RESUMO
Multiple myeloma (MM) is an incurable malignancy of the B-cell lineage. Remarkable progress has been made in the treatment of MM with anti-CD38 monoclonal antibodies such as daratumumab and isatuximab, which can kill MM cells by inducing complement-dependent cytotoxicity (CDC). We showed that the CDC efficacy of daratumumab and isatuximab is limited by membrane complement inhibitors, including CD46 and CD59, which are upregulated in MM cells. We recently developed a small recombinant protein, Ad35K++, which is capable of transiently removing CD46 from the cell surface. We also produced a peptide inhibitor of CD59 (rILYd4). In this study, we tested Ad35K++ and rILYd4 in combination with daratumumab and isatuximab in MM cells as well as in cells from two other B-cell malignancies. We showed that Ad35K++ and rILYd4 increased CDC triggered by daratumumab and isatuximab. The combination of both inhibitors had an additive effect in vitro in primary MM cells as well as in vivo in a mouse xenograft model of MM. Daratumumab and isatuximab treatment of MM lines (without Ad35K++ or rILYd4) resulted in the upregulation of CD46/CD59 and/or survival of CD46high/CD59high MM cells that escaped the second round of daratumumab and isatuximab treatment. The escape in the second treatment cycle was prevented by the pretreatment of cells with Ad35K++. Overall, our data demonstrate that Ad35K++ and rILYd4 are efficient co-therapeutics of daratumumab and isatuximab, specifically in multi-cycle treatment regimens, and could be used to improve treatment of multiple myeloma.
Assuntos
Antineoplásicos , Mieloma Múltiplo , Humanos , Camundongos , Animais , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/patologia , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados/uso terapêutico , Antineoplásicos/uso terapêutico , ADP-Ribosil Ciclase 1/metabolismo , Antígenos CD59/uso terapêutico , Proteína Cofatora de Membrana/metabolismoRESUMO
Rheumatoid arthritis (RA) is typified by persistent joint inflammation, which leads to the deterioration of bone and cartilage and a reduction in overall quality of life. The global prevalence of pain as a primary symptom in RA is influenced by the interplay between inflammation and its resolution. The identification of a family of lipid mediators known as specialized pro-resolving mediators (SPM)s has contributed to the progress of our comprehension of inflammatory conditions. SPMs have been observed to trigger the process of inflammation resolution, thereby reinstating the homeostasis of the inflammatory response. Autacoids are synthesized through the stereo-selective transformation of essential fatty acids, resulting in molecules dynamically modulated during inflammation and possessing strong immunoregulatory properties. This review delves into the available evidence that supports the involvement of certain SPM as protective lipids, biomarkers with potential, and therapeutic targets in the context of RA.
Assuntos
Artrite Reumatoide , Ácidos Docosa-Hexaenoicos , Humanos , Antígenos CD59/uso terapêutico , Qualidade de Vida , Mediadores da Inflamação , Inflamação/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêuticoRESUMO
OBJECTIVES: To describe the clinical phenotype of eight children diagnosed with CD59 deficiency and their ultimate neurological outcome. METHODS: The data of our cases were extensively reviewed both clinical and ancillary tests; investigations included: neuroimaging, neurophysiological studies, and laboratory tests. RESULTS: All patients presented during early infancy with Guillain-Barre syndrome later they suffered repeated relapses leading to the diagnosis of chronic axonal neuropathy. Recurrent stroke and acute necrotizing encephalopathy were described, 2 patients in each group. One girl developed acute disseminated encephalomyelitis while one boy developed acute transverse myelitis. Overt hemolytic anemia requiring blood transfusion reported in six patients. CONCLUSION: Inherited CD59 deficiency is an autosomal recessive disorder which can have devastating neurological consequences. First line immunotherapy including intravenous immunoglobin, corticosteroids, and plasma exchange may have transient beneficial effect. Reports of targeted therapy with eculizumab might be lifesaving. Genetic counseling is crucial.
