RESUMEN
Susceptible-strain animals immunized with P2 peptide could generate the disease of experimental autoimmune neuritis (EAN) with inflammation and demyelination of peripheral nerve. A myriad of transcription factors and inflammatory cytokines have been found to participate in this process; however, the roles of toll-like receptors (TLRs) are poorly understood in EAN. The aim of this study is to explore the role of TLR9 in the pathogenesis of EAN. The EAN was induced in Lewis rat by immunization with P2(53-78) and complete Freund's adjuvant. CpG oligodeoxynucleotides (ODN) (cODN), a suppressive ODN (sODN) and a control non-specific ODN (nODN) were respectively administered to explore the role of TLR9 in EAN both in vivo and vitro. Following immunization up to the peak phase of EAN, EAN rats inoculated with sODN had remarkably better clinical score of EAN and expressed a significantly inhibited TLR9 signaling pathway. Our study suggests that TLR9 may be involved in the pathogenesis of EAN.
Asunto(s)
Síndrome de Guillain-Barré/inmunología , Neuritis Autoinmune Experimental/inmunología , Oligodesoxirribonucleótidos Antisentido/administración & dosificación , Receptor Toll-Like 9/metabolismo , Animales , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Susceptibilidad a Enfermedades , Adyuvante de Freund/administración & dosificación , Síndrome de Guillain-Barré/terapia , Humanos , Inmunización , Masculino , Proteína P2 de Mielina/administración & dosificación , Neuritis Autoinmune Experimental/inducido químicamente , Neuritis Autoinmune Experimental/genética , Neuritis Autoinmune Experimental/fisiopatología , Neuritis Autoinmune Experimental/terapia , Oligodesoxirribonucleótidos Antisentido/efectos adversos , Fragmentos de Péptidos/administración & dosificación , Ratas , Ratas Endogámicas Lew , Receptor Toll-Like 9/genéticaRESUMEN
Experimental autoimmune neuritis (EAN) is a CD4(+) T cell-mediated, inflammatory demyelinating disease of the peripheral nervous system (PNS) that serves as a model for Guillain-Barré syndrome (GBS) in humans. The facial nerve paralysis is relatively commonly found in GBS patients. Here, EAN was established in Lewis rats by immunization with P2 peptide 57-81, a purified component of peripheral nerve myelin, and Freund's complete adjuvant (FCA). To study whether the facial nerves are involved in the pathogenic process during the EAN course, we observed the clinical and pathological changes as well as cytokine production in facial nerves on Day 14 postimmunization (p.i.), i.e. at height of clinical EAN. As a result, all rats immunized with P2 peptide 57-81 developed severe EAN on Day 14 p.i., but none of the rats manifested clinical signs of facial nerve paralysis. Additionally, only mild inflammatory cell infiltration and proinflammatory cytokine, interferon-gamma (IFN-gamma) and tumour necrosis factor (TNF-alpha) production as well as devoid demyelination were seen in facial nerves of the EAN rats. On the contrary, severe inflammation and demyelination as well as upregulated IFN-gamma and TNF-alpha production were observed in sciatic nerves of the same EAN rats. The underlying mechanism for the difference of the local manifestation of the disease process of EAN remains to be resolved.
Asunto(s)
Citocinas/biosíntesis , Enfermedades Desmielinizantes/inmunología , Enfermedades Desmielinizantes/patología , Nervio Facial/inmunología , Nervio Facial/patología , Mediadores de Inflamación/metabolismo , Neuritis Autoinmune Experimental/inmunología , Neuritis Autoinmune Experimental/patología , Animales , Movimiento Celular/inmunología , Enfermedades Desmielinizantes/fisiopatología , Nervio Facial/metabolismo , Masculino , Proteína P2 de Mielina/administración & dosificación , Vaina de Mielina/fisiología , Neuritis Autoinmune Experimental/fisiopatología , Ratas , Ratas Endogámicas Lew , Nervio Ciático/inmunología , Nervio Ciático/metabolismo , Nervio Ciático/patología , Nervio Ciático/fisiopatologíaRESUMEN
The expression of major histocomatibility complex class II in vitro and in vivo by Schwann cells indicates a potential facultative role of Schwann cells in the presentation of antigen to neuritogenic T cells during inflammatory demyelinating neuropathies. Using a T cell proliferation assay, this study demonstrated that processing and presentation of endogenous and exogenous antigen by Schwann cells influences T cell proliferation. Statistical analysis of proliferation and its relation to processing and presentation of antigen by Schwann cells had not been previously addressed. Different combinations of factors including treatment of cultures (untreated, irradiated or fixed), concentration of exogenous antigen (0 or 40 microg/ml), the presence of interferon-gamma and the timing of exogenous antigen addition influence the proliferation P2-specific, non-mammalian protein ovalbumin-specific T cell lines and naive T cells.
