ABSTRACT
Involvement of lower gastrointestinal tract (LGI) occurs in 60% of patients with graft-versus-host-disease (GVHD). Complement components C3 and C5 are involved in GVHD pathogenesis. In this phase 2a study, we evaluated the safety and efficacy of ALXN1007, a monoclonal antibody against C5a, in patients with newly diagnosed LGI acute GVHD receiving concomitant corticosteroid. Twenty-five patients were enrolled; one was excluded from the efficacy analysis based upon negative biopsy. Most patients (16/25, 64%) had acute leukemia; 52% (13/25) had an HLA-matched unrelated donor; and 68% (17/25) received myeloablative conditioning. Half the patients (12/24) had a high biomarker profile, Ann Arbor score 3; 42% (10/24) had high-risk GVHD per Minnesota classification. Day-28 overall response was 58% (13/24 complete response, 1/24 partial response), and 63% by Day-56 (all complete responses). Day-28 overall response was 50% (5/10) in Minnesota high-risk and 42% (5/12) in high-risk Ann Arbor patients, increasing to 58% (7/12) by Day-56. Non-relapse mortality at 6-months was 24% (95% CI 11-53). The most common treatment-related adverse event was infection (6/25, 24%). Neither baseline complement levels (except for C5), activity, nor inhibition of C5a with ALXN1007 correlated with GVHD severity or responses. Further studies are needed to evaluate the role of complement inhibition in GVHD treatment.
Subject(s)
Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Humans , Complement Inactivating Agents/therapeutic use , Complement C5a/therapeutic use , Prospective Studies , Hematopoietic Stem Cell Transplantation/adverse effects , Graft vs Host Disease/etiology , Lower Gastrointestinal Tract/pathologyABSTRACT
INTRODUCTION: Complement C5a is an important component of the innate immune system. An increasing number of reports have revealed the relevance of C5a in tumor progression; however, its exact role in metastatic renal cell carcinoma (mRCC) remains unknown. METHODS: We evaluated C5a expression in tumor tissue microarrays of 231 mRCC patients and analyzed the relationship between C5a levels and clinical outcomes, and the expression of epithelial-mesenchymal transition (EMT)-related proteins, programmed cell death protein 1 (PD-1), and programmed cell death-ligand 1 (PD-L1). In-vitro functional experiments using exogenous C5a stimulation and C5a silencing in renal cell carcinoma cells were used to validate the tissue findings. RESULTS: High C5a expression was associated with poor therapeutic responses, poor overall and progression-free survival, and high expression of EMT-related proteins and PD-1/PD-L1 in mRCC patients. Exogenous C5a promoted proliferation, migration, and invasion of renal cell carcinoma cells, and induced the expression of EMT-related proteins and PD-1/PD-L1. Conversely, C5a silencing inhibited migration and invasion of renal cell carcinoma cells and decreased the expression of EMT-related proteins and PD-1/PD-L1. CONCLUSIONS: Our findings indicate that elevated C5a expression is associated with poor outcomes in patients with mRCC, and this effect may be partly attributed to the ability of C5a to promote EMT and PD-1/PD-L1 expression. C5a may be a potential novel target for the treatment of mRCC.
Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Humans , Carcinoma, Renal Cell/pathology , Programmed Cell Death 1 Receptor , Kidney Neoplasms/genetics , Kidney Neoplasms/pathology , B7-H1 Antigen/metabolism , Complement C5a/genetics , Complement C5a/pharmacology , Complement C5a/therapeutic use , Epithelial-Mesenchymal Transition/geneticsABSTRACT
Emerging evidence has documented that multisystem organ failure in coronavirus disease 2019 (COVID-19) patients is strongly associated with various coagulopathies. Treatments for COVID-19-associated coagulopathy are still a clinical challenge. An advancement in the knowledge of mechanisms of the excessive or inappropriate activation of the complement cascade involved in the genesis of COVID-19-associated coagulopathy might be a fundamental approach for developing novel classes of anticoagulant drugs. In this context, there is emerging evidence indicating that C5a, a component of the complement system, and its receptors (C5aRs) play a critical role in the genesis of the COVID-19-associated hypercoagulable state. Thus, this review describes the mechanisms by which C5a/C5aR signaling participates in the cascade of events involved in the pathophysiology of COVID-19-associated coagulopathy. Furthermore, it highlights the current possibilities for the development of a novel therapeutic approach for COVID-19 patients that targets C5a/C5aRs signaling.