Assuntos
Anemia Hemolítica , Síndrome de Guillain-Barré , Humanos , Recidiva Local de Neoplasia , Anemia Hemolítica/genética , Hemoglobinúria/genética , Antígenos CD59/genética , Antígenos CD59/uso terapêuticoRESUMO
Data from human dry and wet age-related macular degeneration (AMD) eyes support the hypothesis that constant 'tickover' of the alternative complement pathway results in chronic deposition of the complement membrane attack complex (MAC) on the choriocapillaris and the retinal pigment epithelium (RPE). Sub-lytic levels of MAC lead to cell signaling associated with tissue remodeling and the production of cytokines and inflammatory molecules. Lytic levels of MAC lead to cell death. CD59 is a naturally occurring inhibitor of the assembly of MAC. CD59 may thus be therapeutically efficacious against the pathophysiology of dry and wet AMD. The first gene therapy clinical trial for geographic atrophy - the advanced form of dry AMD has recently completed recruitment. This trial is studying the safety and tolerability of expressing CD59 from an adeno-associated virus (AAV) vector injected once into the vitreous. A second clinical trial assessing the efficacy of CD59 in wet AMD patients is also under way. Herein, the evidence for the role of MAC in the pathophysiology of dry as well as wet AMD and the scientific rationale underlying the use of AAV- delivered CD59 for the treatment of dry and wet AMD is discussed.
Assuntos
Antígenos CD59/uso terapêutico , Complexo de Ataque à Membrana do Sistema Complemento/fisiologia , Terapia Genética , Atrofia Geográfica/fisiopatologia , Degeneração Macular Exsudativa/fisiopatologia , Animais , Antígenos CD59/genética , Ensaios Clínicos como Assunto , Dependovirus/genética , Vetores Genéticos , Atrofia Geográfica/terapia , Humanos , Injeções Intravítreas , Degeneração Macular Exsudativa/terapiaRESUMO
The resolution of acute inflammation, once thought to be a passive process, is now recognized as an active one. The productions of endogenous special proresolving mediators (SPMs) are involved in this process. SPMs, including lipoxins, resolvins, protectins, and maresins, are endogenous lipid mediators generated from ω-6 arachidonic acid or ω-3 poly-unsaturated fatty acids during the resolution phase of acute inflammation. They have potent anti-inflammatory and proresolving actions in various inflammatory disorders. Due to the potent proresolving and anti-inflammatory effects, SPMs are also used for pain relief. This review focuses on the mechanisms by which SPMs act on their respective G-protein-coupled receptors in immune cells and nerve cells to normalize pain via regulating inflammatory mediators, transient receptor potential ion channels, and central sensitization. SPMs may offer novel therapeutic approaches for preventing and treating pain conditions associated with inflammation.
Assuntos
Anti-Inflamatórios/uso terapêutico , Inflamação/complicações , Dor/tratamento farmacológico , Dor/etiologia , Animais , Antígenos CD59/uso terapêutico , Ácidos Docosa-Hexaenoicos/uso terapêutico , Humanos , Inflamação/tratamento farmacológico , Lipoxinas/uso terapêuticoRESUMO
PURPOSE OF REVIEW: The literature regarding the use of fish oils in the critically ill to limit the inflammatory and catabolic response have been inconsistent. The objective of this manuscript is to review a newly discovered class of specialized proresolving molecules (SPMs), which could help elucidate the discrepancies reported in the critical care literature regarding the anti-inflammatory benefits of fish oil/ω-3 fatty acids. RECENT FINDINGS: Although use of fish oil has traditionally been thought to reduce or limit the inflammatory process in the critical ill, a new class of endogenously produced highly active lipid mediators derived from arachidonic acid and ω-3 fatty acids (lipoxins, resolvins, protectins, and maresins) have been shown to actively enhance resolution of inflammation. These SPMs stimulate the cardinal signs of resolution of inflammation, which include the cessation of leukocytic infiltration, a countering of the effects of proinflammatory mediators, stimulation of the uptake of apoptotic neutrophils, promotion of the clearance of necrotic cellular debris, and enhancement of the host's ability to eliminate microbial invasion. SUMMARY: By actively turning off inflammation (instead of simply attenuating its natural course), SPMs have shown more consistent effects in decreasing pain and risk of sepsis, increasing epithelialization and wound healing, promoting tissue regeneration, potentiating the effects of antibiotics, and enhancing adaptive immunity.
