Schistosoma mansoni infection or associated products are able to down-modulate the type 1 CD4+ T cell inflammatory response characteristic of autoimmune diseases. In this study, we evaluated how S. mansoni antigens altered the immune response that was induced by the soluble Leishmania antigen (SLA) from cutaneous leishmaniasis (CL) patients. Cytokines were measured from the supernatants of peripheral blood mononuclear cell cultures stimulated with SLA. This was performed using the sandwich enzyme linked immunosorbent assay technique in the presence or absence of S. mansoni recombinant antigens Sm29, SmTSP-2 and PIII. The addition of S. mansoni antigens to the cultures resulted in the reduction of interferon gamma (IFN-γ) levels in 37-50% of patients. Although to a lesser extent, the antigens were also able to decrease the production of tumour necrosis factor-alpha (TNF-α). We compared patients that either had or did not have reduction in IFN-γ and TNF-α production in cultures stimulated with SLA in the presence of S. mansoni antigens. We found that there was no significant difference in the levels of interleukin (IL)-10 and IL-5 in response to S. mansoni antigens between the groups. The antigens used in this study down-modulated the in vitro proinflammatory response induced by SLA in a group of CL patients through a currently undefined mechanism.

CONTEXTO E OBJETIVO: A função da resposta imunológica e das citocinas pró-inflamatórias na patogênese da dor crônica tem tido interesse crescente. Para avaliar se há correlação entre hérnia de disco e aumento de citocinas, foi medida a concentração de citocinas em pacientes com lombalgia crônica e em indivíduos sadios. TIPO DE ESTUDO E LOCAL: Estudo transversal analítico realizado na Clínica de Dor da Universidade Federal da Bahia (UFBA). MÉTODO: As concentrações de citocinas foram medidas pela técnica de ELISA (enzyme linked immunosorbent assay) em 23 pacientes com lombalgia (G1) e 10 sadios (G2). RESULTADOS: As concentrações de fator-alfa de necrose tumoral [TNF-alpha] (G1 = 5.6 ± 2.3 pg/ml; G2 = 1.6 ± 0.5 pg/ml; P = 0,01) e interleucina-6 [IL-6] (G1 = 4.1 ± 3.0 pg/ml; G2 = 0.9 ± 0.4 pg/ml; P = 0,01) foram maiores no G1. Não houve diferença estatisticamente significante para interleucina-1 [IL-1] (G1 = 0.5 ± 0.3 pg/ml; G2 = 0.5 ± 0.1 pg/ml; P = 1) e receptor solúvel do factor de necrose tumoral [sTNF-R] (G1 = 572 pg/ml ± 36; G2 = 581 ± 50 pg/ml; P = 0,87). CONCLUSÃO: Os pacientes com lombalgia crônica por hérnia de disco apresentam concentrações maiores de TNF-alpha e IL-6, mas não de IL-1 ou sTNF-R.

CONTEXT AND OBJECTIVE: The role of immune response and proinflammatory cytokines in the pathogenesis of chronic pain has been of growing interest. In order to evaluate whether there is any association between disc herniation and elevated cytokine levels, we measured cytokine levels in patients with chronic low back pain and in healthy subjects. DESIGN AND SETTING: Analytical cross-sectional study at the Pain Clinic of Universidade Federal da Bahia (UFBA). METHODS: Cytokine levels were measured using the enzyme-linked immunosorbent assay (ELISA) technique on 23 patients with low back pain (G1) and on 10 healthy subjects (G2). RESULTS: The levels of tumor necrosis factor-alpha [TNF-alpha] (G1 = 5.6 ± 2.3 pg/ml; G2 = 1.6 ± 0.5 pg/ml; P = 0.01) and interleukin-6 [IL-6] (G1 = 4.1 ± 3.0 pg/ml; G2 = 0.9 ± 0.4 pg/ml; P = 0.01) were higher in G1. There were no statistically significant differences in relation to interleukin-1 [IL-1] (G1 = 0.5 ± 0.3 pg/ml; G2 = 0.5 ± 0.1 pg/ml; P = 1) or soluble tumor necrosis factor receptor [sTNF-R] (G1 = 572 pg/ml ± 36; G2 = 581 ± 50 pg/ml; P = 0.87). CONCLUSION: The patients with chronic low back pain due to disc herniation presented higher levels of TNF-alpha and IL-6, but not of IL-1 or sTNF-R.