Subject(s)
COVID-19/therapy , Complement C5a/physiology , Complement C5a/therapeutic use , Thrombophilia/therapy , Animals , COVID-19/blood , COVID-19/complications , COVID-19/epidemiology , Complement Activation/physiology , Humans , SARS-CoV-2/immunology , SARS-CoV-2/pathogenicity , Signal Transduction , Thrombophilia/epidemiology , Thrombophilia/etiologyABSTRACT
BACKGROUND: Complement system is theoretically believed to halt the progression of tumor by the activity of C5a/CD88. Protein C5a is a potent pro.inflammatory mediator that activates the complement system by binding to its receptor. OBJECTIVES: The purpose of this study is to determine the expression of the anaphylatoxin C5a receptor on 4T1 cell line and to study the viability of the cells after being treated with the C5a peptides. MATERIALS AND METHODS: The cells 4T1 had undergone immunofluorescence staining, conventional polymerase chain reaction (PCR) and real-time PCR for the expression of determination part. Whereas Alamar Blue and MTT assays were conducted for the viability study of the cells. RESULTS: The cells showed positive result in expressing the receptor of the C5a through immunostaining and PCR. The CT value recorded at initial dilution was 22.24. In cell viability assay, the cell was treated with C5a peptides, PMX205 and EP54. The purpose of this treatment was to see whether C5a had a direct effect on the cell itself using both assays. The result showed that PMX205, which is an antagonist, gave more effects towards the cell as compared with the treatment of EP54. CONCLUSION: This experiment shows the presence of C5a receptor on 4T1 cell line. We believe that the antagonist peptide is eligible to be used widely in cancer immunotherapy field; but in vivo studies need to be carried out first in the future, as it will determine how these drugs affect the tumor cell growth.
Subject(s)
Complement C5a/therapeutic use , Mammary Neoplasms, Experimental/drug therapy , Peptide Fragments/therapeutic use , Peptides, Cyclic/therapeutic use , Receptor, Anaphylatoxin C5a/analysis , Animals , Cell Line, Tumor , Cell Survival , Female , Fluorescent Antibody Technique , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/pathology , Mice , Polymerase Chain Reaction , Receptor, Anaphylatoxin C5a/agonists , Receptor, Anaphylatoxin C5a/antagonists & inhibitorsABSTRACT
The specific role of C5a in cancer, especially in melanoma, has yet to be determined. Differential effects of C5a could be cancer specific. In the host defense system, C5a functions to protect the body from harmful entities via a plethora of mechanisms. Yet, C5a may also serve to potentiate cancerous process. C5a facilitates cellular proliferation and regeneration by attracting myeloid-derived suppressor cells and supporting tumor promotion. In this article, we critically reviewed the properties, mechanisms of action and functions of C5a, with particular emphasis on cancer inhibition and promotion, and clinical application of such knowledge in better management of patients with cancer. Outstanding questions and future directions in regard to the function of C5a in melanoma and other cancers are discussed.
Subject(s)
Complement C5a , Melanoma , Cell Proliferation/drug effects , Complement C5a/adverse effects , Complement C5a/immunology , Complement C5a/therapeutic use , Humans , Melanoma/drug therapy , Melanoma/immunology , Melanoma/pathology , Myeloid Cells/immunologyABSTRACT
We have recently found that central PGD(2) exhibits anxiolytic-like activity. Here we show that complement C5a exhibits anxiolytic-like activity via the PGD(2) system. Centrally administered C5a had anxiolytic-like activity at a dose of 0.3 pmol/mouse in the elevated plus-maze test in mice. C5a-induced anxiolytic-like activity was inhibited by indomethacin, a cyclooxygenase inhibitor, or BWA868C, an antagonist of DP(1) receptor for PGD(2), respectively. The anxiolytic effect of C5a was also blocked by SCH58261 or bicuculline, antagonists of adenosine A(2A) and GABA(A) receptors, respectively, which were activated downstream of PGD(2)-DP(1) receptor. These results suggest that C5a exhibits anxiolytic-like activity via the PGD(2)-DP(1) receptor system coupled to the activation of adenosine A(2A) and GABA(A) receptors.
