RESUMO
We studied the effect of ß1i-specific peptidyl aldehyde IPSI-001 on proteasome from mammalian cells. In concentrations <1 µM, this agent effectively suppressed immunoproteasome, but only slightly reduced chymotrypsin-like activity of constitutive proteasome. Intraperitoneal administration of this inhibitor to C3H/He mice in a dose of 100 mg/kg induced no significant physiological or behavioral changes, which attested to its considerable therapeutic potential in the treatment of autoimmune neurodegenerative pathologies.
Assuntos
Dipeptídeos/farmacologia , Complexo de Endopeptidases do Proteassoma/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Linhagem Celular , Humanos , Camundongos , Camundongos Endogâmicos C3H , Proteína Básica da Mielina/metabolismoRESUMO
Deimination of myelin basic protein (MBP) by peptidylarginine deiminase (PAD) prevents its binding to the proteasome and decelerates its degradation by the proteasome in mammalian cells. Potential anticancer drug tetrazole analogue of chloramidine 2, at concentrations greater than 1 µM inhibits the enzymatic activity of PAD in vitro. The observed acceleration of proteasome hydrolysis of MBP to antigenic peptides in the presence of PAD inhibitor may increase the efficiency of lesion of the central nervous system by cytotoxic lymphocytes in multiple sclerosis. We therefore suggest that clinical trials and the introduction of PAD inhibitors in clinical practice for the treatment of malignant neoplasms should be performed only after a careful analysis of their potential effect on the induction of autoimmune neurodegeneration processes.
Assuntos
Compostos de Bifenilo/farmacologia , Hidrolases/antagonistas & inibidores , Hidrolases/metabolismo , Proteína Básica da Mielina/metabolismo , Fármacos Neuroprotetores/farmacologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Tetrazóis/farmacologia , Animais , Bortezomib/farmacologia , Bovinos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Células HEK293 , Humanos , Hidrolases/genética , Hidrólise , Camundongos Endogâmicos C3H , Esclerose Múltipla/tratamento farmacológico , Esclerose Múltipla/metabolismo , Músculo Esquelético/enzimologia , Proteína Básica da Mielina/genética , Ligação Proteica/fisiologia , Desiminases de Arginina em Proteínas , Proteólise , Coelhos , TransfecçãoRESUMO
In this study, we performed transcriptome profiling of oligodendrocyte culture of mice treated with the remyelinating therapeutic agent benztropine in the presence and absence of interferon gamma (IFNγ). The results of this work are important for understanding the expression profile of oligodendrocytes under conditions of systemic inflammation in the central nervous system in multiple sclerosis as well as the mechanisms of cellular response to benztropine in light of its possible use for the treatment of multiple sclerosis.
Assuntos
Oligodendroglia/imunologia , RNA Mensageiro/imunologia , Transcriptoma/imunologia , Animais , Anti-Inflamatórios/farmacologia , Benzotropina/farmacologia , Células Cultivadas , Perfilação da Expressão Gênica , Fatores Imunológicos/farmacologia , Interferon gama/farmacologia , Camundongos Endogâmicos C3H , Oligodendroglia/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/imunologia , Transcriptoma/efeitos dos fármacosRESUMO
Multiple Sclerosis (MS) is a serve autoimmune neurodegenerative disease. Development of innovative approaches of MS treatment is of a high priority in the modern immunology and pharmacy. In the present study we showed high therapeutic efficiency of immunodominant peptides of myelin basic protein (MBP) incorporated into the monolayer mannosylated liposomes on the development of experimental autoimmune encephalomyelitis (EAE) in DA rats. MBP is a component ofoligodendrocytes' membrane, which form axonal sheath, and is one of the major autoantigens in MS. We analyzed binding pattern ofanti-MBP autoantibodies from MS patients using previously designed MBP epitope library. Utilizing the same approach we investigated pool of anti-MBP antibodies from SJL/J and C57/BL6 mice and DA rats with induced EAE. The most relevant rodent model to MS was EAE in DA rats according to the autoantibodies' binding pattern. We selected three immunodominant MBP fragments encapsulated in monolayer mannosylated liposomes for the following treatment of verified DA rodent model. MBP fragment 46-62 was the most effective in reducing of the first EAE attack, whereas MBP 124-139 and 147-160 inhibited development of pathology during remission stage. Simultaneous administration of these peptides in liposomes significantly decreased level of anti-MBP antibodies. Synergetic therapeutic effect of MBP fragments reduced integral disease score by inhibiting first EAE wave and subsequent remission, thus, our findings disclosure novel approaches for efficient treatment of Multiple Sclerosis.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Epitopos Imunodominantes/administração & dosagem , Esclerose Múltipla/tratamento farmacológico , Proteína Básica da Mielina/administração & dosagem , Nanocápsulas/administração & dosagem , Fragmentos de Peptídeos/administração & dosagem , Adulto , Sequência de Aminoácidos , Animais , Autoanticorpos/imunologia , Cobaias , Humanos , Epitopos Imunodominantes/imunologia , Epitopos Imunodominantes/uso terapêutico , Lipossomos , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteína Básica da Mielina/imunologia , Proteína Básica da Mielina/uso terapêutico , Nanocápsulas/química , Fragmentos de Peptídeos/imunologia , Fragmentos de Peptídeos/uso terapêutico , Ratos , Ratos EndogâmicosRESUMO
Proteolytic degradation of autoantigens is of prime importance in current biochemistry and immunology. The most fundamental issue in this field is the functional role of peptides produced when the specificity of hydrolysis changes during the shift from health to disease and from normal state to pathology. The identification of specific peptide fragments in many cases proposes the diagnostic and prognostic criterion in the pathology progression. The aim of this work is comparative study of the degradation peculiarities of one of the main neuroantigen, myelin basic protein by proteases, activated during progress of pathological demyelinating process, and by proteasome of different origin. The comparison of specificity of different studied biocatalysts gives reason to discuss the critical change in the set of myelin basic protein fragments capable to be presented by major histocompatibility complex class I during neurodegeneration, which can promote the progress of autoimmune pathological process.
Assuntos
Proteína Básica da Mielina/metabolismo , Peptídeo Hidrolases/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Sequência de Aminoácidos , Animais , Calpaína/metabolismo , Catepsina D/metabolismo , Linhagem Celular , Cricetinae , Cricetulus , Humanos , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Fragmentos de Peptídeos/metabolismo , Proteômica , Tripsina/metabolismoRESUMO
Organophosphate toxins (OPs) are the most toxic low-molecular compounds. The extremely potent toxicity of OPs is determined by their specificity toward the nerve system. Human butyrylcholinesterase (hBChE) is a natural bioscavenger against a broad spectrum of OPs, which makes it a promising candidate for the development of DNA-encoded bioscavengers. The high values of the protective index observed for recombinant hBChE (rhBChE) make it appropriate for therapy against OP poisoning, especially in the case of highly toxic warfare nerve agents. Nevertheless, large-scale application of biopharmaceuticals based on hBChE is restricted due to its high cost and extremely rapid elimination from the bloodstream. In the present study, we examine two approaches for long-acting rhBChE production: I) chemical polysialylation and II) in-vivo tetramerization. We demonstrate that both approaches significantly improve the pharmacokinetic characteristics of rhBChE (more than 5 and 10 times, respectively), which makes it possible to use rhBChE conjugated with polysialic acids (rhBChE-CAO) and tetrameric rhBChE (4rhBChE) in the treatment of OP poisonings.