JUSTIFICATIVA E OBJETIVOS: As citocinas pró-inflamatórias têm função importante na fisiopatologia das síndromes dolorosas neuropáticas. O objetivo desse estudo foi avaliar os níveis plasmáticos de citocinas pró-inflamatórias antes e após o tratamento com tramadol em pacientes com hérnia discal e síndrome do túnel do carpo e compará-los com indivíduos normais. MÉTODO: Investigou-se 38 pacientes com dor neuropática por hérnia discal ou síndrome do túnel do carpo. Todos os pacientes foram tratados com tramadol de liberação controlada (100 mg em 12h) durante 10 dias. Realizaram-se coletas de sangue venoso (5 mL), no período matutino, antes do tratamento e no 11º dia e as amostras foram armazenadas até análise (-70ºC). Foram utilizados testes enzimáticos ELISA para dosagem de citocinas plasmáticas (TNF-±, IL-1, IL-6) e receptores sTNF-R1, (R & D Systems). Realizou-se dosagem de citocinas em soro de 10 voluntários sadios. RESULTADOS: A concentração de TNF-± antes (5,8 ± 2,8 pg.mL-1) foi significativamente maior que após o tramadol (4,8 ± 2,1 pg.mL-1; p = 0,04, Teste Mann-Whitney). Não houve diferença significativa de IL-1², IL-6 e sTNF-R1 antes e após o tratamento. As concentrações plasmáticas de TNF-± (sadios: 1,4 ± 0,5; pacientes com dor: 5,8 ± 2,8 pg.mL-1; p = 0.01) e IL-6 (sadios: 1,2 ± 0,8; pacientes com dor: 3,5 ± 2,6 pg.mL-1; p = 0,01) foram significativamente maiores nos pacientes com dor neuropática que nos voluntários, Teste de Mann-Whitney. CONCLUSÕES: Nos pacientes com hérnia discal e síndrome do túnel do carpo as concentrações plasmáticas de TNF-± e IL-6 foram maiores que em voluntários sadios, não havendo diferença das concentrações de sTNF-R e IL-1². Houve redução da concentração plasmática de TNF-± após tratamento com tramadol (100 mg em 12h), mas não de IL-6, sTNF-R e IL-1².

BACKGROUND AND METHODS: Proinflammatory cytokines play an important role in the pathophysiology of neuropathic pain syndromes. The objective of this study was to evaluate plasma levels of proinflammatory cytokines before and after treatment with tramadol in patients with herniated intervertebral disks and carpal tunnel syndrome, and to compare them with normal individuals. METHODS: Thirty-eight patients with neuropathic pain secondary to herniated intervertebral disks or carpal tunnel syndrome participated in this study. All patients were treated with controlled release tramadol (100 mg every 12 hours) for 10 days. Venous blood (5 mL) was collected in the morning, before treatment and on the 11th day, and stored (-70° C) until analysis. ELISA was used to determine the plasma levels of cytokines (TNF-±, IL-1, IL-6) and receptors sTNF-R1 (R & D Systems). Plasma levels of cytokines of 10 healthy volunteers were also determined. RESULTS: The concentration of TNF-± before (5.8 ± 2.8 pg.mL-¹) was significantly higher than after treatment with tramadol (4.8 ± 2.1 pg.mL-1; p = 0.04, Mann-Whitney test). The levels of IL-1², IL-6, and sTNF-R1 before and after treatment with tramadol showed no significant differences. Plasma levels of TNF-± (healthy individuals: 1.4 ± 0.5; pain patients: 5.8 ± 2.8 pg.mL-1; p = 0.01) and IL-6 (healthy individuals: 1.2 ± 0.8; pain patients: 3.5 ± 2.6 pg.mL-1; p = 0.01) were significantly higher in patients with neuropathic pain, Mann-Whitney Test. CONCLUSIONS: In patients with herniated intervertebral disks and carpal tunnel syndrome, plasma levels of TNF-± and IL-6 were higher than in healthy volunteers, while differences in the concentrations of sTNF-R and IL-1² were not observed. Plasma levels of TNF-±, but not of IL-6, sTNF-R, and IL-1², decreased after treatment with tramadol (100 mg every 12 hours).

