RESUMO
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged during the last months of 2019, spreading throughout the world as a highly transmissible infectious illness designated as COVID-19. Vaccines have now appeared, but the challenges in producing sufficient material and distributing them around the world means that effective treatments to limit infection and improve recovery are still urgently needed. This review focuses on the relevance of different glycobiological molecules that could potentially serve as or inspire therapeutic tools during SARS-CoV-2 infection. As such, we highlight the glycobiology of the SARS-CoV-2 infection process, where glycans on viral proteins and on host glycosaminoglycans have critical roles in efficient infection. We also take notice of the glycan-binding proteins involved in the infective capacity of virus and in human defense. In addition, we critically evaluate the glycobiological contribution of candidate drugs for COVID-19 therapy such as glycans for vaccines, anti-glycan antibodies, recombinant lectins, lectin inhibitors, glycosidase inhibitors, polysaccharides, and numerous glycosides, emphasizing some opportunities to repurpose FDA-approved drugs. For the next-generation drugs suggested here, biotechnological engineering of new probes to block the SARS-CoV-2 infection might be based on the essential glycobiological insight on glycosyltransferases, glycans, glycan-binding proteins, and glycosidases related to this pathology.
Assuntos
Antivirais/uso terapêutico , COVID-19/prevenção & controle , Reposicionamento de Medicamentos , Inibidores de Glicosídeo Hidrolases/uso terapêutico , Glicosiltransferases/antagonistas & inibidores , Proteínas Virais/antagonistas & inibidores , Anticorpos Neutralizantes/uso terapêutico , Antivirais/química , COVID-19/epidemiologia , COVID-19/imunologia , COVID-19/virologia , Desenho de Fármacos , Descoberta de Drogas , Expressão Gênica , Glicômica/métodos , Glicosaminoglicanos/química , Glicosaminoglicanos/imunologia , Glicosaminoglicanos/metabolismo , Glicosiltransferases/química , Glicosiltransferases/genética , Glicosiltransferases/imunologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Lectinas/química , Lectinas/imunologia , Lectinas/metabolismo , Polissacarídeos/química , Polissacarídeos/imunologia , Polissacarídeos/metabolismo , SARS-CoV-2/química , SARS-CoV-2/efeitos dos fármacos , SARS-CoV-2/genética , SARS-CoV-2/imunologia , Transdução de Sinais , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/imunologiaRESUMO
Atherogenesis is associated with the early retention of low-density lipoproteins (LDL) in the arterial intima by interaction with glycosaminoglycan (GAG)-side chains of proteoglycans. Retained LDL undergo reactive oxygen species-mediated oxidation. Oxidized LDL trigger oxidative stress (OS) and inflammation, contributing to atherosclerosis development. Recently, we reported the preventive anti-atherogenic properties of the chimeric mouse/human monoclonal antibody (mAb) chP3R99-LALA, which were related to the induction of anti-chondroitin sulfate antibody response able to inhibit chondroitin sulfate dependent LDL-enhanced oxidation. In the present work, we aimed at further investigating the impact of chP3R99-LALA mAb vaccination on progressive atherosclerosis in apolipoprotein E-deficient mice (apoE(-/-)) fed with a high-fat high-cholesterol diet receiving 5 doses (50 µg) of the antibody subcutaneously, when ~5% of the aortic area was covered by lesions. Therapeutic immunization with chP3R99-LALA mAb halted atherosclerotic lesions progression. In addition, aortic OS was modulated, as shown by a significant (p<0.05) reduction of lipid and protein oxidation, preservation of antioxidant enzymes activity and reduced glutathione, together with a decrease of nitric oxide levels. chP3R99-LALA mAb immunization also regulated aortic NF-κB activation, diminishing the proinflammatory IL1-ß and TNF-α gene expression as well as the infiltration of macrophages into the arterial wall. The therapeutic immunization of apoE(-/-) with progressive atheromas and persistent hypercholesterolemia using chP3R99-LALA mAb arrested further development of lesions, accompanied by a decrease of aortic OS and NF-κB-regulated pro-inflammatory cytokine gene expression. These results contribute to broaden the potential use of this anti-GAG antibody-based immunotherapy as a novel approach to target atherosclerosis at different phases of progression.