Subject(s)
Anti-Anxiety Agents/therapeutic use , Complement C5a/therapeutic use , Receptor, Adenosine A2A/metabolism , Receptors, GABA-A/metabolism , Receptors, Immunologic/metabolism , Receptors, Prostaglandin/metabolism , Animals , Bicuculline/pharmacology , Hydantoins/pharmacology , Indomethacin/pharmacology , Male , Mice , Receptors, Immunologic/antagonists & inhibitors , Receptors, Prostaglandin/antagonists & inhibitorsABSTRACT
To investigate the role of C5a generated on complement activation in brain, the lupus model, MRL/lpr mice were treated with C5a receptor(R) antagonist (ant). Neutrophil infiltration, ICAM, TNF-alpha and iNOS mRNA expression, neuronal apoptosis and the expression of p-JNK, pSTAT1 and p-Erk were reduced and p-Akt increased on C5aR inhibition in MRL/lpr brains. MRL/lpr serum caused increased apoptosis in neurons showing that lupus had a direct effect on these cells. C5aRant pretreatment prevented the lupus serum induced loss of neuronal cells. Our findings demonstrate for the first time that C5a/C5aR signaling plays an important role in the pathogenesis of CNS lupus.
Subject(s)
Brain/metabolism , Complement C5a/therapeutic use , Lupus Vasculitis, Central Nervous System/drug therapy , Lupus Vasculitis, Central Nervous System/pathology , Receptor, Anaphylatoxin C5a/metabolism , Animals , Annexin A5/metabolism , Apoptosis/drug effects , Brain/cytology , Cell Adhesion Molecules/genetics , Cell Adhesion Molecules/metabolism , Cells, Cultured , Chemokine CXCL2/genetics , Chemokine CXCL2/metabolism , Disease Models, Animal , Embryo, Mammalian , In Situ Nick-End Labeling/methods , Intercellular Adhesion Molecule-1/metabolism , Male , Mice , Mice, Inbred MRL lpr , Neurons/drug effects , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , RNA, Messenger/metabolism , Receptor, Anaphylatoxin C5a/genetics , Signal Transduction/drug effects , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Up-Regulation/drug effectsABSTRACT
Toll-like receptors (TLRs) and complement are 2 major components of innate immunity that provide a first-line host defense and shape the adaptive immune responses. We show here that coincidental activation of complement and several TLRs in mice led to the synergistic production of serum factors that promoted T-helper cell 17 (Th17) differentiation from anti-CD3/CD28 or antigen-stimulated T cells. Although multiple TLR-triggered cytokines were regulated by complement, Th17 cell-promoting activity in the serum was correlated with interleukin (IL)-6 induction, and antibody neutralization of IL-6 abrogated the complement effect. By using both in vitro and in vivo approaches, we examined in more detail the mechanism and physiologic implication of complement/TLR4 interaction on Th17-cell differentiation. We found that the complement effect required C5a receptor, was evident at physiologically relevant levels of C5a, and could be demonstrated in cultured peritoneal macrophages as well as in the setting of antigen immunization. Importantly, despite an inhibitory effect of complement on IL-23 production, complement-promoted Th17 cells were functionally competent in causing autoimmunity in an adoptive transfer model of experimental autoimmune encephalomyelitis. Collectively, these data establish a link between complement/TLR interaction and Th17-cell differentiation and provide new insight into the mechanism of action of complement in autoimmunity.
