Avaliação da expressão de interleucina 1 beta (IL-1β) e antagonista do receptor de interleucina 1 (IL-1Ra) em pacientes com hanseníase

A hanseníase é uma doença infectocontagiosa espectral que acompanha-se por uma série de eventos imunológicos desencadeados pela resposta do hospedeiro frente ao agente etiológico, o Mycobacterium leprae. Evidências sugerem que a indução e manutenção da resposta imune/inflamatória na hanseníase estão vinculadas a interações de múltiplas células e fatores solúveis, particularmente através da ação de citocinas. Nesse estudo, foram mensurados níveis de IL-1β e IL-1Ra de 37 casos novos de hanseníase acompanhados ao longo do tratamento e 30 controles sadios pelo teste ELISA. A coleta de sangue periférico foi realizada em quatro tempos para os casos de hanseníase (pré-tratamento com PQT, 2ª dose, 6ª dose e pós-PQT) e em único momento para os controles. Na comparação dos níveis das moléculas de casos no pré-PQT e controles, houve diferença estatisticamente significativa somente para IL-1β. Nossos resultados sugerem a participação dessa citocina no processo imune/inflamatório.

Aplicação de sistemas moleculares na detecção rápida de MDR-TB e XDR-TB

Mycobacterium tuberculosis é o agente etiológico da tuberculose em humanos e segundo as estimativas da Organização Mundial da Saúde, um terço da população mundial está infectada com esta bactéria, calculando-se que no ano de 2008 aproximadamente 9,4 milhões de pessoas contraíram tuberculose activa. Associada a esta tendência, encontra-se o aumento alarmante da incidência de tuberculose resistente aos antibióticos, mais propriamente da tuberculose multirresistente e extensivamente resistente. Nesta Dissertação estudaram-se três sistemas de detecção molecular (INNO-LiPA Rif. TB, Innogenetics, Ghent, Bélgica, MTBDRplus e MTBDRsl, GenoType, GmbH, Nehren, Alemanha), que permitem detectar o complexo M. tuberculosis e as mutações mais comuns associadas à resistência aos antibióticos de primeira e segunda linha. Para o efeito, na primeira parte do trabalho os três sistemas em estudo foram testados em 21 isolados clínicos pertencentes à colecção do Laboratório de Micobactérias do Instituto de Higiene e Medicina Tropical (IHMT, UNL), com o propósito de avaliar a sua capacidade para identificar o complexo M. tuberculosis e detectar mutações ligadas à resistência aos fármacos de primeira e segunda linha. Na segunda parte do trabalho, os sistemas INNO-LiPA Rif. TB e MTBDRplus foram testados em 33 amostras respiratórias com baciloscopia positiva, com o propósito de aferir a “performance” destes sistemas para a detecção directa de tuberculose multirresistente em amostras respiratórias. Na primeira parte do trabalho os três sistemas em estudo apresentaram elevada sensibilidade na identificação do complexo M. tuberculosis em culturas, bem como na detecção de mutações ligadas à resistência aos antibióticos de primeira linha, com excepção do etambutol. No que diz respeito à detecção da resistência aos antibióticos de segunda linha, não foi possível calcular os valores de sensibilidade e especificidade. Na segunda parte do trabalho, o INNO-LiPA Rif. TB demonstrou ser o sistema mais robusto para a análise directa de amostras respiratórias com baciloscopia positiva, para um diagnóstico precoce de tuberculose e detecção de resistência à rifampicina. O MTBDRplus não se mostrou uma alternativa viável ao INNO-LiPA Rif. TB, pois apresentou baixa sensibilidade para a identificação do complexo M. tuberculosis e vários problemas no passo de amplificação. O MTBDRsl não foi testado, por não ter sido detectada nenhuma amostra multirresistente.

