RESUMO
Theoretically, the immunization of experimental animals with an anti-idiotype antibody may elicit antibodies that recognize epitopes like the original idiotype; this is archived via internal images. Using this strategy, we attempted to produce anti-epithelial antibodies in Balb/c mice immunized with a pemphigus anti-idiotypic determinant. First, when an anti-idiotype antibody was produced in rabbits by immunization with pemphigus immunoglobulin G (IgG), the anti-idiotypic activity was tested successfully. The anti-idiotype IgG was digested with pepsin and purified by gel filtration chromatography to obtain F(ab')(2) fragments, which were used to immunize Balb/c mice. A control group was immunized with normal IgG. The experimental animals immunized with anti-idiotype F(ab')(2) fragments developed anti-epithelial antibodies in the following two months. The elicited antibodies had anti-desmoglein 1 specificity. Additionally, the skin biopsies of these animals exhibited antibody deposition along intercellular spaces of epidermis, and 25% of them developed blisters. Sera and skin biopsies of control Balb/c mice group were negative. In conclusion, the immunization with pemphigus anti-idiotype antibody may elicit anti-epithelial antibodies via internal images. This experimental approach can be used to understand the pathogenic mechanisms of pemphigus.
Assuntos
Acantólise/imunologia , Anticorpos Anti-Idiotípicos/metabolismo , Desmogleína 1/imunologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Vacinas/farmacologia , Acantólise/patologia , Animais , Anticorpos Anti-Idiotípicos/imunologia , Especificidade de Anticorpos/imunologia , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Modelos Animais de Doenças , Células Epiteliais/patologia , Fragmentos Fab das Imunoglobulinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Vacinação/métodos , Vacinas/imunologiaRESUMO
Fas ligand (L) is a membrane protein from the tumor necrosis factor (TNF) family. It induces apoptosis upon contact with its Fas/CD95/APO1 receptor. Trimerization of FasL on the surface of effector cells is essential in the binding of the Fas trimer of the target cells. The receptor then recruits an adaptor and caspase-like proteins which lead apoptosis. This paper reports on the fate of FasL in HEp-2 cells committed to apoptosis by induction with campthotecin. Our main results demonstrated that in non-apoptotic cells, FasL aggregates in the cytoplasm forming trimers of 120 kDa. Apoptosis increases the trimeric FasL species, but also induces its dissociation into monomers of 35 kDa. In conclusion, camptothecin appears to perturb the Fas and FasL segregation in the cytoplasm by promoting the transit of FasL to the cell surface, thus fostering a process of autocrine or paracrine apoptosis. FasL is trimerized prior to Fas/FasL complex formation, and after apoptosis, FasL undergoes an intense turnover.
Assuntos
Apoptose/efeitos dos fármacos , Camptotecina/farmacologia , Proteína Ligante Fas/química , Proteína Ligante Fas/metabolismo , Clorometilcetonas de Aminoácidos/farmacologia , Caspase 3/genética , Linhagem Celular Tumoral , Fragmentação do DNA/efeitos dos fármacos , Desoxirribonucleases/genética , Proteína Ligante Fas/genética , Humanos , Marcação In Situ das Extremidades Cortadas , Proteínas de Ligação a Poli-ADP-Ribose , Estrutura Quaternária de Proteína , Transporte Proteico/efeitos dos fármacos , Frações Subcelulares/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacos , Receptor fas/genéticaRESUMO
La talidomida es un medicamento útil para el tratamiento de la lepra lepromatosa reaccional y se conoce parcialmente su mecanismo de acción. Para entender mejor las propiedades de este fármaco se estudió la función fagocítica de los polimorfonucleares en presencia de talidomida. Los resultados indican que este fármaco disminuye de manera significativa la fagocitosis in vitro y, probablemente mediante el mismo mecanismo, ayuda a disminuir el proceso inflamatorio de la reacción leprosa y otras enfermedades inflamatorias de la piel