RESUMO
During the last few decades, several efforts have been made towards developing biocompatible materials. Among them, peptide amphiphiles (PAs) constitute a novel nanotechnological strategy used in the field of biomedicine since they can provide tissue- specific binding and localization. PAs possess several regions combining hydrophobic and hydrophilic areas that are able to self-assemble in aqueous media, forming different tertiary nanostructures able to interact with cellular membranes. Moreover, these molecules can be tuned by incorporating collagen, lipids, or fluorescent markers. In addition, they can also be used as carriers in order to encapsulate active compounds for drug delivery showing promising features in this area. In this review, the self-assembled structures of PAs as well as their pharmacological applications have been summarized. Furthermore, their use as drug delivery systems has been highlighted and the latest advances in this field have been reviewed.
Assuntos
Nanoestruturas , Peptídeos , Humanos , Preparações Farmacêuticas , Peptídeos/farmacologia , Peptídeos/química , Nanoestruturas/química , Sistemas de Liberação de Medicamentos , Materiais BiocompatíveisRESUMO
BACKGROUND: Autoantibodies are critical elements in RA pathogenesis and clinical assessment. The anti-malondialdehyde-acetaldehyde (anti-MAA) antibodies are potentially useful because of their claimed high sensitivity for all RA patients, including those lacking RF and anti-CCP antibodies. Therefore, we aimed to replicate these findings. METHODS: We independently attempted replication in Santiago and Barcelona using sera from 517 and 178 RA patients and 272 and 120 healthy controls, respectively. ELISA protocols for anti-MAA antibodies included five antigens (human serum albumin in three formulations, fibrinogen, and a synthetic peptide) and assays for the IgG, IgM, and IgA isotypes. We integrated our results with information found by searching the Web of Science for reports of anti-MAA antibodies in RA. The available patients (4989 in 11 sets) were included in a meta-analysis aimed at heterogeneity between studies. Factors accounting for heterogeneity were assessed with meta-regression. RESULTS: The sensitivity of anti-MAA antibodies in our RA patients was low, even in seropositive patients, with the percentage of positives below 23% for all ELISA conditions. Our results and bibliographic research showed IgG anti-MAA positive patients ranging from 6 to 92%. The extreme between-studies heterogeneity could be explained (up to 43%) in univariate analysis by sex, African ethnicity, the site of study, or recruitment from the military. The best model, including African ancestry and smoking, explained a high heterogeneity fraction (74%). CONCLUSION: Anti-MAA antibody sensitivity is extremely variable between RA patient collections. A substantial fraction of this variability cannot be attributed to ELISA protocols. On the contrary, heterogeneity is determined by complex factors that include African ethnicity, smoking, and sex.
Assuntos
Acetaldeído , Artrite Reumatoide , Humanos , Malondialdeído , Autoanticorpos , Imunoglobulina G , Fator Reumatoide , Peptídeos CíclicosRESUMO
Background: Corneal neovascularization is a sight-threatening disease. It can be treated using antiangiogenic and anti-inflammatory compounds. Therefore, atorvastatin (ATV) constitutes a suitable candidate to be administered topically. To attain suitable efficacy, ATV can be encapsulated into custom-developed nanocarriers such as peptide amphiphiles. Methods: Three peptide amphiphiles bearing one, two or four C16-alkyl groups (mC16-Tat47-57, dC16-Tat47-57 and qC16-Tat47-57) were synthesized, characterized and loaded with ATV. Drug release and ocular tolerance were assessed as well as anti-inflammatory and antiangiogenic properties. Results: ATV-qC16-Tat47-57 showed higher encapsulation efficiency than mC16-Tat47-57 and dC16-Tat47-57 and more defined nanostructures. ATV-qC16-Tat47-57 showed ATV prolonged release with suitable ocular tolerance. Moreover, ATV-qC16-Tat47-57 was antiangiogenic and prevented ocular inflammation. Conclusion: ATV-qC16-Tat47-57 constitutes a promising topical medication against corneal neovascularization.