Identificação laboratorial de micobactérias em amostras respiratórias de pacientes HIV-positivos com suspeita de tuberculose

Foram analisados retrospectivamente os registros (2000 a 2004) do Laboratório de Microbiologia do Instituto Adolfo Lutz de Santos, SP referentes a pacientes infectados pelo virus da imunodeficiência humana com suspeita de tuberculose pulmonar. Foram encaminhadas 1.321 amostras com finalidade de diagnóstico, correspondendo a 880 casos suspeitos de tuberculose em 693 pacientes. Cento e trinta e quatro baciloscopias foram positivas e em 188 culturas houve crescimento de micobactérias, correspondendo a 161 casos confirmados. Houve identificação de Mycobacterium tuberculosis em 126 (78,3%) e micobactérias não tuberculosas em 39 (24,2%). Em quatro casos, houve concomitância de Mycobacterium tuberculosis e micobactérias não tuberculosas (porém em amostras distintas). O perfil de sensibilidade às drogas antituberculose revelou 18 (14,3%) casos de resistência a pelo menos um medicamento. Estes resultados reforçam a necessidade de submeter à rotina laboratorial completa - baciloscopia, cultura com identificação e testes de sensibilidade às drogas - as amostras respiratórias de pacientes soropositivos para o vírus da imunodeficiência humana com suspeita de tuberculose para direcionamento terapêutico adequado.

HTLV-1 associated myelopathy diagnosed during lepromatous leprosy reaction treatment: a case report

Leprosy and human T cell lymphotropic virus type 1 infection are prevalent in Brazil. Coinfection by Mycobacterium leprae and HTLV-1 is reviewed and a case is reported. A 59 year-old woman was followed and HTLV-1 associated myelopathy was diagnosed during leprosy treatment. The clinical and neurological aspects of this unusual association were initially reviewed. Immunological markers and the possible prognoses due to the association of the diseases were discussed. The unexpected association of leprosy and HTLV-1 associated myelopathy may occur in endemic areas and causes difficulties in determining the correct diagnosis and adequate management of the neurological manifestations.

Estudo clínico-radiológico de crianças e adolescentes indígenas Suruí, Região Amazônica

INTRODUÇÃO: Entre os Suruí de Rondônia foram registradas incidências médias de TB > 2.500/100.000 habitantes, entre 1991-2002. Aproximadamente 50% desses casos foram notificados em < 15 anos. MÉTODOS: Trata-se de um estudo clínico-epidemiológico que teve como objetivo descrever as características clínico-radiológicas em crianças e adolescentes identificados como contatos de doentes de TB. Além disto, aplicar o sistema de pontuação para o diagnóstico de TB na infância e verificar se as condutas adotadas no nível local foram concordantes com as diretrizes nacionais. RESULTADOS: Foram analisados 52 Rx de 37 indígenas. Deste conjunto, 48,1% foram normais e 51,9% anormais. Alguns dos Rx apresentaram duas ou mais alterações, totalizando 36 eventos independentes. Observou-se infiltrados (38,9%), calcificações (38,9%), cavitações (11,1%) e atelectasias/derrame pleural (11,1%). Nas imagens anormais, 22,2% eram TB provavelmente ativa e 33,3% sequelas. A confrontação com as diretrizes constatou 52,6% de condutas discordantes. CONCLUSÕES: A presença da infecção tuberculosa latente (ITBL) e TB ativa, entre crianças e adolescentes, são indicadores de transmissão ativa e continuada do Mycobacterium tuberculosis. Os Rx mostrando alta frequência de infiltrados e calcificações é compatível com primo-infecção em idade precoce. Entretanto, essas alterações não são diferentes daquelas observadas entre outros grupos, não sugerindo comprometimento imunológico. As discordâncias apontadas indicam que o momento ideal para o tratamento da ITBL passou despercebido. Conclui-se que é fundamental a utilização do sistema de pontuação para o correto diagnóstico de TB na infância, assim como a realização de baciloscopia e cultura de escarro em adolescentes capazes de expectorar.