Assuntos
Óleos de Peixe/uso terapêutico , Inflamação/tratamento farmacológico , Unidades de Terapia Intensiva , Imunidade Adaptativa/efeitos dos fármacos , Anti-Inflamatórios/uso terapêutico , Ácido Araquidônico/uso terapêutico , Antígenos CD59/uso terapêutico , Estado Terminal , Ácidos Docosa-Hexaenoicos/uso terapêutico , Humanos , Lipoxinas/farmacologia , Dor/tratamento farmacológico , Regeneração/efeitos dos fármacos , Cicatrização/efeitos dos fármacosRESUMO
Periodontal diseases are comprised of a group of inflammatory conditions that result in the destruction of the supporting structures of the dentition. Emphasis has traditionally been placed on the deleterious actions of lipid mediators, such as prostanoids and leukotrienes, in propagating the inflammatory response and enhancing tissue destruction. Recently, the emerging understanding of the molecular basis of inflammation has elucidated that return of tissue homeostasis, triggered as part of a normal inflammatory response i.e. resolution of inflammation is an active, agonist-mediated, well-orchestrated phenomenon. The naturally-occurring pro-resolution lipid mediators, lipoxins, resolvins, protectins, maresins etc. have been identified as a novel genus of potent and stereoselective players that counter-regulate excessive acute inflammation and stimulate molecular and cellular events that define resolution. In this Review, we provide an update and overview of newly identified mediators that play pivotal roles in resolution and focus on the emerging appreciation of the endogenous pathways and mediators that control timely resolution which can be exploited as novel drug targets to extend the pharamaceutical armamentarium to combat chronic inflammation, thus controlling periodontal inflammation and the associated systemic inflammatory effects on the body, in general.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Mediadores da Inflamação/antagonistas & inibidores , Antagonistas de Leucotrienos/uso terapêutico , Terapia de Alvo Molecular , Periodontite/tratamento farmacológico , Periodonto/efeitos dos fármacos , Antagonistas de Prostaglandina/uso terapêutico , Animais , Antígenos CD59/metabolismo , Antígenos CD59/uso terapêutico , Desenho de Fármacos , Ácidos Graxos Ômega-3/química , Ácidos Graxos Ômega-3/uso terapêutico , Humanos , Mediadores da Inflamação/metabolismo , Lipoxinas/metabolismo , Lipoxinas/uso terapêutico , Periodontite/imunologia , Periodontite/metabolismo , Periodontite/prevenção & controle , Periodonto/imunologia , Periodonto/metabolismoRESUMO
Inflammation is a fundamental biologic process evolutionally preserved by a germ line code. The interplay of the epigenetic with the environment directs the code to temporally distinct inflammatory responses, which can be acute or chronic. The aim of this study is to present new aspects regarding the resolution of inflammation. Acute inflammation normally resolves by mechanisms still somewhat elusive. Current evidence suggests that an active coordinated program initiated the first few hours after the inflammatory response begins and its failure lead to chronic inflammation. This process is essential for appropriate host responses, tissue protection and the return to homeostasis. Prostaglandins and leukotrienes are lipid mediators that play important roles in host defense and acute inflammation. Granulocytes promote the switch of arachidonic acid-derived prostaglandins and leukotrienes to lipoxins, active antiinflammatory and pro-resolution mediators. The apoptosis of the neutrophils coincides with the biosynthesis of resolvins and protectins from omega-3 polyunsaturated fatty acids and releases anti-inflammatory and reparative cytokines. This information could lead to new treatments for inflammatory diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Mediadores da Inflamação/imunologia , Inflamação/tratamento farmacológico , Inflamação/imunologia , Anti-Inflamatórios/uso terapêutico , Antígenos CD59/farmacologia , Antígenos CD59/uso terapêutico , Doença Crônica , Ácidos Docosa-Hexaenoicos/farmacologia , Ácidos Docosa-Hexaenoicos/uso terapêutico , Glucocorticoides/farmacologia , Glucocorticoides/uso terapêutico , Humanos , Inflamação/metabolismo , Inflamação/prevenção & controle , Lipoxinas/farmacologia , Lipoxinas/uso terapêutico , Superóxido Dismutase/farmacologia , Superóxido Dismutase/uso terapêutico , Resultado do TratamentoRESUMO