Assuntos
Anticorpos Monoclonais/farmacologia , Aterosclerose/patologia , Sulfatos de Condroitina/antagonistas & inibidores , Glicosaminoglicanos/antagonistas & inibidores , Vacinação/métodos , Animais , Apolipoproteínas E/deficiência , Sulfatos de Condroitina/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Glicosaminoglicanos/imunologia , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Knockout , Oxirredução , Reação em Cadeia da Polimerase , Proteínas Recombinantes de Fusão/imunologiaRESUMO
Retained low-density lipoproteins (LDL) by arterial glycosaminoglycans (GAG) are more susceptible to reactive oxygen species-mediated oxidation, contributing to oxidative stress and atherosclerosis. Recently, we reported the properties of the chimeric mouse/human monoclonal antibody chP3R99-LALA to bind sulfated GAG, to inhibit LDL-chondroitin sulfate binding, and to avoid LDL oxidation in vitro. Here, we hypothesized that chP3R99-LALA treatment might reduce aortic oxidative stress in a therapeutic setting. Redox biomarkers and serum lipids were determined by spectrophotometric methods. Subcutaneous administration of five doses (100 µg) of chP3R99-LALA, after Lipofundin administration (2 mL/kg/day, i.v.) during 8 days, reduced atherosclerotic lesion development, which was not associated with a serum lipid modulation. In contrast, the treatment with chP3R99-LALA reduced (p < 0.05) malondialdehyde and protein oxidation, induced a restoration of reduced glutathione level, of the superoxide dismutase and catalase activities and of endothelial nitric oxide level. Thus, the antiatherogenic effect of chP3R99-LALA treatment seems to be associated with a reduction of aortic oxidative stress. These results contribute in understanding the molecular mechanisms associated with chP3R99-LALA atheroprotection and support the use of anti-GAG antibody-based immunotherapy as a potential tool to treat the atherosclerosis.
Assuntos
Anticorpos Anti-Idiotípicos/administração & dosagem , Aorta/efeitos dos fármacos , Aterosclerose/imunologia , Glicosaminoglicanos/imunologia , Animais , Aorta/imunologia , Aorta/patologia , Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Modelos Animais de Doenças , Glicosaminoglicanos/metabolismo , Humanos , Lipoproteínas LDL/imunologia , Lipoproteínas LDL/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Coelhos , Espécies Reativas de Oxigênio/metabolismoRESUMO
OBJECTIVE: The pathogenesis of atherosclerosis is associated with the early retention of low-density lipoproteins that are trapped in the extracellular matrix of the arterial intima by interaction with glycosaminoglycan side chains of proteoglycans. Mutant mouse/human chimeric antibodies of the murine monoclonal antibody P3, which react with N-glycolyl-containing gangliosides and sulfated glycosaminoglycans, were tested for their potentially antiatherogenic properties through the induction of an idiotypic antibody network that may specifically interfere with the binding of low-density lipoproteins to proteoglycan side chains, low-density lipoprotein modification, and foam cell formation. METHODS AND RESULTS: Apolipoprotein E-deficient mice fed a high-fat, high-cholesterol diet received 5 to 6 doses of chP3R99 or chP3S98 mutant antibodies, showing high and low reactivity, respectively, against their respective antigens. Both chimeric antibodies elicited an immunodominant anti-idiotypic response in the absence of adjuvant. A striking (40%-43%) reduction (P<0.01) in total lesion areas was observed in 18-week-old mice immunized with chP3R99, but not chP3S98, compared with PBS-treated mice. The antiatherosclerotic effect was associated with increased mice sera reactivity against heparin and sulfated glycosaminoglycans, including chondroitin and dermatan sulfate. In addition, purified IgG from chP3R99-immunized mice blocked the retention of apolipoprotein B-containing lipoproteins within the arterial wall of apolipoprotein E(-/-) mice. CONCLUSIONS: The present study supports use of active immunization and the mounting of an idiotypic antibody network response against glycosaminoglycans as a novel approach to target atherosclerosis.