Subject(s)
Complement Activation , Inflammation/immunology , Interleukin-6/physiology , T-Lymphocytes, Helper-Inducer/cytology , Toll-Like Receptors/physiology , Adoptive Transfer , Animals , CD55 Antigens/genetics , Cells, Cultured/cytology , Cells, Cultured/metabolism , Complement C5a/physiology , Complement C5a/therapeutic use , Cytokines/blood , Cytokines/physiology , Encephalomyelitis, Autoimmune, Experimental/etiology , Encephalomyelitis, Autoimmune, Experimental/immunology , Immunity, Innate , Immunization , Inflammation/chemically induced , Interleukin-6/biosynthesis , Interleukin-6/blood , Interleukin-6/deficiency , Interleukin-6/genetics , Lymphopoiesis , Macrophages, Peritoneal/cytology , Macrophages, Peritoneal/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Receptor, Anaphylatoxin C5a/deficiency , Receptor, Anaphylatoxin C5a/physiology , Recombinant Proteins/therapeutic use , Specific Pathogen-Free Organisms , T-Lymphocytes, Helper-Inducer/classification , T-Lymphocytes, Helper-Inducer/immunology , Toll-Like Receptor 4/physiologyABSTRACT
BACKGROUND: The instant blood-mediated inflammatory reaction, characterized by activation of both the coagulation and complement cascades, is a serious obstacle to successful islet engraftment. No attractive protocol is clinically available as yet. The objective of the present study was to examine whether complementary peptide against an active region of C5a in combination with a clinically available anticoagulant could provide an effective protocol for suppression of the instant blood-mediated inflammatory reaction. METHODS: Three islet equivalents per gram of syngeneic rat grafts were transplanted intraportally into 6 pairs of rats with streptozotocin-induced diabetes. Islets from the same donor were transplanted into each pair. In each pair, one rat was treated with C5a inhibitory peptide in addition to continuous intravenous infusion of gabexate mesilate and the other rat, injected with equivalent amount of saline solution, served as the control. In addition, 6 rats that received transplants from irrelevant donors were treated with the same dose of gabexate mesilate. We evaluated the cure rate, time to normoglycemia, liver insulin concentration in recipients, and results of in vivo glucose tolerance tests. RESULTS: The cure rate was remarkably improved and the time to normoglycemia in cured animals was significantly shortened with C5a inhibitor plus gabexate treatment. In six rats that received only gabexate mesilate, normoglycemia was not restored during the study. CONCLUSIONS: These data suggest that C5a inhibitory peptide combined with gabexate mesilate could be an attractive drug candidate without adverse effects to control the detrimental innate immune responses induced in clinical islet transplantation.
Subject(s)
Complement C5a/therapeutic use , Diabetes Mellitus, Experimental/surgery , Gabexate/therapeutic use , Graft Survival/drug effects , Inflammation/prevention & control , Animals , Blood Glucose/drug effects , Blood Glucose/metabolism , Disease Models, Animal , Inflammation/blood , Islets of Langerhans Transplantation/adverse effects , Rats , Serine Proteinase Inhibitors/therapeutic useABSTRACT
Amyotrophic lateral sclerosis (ALS) is one of the major forms of motor neuron disease (MND), a group of degenerative disorders causing progressive motor neuron death leading to eventual paralysis and death. The pathogenesis of MND is poorly understood and may include genetic and/or environmental factors, with a common end-stage outcome. The majority of cases are sporadic, with a small percentage of familial cases identified. Mutations in the copper/zinc superoxide dismutase (SOD1) enzyme are frequent in familial ALS, and have allowed for the development of transgenic SOD1 rodent models of ALS. There has been evidence for immune system involvement in the disease, and activated components of the classical complement pathway have been observed in the serum, cerebrospinal fluid and neuronal tissue of diseased individuals. Furthermore, motor neurons and spinal cord tissue from SOD1 transgenic mice show an upregulation in C1q mRNA transcript and protein, in some cases prior to disease onset. Our laboratory has preliminary data indicating a specific pathogenic role for the activation fragment of complement C5 (C5a) in this disease. Using selective C5a receptor antagonists, we dosed SOD1 transgenic rats and observed an extension in survival and reduced motor symptoms compared to untreated rats. Collectively, these clinical and experimental findings suggest that targeting complement using specific inhibitors may represent a novel therapeutic approach to treating MND. Further experimental and clinical studies are required to validate this hypothesis. This review will summarize the clinical and experimental evidence to date implicating complement in the pathogenesis of MND.
Subject(s)
Complement C5a , Disease Models, Animal , Motor Neuron Disease/metabolism , Receptor, Anaphylatoxin C5a/antagonists & inhibitors , Superoxide Dismutase/genetics , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/metabolism , Amyotrophic Lateral Sclerosis/pathology , Animals , Complement Activation , Complement C5a/antagonists & inhibitors , Complement C5a/immunology , Complement C5a/therapeutic use , Motor Neuron Disease/genetics , Motor Neuron Disease/pathology , Motor Neurons/metabolism , Motor Neurons/pathology , RNA, Messenger/metabolism , Rats , Rats, Transgenic , Spinal Cord/enzymology , Spinal Cord/metabolism , Spinal Cord/pathology , Superoxide Dismutase/metabolismABSTRACT
A novel method to improve targeting and presentation of poorly immunogenic tumor-related antigens was investigated. This was performed with a molecular adjuvant constructed by covalently linking a response selective peptide agonist of C5a (YSFKDMP(MeL)aR) to known melanoma tumor-related antigens. C57Bl/6J mice were injected subcutaneously with bone marrow derived dendritic cells (DCs) pulsed with a melanoma epitope (TRP2-P2/Agonist), melanoma epitope tyrosinase (TYR/Agonist), a nonfunctional reverse conformation C5a agonist bound to TYR(reverse peptide) or DMSO-PBS vehicle. Mice were injected with the pulsed DCs and cytokines IL-2 and GMCSF three times prior to subcutaneous challenge with B16-F10 melanoma cells. All groups subsequently received DC vaccine boosters twice per week. Tumor growth was reduced and survival enhanced in mice immunized with the combination of TRP2-P2/Agonist and TYR/Agonist compared to mice receiving reverse peptide or vehicle.
