Moneyed by Hansa Medical; ClinicalTrials.gov numbers, NCT02224820, NCT02426684, as well as NCT02475551. ). Find out more( exterior web link)’. Enzymatic inactivation of endogenous IgG by IdeS improves restorative antibody efficiency. Järnum et al., Mol Cancer Ther. 2017 May 22. pii: molcanther.0108.2017. Abstract. Endogenous plasma IgG sets an immunological threshold that determines the activity of tumor-directed healing antibodies. Saturation of mobile antibody receptors by endogenous antibody restrictions antibody-dependent cell-mediated cytotoxicity( ADCC) and antibody dependent cellular phagocytosis (ADCP). Below we show how chemical bosom of IgG using the bacterial enzyme IdeS can be made use of to clear both low and high fondness Fcγ-receptors and also clear the entire endogenous antibody swimming pool. Using artificial insemination versions, tumor pet versions in addition to ex vivo analysis of sera accumulated during a previous medical trial with IdeS, we demonstrate how cleaning of competing plasma antibody levels with IdeS unclogs mobile antibody receptors. We show that therapeutic antibodies versus breast cancer( trastuzumab), colon cancer cells( cetuximab) and also lymphomas (rituximab and alemtuzumab) can be potentiated when endogenous IgG is eliminated. On the whole, IdeS is revealed to be a powerful tool to reboot the human antibody collection and also tocreate a window to preferentially load therapeutic antibodies onto effector cells and also thereby produce an armada of committed growth seeking immune cells. IgG-degrading enzyme of Streptococcus pyogenes( IdeS )prevents disease development and also helps with improvement in a bunny version of Guillain-Barré disorder. Wang et al., Exp Neurol. 2017 May; 291:134 -140. Abstract. Autoantibodies binding to peripheral nerves adhered to by enhance deposition and also membrane strike facility development leads to nerve damages in Guillain-Barré disorder (GBS). Techniques to eliminate the pathogenic autoantibodies or block the enhance deposition benefit most individuals with GBS. Immunoglobulin G-degrading enzyme of Streptococcus pyogenes( IdeS) is a cysteine protease which cleaves IgG antibodies right into F (abdominal muscle’) 2 and Fc fragments. In this research, utilizing a bunny design of axonal GBS, severe motor axonal neuropathy (AMAN), we demonstrated that IdeS treatment significantly lowered the interruption of Nav channels in addition to activated C3 deposition at the former spine origin nodes of Ranvier in AMAN rabbits. IdeS dramatically promoted the professional recovery of AMAN rabbits and also there were considerable reduced frequencies of axonal degeneration in former spinal origins of AMAN rabbits with IdeS treatment contrasted to the saline controls. Our information support that IdeS treatment is an encouraging healing method for GBS. The Microbial Enzyme IdeS Cleaves the IgG-Type of B Cell Receptor( BCR), Abolishes BCR-Mediated Cell Signaling and also Prevents Memory B Cell Activation. Jarnum et alia, J Immunol. 2015 Nov 9. pii: 1501929.

Abstract.