Corneal neovascularization is an eye disease that affects over 1 million people every year and can lead to blindness. It is caused by inflammation and the unwanted formation of blood vessels in the eye. Current treatments for this disease are not fully effective. Atorvastatin (ATV) is one drug that has been partially successful at treating corneal neovascularization, but it does not stay in the eye long enough and does not mix well with the water-based environment of the eye. To overcome this, ATV was combined with three specially designed nanocarriers. These nanocarriers were peptides, short stretches of protein. They were designed to be amphiphilic, meaning that one section is hydrophilic (literally meaning 'water loving') and one section is hydrophobic ('water hating'). These peptide nanocarriers allowed ATV to stay in the water-based environment of the eye longer. The peptide with the most hydrophobic chains (qC16-Tat47-57) was able to carry more ATV than the other peptides and produced particles of a desired shape. ATV-qC16-Tat47-57 nanocarriers were found to release slowly. These nanocarriers were also found to prevent the development of new blood vessels on a membrane in a hen's egg used to mimic the eye. There was also no sign of irritation on this membrane or in the eyes of New Zealand rabbits. These results show ATV-qC16-Tat47-57 has a prolonged therapeutic effect, prevents the formation of new blood vessels and is tolerated in the eye. ATV-qC16-Tat47-57 is therefore potentially a more effective alternative to ATV treatment alone.
Assuntos
Neovascularização da Córnea , Humanos , Neovascularização da Córnea/tratamento farmacológico , Atorvastatina/farmacologia , Atorvastatina/uso terapêutico , Olho , Liberação Controlada de Fármacos , PeptídeosRESUMO
Amyloid ß-peptide (Aß) misfolding into ß-sheet structures triggers neurotoxicity inducing Alzheimer's disease (AD). Molecules able to reduce or to impair Aß aggregation are highly relevant as possible AD treatments since they should protect against Aß neurotoxicity. We have studied the effects of the interaction of dynorphins, a family of opioid neuropeptides, with Aß40 the most abundant species of Aß. Biophysical measurements indicate that Aß40 interacts with Big Dynorphin (BigDyn), lowering the amount of hydrophobic aggregates, and slowing down the aggregation kinetics. As expected, we found that BigDyn protects against Aß40 aggregates when studied in human neuroblastoma cells by cell survival assays. The cross-interaction between BigDyn and Aß40 provides insight into the mechanism of amyloid pathophysiology and may open up new therapy possibilities.
RESUMO
Rheumatoid arthritis (RA) is characterized by the presence of autoantibodies that are of paramount importance for the diagnosis and prognosis of the disease and have been implicated in its pathogenesis. Proteins resulting from post-translational modifications (PTMs) are capable of triggering autoimmune responses important for the development of RA. In this work, we investigate serum antibody reactivity in patients with an established RA against a panel of chimeric peptides derived from fibrin and filaggrin proteins and bearing from one to three PTMs (citrullination, carbamylation and acetylation) by home-designed ELISA tests (anti-AMPA autoantibodies). The role of anti-AMPAs as biomarkers linked to the presence of a more severe RA phenotype (erosive disease with radiological structural damage) and to the presence of interstitial lung disease (ILD), a severe extra-articular manifestation in RA patients entailing a high mortality, was also analyzed. In general, the association with the clinical phenotype of RA was confirmed with the different autoantibodies, and especially for IgA and IgM isotypes. The prevalence of severe joint damage was only statistically significant for the IgG isotype when working with the peptide bearing three PTMs. Furthermore, the median titers were significantly higher in patients with RA-ILD, a finding not observed for the IgG isotype when working with the single- and double-modified peptides.
