RESUMEN
OBJECTIVE: To analyze cytokine gene expression in keratinocytes from patients with systemic lupus erythematosus (SLE). INTRODUCTION: Keratinocytes represent 95% of epidermal cells and can secrete several cytokines. METHODS: Keratinocytes were obtained by laser microdissection from 21 patients with SLE (10 discoid and 11 acute lesions) at involved and uninvolved sites. All patients were receiving a low/moderate prednisone dose and 18 were receiving chloroquine diphosphate. IL-2, IL-5, TNF-α and IFN-γ gene expression was evaluated by real-time PCR and expressed as the ratio (R) to a pool of skin samples from 12 healthy volunteers. RESULTS: Heterogeneity in cytokine gene expression was found among patients with SLE. Eighteen of 38 valid SLE samples (47%) presented overexpression (R>1) of at least one cytokine. Lesional skin samples tended to show higher cytokine expression than samples from uninvolved skin (p = 0.06). IL-5 and IFN-γ were the most commonly overexpressed cytokines. Samples with cytokine overexpression corresponded to more extensive and severe lesions. Prednisone dose did not differ between samples without cytokine overexpression (15.71±3.45 mg/day) and those with overexpressed cytokines (12.68±5.41 mg/day) (p = 0.216). Samples from all patients not receiving diphosphate chloroquine had at least one overexpressed cytokine. CONCLUSIONS: The heterogeneous keratinocyte cytokine gene expression reflects the complex immunological and inflammatory background in SLE. Patients with severe/extensive skin lesions showed a higher frequency of cytokine gene overexpression. Increased IFN-γ and IL-5 expression suggests that Th1 and Th2 cells are involved in SLE skin inflammation. The possibility that prednisone and antimalarial drugs may have contributed to low cytokine gene expression in some samples cannot be ruled out.
Asunto(s)
Citocinas/genética , Expresión Génica/genética , Queratinocitos/metabolismo , Lupus Eritematoso Sistémico/genética , Enfermedades de la Piel/genética , Adulto , Citocinas/metabolismo , Femenino , Humanos , Lupus Eritematoso Sistémico/metabolismo , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Enfermedades de la Piel/metabolismo , Adulto JovenRESUMEN
OBJECTIVE: To analyze cytokine gene expression in keratinocytes from patients with systemic lupus erythematosus (SLE). INTRODUCTION: Keratinocytes represent 95 percent of epidermal cells and can secrete several cytokines. METHODS: Keratinocytes were obtained by laser microdissection from 21 patients with SLE (10 discoid and 11 acute lesions) at involved and uninvolved sites. All patients were receiving a low/moderate prednisone dose and 18 were receiving chloroquine diphosphate. IL-2, IL-5, TNF-α and IFN-γ gene expression was evaluated by real-time PCR and expressed as the ratio (R) to a pool of skin samples from 12 healthy volunteers. RESULTS: Heterogeneity in cytokine gene expression was found among patients with SLE. Eighteen of 38 valid SLE samples (47 percent) presented overexpression (R>1) of at least one cytokine. Lesional skin samples tended to show higher cytokine expression than samples from uninvolved skin (p = 0.06). IL-5 and IFN-γ were the most commonly overexpressed cytokines. Samples with cytokine overexpression corresponded to more extensive and severe lesions. Prednisone dose did not differ between samples without cytokine overexpression (15.71±3.45 mg/day) and those with overexpressed cytokines (12.68±5.41 mg/day) (p = 0.216). Samples from all patients not receiving diphosphate chloroquine had at least one overexpressed cytokine. CONCLUSIONS: The heterogeneous keratinocyte cytokine gene expression reflects the complex immunological and inflammatory background in SLE. Patients with severe/extensive skin lesions showed a higher frequency of cytokine gene overexpression. Increased IFN-γ and IL-5 expression suggests that Th1 and Th2 cells are involved in SLE skin inflammation. The possibility that prednisone and antimalarial drugs may have contributed to low cytokine gene expression in some samples cannot be ruled out.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Citocinas/genética , Expresión Génica/genética , Queratinocitos/metabolismo , Lupus Eritematoso Sistémico/genética , Enfermedades de la Piel/genética , Citocinas/metabolismo , Lupus Eritematoso Sistémico/metabolismo , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Enfermedades de la Piel/metabolismoRESUMEN
Embora existam muitos estudos mostrando bons resultados com o uso do metotrexato (MTX) no tratamento da artrite reumatóide (AR), não há na literatura nenhum ensaio clínico prospectivo, randômico e controlado utilizando o MTX para tratar LES. O objetivo primário do estudo foi avaliar a capacidade do MTX em controlar atividade leve a moderada do LES. Os objetivos secundários foram avaliar a capacidade do MTX em reduzir a necessidade do corticosteróide, assim como os efeitos adversos do MTX. O estudo foi prospectivo, randômico, controlado e duplo-cego. Participaram do estudo 41 pacientes com LES, com tempo médio de doença de 82,5 meses. Vinte pacientes receberam MTX (15mg a 20mg/semana) (grupo MTX) e 21 receberam placebo (grupo PL). A dose de prednisona foi mantida, aumentada ou reduzida após o primeiro mês, de acordo com avaliação clínica e laboratorial. A dose da prednisona, escores do SLEDAI, escore da escala visual analógica para dor articular e testes laboratoriais foram anotados mensalmente. Ambos os grupos eram homogêneos e comparáveis no início do estudo quanto às manifestações clínicas e laboratoriais. Duas pacientes do grupo PL saíram do estudo por apresentar grave atividade da doença, e duas do grupo MTX foram retiradas do estudo devido a efeitos adversos (uma paciente com tuberculose pulmonar e outra com urticária e dispepsia intensa). Trinta e sete pacientes (18 MTX e 19 PL) terminaram os seis meses de estudo. Ao final do estudo, observou-se que 16 pacientes do grupo PL e somente um do grupo MTX apresentaram manifestações articulares (p<0,001). Os valores da escala analógica visual para dor articular foram significantemente maiores no grupo PL do que no grupo MTX a partir do primeiro mês de estudo. Dezesseis pacientes do grupo PL e somente um do grupo MTX apresentaram lesões cutâneas após seis meses de tratamento (p<0,001). Ao término do estudo quatro pacientes do grupo MTX e 11 do grupo PL apresentaram hipocomplementemia (p<0,001). Quando comparados os pacientes do grupo MTX e PL durante o estudo, os escores médios do SLEDAI no grupo PL foram significantemente maiores do que no grupo MTX nos meses 3, 4, 5 e 6. Ao longo do estudo foi possível reduzir a dose da prednisona em 13 pacientes do grupo MTX e somente em um paciente do grupo PL (p<0,001). Quatorze pacientes do grupo MTX (70 por cento) apresentaram efeitos adversos, principalmente síndrome dispéptica...