Ag binding to the BCR is an essential action in B cell development and also activation, starting a waterfall of signaling occasions inevitably leading to expansion, differentiation, or cell death. A bacterial enzyme, IgG-degrading enzyme of Streptococcus pyogenes (IdeS), was shown to particularly cleave IgG particles listed below the joint region of soluble IgG and when IgG is bound to Ag, causing one F( abdominal muscle’) 2 particle and also one homodimeric Fc piece. Whether IdeS can likewise cleave the IgG molecule when it is present in the BCR connected to the B cell membrane in a complex with CD79a and CD79b is unidentified. In this article, we offer human artificial insemination and ex vivo data showing that IdeS cleaves the IgG present in the BCR complicated as well as extremely effectively obstructs Ag binding to the BCR. As a consequence of IdeS cleaving the BCR, indicating waterfalls downstream of the BCR are obstructed, as well as memory B cells are briefly silenced, stopping them from responding to antigenic excitement as well as their change into Ab-producing cells. Streptococcal IdeS: restorative capacity for Guillain-Barré disorder. Takahashi et al., Sci Rep. 2015 Jul 21; 5:10809. Abstract. Plasma exchange and intravenous immunoglobulin are effective in dealing with Guillain– Barré syndrome (GBS) probably because the previous eliminates IgG autoantibodies as well as complement as well as the last prevents enhance activation succeeding to the autoantibody binding to outer nerve antigens. IgG derogatory enzyme of Streptococcus pyogenes( IdeS) can cleave the pathogenic autoantibodies right into F (abdominal’) 2 and also Fc. The function of this research study is to show whether IdeS has unique restorative potential for GBS. Product with anti-ganglioside IgG antibodies from 15 clients with GBS or Miller Fisher syndrome were used. We evaluated whether IdeS cleaved the anti-ganglioside IgG antibodies as well as inhibited deposition of turned on enhance component on ELISA plates. IdeS successfully cleaved IgG and also obstructed enhance activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies. IdeS has therapeutic potential for GBS and also related conditions. Total Elimination of Extracellular IgG Antibodies in a Randomized Dose-Escalation Stage I Research Study with the Microbial Enzyme IdeS– A Novel Healing Possibility. Winstedt et al. PLOS ONE, July 15, 2015. Abstract. IdeS is a streptococcal protease that cleaves IgG antibodies into F( abdominal muscle’) 2 and Fc fragments with a special degree of specificity, consequently supplying a novel treatment possibility of IgG-driven autoimmune problems and also antibody moderated transplant being rejected. Right here we report the arise from a very first in man, double blinded and also randomized research study with single rising dosages of IdeS in healthy, male subjects. Twenty healthy topics were given intravenous solitary rising doses of IdeS. With outstanding efficacy IdeS cleaved the entire plasma IgG-pool just minutes after dosing. IgG got to nadir 6-24 hrs after dosing and then slowly recuperated. The half-life of IdeS was 4.9( ± 2.8) hrs at 0.24 mg/kg with the main portion eliminated throughout 1 day.

Currently 2 hrs after IdeS-dosing, the phagocytic capacity of IgG/IgG-fragments was minimized to history degrees. Notably, IdeS has the capacity to suspend Fc-mediated effector function in vivo, was considered safe without any serious negative events, and also without dosage limiting poisoning in this study. The total, fast, but momentary elimination of IgG gives a new powerful healing opportunity in IgG-mediated pathogenic problems.

Successful treatment of speculative glomerulonephritis with IdeS and EndoS, IgG-degrading streptococcal enzymes. Yang et al. NEPHROLOGY DIALYSIS TRANSPLANTATION, Vol. 25, Issue 8, 2479-2486, March 2010.

Abstract.

BACKGROUND: Anti-glomerular cellar membrane layer (anti-GBM) disease frequently results in end-stage kidney failing regardless of treatment with plasma exchange as well as immunosuppressive medications. The newly uncovered streptococcal enzymes IgG-degrading enzyme of S.pyogenes( IdeS) and also endoglycosidase S( EndoS) act with exceptional uniqueness on flowing IgG. In this research, we explore their capacity in vivo to stop damage mediated by kidney-bound antibodies in a mouse model of anti-GBM illness. TECHNIQUES: Anti-GBM illness was caused in mice by shot of subnephritogenic dosages of rabbit anti-mouse GBM, adhered to a week later on by injection of monoclonal mouse anti-rabbit IgG antibodies. By supervising IdeS or EndoS as combination partners with GST between these antibody shots, we evaluated their ability to stop damages by acting upon kidney-bound bunny anti-GBM. Control animals obtained placebo injections. RESULTS: All pets in the favorable control groups established extreme albuminuria instantly after the 2nd antibody shot (mean, 2.51 mg/24 h; range, 0.13-8.20). This was considerably lessened by EndoS( 1.3 +/- 1.3 mg/24 h) as well as entirely stopped by IdeS (0.017 +/- 0.014 mg/24 h). Immunofluorescence researches showed that IdeS treatment effectively removed the Fc fragments of the bunny IgG. This was accompanied by a considerable reduction of the deposition of the complement parts C3 and C1q, and also this decreased the employment of leukocytes to the glomeruli. FINAL THOUGHT: IdeS degrades IgG bound to the GBM in vivo, therefore preventing kidney damages in this pet version. Most likely, IdeS would certainly deteriorate both circulating and also kidney-bound anti-GBM in patients with Goodpasture’s illness. Whether this would cause a stop in condition development and