Assuntos
Artrite Reumatoide/metabolismo , Autoanticorpos/sangue , Peptídeos/imunologia , Processamento de Proteína Pós-Traducional , Acetilação , Artrite Reumatoide/sangue , Artrite Reumatoide/complicações , Artrite Reumatoide/imunologia , Biomarcadores/sangue , Citrulinação , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Humanos , Doenças Pulmonares Intersticiais/complicações , Peptídeos/metabolismo , Carbamilação de ProteínasRESUMO
In a previous work, we defined a novel HIV-1 fusion inhibitor peptide (E1P47) with a broad spectrum of activity against viruses from different clades, subtypes, and tropisms. With the aim to enhance its efficacy, in the present work we address the design and synthesis of several peptide amphiphiles (PAs) based on the E1P47 peptide sequence to target the lipid rafts of the cell membrane where the cell-cell fusion process takes place. We report the synthesis of novel PAs having a hydrophobic moiety covalently attached to the peptide sequence through a hydrophilic spacer of polyethylene glycol. Characterization of self-assembly in condensed phase and aqueous solution as well as their interaction with model membranes was analyzed by several biophysical methods. Our results demonstrated that the length of the spacer of polyethylene glycol, the position of the peptide conjugation as well as the type of the hydrophobic residue determine the antiviral activity of the construct. Peptide amphiphiles with one alkyl tail either in C-terminus (C-PAmonoalkyl) or in N-terminus (N-PAmonoalkyl) showed the highest anti-HIV-1 activities in the cellular model of TZM-bl cells or in a preclinical model of the human mucosal tissue explants.
Assuntos
HIV-1 , Peptídeos , Interações Hidrofóbicas e Hidrofílicas , NanofibrasRESUMO
BACKGROUND: Blocking of the Tumor Necrosis Factor (TNF) activity is a successful therapeutic approach for 50-60% of rheumatoid arthritis (RA) patients. However, there are yet no biomarkers to stratify patients for anti-TNF therapy. Rheumatoid factor (RF) and anti-cyclic-citrullinated antibodies (anti-CCP) have been evaluated as biomarkers of response but the results have shown limited consistency. Anti-carbamylated protein (anti-CarP) and anti-peptidylarginine deiminase type 4 (anti-PAD4) antibodies have been much less studied. Despite being linked to common immune processes, the interaction between these markers has not been evaluated yet. Our aim was to analyze the interaction between these four antibodies in relation to the response to anti-TNF therapy. METHODS: For this objective, a prospective cohort of n = 80 RA patients starting anti-TNF therapy was recruited. Serum determinations at baseline were performed for RF, anti-CCP, anti-CarP and anti-PAD4 antibodies using enzyme-linked immunosorbent assays (ELISA). The clinical response to anti-TNF therapy was determined at week 12 using the change in DAS28 score. Association was performed using multivariate linear regression adjusting for baseline DAS28, sex and age. RESULTS: The interaction between pairs of antibodies was tested by the addition of an interaction term. We found two highly significant antibody interactions associated with treatment response: anti-CarP with anti-PAD4 (p = 0.0062), and anti-CCP with RF (p = 0.00068). The latter antibody interaction was replicated in an independent retrospective cohort of RA patients (n = 199, p = 0.04). CONCLUSIONS: The results of this study suggest that antibody interaction effects are important factors in the response to anti-TNF therapy in RA.
Assuntos
Artrite Reumatoide , Autoanticorpos , Inibidores do Fator de Necrose Tumoral/uso terapêutico , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/tratamento farmacológico , Humanos , Peptídeos Cíclicos , Estudos Prospectivos , Estudos Retrospectivos , Fator ReumatoideRESUMO
BACKGROUND: Vaginal drug delivery offers an opportunity for effective microbicide treatments. However, the physiological characteristics of the vagina represent a challenge when preparing microbicide formulations for local delivery that avoid systemic effects. OBJECTIVE: To review recent advances in vaginal microbicides placing special emphasis on nanocarriers that encapsulate peptide microbicides. The review includes an overview of local treatment and prevention strategies for bactericidal, fungal, protozoal and viral vaginal infections that use peptide microbicides. METHODS: Scientific literature and clinical trials published in the PubMed database since 2015 are analyzed. RESULT: Peptide microbicides are used for vaginal drug delivery as a means of prevention and treatment of several infectious diseases, especially sexually transmitted infections, and it is clear that such peptide-based microbicides constitute a suitable strategy. Their combination with nanotechnological drug delivery systems improves local peptide administration while avoiding their degradation and adverse effects. To date, only a few polymeric and lipid-based nanocarriers have been used for peptide microbicide vaginal delivery. CONCLUSION: The combination of nanotechnology and peptide microbicides has emerged over recent years, and different platforms are being used for vaginal peptide delivery. Nanostructured systems for peptide delivery via vaginal rings or hydrogels are increasingly used as platforms capable of providing suitable drug delivery. However, there is still a lack of knowledge about nanocarriers aimed at encapsulating peptides for local microbicide delivery.