JUSTIFICATIVA Y OBJETIVOS: Las interleucinas proinflamatorias tienen una función importante en la fisiopatología de los síndromes dolorosos neuropáticos. El objetivo de este estudio, fue evaluar los niveles plasmáticos de interleucinas proinflamatorias antes y después del tratamiento con tramadol en pacientes con hernia de disco y síndrome del túnel del carpo, y compararlos con individuos normales. MÉTODO: Se investigaron 38 pacientes con dolor neuropático por hernia de disco o síndrome del túnel del carpo. Todos los pacientes fueron tratados con tramadol de liberación controlada (100 mg en 12h) durante 10 días. Se realizaron muestras de sangre venosa (5 mL), por la mañana, antes del tratamiento y en el 11º día, y las mismas se almacenaron para ser analizadas (-70ºC). Se utilizaron test enzimáticos ELISA para la dosificación de las interleucinas plasmáticas (TNF-±, IL-1, IL-6) y receptores sTNF-R1, (R & D Systems). Se realizó la dosificación de interleucinas en suero de 10 voluntarios sanos. RESULTADOS: La concentración de TNF-± antes (5,8 ± 2,8 pg.mL-1) fue significativamente mayor que después del tramadol (4,8 ± 2,1 pg.mL-1; p = 0,04, Test de Mann-Whitney). No hubo diferencia significativa de IL-1², IL-6 y sTNF-R1 antes y después del tratamiento. Las concentraciones plasmáticas de TNF-± (sanos: 1,4 ± 0,5; pacientes con dolor: 5,8 ± 2,8 pg.mL-1; p = 0.01) y IL-6 (sanos: 1,2 ± 0,8; pacientes con dolor: 3.5 ± 2,6 pg.mL-1; p = 0,01) fueron significativamente mayores en los pacientes con dolor neuropático que en los voluntarios, test de Mann-Whitney. CONCLUSIONES: En los pacientes con hernia discal y síndrome del túnel del carpo, las concentraciones plasmáticas de TNF-± y IL-6, fueron más elevadas que en los voluntarios sanos, no habiendo ninguna diferencia en las concentraciones de sTNF-R y IL-1². Hubo una reducción de la concentración plasmática de TNF-±; después del tratamiento con tramadol (100 mg en 12h), pero no de IL-6 sTNF-R y IL-1².

A candidíase recorrente cutânea ou mucosa é caracterizada pela ocorrência de, no mínimo, 4 episódios de candidíase no período de um ano. Não são conhecidos os fatores que levam à recorrência desta infecção. O presente estudo avaliou a resposta linfoproliferativa e a produção de IFN-g de pacientes com candidíase recorrente. Os índices de estimulação da resposta linfoproliferativa em culturas de células de pacientes com candidíase recorrente estimuladas com antígeno de Candida albicans, PPD e TT foram respectivamente de 6±8, 17±20 e 65±30. A adição de anticorpo monoclonal anti-IL-10 às culturas de células de 6 pacientes aumentou a resposta linfoproliferativa de 735±415 para 4143±1746 cpm. A produção de IFN-g em culturas de células estimuladas com antígeno de Candida, foi 162±345pg/ml. Pacientes com candidíase recorrente apresentam uma deficiência na resposta linfoproliferativa e na produção de IFN-g, podendo a resposta imune celular ao antígeno de Candida ser restaurada parcialmente através da neutralização da IL-10 in vitro.

Recurrent cutaneous or mucosal candidiasis is characterized by the occurrence of at least four candidiasis episodes within a one-year period. The factors involved in recurrence of infection are still unknown. In the present study the lymphoproliferative response and the IFN-g production by candidiasis patients were evaluated. The stimulation index of mononuclear cell cultures of candidiasis patients stimulated with Candida albicans antigen, PPD and TT were 6±8, 17±20 and 65±30, respectively. The addition of monoclonal antibody anti-IL-10 to Candida albicans antigen stimulated cultures raised the lymphoproliferative response from 735±415 to 4143±1746 cpm. The IFN-g production by cells of candidiasis patients stimulated with Candida albicans antigen was 162±345pg/ml. Candidiasis patients have an impairment in the lymphoproliferative response specific to C. albicans antigen and on IFN-g production and the lymphoproliferative response can be partially restored, in vitro, by IL-10 neutralization.