AIM: Periodontitis is an inflammatory disease initiated by microbial biofilm. The host response to the biofilm destroys the periodontium mediated by an overly robust inflammatory response in susceptible individuals. Whether the excessive host response is genetic, epigenetic or mediated by environment is unknown. New pathways of resolution of inflammation have been discovered. Resolution of inflammation is an active, agonist-mediated, programmed return to tissue homeostasis. MATERIALS AND METHODS: Various computer-based search engines were employed to identify papers relevant to resolution of inflammation. RESULTS: Recent data suggest that chronic inflammatory periodontal disease may be a failure of resolution pathways as well as overexpression of proinflammatory pathways. In this review, the biology of resolution of inflammation will be examined in normal tissues and periodontal disease. Anti-inflammatory pharmacologic agents [non-steroidal anti-inflammatory drugs (NSAIDs)] have been shown to prevent and slow the progression of periodontitis in animals and humans. However, the side effect profile of NSAIDS or other inhibitors or receptor antagonists preclude their use in periodontal therapy. CONCLUSION: The isolation and characterization of proresolving lipid mediators that are receptor agonists has opened a new area of research for potential therapeutic agents for the management of inflammatory periodontitis.
Assuntos
Ácidos Docosa-Hexaenoicos/uso terapêutico , Lipoxinas/uso terapêutico , Periodontite/prevenção & controle , Animais , Anti-Inflamatórios/uso terapêutico , Biofilmes , Antígenos CD59/uso terapêutico , Homeostase/fisiologia , Humanos , Mediadores da Inflamação/fisiologia , Periodontite/microbiologia , Periodontite/terapiaRESUMO
PURPOSE OF REVIEW: To report recent data on the potential role of omega-3 fatty acids, in particular docosahexaenoic acid and its derivatives, in the treatment of dry eye syndrome. RECENT FINDINGS: Dietary supplementation with polyunsaturated fatty acids yields positive results in the improvement of dry eye signs and symptoms. Although several studies have shown this, evidence is still lacking as to which fatty acid or what combination constitutes the most effective treatment. Studies show that treatment with alpha-linoleic acid reduces dry eye-induced inflammation. Eicosapentaenoic acid and docosahexaenoic acid derivatives, particularly resolvin E1 (RvE1) and neuroprotectin D1, appear to be responsible for docosahexaenoic acid's anti-inflammatory effect. This is supported in a study in which topical RvE1 resulted in decreased inflammation in a mouse dry eye model. Topical administration of pigment epithelium-derived factor in combination with docosahexaenoic acid accelerates the regeneration of corneal nerves after their damage during corneal surgery, promoting the return of sensitivity and reducing the signs of dry eye. This combined treatment also reduces objective signs of dry eye, such as rose bengal staining. SUMMARY: No firm recommendations can be made regarding optimal dietary supplementation of essential fatty acids that benefit dry eye patients. On the basis of animal data and preliminary human studies, docosahexaenoic acid and its derivatives appear to be a safe, effective topical treatment for dry eye patients. This may result from their role in the resolution of inflammation and the regeneration of damaged corneal nerves.