Class-I human leukocyte alleles in leprosy patients from Southern Brazil

INTRODUCTION: The present study was designed to investigate a possible role of HLA (histocompatibility leucocyte antigen) class-I alleles (HLA-A, -B, and -C) in leprosy patients from Southern Brazil. METHODS: Two hundred and twenty-five patients with leprosy and 450 individuals for the control group were involved in this research. HLA genotyping was performed through PCR-SSO protocols (One Lambda, USA); the frequency of these alleles was calculated in each group by direct counting, and the frequencies were then compared. RESULTS: There was an association between HLA-A*11 (6.9% vs 4.1%, p=0.0345, OR=1.72, 95% CI=1.05-2.81), HLA-B*38 (2.7% vs. 1.1%, p=0.0402, OR=2.44, 95% CI=1.05-5.69), HLA-C*12 (9.4% vs. 5.4%, p=0.01, OR=1.82, 95% CI=1.17-2.82), and HLA-C*16 (3.1% vs. 6.5%, p=0.0124, OR=0.47, 95% CI=0.26-0.85) and leprosy per se. In addition, HLA-B*35, HLA-C*04, and HLA-C*07 frequencies were different between lepromatous (LL) and tuberculoid (TT) patients. However, after adjusting for the number of alleles compared, Pc values became nonsignificant. CONCLUSIONS: Although our results do not support the previous findings that HLA class-I alleles play a role in leprosy pathogenesis, we suggest new studies because of the importance of the association between the HLA and KIR in the innate immune response to leprosy.

Epidemiological, clinical, and operational aspects of leprosy patients assisted at a referral service in the state of Maranhão, Brazil

INTRODUCTION: Leprosy is an infectious disease caused by Mycobacterium leprae. The aim of this study was to describe the epidemiological, clinical, and operational aspects of leprosy carriers. METHODS: A cross-sectional study leprosy patients assisted in São Luis, MA, was performed. RESULTS: Of the 85 cases analyzed, 51.7% were male participants, and 60% were brown. Concerning the age, 54.8% of women were between 35 and 49 years, and 57.6% of men were between 20 and 34 years. Lepromatous leprosy was found in 42.3% of cases, and the multibacillary form was found in 72.9%. The skin smear was positive in 42.3%. The occurrence of reaction was found in 43.5% of cases, and 83.5% had no Bacillus Calmette-Guérin scar. Leprosy in the family was reported by 44.7% of the patients. Most of the individuals (96.4%) lived in houses made of brick with more than three rooms (72.6%) and two persons per room (65.1%). Concerning the level of education, 41.4% of women and 34.1% of men had more than one to three years of education. The most evaluated age group in the beginning of the treatment was that of 35 to 49 years with a Grade 0 incapability (64.5%), and that in the end was the age group of 20 to 34 (29.9%) with Grade 0, 30.7% Grade 1, and 11.5% Grade 2. CONCLUSIONS: The frequency of multibacillary forms found in this study and the cases in family members point out delayed diagnoses. Thus, early diagnosis and appropriate treatment are important in decreasing the outcome of disabilities.

Abnormal spontaneous interleukin 8 receptor expression: a brief report of two cases

Interleukin 8 (CXCL8) is an autocrine chemokine specific for the chemoattraction and activation of granulocytes, NKT cells and T lymphocytes. Patients with tuberculosis and latent Mycobacterium tuberculosis infection were assessed for the spontaneous expression of CXCR1 (CD128) and CXCR2 on lymphocytes and monocytes. Compared with ex vivo profiles, increased spontaneous CXCR2 expression and normal CXCR1 expression were found on lymphocytes in two out of 59 individuals. Monocytes showed normal ex vivo profiles for both receptors. After stimulation with purified protein derivative, the in vitro levels of CXCL8 were below the median levels of all patients with prior tuberculosis. Spontaneous CXCR2 modulation did not cause notable variation in the in vitro levels of CXCL8.

