RESUMEN
Viral and nonviral gene therapy vectors have been successfully employed to deliver inflammatory cytokine inhibitors (anticytokines), or anti-inflammatory cytokines, such as transforming growth factor beta-1 (TGF-beta 1), which protect against experimental autoimmune diseases. These vectors carry the relevant genes into a variety of tissues, for either localised or systemic release of the encoded protein. Administration of cDNA encoding soluble IFN-gamma receptor (IFN-gamma R)/IgG-Fc fusion proteins, soluble TNF-alpha receptors, or IL-1 receptor antagonist (IL-1ra), protects against either lupus, various forms of arthritis, autoimmune diabetes, or other autoimmune diseases. These inhibitors, unlike many cytokines, have little or no toxic potential. Similarly, TGF-beta 1 gene therapy protects against numerous forms of autoimmunity, though its administration entails more risk than anticytokine therapy. We have relied on the injection of naked plasmid DNA into skeletal muscle, with or without enhancement of gene transfer by in vivo electroporation. Expression plasmids offer interesting advantages over viral vectors, since they are simple to produce, non-immunogenic and nonpathogenic. They can be repeatedly administered and after each treatment the encoded proteins are produced for relatively long periods, ranging from weeks to months. Moreover, soluble receptors which block cytokine action, encoded by gene therapy vectors, can be constructed from non-immunogenic self elements that are unlikely to be neutralised by the host immune response (unlike monoclonal antibodies [mAbs]), allowing long-term gene therapy of chronic inflammatory disorders.
Asunto(s)
Enfermedades Autoinmunes/terapia , Citocinas/antagonistas & inhibidores , Citocinas/genética , Terapia Genética , Animales , HumanosRESUMEN
The cyclin kinase inhibitor protein p21 affects multiple processes relevant to the immune system, including cell cycle progression, replicative senescence, hemopoietic stem cell quiescence, and apoptosis. Therefore, malfunction of this protein may be a contributor to the pathogenesis of systemic autoimmunity. Here, we report that mixed background p21-deficient 129/Sv x C57BL/6 mice showed increased in vitro and in vivo T cell cycling and activation, moderate hypergammaglobulinemia and, at low penetrance, anti-chromatin autoantibodies. Homeostatic anti-self MHC/peptide ligand-induced proliferation of p21-deficient T cells was also enhanced. However, lymphoid organ enlargement was very mild, presumably due to increased apoptosis of the rapidly dividing cells. Moreover, the older p21-deficient mice had kidney pathology representing a similar, but slightly more advanced, state than that seen in the control mice. The timing and severity of the above serologic, cellular, and histologic manifestations in p21-deficient mice were unaffected by gender. Thus, p21 deficiency significantly enhances T cell activation and homeostatic proliferation, and can induce mild autoimmune manifestations at a low incidence without gender bias, but does not in itself generate the full spectrum of lupus-like disease.
Asunto(s)
Enfermedades Autoinmunes/etiología , Ciclinas/fisiología , Animales , Apoptosis , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Autoinmunidad , Células Cultivadas , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Ciclinas/genética , Femenino , Hipergammaglobulinemia/sangre , Inmunofenotipificación , Riñón/patología , Activación de Linfocitos , Subgrupos Linfocitarios/clasificación , Tejido Linfoide/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Superantígenos/inmunología , Tasa de Supervivencia , Linfocitos T/inmunologíaRESUMEN
The neonatal Ab and TCR repertoires are much less diverse, and also very different from, the adult repertoires due to the delayed onset of terminal deoxynucleotidyl transferase (TdT) expression in ontogeny. TdT adds nontemplated N nucleotides to the junctions of Igs and TCRs, and thus its absence removes one of the major components of junctional diversity in complementarity-determining region 3 (CDR3). We have generated TdT-deficient MRL/lpr, Fas-deficient (MRL-Fas(lpr)) mice, and show that they have an increased lifespan, decreased incidence of skin lesions, and much lower serum levels of anti-dsDNA, anti-chromatin, and IgM rheumatoid factors. The generalized hypergammaglobulinemia characteristic of MRL-Fas(lpr) mice is also greatly reduced, as is the percentage of CD4(-)CD8(-)B220(+) (double-negative) T cells. IgG deposits in the kidney are significantly reduced, although evidence of renal disease is present in many mice at 6 mo. CDR3 regions of both IgH and TCR from peripheral lymphocytes of MRL-Fas(lpr) mice are shorter in the absence of TdT, and there is a paucity of arginines in the IgH CDR3 regions of the MRL-Fas(lpr) TdT(-/-) mice. Because the amelioration of symptoms is so widespread, it is likely that the absence of N regions has more of an affect than merely decreasing the precursor frequency of anti-dsDNA B cells. Hence, either the T or B cell repertoires, or more likely both, require N region diversity to produce the full spectrum of autoimmune lupus disease.