Assuntos
Ácidos Docosa-Hexaenoicos/uso terapêutico , Síndromes do Olho Seco/tratamento farmacológico , Ácidos Graxos Ômega-3/uso terapêutico , Animais , Antígenos CD59/uso terapêutico , Suplementos Nutricionais , Modelos Animais de Doenças , Humanos , Camundongos , Resultado do TratamentoRESUMO
BACKGROUND: Influenza is a respiratory disease that seriously threatens human health. In fact, influenza virus itself does not make critical contribution to mortality induced by influenza, but "cytokine storm" produced by the excessive immune response triggered by the virus can result in inflammatory reaction of lung tissues and fatal lung tissue injury, and thus increase influenza mortality. Therefore, besides antiviral drugs, immunosuppression drugs should also be included in infection treatment. PRESENTATION OF THE HYPOTHESIS: Complement is the center of inflammatory reaction. If complement system is over activated, the body will have strong inflammatory reaction or tissue injury, resulting in pathological process. Many studies have proved that, inflammatory injury of lung tissues caused by influenza virus is closely related to complement activation. Therefore, inhibiting complement activation can significantly reduce inflammatory injury in lung tissues. As complement is both a physiological defense and pathological damage medium, systematic inhibition may result in side effects including infection. Therefore, we design targeting complement inhibitors for complement activation sites, i.e. with CR2 as targeting vector, complement inhibitors like CD59 and Crry are targeted to inflammatory sites to specially inhibit the complement activation in local injury, thus local inflammatory reaction is inhibited. TESTING THE HYPOTHESIS: CR2-CD59 and CR2-Crry targeting complement inhibitors are fusion-expressed, and their biological activity is examined via in vivo and in vitro tests. CR2 targeting complement inhibitors are used to treat mouse influenza viral pneumonia model, with PBS treatment group as the control. The survival and lung tissue injury of the mice is observed and the effect of CR2 targeting complement inhibitors on pneumonia induced by influenza virus is evaluated. IMPLICATIONS OF THE HYPOTHESIS: CR2 targeting complement inhibitors are expected to be ideal drugs for viral pneumonia.
Assuntos
Imunossupressores/uso terapêutico , Influenza Humana/imunologia , Influenza Humana/patologia , Pulmão/patologia , Orthomyxoviridae/imunologia , Receptores de Complemento 3d/antagonistas & inibidores , Animais , Produtos Biológicos/genética , Produtos Biológicos/uso terapêutico , Antígenos CD59/genética , Antígenos CD59/uso terapêutico , Humanos , Influenza Humana/tratamento farmacológico , Pulmão/virologia , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Orthomyxoviridae/tratamento farmacológico , Infecções por Orthomyxoviridae/patologia , Receptores de Complemento/genética , Receptores de Complemento/uso terapêutico , Receptores de Complemento 3b , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/uso terapêutico , Análise de SobrevidaRESUMO
Age-related macular degeneration (AMD) is the major reason of blindness of the elderly all over the world. AMD is characterized by a progressive loss of central vision attributable to degenerative and neovascular changes in the macula, the highly specialized region of the retina responsible for sharp and color visual acuity. Degeneration and cell death of retinal pigment epithelial cells (RPE) cause secondarily adverse effects on neural retina leading to visual loss. Most AMD patients cannot benefit from any treatment modalities. The prevalence of AMD is rising as a consequence of the aging of the population. As the RPE cells age, they are subject to continued oxidative stress and this believed to induce inflammation and progression of AMD. Interestingly, many clinical trials have revealed that dietary intakes of omega-3 fatty acids can reduce the risk of both early and late AMD, although their molecular targets in cellular signaling in AMD pathology are not understood. Recently, it has been proposed that the omega-3 fatty acid metabolites, resolvins and protectins, function as endogenous anti-inflammatory compounds. In this review, we propose that resolvins and protectins mediate their beneficial effects by preventing NF-kappaB signaling and this that may represent a new target for regulating the inflammatory responses in AMD.