Para avaliar se os níveis séricos de IFN-gama e TNF-alfa podem ser usados como marcadores da resposta terapêutica na leishmaniose cutânea, 54 pacientes com história de uma única lesão ulcerada com duração de até 30 dias foram arrolados para o estudo. IFN-gama e TNF-alfa foram mensurados por ELISA no sobrenadante de cultura de linfócitos, antes e 60 dias após início da terapia. Cicatrização completa da lesão 60 dias após o tratamento foi considerada critério de cura. Os níveis de IFN-gama e TNF-alfa foram similares em ambos os grupos antes da terapia. Houve tendência de aumento dos níveis de IFN-gama nos pacientes curados aos 60 dias, os valores não alcançam significância estatística. Em ambos os grupos de pacientes, os níveis iniciais de TNF-alfa foram semelhantes antes da terapia e caíram significantemente após tratamento independente da cura ou manutenção da lesão ativa.

To evaluate if IFN-gamma and TNF-alpha levels could be used as markers of therapeutic response in cutaneous leishmaniasis, 54 patients with history of one ulcerated cutaneous lesion, with up to 30 days onset, were enrolled in the study. IFN-gammaand TNF-alpha were measured by ELISA in lymphocyte cultures supernatant before and 60 days after initiating therapy. Cure was considered to be a complete healing of lesion 60 days after treatment. IFN-gamma and TNF-alpha levels were similar in both groups of patients before therapy. There was a tendency to increase IFN-gamma levels in patients that were cured in 60 days, however the values did not reach statistical significance. In both groups of patients, TNF-alpha levels were similar before therapy and fell significantly after treatment, irrespective of cure or maintenance of active lesion.

O teste de Montenegro, utilizado no diagnóstico da leishmaniose tegumentar, tem sido recentemente considerado marcador de imunogenicidade após vacinação. Neste estudo, avaliou-se o poder sensibilizante deste teste e a produção de y-IFN in vitro, em indivíduos não expostos a Leishmania. Utilizaram-se para o teste antígenos de L. amazonensis produzido em nosso laboratório (grupo I) ou produzido pela FIOCRUZ-RJ (grupo II). No dia 30, 33% dos indivíduos do grupo I e 42% do grupo II converteram o teste para positivo, comparando-se com 67% do grupo I e 50% do grupo II no dia 90. y-IFN foi detectado em 56% dos indivíduos do grupo I e 17% do grupo II no dia 30 (169±309 e 11±36pg/ml) e em 67% do grupo I e 58% do grupo II no dia 360 (69±107 e 18±20pg/ml). Estes dados demonstram que o teste de Montenegro induz, além de hipersensibilidade tardia, a produção de y-IFN.

The Montenegro skin test, used to diagnose cutaneous leishmaniasis, is now being considered to detect immunogenicity after vaccination. In this study, we evaluated the ability of this test to induce immune response and IFN-g production in subjects not previously exposed to Leishmania. The Montenegro skin test was performed using antigens of L. amazonensis produced by our laboratory (group I) or by FIOCRU-RJ (group II). At day 30, 33% of the subjects from group I and 42% from group II were positive, compared to 67% from group I and 50% from group II at day 90. IFN-y was detected in 56 % of subjects from group I and 17% from group II at day 30 (169±309 and 11±36pg/ml) and in 67% from group I and 58% from group II by day 360 (69±107 and 18±20pg/ml). These data demonstrate that the Montenegro skin test induces not only a delayed hypersensitivity reaction, but also IFN-y production.

In areas of Leishmania chagasi transmission the ability to control leishmania infection is associated with IFN-g production. In visceral leishmaniasis down-regulation of T cell responses is mediated by interleukin-10 (IL-10). In this study we evaluated the lymphoproliferative response, IFN-g and IL-10 production on lymphocyte cultures stimulated with recombinant leishmania antigens in subjects with asymptomatic L. chagasi infection. There was a statistically significant difference in the lymphoproliferative response of the subjects with asymptomatic infection as compared to patients with visceral leishmaniasis and healthy subjects with respect to crude antigens (p<0.01), gp-63 (p<0.05) and hsp-70 (p<0.01), as well as between asymptomatic L. chagasi infected subjects and patients with visceral leishmaniasis with respect to the response to all antigens tested. The IFN-g production observed in the group with asymptomatic infection with all the three recombinant antigens tested was higher (p<0.01) than that observed in patients with visceral leishmaniasis and in healthy subjects. Furthermore, in individuals with asymptomatic infection, IL-10 levels in cultures stimulated with recombinant antigens were very low. This study shows that lymphocytes from individuals with asymptomatic L. chagasi infection are able to recognize recombinant leishmania antigens with production of a cytokine that is associated with leishmania killing.