a much better diagnosis continues to be to be determined. Healing cleavage of IgG: new avenues for dealing with inflammation. Nandakumar et al. PATTERN IN IMMUNOLOGY, Vol. 29, Problem 4, Apr 2008. Abstract. Autoantibodies creating in people add to the pathogenesis of numerous diseases, and injected therapeutic antibodies can additionally set off damaging side effects. An effective and also quick removal of these antibodies are for that reason seriously required. Antibody elimination by plasmapheresis and immunoadsorption are frequently used approaches however have their very own constraints. Bacterial enzymes that can cleave IgG molecules or get rid of carbohydrate moieties to alleviate their immunogenicity or effector functions in vivo supply new opportunities for medicine development. Recent discoveries highlight the opportunity of cleaving or modifying IgG in vivo by injection of enzymes. Such a method opens new healing opportunities not only for the control of pathogenic antibody-mediated inflammatory diseases yet additionally allograft being rejected or the treatment of side-effects of ‘biologicals’ such as monoclonal antibodies. Cleaving antibodies in vivo with a microbial enzyme- an unique healing concept. Johansson et al. PLOS ONE, Feb 2008, Vol 3, Issue 2. Abstract. HISTORY: IdeS, a proteinase from Streptococcus pyogenes, cleaves immunoglobulin( Ig) G antibodies with

a special degree of uniqueness. Pathogenic IgG antibodies make up an essential scientific trouble contributing to the pathogenesis of a number of autoimmune problems and intense transplant rejection. To be able to successfully remove such antibodies is for that reason a crucial clinical difficulty. METHODOLOGY/PRINCIPAL SEARCHINGS FOR: IdeS was discovered to particularly and efficiently cleave IgG in human blood artificial insemination (20 µg of IdeS triggered a total destruction of IgG in one ml of human whole blood in 15 mins) and to clear IgG from the blood stream of bunnies in vivo( no IgG was identified six hours complying with an intravenous injection of 5 mg of IdeS) without any adverse effects. In a mouse model of immune thrombocytopenic purpura (ITP), polyclonal IgG antibodies against platelet surface antigens were utilized to generate a lethal disease. These profoundly thrombocytopenic pets were treated and treated by a solitary injection of IdeS. CONCLUSIONS/SIGNIFICANCE: Novel info is offered concerning the IgG-cleaving activity of IdeS artificial insemination and in vivo. The highly details and also fast removal of IgG in vivo, the significant impact in a computer mouse model of ITP, as well as the absence of adverse effects in the treated pets, show that IdeS can likewise be utilized to treat IgG-driven conditions in human beings. Blocking of speculative arthritis by bosom of IgG antibodies in vivo. Nandakumar et al. JOINT INFLAMMATION & RHEUMATISM, Vol. 56, Issue 10, pp. 3253-3260, Oct. 2007. Abstract. GOAL: To explore whether IgG-degrading enzyme of Streptococcus pyogenes( IdeS), a microbial cysteine endopeptidase that cleaves human IgG in the joint area, can be made use of for blocking the advancement of arthritis. METHODS: Recombinant IdeS was cleansed as well as examined for uniqueness versus computer mouse IgG. IdeS was infused intravenously right into computer mice with collagen antibody-induced arthritis( CAIA), collagen-induced joint inflammation( CIA ), or slipping back CIA, as well as its results on joint inflammation growth and severity were examined. OUTCOMES: IdeS effectively cleaved mouse IgG2a/c and IgG3 in vitro. Also at reduced dosage (10 microg), IdeS especially cleaved IgG2a in vivo with no evident adverse effects. IdeS therapy efficiently obstructed CAIA caused by IgG2a antibodies. No impact was observed when arthritis was caused with IgG2b anti-type II collagen antibodies; given that IdeS does not cleave IgG2b, this indicated that IgG bosom was the system of action. IdeS therapy reduced the severity of arthritis if provided within 24 hr after the beginning of scientific joint inflammation, yet did not block continuous serious joint inflammation. IdeS therapy also dramatically prevented an antibody-induced regression in mice that had chronic joint inflammation, as well as delayed the beginning as well as lowered the intensity of joint inflammation in classic CIA. FINAL THOUGHT: IdeS has therapeutic capacity in IgG antibody-mediated autoimmune joint inflammation, representing a new as well as distinct methods of obstructing pathogenic antibodies. Chemical characterization of the streptococcal endopeptidase, IdeS, reveals that it is a cysteine protease with strict uniqueness for IgG bosom due to exosite binding. Vincents et al. BIOCHEMISTRY AND BIOLOGY 2004, 43, 15540-15549.

Abstract.

Streptococcus pyogenes, a vital virus in humans, secretes an IgG details endopeptidase called IdeS. To elucidate the mechanism that is accountable for this uniqueness, we have here defined the activity of IdeS carefully. Both gamma chains of human IgG or its Fc piece were cleaved in the joint area after Gly236 by IdeS, however other healthy proteins or artificial peptides consisting of sequences such as the P( 4)- P (1) section in the IgG bosom site, or long peptides looking like the IgG hinge, were not hydrolyzed whatsoever. This is likely because of a 2nd binding website engaging with the Fc part of IgG. The absence of IdeS task on peptide substrates necessitated the development of an assay with IgG as the substratum for kinetic research studies.