Assuntos
Envelhecimento/fisiologia , Ácidos Graxos Ômega-3/metabolismo , Degeneração Macular/prevenção & controle , NF-kappa B/antagonistas & inibidores , Antígenos CD59/uso terapêutico , Ácidos Docosa-Hexaenoicos/uso terapêutico , Humanos , Degeneração Macular/genética , NF-kappa B/genética , NF-kappa B/fisiologia , Estresse Oxidativo , Transdução de Sinais/genéticaRESUMO
Asthma pathobiology is remarkable for chronic airway inflammation that fails to spontaneously resolve. No curative therapy is currently available. A growing body of evidence indicates that, in health, inflammation resolution is an active process orchestrated by specific chemical mediators that are elaborated to restore tissue homeostasis. Activated cell membranes release polyunsaturated fatty acids from phospholipids for enzymatic conversion to biologically active mediators with profound regulatory effects on innate and adaptive immunity. Some of these mediators carry anti-inflammatory and pro-resolving actions that are transduced in a cell-type specific manner via specific recognition sites that initiate regulatory intracellular signals, such as presqualene diphosphate remodeling, to limit pro-phlogistic cell activation. Some of these counter-regulatory lipid mediators have been identified in the airway during asthma and defects in their production are associated with disease severity. In this review, we describe the biosynthesis and bioactions of pro-resolving chemical mediators and provide examples of select mediators and their structural analogs with particular relevance to asthma.
Assuntos
Ácido Eicosapentaenoico/análogos & derivados , Lipoxinas/metabolismo , Hipersensibilidade Respiratória/metabolismo , Animais , Biomimética/tendências , Antígenos CD59/metabolismo , Antígenos CD59/uso terapêutico , Dieta , Ácido Eicosapentaenoico/genética , Ácido Eicosapentaenoico/metabolismo , Retroalimentação Fisiológica , Óleos de Peixe/metabolismo , Cobaias , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/isolamento & purificação , Inibidores de Hidroximetilglutaril-CoA Redutases/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Metabolismo dos Lipídeos/imunologia , Lipoxinas/genética , Camundongos , Fosfatos de Poli-Isoprenil/metabolismo , Fosfatos de Poli-Isoprenil/uso terapêutico , Hipersensibilidade Respiratória/dietoterapia , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/imunologia , Transdução de Sinais/imunologiaRESUMO
PURPOSE: Complement-mediated damage to the retinal pigment epithelium (RPE), Bruch membrane, and choroid has been associated with pathogenesis in age-related macular degeneration (AMD). The terminal step of complement activation involves lysis of cells by the insertion of the membrane attack complex (MAC) in the plasma membrane. The hypothesis that local overexpression of human CD59 (hCD59) delivered by an adenovirus (Ad) vector to primary murine RPE cells in vitro, RPE in vivo, or cornea ex vivo protects those cells from human MAC deposition and lysis was tested. METHODS: A humanized model of MAC deposition on murine cells and murine ocular tissues including RPE and cornea was developed to permit testing of human complement regulators in mice. A recombinant adenovirus-expressing hCD59 was generated, and this virus was injected into the subretinal space of adult mice. Subsequently, eyecups from these mice were exposed to human serum, and the levels of MAC deposition on the RPE were quantified. hCD59 was also expressed on murine cornea ex vivo and in murine hepatocytes, and primary RPE cells in vitro and levels of human MAC deposition and cell lysis were measured. RESULTS: Adenovirus-mediated delivery of hCD59 to the RPE, cornea, or cells in culture protects those cells from human MAC deposition and MAC-mediated damage and vesiculation. CONCLUSIONS: The humanized model of MAC deposition on murine ocular tissues allows testing of human complement regulators that may have potential in the treatment of AMD or other diseases associated with complement activation.