Subject(s)
Cancer Vaccines/immunology , Cancer Vaccines/therapeutic use , Complement C5a/agonists , Dendritic Cells/metabolism , Dendritic Cells/transplantation , Disease Models, Animal , Melanoma, Experimental/prevention & control , Animals , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Antigens, Neoplasm/metabolism , Cancer Vaccines/administration & dosage , Complement C5a/genetics , Complement C5a/therapeutic use , Dendritic Cells/immunology , Growth Inhibitors/administration & dosage , Growth Inhibitors/metabolism , Growth Inhibitors/therapeutic use , Humans , Interleukin-2/metabolism , Intramolecular Oxidoreductases/administration & dosage , Intramolecular Oxidoreductases/metabolism , Intramolecular Oxidoreductases/therapeutic use , Melanoma, Experimental/genetics , Melanoma, Experimental/immunology , Mice , Mice, Inbred C57BLABSTRACT
Complement fragment (C)5a is a 74 residue pro-inflammatory polypeptide produced during activation of the complement cascade of serum proteins in response to foreign surfaces such as microorganisms and tissue damaged by physical or chemical injury. C5a binds to at least two seven-transmembrane domain receptors, C5aR (C5R1, CD88) and C5L2 (gpr77), expressed ubiquitously on a wide variety of cells but particularly on the surface of immune cells like macrophages, neutrophils and T cells. C5aR is a classical G protein-coupled receptor that signals through G alpha i and G alpha 16, whereas C5L2 does not appear to couple to G proteins and has no known signalling activity. Although C5a was first described as an anaphylatoxin and later as a leukocyte chemoattractant, the widespread expression of C5aR suggested more general functionality. Our understanding of the physiology of C5a has improved significantly in recent years through exploitation of receptor knockout and knocking mice, C5 and C5a antibodies, soluble recombinant C5a and C5a analogues and newly developed receptor antagonists. C5a is now also implicated in non-immunological functions associated with developmental biology, CNS development and neurodegeneration, tissue regeneration, and haematopoiesis. Combined receptor mutagenesis, molecular modelling, structure-activity relationship studies and species dependence for ligand potency on C5aR have been helpful for identifying ligand binding sites on the receptor and for defining mechanisms of receptor activation and inactivation. This review will highlight major developments in C5a receptor research that support C5aR as an important therapeutic target. The intriguing possibilities raised by the existence of a non-signalling C5a receptor are also discussed.
Subject(s)
Complement C5a/pharmacology , Receptor, Anaphylatoxin C5a/antagonists & inhibitors , Receptor, Anaphylatoxin C5a/physiology , Amino Acid Sequence , Animals , Complement C5a/chemistry , Complement C5a/therapeutic use , Humans , Models, Biological , Molecular Sequence Data , Molecular Structure , Protein Binding , Receptor, Anaphylatoxin C5a/geneticsABSTRACT
Some in vivo activities of two complement C5a agonist analogues have been evaluated by measuring changes in blood pressure and neutropenia in the rat and comparing the results with their receptor affinities in peritoneal macrophages and polymorphonuclear leucocytes (PMNs). In vitro C5a receptor (C5aR) binding experiments showed that YSFKPMPLaR and YSFKD(NMeNle)PlaR had similar affinities for the macrophage C5aR (IC50 0.2, 0.1 microM respectively). In PMNs, the affinity of YSFKPMPLaR (IC50 0.1 microM) was similar to that in macrophages, whereas the affinity of YSFKD(NMeNle)PLaR for the PMN C5aR was >100 microM. Given i.v., YSFKD(NMeNle)PLaR had similar activity to YSFKPMPLaR on blood pressure but did not cause neutropenia. These results demonstrate selectivity of a new C5a agonist in vitro, which is paralleled in vivo. The results suggest the possibility of developing selective agonists of C5a for in vivo use in humans.