Asunto(s)
Enfermedades Autoinmunes/enzimología , Regiones Determinantes de Complementariedad/genética , ADN Nucleotidilexotransferasa/deficiencia , Lupus Eritematoso Sistémico/enzimología , Animales , Anticuerpos Antinucleares/sangre , Anticuerpos Antinucleares/genética , Enfermedades Autoinmunes/genética , Cromatina/inmunología , Cruzamientos Genéticos , ADN/sangre , ADN Nucleotidilexotransferasa/genética , Modelos Animales de Enfermedad , Reordenamiento Génico de Cadena Pesada de Linfocito B , Reordenamiento Génico de la Cadena beta de los Receptores de Antígenos de los Linfocitos T , Hiperplasia , Lupus Eritematoso Sistémico/complicaciones , Lupus Eritematoso Sistémico/genética , Lupus Eritematoso Sistémico/patología , Nefritis Lúpica/enzimología , Nefritis Lúpica/genética , Recuento de Linfocitos , Subgrupos Linfocitarios , Trastornos Linfoproliferativos/genética , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos MRL lpr , Ratones Endogámicos NZB , Ratones Noqueados , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Factor Reumatoide/análisis , Piel/patología , Organismos Libres de Patógenos Específicos , Subgrupos de Linfocitos T/inmunologíaRESUMEN
Male BXSB mice develop an early life, severe lupus-like disease largely attributed to an undefined Y-chromosome-associated autoimmunity accelerator, termed YAA: Although the exact disease pathogenesis is uncertain, indirect evidence suggests that T cells play an important role in the male BXSB disease. We have developed TCR alpha-chain gene-deleted BXSB mice to directly examine the role of alphabeta+ T cells and the mode by which Yaa promotes disease in this strain. All disease parameters, including hypergammaglobulinemia, autoantibody production, glomerulonephritis, and the unique monocytosis of BXSB males, were severely reduced or absent in the alphabeta+ T cell-deficient mice. Adoptively transferred CD4+ T cells of either male or female BXSB origin showed equal homeostatic proliferation in alphabeta+ T cell-deficient male recipients. Moreover, deficient male mice eventually developed equally severe lupus-like disease after adoptive transfer and homeostatic expansion of T cells from wild-type BXSB males or females. The results directly demonstrate that the Yaa-mediated disease requires alphabeta+ T cells that are not, in themselves, abnormal in either composition or properties, but are engaged by a Yaa-encoded abnormality in a non-T cell component. In addition, homeostatic anti-self proliferation of mature T cells derived from a small number of precursors can induce systemic autoimmunity in an appropriate background.