IdeS revealed a sigmoidal velocity curve at physiological IgG concentrations, and a declining enzyme price at greater IgG concentrations. This irregular velocity contour suggests item restraint and/or allosteric control, which once more shows the presence of an exosite associated with substrate binding. The pseudoequilibrium consistent for IdeS hydrolysis of IgG was 90 microM. The enzyme exhibited task in the pH series of 5.1-7.6, with an optimum at pH 6.6. IdeS was secure above pH 10 but not at acidic pH. It showed an activity maximum around 37 levels C and a decreased thermal security at 42 degrees C. Iodoacetate and iodoacetamide prevented IdeS, as expected for a cysteine protease, and biochemical proof confirmed this classification.

E-64 as well as hen cystatin, certain preventions of family C1 and C13 cysteine proteases, were without effect on enzyme activity, as were class certain serine, aspartic, and also metallo protease preventions. No substantial similarities were found in protein sequence comparisons with recognized enzyme households, suggesting that IdeS stands for a novel family of cysteine proteases. Framework of the streptococcal endopeptidase IdeS, a cysteine proteinase with rigorous uniqueness for IgG.

Wenig et al. PNAS Dec. 2004, Vol. 101, No. 50, pp. 17371-17376. Abstract. Pathogenic germs have actually established facility as well as varied virulence devices that compromise or disable the host immune defense system. IdeS (IgG-degrading enzyme of Streptococcus pyogenes) is a produced cysteine endopeptidase from the human microorganism S. pyogenes with a very high degree of substrate specificity, catalyzing a single proteolytic cleavage at the reduced hinge of human IgG. This proteolytic destruction promotes inhibition of opsonophagocytosis as well as interferes with the killing of group A Streptococcus. We have figured out the crystal structure of the catalytically non-active mutant IdeS-C94S by x-ray crystallography at 1.9-A resolution. Regardless of minimal sequence homology to known proteinases, the core of the structure looks like the canonical papain layer although with significant insertions as well as a distinctive substrate-binding website. For that reason IdeS comes from an one-of-a-kind family members within the CA clan of cysteine proteinases. Based on analogy with prevention facilities of papain-like proteinases, we suggest a version for substratum binding by IdeS. IdeS, a novel streptococcal cysteine protease with distinct uniqueness for immunoglobulin G. Von Pawel Rammingen et al. THE EMBO JOURNAL Vol. 21, No. 7 pp. 1607-1615, 2002. Abstract. Recent work from numerous laboratories has actually demonstrated that proteolytic devices significantly add to the molecular interaction in between Streptococcus pyogenes, an essential human microorganism, and its host. Right here we describe the recognition, purification and also characterization of an unique extracellular cysteine proteinase produced by S.pyogenes. This enzyme, assigned IdeS for Immunoglobulin G-degrading enzyme of S.pyogenes, is distinct from the well-characterized streptococcal cysteine proteinase, SpeB, and also cleaves human IgG in the joint region with a high level of specificity. Hence, various other human proteins, including immunoglobulins M, A, D and also E, are not broken down by IdeS. The enzyme efficiently cleaves IgG antibodies bound to streptococcal surface structures, thereby preventing the murder of S.pyogenes by phagocytic cells. This as well as added monitorings on the distribution and expression of the ideS gene indicate that IdeS stands for a novel and considerable bacterial virulence factor, and also a potential restorative target. HBP-assay. Heparin-Binding Protein Measurement Improves the Prediction of Severe Infection With Organ Disorder in the Emergency Department. Crit Treatment Med. 2015 Nov; 43 (11):2378 -86. Abstract. Objectives: Early identification of people with infection as well as at risk of establishing extreme illness with organ dysfunction continues to be a hard difficulty. We intended to evaluate as well as confirm the heparin-binding protein, a neutrophil-derived conciliator of vascular leak, as a prognostic biomarker for threat of progression to extreme blood poisoning with blood circulation failing in a multicenter setup. Layout: A possible international multicenter cohort research. Establishing: 7 various emergency departments in Sweden, Canada, as well as the USA. People: Grownup people with a thought infection and also a minimum of one of three medical systemic inflammatory reaction syndrome requirements (omitting leukocyte count). Intervention: None. Measurements and also Main Outcomes: Plasma degrees of heparin-binding protein, procalcitonin, C-reactive protein, lactate, and also leukocyte count were identified at admission as well as 12-24 hours after admission in 759 emergency situation department individuals with