Assuntos
Anti-Infecciosos , Infecções por HIV , Administração Intravaginal , Anti-Infecciosos/uso terapêutico , Parto Obstétrico , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/prevenção & controle , Humanos , Nanotecnologia , Peptídeos , GravidezRESUMO
Based on the structure of an HIV-1 entry inhibitor peptide two stapled- and a retro-enantio peptides have been designed to provide novel prevention interventions against HIV transmission. The three peptides show greater inhibitory potencies in cellular and mucosal tissue pre-clinical models than the parent sequence and the retro-enantio shows a strengthened proteolytic stability. Since HIV-1 fusion inhibitor peptides need to be embedded in the membrane to properly interact with their viral target, the structural features were determined by NMR spectroscopy in micelles and solved by using restrained molecular dynamics calculations. Both parent and retro-enantio peptides demonstrate a topology compatible with a shared helix-turn-helix conformation and assemble similarly in the membrane maintaining the active conformation needed for its interaction with the viral target site. This study represents a straightforward approach to design new targeted peptides as HIV-1 fusion inhibitors and lead us to define a retro-enantio peptide as a good candidate for pre-exposure prophylaxis against HIV-1.
Assuntos
Fármacos Anti-HIV/química , HIV-1/efeitos dos fármacos , Oligopeptídeos/química , Proteínas Estruturais Virais/antagonistas & inibidores , Fármacos Anti-HIV/farmacologia , HIV-1/química , Humanos , Simulação de Acoplamento Molecular , Oligopeptídeos/farmacologia , Ligação Proteica , Proteínas Estruturais Virais/química , Proteínas Estruturais Virais/metabolismoRESUMO
OBJECTIVE: To analyse the association between anti-carbamylated protein antibodies (Anti-CarP) and interstitial lung disease (ILD) in rheumatoid arthritis (RA) patients. METHODS: Cross-sectional study including RA patients fulfilling the 2010 ACR/EULAR criteria. The main population comprised two groups: (1) RA patients diagnosed with RA-ILD (RA-ILD group); (2) RA patients without ILD (non-ILD RA group). Non-ILD RA patients in whom ILD was suspected underwent a diagnostic work-up and, if ILD was diagnosed, were switched to the RA-ILD group. ILD was diagnosed by high-resolution computed tomography and confirmed by a multidisciplinary committee. An independent replication sample was also obtained. Three Anti-CarP IgG autoantibodies against fetal calf serum (Anti-FCS), fibrinogen (Anti-Fib) and chimeric fibrine/filagrine homocitrullinated peptide (Anti-CFFHP) and one Anti-CarP IgA against FCS (Anti-FCS-IgA) were determined by home-made ELISA. Associations between Anti-CarP and ILD were analysed using multivariable logistic regression adjusted by smoking, sex, age, RA disease duration, rheumatoid factor and anticitrullinated protein antibodies. RESULTS: We enrolled 179 patients: 37 (21%) were finally diagnosed with RA-ILD. Anti-CarP specificities were more frequent in RA-ILD patients (Anti-FCS 70% vs 43%; Anti-Fib 73% vs 51%; Anti-CFFHP 38% vs 19%; Anti-CarP-IgA 51% vs 20%, p<0.05 for all comparisons). Serum titers of Anti-CarP were significantly higher in RA-ILD patients. Anti-CarP specificities showed a robust effect towards increasing the odds of ILD in the multivariate analysis (Anti-FCS (OR: 3.42; 95% CI: 1.13 to 10.40), Anti-Fib (OR: 2.85; 95% CI: 0.83 to 9.70), Anti-CFFHP (OR: 3.11; 95% CI: 1.06 to 9.14) and Anti-FCS-IgA (OR: 4.30; 95% CI: 1.41 to 13.04)). Similar findings were observed in the replication sample. CONCLUSIONS: Anti-CarP were strongly associated with ILD. The role of homocitrullination in RA-ILD merits further investigation.