Assuntos
Anticorpos Anti-Idiotípicos/uso terapêutico , Apolipoproteínas E/deficiência , Aterosclerose/prevenção & controle , Glicosaminoglicanos/imunologia , Glicosaminoglicanos/metabolismo , Sulfatos/metabolismo , Animais , Anticorpos Anti-Idiotípicos/imunologia , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Artérias/metabolismo , Aterosclerose/etiologia , Aterosclerose/metabolismo , Colesterol na Dieta/efeitos adversos , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Humanos , Lipoproteínas LDL/metabolismo , Masculino , Camundongos , Camundongos Knockout , Proteínas Mutantes Quiméricas/imunologia , Proteínas Mutantes Quiméricas/uso terapêuticoRESUMO
OBJECTIVE: Subendothelial retention of proatherogenic lipoproteins by proteoglycans is critical in atherosclerosis. The aim of this study was to characterize the recognition and antiatherogenic properties of a chimeric monoclonal antibody (mAb) that reacts with sulfated molecules. METHODS AND RESULTS: chP3R99 mAb recognized sulfated glycosaminoglycans, mainly chondroitin sulfate (CS), by ELISA. This mAb blocked ≈70% of low-density lipoprotein (LDL)-CS association and ≈80% of LDL oxidation in vitro, and when intravenously injected to Sprague-Dawley rats (n=6, 1 mg/animal), it inhibited LDL (4 mg/kg intraperitoneally, 1 hour later) retention and oxidation in the artery wall. Moreover, subcutaneous immunization of New Zealand White rabbits (n=19) with chP3R99 mAb (100 µg, 3 doses at weekly intervals) prevented Lipofundin-induced atherosclerosis (2 mL/kg, 8 days) with a 22-fold reduction in the intima-media ratio (P<0.01). Histopathologic and ultrastructural studies showed no intimal alterations or slight thickening, with preserved junctions between endothelial cells and scarce collagen fibers and glycosaminoglycans. In addition, immunization with chP3R99 mAb suppressed macrophage infiltration in aorta and preserved redox status. The atheroprotective effect was associated with the induction of anti-CS antibodies in chP3R99-immunized rabbits, capable of blocking CS-LDL binding and LDL oxidation. CONCLUSION: These results support the use of anti-sulfated glycosaminoglycan antibody-based immunotherapy as a potential tool to prevent atherosclerosis.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Aterosclerose/prevenção & controle , Sulfatos de Condroitina/antagonistas & inibidores , Glicosaminoglicanos/antagonistas & inibidores , Imunização , Animais , Especificidade de Anticorpos , Aterosclerose/induzido quimicamente , Aterosclerose/imunologia , Aterosclerose/metabolismo , Aterosclerose/patologia , Transporte Biológico , Linhagem Celular , Sulfatos de Condroitina/imunologia , Modelos Animais de Doenças , Combinação de Medicamentos , Ensaio de Imunoadsorção Enzimática , Células Espumosas/imunologia , Células Espumosas/metabolismo , Glicosaminoglicanos/imunologia , Lipoproteínas LDL/metabolismo , Camundongos , Oxirredução , Estresse Oxidativo , Fosfolipídeos , Coelhos , Ratos , Ratos Sprague-Dawley , SorbitolRESUMO
We conducted a study in the southern mountains of the Mexican State of Oaxaca that consisted of isolation of wild Sporothrix schenckii strains obtained from soil samples and investigation of positive reactors to skin test reaction with sprotrichin antigen. The study was conducted by means of recollection of soil samples and processing of these with dilution methods and fungal isolation in ordinary culture media Sabouraud simple Agar with and without antibiotics (SS, SA). Suspected strains underwent dimorphism, melanin formation, and virulence confirmation tests. Investigation of positive reactors to sporotrichin Y (yeast) was also conducted. Three supposed strains were identified due to their reproductive characteristics, melanin production, and virulence. In the community, 144 individuals were studied, of whom 6.25% were positive to sporotichin. Isolation of virulent strains of Sporothrix schenkii from nature (soil) and primoinfection of a percentage of the studied population were confirmed.
