RESUMEN
BACKGROUND: We previously detected antibodies against tyrosine hydroxylase (TH) in 23% of patients with nonsegmental vitiligo and in 19% of patients with alopecia areata (AA). OBJECTIVES: To identify TH epitopes recognized by TH antibodies in patients with vitiligo and AA. METHODS: Recombinant plasmids containing defined fragments of TH cDNA were constructed. The cloned TH cDNA fragments were subsequently translated in vitro to produce a series of [(35) S]-labelled TH protein fragments which were then used in radioimmunoassays to analyse the immunoreactivity of sera from 18 TH antibody-positive patients with vitiligo and so initially define TH epitope domains. Further localization of TH epitopes was investigated by antibody absorption experiments using synthetic TH peptides and nonradiolabelled, in vitro-expressed TH protein fragments. Antibody binding to identified epitopes was confirmed in TH peptide enzyme-linked immunosorbent assays. RESULTS: Analysis of the results obtained indicated the presence of two major antibody-binding sites on TH between amino acids 1 and 14 (epitope 1-14) and between amino acids 61 and 80 (epitope 61-80). Of 18 patients with vitiligo and six with AA, 17 (94%) and five (83%), respectively, had antibodies against epitope 1-14. In addition, 11/18 (61%) vitiligo and 2/6 (33%) AA patient sera displayed immunoreactivity against epitope 61-80. CONCLUSIONS: Two major binding sites for human TH antibodies are located at the N-terminus of the protein. The humoral immune response to TH in vitiligo and AA is heterogeneous in nature in that patients may have antibodies to more than one TH epitope. TH antibodies from patients with vitiligo or AA can recognize identical epitopes.
Asunto(s)
Alopecia Areata/inmunología , Autoanticuerpos/metabolismo , Epítopos de Linfocito B/metabolismo , Inmunoglobulina G/metabolismo , Tirosina 3-Monooxigenasa/inmunología , Vitíligo/inmunología , Adolescente , Adulto , Anciano , Sitios de Unión , Niño , Preescolar , ADN Complementario/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina G/clasificación , Masculino , Persona de Mediana Edad , Radioinmunoensayo , Adulto JovenRESUMEN
Thyroid cells are exposed to complement attack in Graves' disease and Hashimoto's thyroiditis, but are resistant to killing by homologous complement. We have examined the effects of sublethal complement attack on thyroid cells in vitro. Extracellular reactive oxygen metabolites were produced and prostaglandin E2, interleukin-1 alpha, and interleukin-6 were released after complement attack. Cells pretreated with interferon-gamma and interleukin-1 alpha, which increase expression of CD59, were more resistant to these effects of complement. Conversely, blockade of CD59 with monoclonal antibody increased complement-mediated oxygen radical production and release of prostaglandin E2, interleukin-1 alpha, and interleukin-6. The antithyroid drugs methimazole and propylthiouracil abolished or reduced oxygen radical production by complement-attacked thyroid cells and reduced cytokine release. These results suggest that sublethal complement attack in autoimmune thyroid diseases exacerbates tissue injury by causing thyroid cells to release potent phlogistic mediators, although some degree of protection may be afforded in vivo by cytokine-mediated upregulation of CD59. Antithyroid drugs, concentrated within thyroid cells, will prevent the release of these inflammatory molecules, which may in turn explain the amelioration of thyroiditis and remission of Graves' disease after such treatment.
Asunto(s)
Antígenos CD/inmunología , Interferón gamma/inmunología , Interleucina-1/inmunología , Interleucina-6/metabolismo , Glicoproteínas de Membrana/inmunología , Oxígeno/metabolismo , Prostaglandinas E/metabolismo , Glándula Tiroides/inmunología , Antígenos CD59 , Vía Clásica del Complemento/inmunología , Evaluación Preclínica de Medicamentos , Radicales Libres , Humanos , Interleucina-1/metabolismo , Metimazol/farmacología , Propiltiouracilo/farmacología , Glándula Tiroides/efectos de los fármacos , Glándula Tiroides/metabolismo , Factores de TiempoRESUMEN
We have investigated the effect of excess iodide alone, following iodide depletion or in conjunction with a mild thyroid insult (trypan blue) in Buffalo strain rats, which are genetically susceptible to experimental autoimmune thyroiditis. In all three cases a high iodine diet led to enhanced thyroglobulin antibody production, usually accompanied by worsening of the severity of thyroiditis. Moreover, the administration of a normal iodine diet after iodide depletion resulted in thyroglobulin antibody formation. These results provide further support for a role for dietary iodine in modulating experimental autoimmune thyroiditis with implications for human thyroid autoimmunity.
Asunto(s)
Yodo/administración & dosificación , Ratas Endogámicas BUF/metabolismo , Ratas Endogámicas/metabolismo , Animales , Dieta , Femenino , Yodo/deficiencia , Yodo/farmacocinética , Masculino , Ratas , Glándula Tiroides/efectos de los fármacos , Tiroiditis/inducido químicamente , Azul de Tripano/farmacologíaRESUMEN
Several studies have suggested that iodide may increase thyroiditis and autoantibody synthesis. We have investigated the effect of iodide in vitro at physiologically relevant concentrations on immunoglobulin synthesis by normal human peripheral blood lymphocytes stimulated with pokeweed mitogen. Both the number of cells which synthesised IgG and the amount of IgG released into the culture supernatant increased significantly after culture in a medium with added iodide compared to a medium with added chloride. No effect of increasing concentrations of the added iodide from 10(-3) mM to 10 mM was observed. These findings suggest that iodides may have a role in enhancing antibody synthesis which may be important when programmes of iodide supplementation are introduced into areas which are deficient.