Improving tuberculosis control through the partnership between university and the health system

INTRODUCTION: Tuberculosis (TB) control is linked to the availability of qualified methods for microbiological diagnostics; however, microscopy with limited sensitivity is the only method available in many locations. The objective of this study was to evaluate the introduction of culture, drug susceptibility testing (DST), and genotyping in the routine of a Municipal Program of Tuberculosis Control. METHODS: Direct microscopy of sputum and culture in Ogawa-Kudoh were performed on 1,636 samples from 787 patients. DST of positive cultures was performed by resazurin microtiter assay and genotyping by mycobacterial interspersed repetitive units-variable number tandem repeat. RESULTS: A total 91 patients with TB were identified. The culture increased case detection by 32% compared with the microscopy; acquired resistance was 3.3% and the genotyping showed high genetic diversity. CONCLUSIONS: Ogawa-Kudoh contributed significantly to the increase in case detection and is suitable for implementation in poor-resource locations. The acquired resistance rate was lower than that reported in a recent Brazilian survey. The high genetic diversity is possibly related to the high TB prevalence in the population, as well as to early detection and suitable treatment of patients. The interaction between research and health care is important for reorienting the practice, transferring technology, and improving TB control.

Oral coinfection can stress peripheral lymphocyte to inflammatory activity in leprosy

INTRODUCTION: This study evaluated the intracellular profile of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-γ (IFN-γ) in peripheral blood mononuclear cells (PBMCs) from leprosy patients based on oral infections presence to determine whether these coinfections could be associated with pro-inflammatory activity in leprosy. METHODS: Leprosy patients regardless of clinical form and specific leprosy treatment (n=38) were divided into two groups: Group I - leprosy patients with oral infections (n=19), and Group II - leprosy patients without oral infections (n=19). Non-leprosy patients presenting oral infections were assigned to the control Group (n=10). Intracellular IL-2, IL-4, IL-10 and IFN-γ production was evaluated by flow cytometry (FACS) before and 7 days after controlling the oral infection in the Group I, before and 7 days after dental prophylaxis in the Group II, and during oral infection process in control Group. RESULTS: Low percentages of CD3+ lymphocytes bearing IL-2, IL-10 and IFN-γ were observed in the Group I and Group II at baseline and 7 days after therapy or prophylaxis compared to controls. Group I showed reduced percentages of IL-4 at baseline and 7 days after therapy compared to controls, or at baseline of Group II, and the Group II showed reduced percentages of CD3+ cells bearing IL-4 compared to control. An increase of the percentages of CD3+cells bearing IL-4 was observed in the Group I after the oral infections treatment. CONCLUSIONS: The occurrence of oral infections favors the intracellular cytokines expression and, probably, the inflammatory reaction operating as a stimulatory signal triggering the leprosy reactions.

Bone tuberculosis: a case report on child

The authors report a case of a 12-year-old child with a complaint of pain and deformity in the lower thoracic region that had lasted for two years. Clinical, epidemiological and laboratory characteristics associated with images of apparent damage in the T9-T10 and T11-T12 vertebrae taken by radiography of the thoracic spine and nuclear magnetic resonance in addition to the positivity of the molecular test based on the polymerase chain reaction, led to tuberculous spondylitis being diagnosed and specific therapy was started. Culture of vertebral biopsy was positive for Mycobacterium tuberculosis after thirty days.

Rapid detection and differentiation of mycobacterial species using a multiplex PCR system