Asunto(s)
Homeostasis/inmunología , Nefritis Lúpica/inmunología , Activación de Linfocitos , Receptores de Antígenos de Linfocitos T alfa-beta/fisiología , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Cromosoma Y , Traslado Adoptivo , Animales , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Femenino , Inmunoglobulina G/sangre , Riñón/inmunología , Riñón/patología , Leucocitosis/genética , Leucocitosis/inmunología , Nefritis Lúpica/genética , Nefritis Lúpica/mortalidad , Nefritis Lúpica/patología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Activación de Linfocitos/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Monocitos/inmunología , Monocitos/patología , Receptores de Antígenos de Linfocitos T alfa-beta/deficiencia , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Bazo/inmunología , Bazo/patología , Análisis de Supervivencia , Subgrupos de Linfocitos T/patología , Subgrupos de Linfocitos T/trasplante , Cromosoma Y/genéticaRESUMEN
Immuno-gene therapy can be advantageously performed with nonviral approaches. Genes that encode regulatory cytokines or inflammatory cytokine inhibitors can be delivered intramuscularly and expressed for weeks or months. This type of gene transfer into muscle has been shown to ameliorate several autoimmune diseases and is relevant to the development of effective DNA vaccines in autoimmune diseases, infectious diseases and cancer.
Asunto(s)
Técnicas de Transferencia de Gen , Inmunoterapia/métodos , Músculo Esquelético , Animales , Presentación de Antígeno , Autoinmunidad/inmunología , Proteínas del Sistema Complemento/inmunología , Citocinas/genética , Citocinas/inmunología , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/prevención & control , Terapia Genética/métodos , Humanos , Tolerancia Inmunológica/inmunología , Inyecciones Intramusculares , Lupus Vulgar/inmunología , Lupus Vulgar/terapia , Vacunas de ADN/administración & dosificaciónRESUMEN
The classification of T helper cells into type 1 (Th1) and type 2 (Th2) led to the hypothesis that Th1 cells and their cytokines (interleukin [IL]-2, interferon [IFN]-gamma) are involved in cell-mediated autoimmune diseases, and that Th2 cells and their cytokines (IL-4, IL-5, IL-10, IL-13) are involved in autoantibody(humoral)-mediated autoimmune diseases. However, this paradigm has been refuted by recent studies in several induced and spontaneous mouse models of systemic lupus erythematosus, which showed that IFN-gamma is a major effector molecule in this disease. These and additional findings, reviewed here, suggest that these two cross-talking classes of cytokines can exert autoimmune disease-promoting or disease-inhibiting effects without predictability or strict adherence to the Th1-versus-Th2 dualism.
Asunto(s)
Autoinmunidad/fisiología , Interferón gamma/fisiología , Lupus Eritematoso Sistémico/inmunología , Células TH1/inmunología , Células Th2/inmunología , Animales , ADN Complementario/uso terapéutico , Modelos Animales de Enfermedad , Terapia Genética , Fragmentos Fc de Inmunoglobulinas/genética , Fragmentos Fc de Inmunoglobulinas/uso terapéutico , Interleucinas/inmunología , Lupus Eritematoso Sistémico/terapia , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Mutantes , Receptor Cross-Talk , Proteínas Recombinantes de Fusión/uso terapéuticoRESUMEN
IFN-gamma, a pleiotropic cytokine, is a key effector molecule in the pathogenesis of several autoimmune diseases, including lupus. Importantly, deletion of IFN-gamma or IFN-gammaR in several lupus-predisposed mouse strains resulted in significant disease reduction, suggesting the potential for therapeutic intervention. We evaluated whether intramuscular injections of plasmids with cDNA encoding IFN-gammaR/Fc can retard lupus development and progression in MRL-Fas(lpr) mice. Therapy significantly reduced serum levels of IFN-gamma, as well as disease manifestations (autoantibodies, lymphoid hyperplasia, glomerulonephritis, mortality), when treatment was initiated at the predisease stage, particularly when IFN-gammaR/Fc expression was enhanced by electroporation at the injection site. Remarkably, disease was arrested and even ameliorated when this treatment was initiated at an advanced stage. This therapy represents a rare example of disease reversal and makes application of this nonviral gene therapy in humans with lupus (and perhaps other autoimmune/inflammatory conditions) highly promising.