Assuntos
Glicosaminoglicanos/imunologia , Sporothrix/isolamento & purificação , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Se llevó a cabo un estudio en la sierra sur de Oaxaca de aislamiento de cepas silvestres de Sporothrix schenckii a partir de muestras de tierra, así como investigación de reactores positivos a la intradermorreacción con Esporotricina. El estudio se llevó a cabo a partir de la recolección de muestras de tierra y su posterior procesamiento mediante métodos de dilución y aislamiento del hongo en medios habituales de cultivo de Sabouraud simple y Sabouraud con antibióticos (SS y SA). Las cepas presuntivas se sometieron apruebas de dimorfismo, formación de melanina y comprobación de virulencia en animales. Se investigaron los reactores positivos a la Esporotricina L (Levaduriforme). Se identificaron tres cepas presuntivas por sus características reproductivas, formación demelanina y capacidad virulenta. En la comunidad se investigaron a 144 individuos dando positivos a la Esporotricina L el 6.25%. Se comprueba el aislamiento de cepas virulentas de S. schenckii a partir de la naturaleza (suelo) y se comprueba primocontacto con éste en un porcentaje de la población estudiada.
We conducted a study in the southern mountains of the Mexican State of Oaxaca that consisted of isolation of wild Sporothrix schenckii strains obtained from soil samples and investigation of positive reactors to skin test reaction with sprotrichin antigen. The study was conducted by means of recollection of soil samples and processing of these with dilution methods and fungal isolation in ordinary culture media Sabouraud simple Agar with and without antibiotics (SS, SA). Suspected strains underwent dimorfism, melanin formation, and virulence confirmation tests. Investigation of positive reactors to sporotrichin Y (yeast) was also conducted. Three supposed strains were identified due to their reproductive characteristics, melaninproduction, and virulence. In the community; 144 individuals were studied, ofwhom6.25%werepositivetosporotichin. Isolation of virulent strains o/Sporotrhix schenkii from nature (soil) andprimoinfection of a percentage of the studied population were confirmed.
Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Glicosaminoglicanos/imunologia , Sporothrix/isolamento & purificaçãoRESUMO
Since Paracoccidioides brasiliensis and Histoplasma capsulatum are known to be present in similar environments, there have been many epidemiologic investigations regarding the prevalences of these two organisms. However, cross-reactivity can occur in paracoccidioidin and histoplasmin skin tests, and this usually results in the overestimation of the prevalence of P. brasiliensis. The prevalence of infection with P. brasiliensis was evaluated in a cross-sectional study of 298 asymptomatic school children in the Brazilian Amazon region (Mato Grosso State). In this investigation, the reactivity of children to two different P. brasiliensis antigen preparations, paracoccidioidin and a purified 43-kD glycoprotein (gp43), was compared with or without the co-administration of histoplasmin. In the group of individuals receiving paracoccidioidin who had a positive histoplasmin skin test result, the prevalence of exposure to P. brasiliensis was 44% (16 of 36). This reactivity to P. brasiliensis was significantly higher than that observed in other groups, which ranged from 4% to 6% (P < 5 x 10(-4) for each). Overall prevalence was 4.6% (95% confidence interval = 2.5-7.7%). These data suggest that gp43 provides a better estimate of exposure to P. brasiliensis when the co-administration of histoplasmin is desired.