Assuntos
Artrite Reumatoide/epidemiologia , Autoanticorpos/sangue , Doenças Pulmonares Intersticiais/epidemiologia , Doenças Pulmonares Intersticiais/imunologia , Peptídeos Cíclicos/imunologia , Adulto , Idoso , Anticorpos Anti-Idiotípicos/sangue , Artrite Reumatoide/imunologia , Comorbidade , Intervalos de Confiança , Estudos Transversais , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Incidência , Modelos Logísticos , Doenças Pulmonares Intersticiais/diagnóstico , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Valores de Referência , Medição de Risco , Índice de Gravidade de Doença , Análise de SobrevidaRESUMO
OBJECTIVE: To analyze the prevalence of preexisting palindromic rheumatism (PR) in patients with established rheumatoid arthritis (RA) and to evaluate whether these patients have a distinctive clinical and serological phenotype. METHODS: Cross-sectional study in patients with established RA. Preexisting PR was determined using a structured protocol and confirmed by retrospective review of medical records. Demographic, clinical, radiological, immunological, and therapeutic features were compared in patients with and without PR. RESULTS: Included were 158 patients with established RA (78% female) with a mean disease duration since RA onset of 5.1 ± 2.7 years. Preexisting PR was recorded in 29 patients (18%). The median time from the onset of PR to progression to RA was 1.2 years. No between-group differences in demographic features, current disease activity, radiographic erosive disease, or disability were observed. Patients with PR had a higher prevalence of smoking (72% vs 40%). Positive rheumatoid factor, anticitrullinated peptide antibodies, and anticarbamylated protein antibodies were numerically higher in patients with PR. No differences in treatment were observed except for greater hydroxychloroquine (HCQ) use in patients with PR (38% vs 6%). Palindromic flares persisted in a significant proportion of patients during the RA course, including patients in clinical remission or receiving biological disease-modifying antirheumatic drugs. CONCLUSION: Eighteen percent of patients with RA had a history compatible with PR previous to RA onset. No specific clinical or serological phenotype was identified in these patients, although higher HCQ use and smoking prevalence were identified. Palindromic flares may persist during the RA disease course despite treatment.
Assuntos
Antirreumáticos , Artrite Reumatoide , Artrite Reumatoide/diagnóstico , Estudos Transversais , Feminino , Humanos , Masculino , Fenótipo , Estudos RetrospectivosRESUMO
Autosomal dominantly inherited genetic disorders such as corneal dystrophies are amenable to allele-specific gene silencing with small interfering RNA (siRNA). siRNA delivered to the cornea by injection, although effective, is not suitable for a frequent long-term treatment regimen, whereas topical delivery of siRNA to the cornea is hampered by the eye surface's protective mechanisms. Herein we describe an attractive and innovative alternative for topical application using cell-penetrating peptide derivatives capable of complexing siRNA non-covalently and delivering them into the cornea. Through a rational design approach, we modified derivatives of a cell-penetrating peptide, peptide for ocular delivery (POD), already proved to diffuse into the corneal layers. These POD derivatives were able to form siRNA-peptide complexes (polyplexes) of size and ζ-potential similar to those reported able to undergo cellular internalization. Successful cytoplasmic release and gene silencing in vitro was obtained when an endosomal disruptor, chloroquine, was added. A palmitoylated-POD, displaying the best delivery properties, was covalently functionalized with trifluoromethylquinoline, an analog of chloroquine. This modified POD, named trifluoromethylquinoline-palmitoyl-POD (QN-Palm-POD), when complexed with siRNA and topically applied to the eye in vivo, resulted in up to 30% knockdown of luciferase reporter gene expression in the corneal epithelium. The methods developed within represent a valid standardized approach that is ideal for screening of a range of delivery formulations.