Asunto(s)
Presentación de Antígeno , Antígenos/inmunología , Células de Schwann/inmunología , Linfocitos T/inmunología , Animales , División Celular/fisiología , Células Cultivadas , Genes MHC Clase II , Interferón gamma/farmacología , Proteína P2 de Mielina/administración & dosificación , Proteína P2 de Mielina/inmunología , Neuritis Autoinmune Experimental/inmunología , Ratas , Ratas Endogámicas Lew , Células de Schwann/citología , Células de Schwann/efectos de los fármacos , Células de Schwann/metabolismo , Linfocitos T/citología , Linfocitos T/metabolismoRESUMEN
We have developed and described a new method of altering T cell-mediated autoimmune diseases by immunization with the complementary peptide against T cell epitopes. The complementary peptide (denoted NAE 07-06) to the bovine P2 protein, residues 60-70 (denoted EAN 60-70), was tested in the Lewis rat model of experimental allergic neuritis (EAN). Immunization with NAE 07-06 induced polyclonal and monoclonal Abs that inhibited the proliferation of the P2-specific T cell line, stimulated with EAN 60-70, and recognized Vbeta, but not Valpha, of TCRs. Proliferation of T cells treated with anti-NAE 07-06 Abs could be partially restored by treatment with rIL-2, in accordance with an anergy model. A homologous sequence was found between NAE 07-06 and the VDJ junction of the TCR beta-chain from an EAN 60-70-specific T cell line. Rats preimmunized with NAE 07-06 in vivo before EAN induction showed less disease severity clinically and histologically. These data suggest a new therapeutic approach for T cell-mediated autoimmune disorders through the induction of anti-TCR Abs with complementary peptide Ags.
Asunto(s)
Anergia Clonal/inmunología , Proteína P2 de Mielina/inmunología , Neuritis Autoinmune Experimental/prevención & control , Fragmentos de Péptidos/inmunología , Linfocitos T/inmunología , Vacunas Sintéticas/inmunología , Animales , Anticuerpos Monoclonales/metabolismo , Bovinos , Línea Celular , Epítopos de Linfocito T/metabolismo , Femenino , Sueros Inmunes/metabolismo , Inyecciones Subcutáneas , Proteína P2 de Mielina/administración & dosificación , Neuritis Autoinmune Experimental/inmunología , Neuritis Autoinmune Experimental/patología , Fragmentos de Péptidos/administración & dosificación , Conejos , Ratas , Ratas Endogámicas Lew , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismo , Vacunas Sintéticas/administración & dosificaciónRESUMEN
Experimental autoimmune neuritis (EAN) is a CD4+ T cell-mediated autoimmune disorder of the peripheral nervous system (PNS) that can be actively induced in susceptible animal species and strains by active immunization with PNS myelin + Freund's complete adjuvant (FCA). EAN represents an animal model for studying the immunopathogenesis and treatment of Guillain-Barré syndrome (GBS), which is a major inflammatory demyelinating disease of the PNS in humans. Here, we report that treatment by nasal administration of the neuritogenic peptide 57-81 of the PNS myelin component, P2 protein, dose-dependently suppressed EAN severity and shortened clinical EAN. Clinical EAN relapse induced by rechallenge with BPM + FCA was also prevented in EAN rats receiving high dose P2 peptide. P2 peptide induced suppression of EAN was associated with PNS antigen specific T cell hyporesponsiveness reflected by lymphocyte proliferation, numbers of PNS antigen-reactive IFN-gamma secreting and IFN-gamma mRNA expressing lymph node cells, but elevated levels of PNS antigen reactive TGF-beta mRNA secreting cells. Reduced CD4+ T cell and macrophage infiltrations within the PNS were also observed. Based on these observations, nasal autoantigen administration should be further evaluated, considering its possible future use in GBS.
Asunto(s)
Proteína P2 de Mielina/uso terapéutico , Neuritis Autoinmune Experimental/tratamiento farmacológico , Fragmentos de Péptidos/uso terapéutico , Administración Intranasal , Secuencia de Aminoácidos , Animales , Citocinas/genética , Inmunohistoquímica , Datos de Secuencia Molecular , Proteína P2 de Mielina/administración & dosificación , Neuritis Autoinmune Experimental/patología , Fragmentos de Péptidos/administración & dosificación , ARN Mensajero/análisis , Ratas , Ratas Endogámicas LewRESUMEN
Experimental autoimmune neuritis (EAN) is a CD4+ T cell-mediated inflammatory demyelinating disease of the peripheral nervous system (PNS) that serves as a model for Guillain-Barre syndrome (GBS) in humans. Both EAN and GBS are associated with upregulated T and B cells responses to PNS myelin proteins including P2 protein, and by changes of the Th1/Th2 cell balance in favor of Th1. Here we report that EAN can be prevented by the dominant neuritogenic peptide 57-81 of the PNS P2 protein when given nasally before immunization of Lewis rats with bovine PNS myelin (BPM) + Freund's complete adjuvant (FCA). P2 peptide-tolerized rats were also resistant to EAN relapse after challenge with BPM. Tolerance to EAN in rats receiving high dose (60 microg/day/rat) P2 peptide nasally was associated with specific T and B cell anergy. This was characterized by the failure of T cells to proliferate in response to PNS myelin antigens, while responsiveness to phytohemagglutinin was retained. Numbers of BPM- and P2 peptide-reactive interferon-gamma mRNA expressing lymph node cells were reduced, while levels of P2 peptide-reactive interleukin 4 and transforming growth factor-beta mRNA-expressing cells were markedly upregulated on day 18 post immunization in the rats receiving high dose P2 peptide nasally. Tolerance to EAN was also associated with lower CD4+ cell infiltration, low-grade inflammation, or the absence of histological evidence of EAN, as well as with low IL-2 receptor and MHC class II molecule expression within the PNS. This is the first study showing that mucosal tolerance is applicable to EAN and, as an extension, could be considered in GBS.