Assuntos
Antígenos CD59/uso terapêutico , Complexo de Ataque à Membrana do Sistema Complemento/fisiologia , Degeneração Macular/tratamento farmacológico , Epitélio Pigmentado Ocular/fisiologia , Adenoviridae , Adulto , Envelhecimento , Animais , Antígenos CD59/administração & dosagem , Técnicas de Cultura de Células , Linhagem Celular , Embrião de Mamíferos , Citometria de Fluxo , Vetores Genéticos , Humanos , Degeneração Macular/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Biológicos , Epitélio Pigmentado Ocular/citologiaRESUMO
We have shown that membrane attack complex (MAC) formation via the activation of the alternative pathway plays a central role in the laser-induced choroidal neovascularization (CNV). This study was undertaken to understand the role of a complement regulatory protein, CD59, which controls MAC assembly and function, in this model. CNV was induced by laser photocoagulation in C57BL/6 and Cd59a(-/-) mice using an argon laser. Animals from each group were sacrificed on day 1, 3, 5, and 7 postlaser. Retinal pigment epithelium-choroid-scleral tissue was examined to determine the incidence and size of CNV complex, and semiquantitative RT-PCR and Western blot analysis for CD59a was studied. Recombinant soluble mouse CD59a-IgG2a fusion (rsCD59a-Fc) protein was injected via i.p. or intravitreal routes 24 h before laser. Our results demonstrated that CD59a (both mRNA and protein) was down-regulated during laser-induced CNV. Cd59a(-/-) mice developed CNV complex early in the disease process. Increased MAC deposition was also observed in these Cd59a(-/-) mice. Administration of rsCD59a-Fc inhibited the development of CNV complex in the mouse model by blocking MAC formation and also inhibited expression of angiogenic growth factors. These data provide strong evidence that CD59a plays a crucial role in regulating complement activation and MAC formation essential for the release of growth factors that drive the development of laser-induced CNV in mice. Thus, our results suggest that the inhibition of complement by soluble CD59 may provide a novel therapeutic alternative to current treatment.
Assuntos
Antígenos CD59/fisiologia , Neovascularização de Coroide/etiologia , Degeneração Macular/patologia , Animais , Antígenos CD59/análise , Antígenos CD59/genética , Antígenos CD59/uso terapêutico , Neovascularização de Coroide/tratamento farmacológico , Ativação do Complemento , Complexo de Ataque à Membrana do Sistema Complemento , Modelos Animais de Doenças , Regulação para Baixo , Imunoglobulina G/genética , Peptídeos e Proteínas de Sinalização Intercelular , Lasers/efeitos adversos , Degeneração Macular/etiologia , Degeneração Macular/terapia , Camundongos , Camundongos Knockout , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/farmacologiaRESUMO
Paroxysmal nocturnal hemoglobinuria (PNH) results from the expansion of a hematopoietic clone that is deficient in glycosylphosphatidylinositol-anchored molecules. PNH is characterized by chronic hemolysis with acute exacerbations due to the uncontrolled activity of complement on PNH cells, which lack the inhibitor of homologous complement, CD59. Symptoms include severe fatigue, hemoglobinuria, esophageal spasm, erectile dysfunction, and thrombosis. We report the use of a novel synthetically modified recombinant human CD59, rhCD59-P, a soluble protein that attaches to cell membranes. In vitro treatment of PNH erythrocytes with rhCD59-P resulted in levels of CD59 equivalent to normal erythrocytes and effectively protected erythrocytes from complement-mediated hemolysis. The administration of rhCD59-P to CD1 mice resulted in levels of CD59 on erythrocytes, which protected them from complement-mediated lysis. Thus, rhCD59-P corrects the CD59 deficiency in vitro and can bind to erythrocytes in an in vivo murine model, protecting the cells from the activity of human complement, and represents a potential therapeutic strategy in PNH.