RESUMO
Anti-citrullinated peptide/protein antibodies (ACPAs) are the most specific serological biomarkers for rheumatoid arthritis (RA). They have both diagnostic and prognostic value, and are related to more aggressive joint disease in RA. However, a single biomarker cannot differentiate RA subtypes. So, simultaneous analysis of target citrullinated peptides, using a multiplex array based on chimeric peptides composed of several domains of human proteins, could be useful. In this work, eight chimeric peptides and the corresponding native arginine-containing control peptides were obtained by solid-phase peptide synthesis. The study included RA and psoriatic arthritis (PsA) patients attending the Rheumatology Unit of the Hospital Clinic in Barcelona, as well as healthy blood donors (BD) at the same hospital. Our main aim was to explore the diagnostic value of the novel multiplex array compared to a commercial ELISA-based ACPA assay in a serum-saving way. Using the combination of the eight chimeric peptide antigens in the multiplex array, 61.4% of the RA cohort were positive for 3 or more peptides; while, the healthy BD and PsA cohorts did not show any reactivity with the tested peptides. These results indicate that we have developed a highly specific multiplex assay based of chimeric citrullinated peptides derived from filaggrin, fibrin, vimentin and human enolase proteins for the detection of ACPAs in a serum-saving way.
Assuntos
Artrite Reumatoide/diagnóstico , Ensaio de Imunoadsorção Enzimática , Peptídeos Cíclicos/química , Proteínas Recombinantes de Fusão/química , Sequência de Aminoácidos , Anticorpos Antiproteína Citrulinada/sangue , Anticorpos Antiproteína Citrulinada/imunologia , Artrite Reumatoide/sangue , Artrite Reumatoide/imunologia , Proteínas Filagrinas , Humanos , Peptídeos Cíclicos/imunologia , Proteínas Recombinantes de Fusão/imunologiaRESUMO
Novel strategies in the design of HIV-1 fusion/entry inhibitors are based on the construction of dual-targeting fusion proteins and peptides with synergistic antiviral effects. In this work we describe the design of dual-targeting peptides composed of peptide domains of E2 and E1 envelope proteins from Human Pegivirus with the aim of targeting both the loop region and the fusion peptide domains of HIV-1 gp41. In a previous work, we described the inhibitory role of a highly conserved fragment of the E1 protein (domain 139-156) which interacts with the HIV-1 fusion peptide at the membrane level. Here, two different dual-targeting peptides, where this E1 peptide is located on the N- or the C-terminus respectively, have been chemically synthesized and their antiviral activities have been evaluated with HIV pseudotyped viruses from different clades. The study of the functional behaviour of peptides in a membranous environment attending to the peptide recognition of the target sites on gp41, the peptide conformation as well as the peptide affinity to the membrane, demonstrate that antiviral activity of the dual-targeting peptides is directly related to the peptide affinity and its subsequent assembly into the model membrane. The overall results point out to the necessity that fusion inhibitor peptides that specifically interfere with the N-terminal region of gp41 are embedded within the membrane in order to properly interact with their viral target.
Assuntos
Membrana Celular/metabolismo , Inibidores da Fusão de HIV/farmacologia , HIV-1/efeitos dos fármacos , Peptídeos/metabolismo , Peptídeos/farmacologia , Sequência de Aminoácidos , Proteína gp41 do Envelope de HIV/química , Proteína gp41 do Envelope de HIV/metabolismo , Humanos , Micelas , Peptídeos/química , Domínios Proteicos , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Fluorescência , Espectroscopia de Infravermelho com Transformada de Fourier , Triptofano/metabolismoRESUMO
New therapeutic alternatives to fight against the spread of HIV-1 are based on peptides designed to inhibit the early steps of HIV-1 fusion in target cells. However, drawbacks, such as bioavailability, short half-life, rapid clearance, and poor ability to cross the physiological barriers, make such peptides unattractive for the pharmaceutical industry. Here we developed, optimized, and characterized polymeric nanoparticles (NPs) coated with glycol chitosan to incorporate and release an HIV-1 fusion inhibitor peptide (E1) inside the vaginal mucosa. The NPs were prepared by a modified double emulsion method, and optimization was carried out by a factorial design. In vitro, ex vivo, and in vivo studies were carried out to evaluate the optimized formulation. The results indicate that the physicochemical features of these NPs enable them to incorporate and release HIV fusion inhibitor peptides to the vaginal mucosa before the fusion step takes place.