Introduction The early diagnosis of mycobacterial infections is a critical step for initiating treatment and curing the patient. Molecular analytical methods have led to considerable improvements in the speed and accuracy of mycobacteria detection. Methods The purpose of this study was to evaluate a multiplex polymerase chain reaction system using mycobacterial strains as an auxiliary tool in the differential diagnosis of tuberculosis and diseases caused by nontuberculous mycobacteria (NTM) Results Forty mycobacterial strains isolated from pulmonary and extrapulmonary origin specimens from 37 patients diagnosed with tuberculosis were processed. Using phenotypic and biochemical characteristics of the 40 mycobacteria isolated in LJ medium, 57.5% (n=23) were characterized as the Mycobacterium tuberculosis complex (MTBC) and 20% (n=8) as nontuberculous mycobacteria (NTM), with 22.5% (n=9) of the results being inconclusive. When the results of the phenotypic and biochemical tests in 30 strains of mycobacteria were compared with the results of the multiplex PCR, there was 100% concordance in the identification of the MTBC and NTM species, respectively. A total of 32.5% (n=13) of the samples in multiplex PCR exhibited a molecular pattern consistent with NTM, thus disagreeing with the final diagnosis from the attending physician. Conclusions Multiplex PCR can be used as a differential method for determining TB infections caused by NTM a valuable tool in reducing the time necessary to make clinical diagnoses and begin treatment. It is also useful for identifying species that were previously not identifiable using conventional biochemical and phenotypic techniques.

The performance of an in-house nested-PCR technique for pleural tuberculosis diagnoses

Introduction This study evaluated the performance of an in-house nested-PCR system for the detection of the Mycobacterium tuberculosis complex in pleural fluid, blood and urine samples from pleural effusion tuberculosis patients by health services physicians in Pernambuco, Brazil. Methods A prospective double-blind study with 37 hospitalized patients of both sexes, aged over 15, was used to investigate the diagnosis of pleural effusion. The criteria used to define the cases included the demonstration of bacillus in biological samples by smear or culture or by a granulomatous finding in the histopathological examination, associated with an evident response to specific treatments to each clinical situation. Pleural fluid, blood and urine samples were collected and subjected to routine tests and the nested PCR technique to assess for M. tuberculosis amplification. Results In total, 37 pleural effusion patients took part in the study, of whom 19 (51.3%) had tubercular etiologies and 18 (48.7%) had etiologies from other causes. When the pleural fluid, blood and/or urine sample in-house nested-PCR sensitivities were evaluated simultaneously, the results were positive regardless of the biological specimen (the sensitivity was 84.2%); however, when the blood and/or urine samples were analyzed together, the sensitivity was 72.2%. When the pleural fluid samples were evaluated alone, the sensitivity was only 33.3%. Conclusions The performance of the diagnostic pleural tuberculosis nested-PCR was directly related to the diversity of the samples collected from the same patient. Additionally, this study may identify a need to prioritize non-invasive blood and urine collection for this diagnosis.

Antioxidant factors, nitric oxide levels, and cellular damage in leprosy patients

Introduction The immune response caused by Mycobacterium leprae is a risk factor for the development of oxidative stress (OS) in leprosy patients. This study aimed to assess OS in leprosy patients before the use of a multidrug therapy. Methods We evaluated the nitric oxide (NO) concentration; antioxidant capacity; levels of malondialdehyde, methemoglobin and reduced glutathione; and the activity of catalase and superoxide dismutase (SOD) in leprosy patients. Results We observed lower SOD activity in these leprosy patients; however, the NO levels and antioxidant capacity were increased. Conclusions The infectious process in response to M. leprae could primarily be responsible for the OS observed in these patients.

Factors associated with neural alterations and physical disabilities in patients with leprosy in São Luis, State of Maranhão, Brazil

Introduction Leprosy is a chronic infectious disease that is caused by Mycobacterium leprae. The objective of this study was to evaluate the risk factors that are associated with neural alterations and physical disabilities in leprosy patients at the time of diagnosis. Methods A prospective cross-sectional study was conducted on 155 leprosy patients who participated in a program that aimed to eliminate leprosy from São Luis, State of Maranhão. Results Patients who were 31-45 years of age, were older than 60 years of age or had a partner were more likely to have a disability. Patients with partners were 1.14 times more likely (p = 0.025) to have disabilities of the hands. The frequency of disabilities in the feet among the patients with different clinical forms of leprosy was statistically significant. Conclusions The identification of risk factors that are associated with neural alterations and physical disabilities in leprosy patients is important for diagnosing the disease because this approach enables physicians to plan and prioritize actions for the treatment and monitoring of patients.