Assuntos
Antígenos CD59/farmacologia , Ativação do Complemento/efeitos dos fármacos , Proteínas do Sistema Complemento/metabolismo , Eritrócitos/metabolismo , Hemoglobinúria Paroxística/metabolismo , Hemólise/efeitos dos fármacos , Animais , Antígenos CD59/uso terapêutico , Eritrócitos/patologia , Hematopoese , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/patologia , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/patologia , Humanos , Camundongos , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêuticoRESUMO
Hemolytic anemia is a major feature of paroxysmal nocturnal hemoglobinuria (PNH). Intravascular red blood cell (RBC) destruction is caused by increased sensitivity of the abnormal erythrocyte to complement-mediated lysis, due to the GPI absence of a membrane-bound glycosylphosphatidylinositol (GPI)-linked protein, which functions as an inhibitor of reactive lysis (CD59). Both in vivo and in vitro models have suggested the feasibility of cell-to-cell transfer of GPI proteins, and patients with hemolysis could potentially benefit from transfer of CD59 to their deficient erythrocytes. We studied the ability of RBC components prepared from outdated packed RBC collections, as well as high-density lipoprotein (HDL) preparations, rich in CD55 and CD59, to promote protein transfer, as assessed by flow cytometry, immunoblotting, and susceptibility to complement-mediated lysis. By flow cytometry, CD55 and CD59 were present on RBC-derived microvesicles that stained with an antiglycophorin antibody Ab; in addition, soluble CD59 and CD55 were detected by immunoblot in soluble fractions eluated from RBC units stored for more than 35 days, but not in fresh blood. Both commercial HDL preparations and those prepared in our laboratory contained CD55 and CD59, as assayed by immunoblot. When RBC that were deficient (GPI)-anchored protein, obtained from five patients, with PNH were incubated with HDL preparations for 2 to 4 hours, there was significant transfer of both proteins to the cell surface, as demonstrated by flow cytometry. Washed RBC microvesicles, prepared by ultrasonification, also mediated transfer of GPI-linked proteins to deficient RBC. Pretreatment of microvesicles, RBC eluate preparations, and HDL with phosphatidylinositol-specific, phospholipase C, abrogated protein transfer to deficient cells, indicating that increased cell-associated CD55 and CD59 levels were related to insertion of the intact GPI moiety, rather than to simple adhesion. PNH RBC that were exposed to HDL, RBC eluate preparations, or microvesicles demonstrated decreased in vitro complement-mediated hemolysis in the Ham test. Transfer of GPI-linked proteins from soluble preparations containing CD55 and CD59 to PNH erythrocytes is feasible and may have clinical utility.
Assuntos
Antígenos CD55/administração & dosagem , Antígenos CD59/administração & dosagem , Sistemas de Liberação de Medicamentos , Eritrócitos/metabolismo , Hemoglobinúria Paroxística/sangue , Antígenos CD55/imunologia , Antígenos CD55/metabolismo , Antígenos CD55/uso terapêutico , Antígenos CD59/imunologia , Antígenos CD59/metabolismo , Antígenos CD59/uso terapêutico , Eritrócitos/imunologia , Eritrócitos/patologia , Citometria de Fluxo , Glicosilfosfatidilinositóis , Hemoglobinúria Paroxística/tratamento farmacológico , Humanos , Lipoproteínas HDLRESUMO
CD59 regulates complement activation cascade at the final step, inhibiting formation of membrane attack complex (MAC). This protein, being anchored to the cell membrane via glycosyl phosphatidyl inositol (GPI), is expressed ubiquitously on cells which are in contact with body fluids containing components. Recently, MAC formation has been reported to play an important role in pathogenesis of inflammatory diseases such as ischemia or autoimmune diseases. In this review, we describe the structure and biological activities of CD59, the pathogenic role of MAC formation, and discuss application of soluble molecules of CD59 for therapeutic use.