Assuntos
Portadores de Fármacos/química , HIV-1/efeitos dos fármacos , Peptídeos/administração & dosagem , Inibidores de Proteínas Virais de Fusão/administração & dosagem , Administração Intravaginal , Animais , Quitosana/química , Desenho de Fármacos , Feminino , Proteína gp41 do Envelope de HIV/antagonistas & inibidores , Infecções por HIV/prevenção & controle , Infecções por HIV/virologia , HIV-1/fisiologia , Modelos Animais , Mucosa/efeitos dos fármacos , Mucosa/metabolismo , Mucosa/virologia , Nanopartículas/química , Tamanho da Partícula , Peptídeos/química , Peptídeos/farmacocinética , Suínos , Vagina/efeitos dos fármacos , Vagina/metabolismo , Vagina/virologia , Proteínas do Envelope Viral/química , Inibidores de Proteínas Virais de Fusão/química , Inibidores de Proteínas Virais de Fusão/farmacocinética , Internalização do Vírus/efeitos dos fármacosRESUMO
BACKGROUND: The objective of the study was to compare the diagnostic yield of home-made ELISA tests based on synthetic chimeric fibrin/filaggrin citrullinated peptides (CFFCPs) with CCP3 and CCP3.1 commercial tests to detect anti-citrullinated protein/peptide antibodies (ACPAs) in rheumatoid arthritis (RA) patients. The prognostic value is also studied in a cohort of patients with early RA. Moreover, we transfer immunological assays from microtiter plates to microarray formats to allow the simultaneous analysis of several peptide sequences and reduce the volume of serum from patients. METHODS: The diagnostic study includes: 100 RA patients who fulfilled the 1987 ACR criteria; 100 healthy blood donors; 35 patients with SLE according ACR criteria; 35 patients with PsA fulfilling the Wright and Moll criteria and 30 patients with HCV infection. The prognostic value study includes 50 patients with early RA with follow-up data available. All samples are from outpatients attending the Rheumatology Department of the Hospital Clinic of Barcelona. RESULTS: Similar sensitivity, specificity and predictive values for the diagnosis of RA of CCFCPs compared to CCP3/CCP3.1 were obtained. Although a high concordance is observed between anti-CFFCPs and anti-CCP3/CCP3.1 in the early patients that rendered Larsen radiographic progression, CFFCPs could be a better marker of radiographic outcome. Strong correlations between the microarray and ELISA results were found for individual CFFCPs peptides. CONCLUSIONS: The development of multiplexing techniques combining a different spectrum of markers in a single analysis, including CFFCP peptides, could allow a more detailed analysis of the autoantibodies reactivity found in the sera of patients suffering of this heterogeneous disease.