Assuntos
Antígenos de Fungos/análise , Proteínas Fúngicas , Glicosaminoglicanos/análise , Histoplasmina/análise , Paracoccidioides/imunologia , Paracoccidioidomicose/epidemiologia , Adolescente , Brasil/epidemiologia , Criança , Feminino , Proteínas Fúngicas/imunologia , Glicoproteínas/imunologia , Glicosaminoglicanos/imunologia , Histoplasmina/imunologia , Humanos , Testes Intradérmicos , Masculino , Paracoccidioidomicose/diagnóstico , Paracoccidioidomicose/imunologia , Prevalência , Testes CutâneosRESUMO
Antisera were raised in rabbits against acetone-dried yeast-like and mycelium forms of Sporothrix schenckii. These antisera were tested for immunoprecipitation of peptidorhamnomannans isolated from both cell types. Both antisera reacted strongly with S.schenckii peptidorhamnomannans, but the reactions were weak with beta-eliminated peptidopolysaccharides. These antisera did not recognize the Saccharomyces cerevisiae mannoprotein, and reacted poorly with Ceratocystis (Ophiostoma) stenoceras cell wall glycopeptides. Since beta-eliminated glycopeptides were poorly reactive, we investigated the activity of O-glycosidically linked oligosaccharides which were liberated from the peptidorhamnomannans by mild alkaline hydrolysis, using a hapten inhibition test. The rates of inhibition showed that the immunodominant epitopes in O-linked tetra- and pentasaccharides had the following novel structures: alpha-L-Rhap (1-->4)-alpha-D-GlcpA and alpha-L-Rhap(1-->4)-[alpha-L-Rhap(1-->2)]alpha-D-GlcpA . Results suggest that peptidorhamnomannans or synthetic antigens bearing these epitopes should be used for the specific detection of anti-S.schenckii antibodies rather than the long-chain N-linked polysaccharides.
Assuntos
Antígenos de Fungos/análise , Epitopos/análise , Glicosaminoglicanos/imunologia , Sporothrix/imunologia , Especificidade de Anticorpos , Antígenos de Fungos/química , Configuração de Carboidratos , Sequência de Carboidratos , Epitopos/química , Glicosaminoglicanos/química , Haptenos/farmacologia , Humanos , Soros Imunes/imunologia , Técnicas de Imunoadsorção , Dados de Sequência Molecular , Oligossacarídeos/antagonistas & inibidores , Oligossacarídeos/química , Oligossacarídeos/imunologia , Sporothrix/químicaRESUMO
Sporotrichosis and histoplasmosis are deep mycosis with a high incidence in human beings in Brazil. In domestic animals histoplasmosis has been described only in dogs, but the occurrence of sporotrichosis among domestic animals in Brazil has been described in dogs, cats, mules and asses. There is also a case of this disease reported in a chimpanzee (Pan troglodites). The purpose of this research was to perform an epidomiological study of these mycoses using delayed hypersensitivity tests (histoplasmin and sporotrichin) in Latin American wild mammals. This research was assayed using 96 healthy animals at Parque Zoológico de São Paulo, Brazil: Primates: 33 Cebus apella--weeping-capuchin and 16 Callithrix jacchus--marmoset; Procyonidae: 37 Nasua nasua--coatimundi and 10 Felidae (Panthera onca--jaguar; Felis pardalis--ocelot Felis wiedii--margay; Felis tigrina--wild cat). For intradermic tests, the following antigens were used: Sporothrix schenkii cell suspension (sporotrichin, histoplasmin-filtrate), Histoplasma capsulatum cell suspension (histoplasmin), and Histoplasma capsulatum (polysaccharide). The positivity to histoplasmin was 44.79% (Cebidae 15.15%; Callithricidae 6.25%; Procyonidae 86.49% and Felidae 50.00%, respectively). With respect to sporotrichin, 30.21% (Cebidae 6.06%, Callithricidae 0.0%; Procyonidae 64.86% and Felidae 30.00% respectively). The pattern of infection is similar to that shown by human beings and this may suggest that these animals could be involved in the epidemiologic chain of sporotrichosis and histoplasmosis, the second most prevalent human deep mycoses in Brazil. It is important to point out the absence of similar studies in Latin American wild animals.