thought infection. People were specified depending upon the presence of infection and also body organ dysfunction. Plasma examples from 104 emergency division patients with thought sepsis accumulated at an independent center were utilized to confirm the results. Of the 674 individuals identified with an infection, 487 did not have body organ disorder at enrollment. Of these 487 individuals, 141 (29 %) developed organ dysfunction within the 72-hour study period; 78.0 % of the last individuals had an elevated plasma heparin-binding protein level (> 30 ng/mL) prior to development of organ dysfunction (mean, 10.5 human resources). Compared to various other biomarkers, heparin-binding healthy protein was the best predictor of development to organ dysfunction (location under the receiver operating particular curve = 0.80). The efficiency of heparin-binding protein was confirmed in the recognition associate. Final thought: In clients offering at the emergency department, heparin-binding protein is an early indicator of infection-related body organ disorder and a solid predictor of illness progression to extreme blood poisoning within 72 hrs. Heparin-binding protein: a diagnostic biomarker of urinary tract infection in adults. Kjölvmark et al., Open Online forum Infect Dis. 2014 Apr 23; 1 (1). Abstract. BACKGROUND: Urinary tract infections (UTIs) are connected with substantial morbidity and high frequency of antibiotic prescription. Identifying UTI is commonly challenging, particularly in the critically sick patient and also in individuals with unspecific and also moderate symptoms. The common quick tests have restricted value, as well as there is a need for even more reputable diagnostic devices. Heparin-binding protein (HBP) is released from neutrophils and has actually previously been researched as a diagnostic and anticipating biomarker in various bacterial infections. METHODS: This prospective study registered adult people at 2 primary care devices and 2 healthcare facility emergency situation divisions, to explore in urine HBP as a biomarker of UTI. In addition, urine levels of interleukin-6, white blood cells, and nitrite were analyzed and compared with HBP. Based on symptoms of UTI and microbiological findings, patients were classified into different groups, UTI (cystitis and pyelonephritis) and no UTI. RESULTS: Three hundred ninety patients were evaluated. The prevalence of UTI in the study group was 45.4 %. The sensitivity and specificity for HBP in urine as a marker for UTI were 89.2 % and 89.8 %, respectively. The positive and negative predictive values were 90.2 % and 88.8 %, respectively. Heparin-binding protein was the best diagnostic marker for UTI, with an area-under-curve value of 0.94 (95 % confidence interval, 0.93-0.96). Heparin-binding protein was significantly better in distinguishing cystitis from pyelonephritis, compared with the other markers. CONCLUSIONS: An elevated level of HBP in the urine is associated with UTI and may be a useful diagnostic marker in adult patients with a suspected UTI. Elevated urine levels of heparin-binding protein in children with urinary tract infection. Kjölvmark et al., Pediatr Nephrol. 2012 Aug; 27 (8):1301 -8. Abstract. BACKGROUND: Urinary tract infection (UTI) is a common infection diagnosis in children, and efficient diagnosis and treatment are important to avoid serious complications. In this study we investigated whether urinary levels of neutrophil-derived heparin-binding protein (HBP) can be used as a marker of UTI in children. These results were compared to those of dipstick analysis, interleukin-6 (IL-6) analysis in urine, and bacterial culturing. METHODS: Seventy-eight children aged 0-18 years with fever and/or symptoms indicating UTI were enrolled in a prospective consecutive study. Urine samples were cultured and analyzed with dipstick, and concentrations of HBP and IL-6 were measured. RESULTS: Fifteen patients were classified as having UTI, 30 patients had fever but were diagnosed with a non-urinary tract infection, and 33 patients had neither UTI nor fever. Using a urine HBP (U-HBP) cut-off level of 32 ng/mL, the sensitivity and specificity for detecting UTI were 93.3 and 90.3 %, respectively. Receiver operating characteristic curves demonstrated that U-HBP levels were a higher specificity indicator of UTI than urine white blood cell counts or urine IL-6 levels; they also showed a higher sensitivity than the results of the urine nitrite test. All patients with significant growth of clinically relevant bacteria had elevated U-HBP levels. CONCLUSION: The results indicate that rapid analysis of U-HBP can provide helpful guidance in the management of children with suspected UTI. Elevated plasma levels of heparin-binding protein in intensive care unit patients with severe sepsis and septic shock. Linder et al., Crit Care. 2012 May 21; 16 (3): R90.

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