Assuntos
Anticorpos Antiproteína Citrulinada/sangue , Artrite Reumatoide/diagnóstico , Biomarcadores/sangue , Citrulina/imunologia , Peptídeos Cíclicos/imunologia , Anticorpos Antiproteína Citrulinada/imunologia , Citrulina/química , Ensaio de Imunoadsorção Enzimática/métodos , Fibrina/química , Fibrina/imunologia , Proteínas Filagrinas , Humanos , Proteínas de Filamentos Intermediários/química , Proteínas de Filamentos Intermediários/imunologia , Peptídeos Cíclicos/química , Prognóstico , Análise Serial de Proteínas/métodos , Domínios ProteicosRESUMO
Despite the great effort to decrease the HIV infectivity rate, current antiretroviral therapy has several weaknesses; poor bioavailability, development of drug resistance and poor ability to access tissues. However, molecules such as peptides have emerged asa new expectative to HIV eradication. The vaginal mucosa is the main spreading point of HIV. There are natural barriers such as the vaginal fluid which protects the vaginal epithelium from any foreign agents reaching it. This work has developed and characterized Nanoparticles (NPs) coated with glycol chitosan (GC), loaded with an HIV-1 inhibitor peptide (E2). In vitro release and ex vivo studies were carried out using the vaginal mucosa of swine and the peptide was determined by HPLC MS/MS validated method. Moreover, the peptide was labeled with 5(6)-carboxyfluoresceine and entrapped into the NPs to carried out in vivo studies and to evaluate the NPs penetration and toxicity in the vaginal mucosa of the swine. The mean size of the NPs, ξ and the loading percentage were fundamental features for to reach the vaginal tissue and to release the peptide within intercellular space. The obtained results suggesting that the fusion inhibitor peptides loaded into the NPs coated with GC might be a new way to fight the HIV-1, due to the formulation might reach the human epithelial mucosa and release peptide without any side effects.
Assuntos
Fármacos Anti-HIV/administração & dosagem , Vírus GB C/metabolismo , Mucosa/metabolismo , Nanopartículas/administração & dosagem , Peptídeos/administração & dosagem , Vagina/metabolismo , Animais , Fármacos Anti-HIV/química , Disponibilidade Biológica , Quitosana/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Feminino , Infecções por HIV/tratamento farmacológico , HIV-1/efeitos dos fármacos , Nanopartículas/química , Peptídeos/química , Polímeros/química , Suínos , Espectrometria de Massas em Tandem/métodosRESUMO
Previous studies support the hypothesis that the envelope GB virusâ C (GBV-C) E1 protein interferes the HIV-1 entry and that a peptide, derived from the regionâ 139-156 of this protein, has been defined as a novel HIV-1 entry inhibitor. In this work, we firstly focus on the characterization of the structural features of this peptide, which are determinant for its anti-HIV-1 activity and secondly, on the study of its interaction with the proposed viral target (i.e., the HIV-1 fusion peptide). We report the structure of the peptide determined by NMR spectroscopy in dodecylphosphocholine (DPC) micelles solved by using restrained molecular dynamics calculations. The acquisition of different NMR experiments in DPC micelles (i.e., peptide-peptide titration, diffusion NMR spectroscopy, and addition of paramagnetic relaxation agents) allows a proposal of an inhibition mechanism. We conclude that a 18-mer peptide from the non-pathogenic E1 GBV-C protein, with a helix-turn-helix structure inhibits HIV-1 by binding to the HIV-1 fusion peptide at the membrane level, thereby interfering with those domains in the HIV-1, which are critical for stabilizing the six-helix bundle formation in a membranous environment.
Assuntos
HIV-1/metabolismo , Micelas , Peptídeos/metabolismo , Proteínas do Envelope Viral/metabolismo , Sequência de Aminoácidos , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacologia , Concentração Inibidora 50 , Simulação de Dinâmica Molecular , Ressonância Magnética Nuclear Biomolecular , Peptídeos/química , Fosforilcolina/análogos & derivados , Fosforilcolina/química , Ligação Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/farmacologia , Internalização do Vírus/efeitos dos fármacosRESUMO
The work reports the design and synthesis of a chimeric peptide that is composed of the peptide sequences of two entry inhibitors which target different sites of HIV-1 gp41. The chimeric peptide offers the advantage of targeting two gp41 regions simultaneously: the fusion peptide and the loop both of which are membrane active and participate in the membrane fusion process. We therefore use lipid raft-like liposomes as a tool to specifically direct the chimeric inhibitor peptide to the membrane domains where the HIV-1 envelope protein is located. Moreover, the liposomes that mimic the viral membrane composition protect the chimeric peptide against proteolytic digestion thereby increasing the stability of the peptide. The described liposome preparations are suitable nanosystems for managing hydrophobic entry-inhibitor peptides as putative therapeutics.