Different immunosuppressive mechanisms in multi-drug-resistant tuberculosis and non-tuberculous mycobacteria patients.

Previous studies have demonstrated that cells from both multi-drug-resistant tuberculosis (MDR-TB) and non-tuberculous mycobacteria (NTM) patients respond poorly to mycobacterial antigens in vitro. In the present study, we compared the in vitro response of cells isolated from sensitive TB (NR-TB)-, MDR-TB- and NTM-infected patients. Analysis of T cell phenotype ex vivo revealed that both MDR-TB and NTM patients present an increased percentage of CD4(+) CD25(+-) forkhead box protein 3 (FoxP3)(+) and CD4(+) CD25(+) CD127(-) regulatory T (T(reg) ) cells when compared to NR-TB. Increased numbers of T(reg) cells and interleukin (IL)-10 serum levels were detected in MDR-TB, whereas elevated serum transforming growth factor (TGF)-ß was found in the NTM group. Cells of MDR-TB patients stimulated with early secretory antigenic target (ESAT)-6, but not purified protein derivative (PPD), showed a lower frequency of CD4(+) /interferon (IFN)-γ(+) T cells and enhanced CD4(+) CD25(+) FoxP3(+) , CD4(+) CD25(+) CD127(-) and CD4(+) CD25(+) IL-10(+) T cell population. In addition, increased IL-10 secretion was observed in cultured MDR-TB cells following ESAT-6 stimulation, but not in NR-TB or NTM patients. In vitro blockade of IL-10 or IL-10Rα decreased the CD4(+) CD25(+) FoxP3(+) frequencies induced by ESAT-6 in MDR-TB, suggesting a role of IL-10 on impaired IFN-γ responses seen in MDR-TB. Depletion of CD4(+) CD25(+) T lymphocytes restored the capacity of MDR-TB T cells to respond to ESAT-6 in vitro, which suggests a potential role for T(reg) /T regulatory 1 cells in the pathogenesis of MDR-TB. Together, our results indicate that although the similarities in chronicity, NTM- and MDR-TB-impaired antigenic responses involve different mechanisms.

Long-term culture of multibacillary leprosy macrophages isolated from skin lesions: a new model to study Mycobacterium leprae-human cell interaction.

BACKGROUND: Leprosy is characterized by a disease spectrum having two polar clinical forms dependent on the presence or not of cell-mediated immunity. In the tuberculoid forms, granuloma-activated macrophages kill Mycobacterium leprae in conjunction with a Th1 response while, in multibacillary (MB) lesions, M. leprae nonactivated macrophages infiltrate the nerves and internal organs together with a Th2 response. The functional properties and activation pathways of macrophages isolated from patients with MB leprosy remain only partially understood. OBJECTIVES: To establish an ex vivo methodology capable of evaluating the activation pathways, grade and fate of cultured macrophages isolated from MB lesions. METHODS: Skin biopsies from patients with borderline tuberculoid, bordeline lepromatous and lepromatous leprosy (LL) were characterized by immunohistochemistry and transcriptional analysis. To isolate inflammatory cells, a portion of the samples was submitted to enzymatic digestion. These same cells, maintained in culture for a minimum 7-day period, were characterized morphologically and via flow cytometry at different culture time points. Cytokine [interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-10] mRNA levels were quantified by real-time polymerase chain reaction and protein secretion in the culture supernatants was measured by enzyme-linked immunosorbent assay and the nitric oxide levels by Griess reagent. RESULTS: RNA expression in tuberculoid and MB lesions showed the profile expected of characteristic Th1 and Th2 responses, respectively. The inflammatory cells in all biopsies were successfully isolated. Although the number of cells varied between biopsies, it was highest in LL biopsies. The frequency of isolated CD14+ and CD3+ cells measured by flow cytometry correlated with the percentages of macrophages and lymphocytes in the lesions. Throughout the culture period, CD68+ macrophages showed morphological changes. A progressive increase in cell number and reduction of infected cells were perceptible in the cultures. In contrast to the biopsies, TNF-alpha, IFN-gamma and IL-10 expression in the tuberculoid and MB leprosy cells in 24-h culture and the cytokine levels in the supernatants did not differ significantly. During the culture period, cytokine expression in the MB cells progressively declined, whereas, from days 1 to 7, nitrite levels progressively increased. After day 40, the remaining macrophages were able to ingest fluorescein isothiocyanate-labelled M. leprae. These data need to be confirmed. CONCLUSIONS: This study confirmed the feasibility of obtaining ex vivo macrophages from leprosy lesions and keeping them in long-term culture. This procedure may open new pathways to studying the interaction between M. leprae and human macrophages, which might, in turn, lead to the development of therapeutic tools capable of overcoming the specific anergy found in patients with MB leprosy.

Prevalência de teníase no município de Pedra Branca Estado do Ceará, Brasil / Prevalence of taeniasis in municipal district Pedra Branca of Ceará State, Brazil

A teníase é reconhecida como importante problema de saúde pública em regiões onde as condições sanitárias são precárias e em países industrializados que recebem imigrantes de regiões onde a infecção é endêmica. O homem, responsável pela manutenção do ciclo parasitário, é o único hospedeiro da forma adulta dos parasitos Taenia solium e Taenia saginata. Apesar da importância do porco no ciclo de vida da Taenia solium, pouca atenção é dada em relação à infecção suína e humana. Estudo epidemiológico realizado no município de Pedra Branca no período de 1997 a 1998 demonstrou alto índice de carne de porco com cisticercose. O objetivo do presente estudo foi investigar a prevalência de teníase na população de Pedra Branca a fim de identificar os casos de teníase. Para isso empregou-se o método de Kato modificado por Katz por ser de fácil execução e de nos fornecer preparações quase permanentes. Nesse estudo, foram analisadas 831 amostras, de indivíduos de ambos os sexos, na faixa etária de 02 a 70 anos, com 54 (6,5%) de positivas. Os resultados mostraram a prevalência de diferentes espécies de helmintos, onde a maior freqüência foi de Ascaris lumbricoides 19 (2,30%), seguido de ancilostomideos 12 (1,44%) e em terceiro de Taenia sp 10 (1,20%). A prevalência de teníase foi bastante significativa levando-se em consideração que o método utilizado não é específico para o diagnóstico de teníase.

Taeniasis are recognized as important public health problem in many developed countries for regions with bad sanitary conditions as well as for industrial countries in the cases of emigration from the regions with endemic infection. Human intestin is responsible for realization of the parasite cycle and is unique accommodation for adult parasite form: Taenia solium and Taenia saginata In spite of importance of pigs in the life cycle of Taenia solium, it was small attention to relation between human and pig infection. Epidemiological study accomplished in Pedra Branca in the period 1997–1998 demonstrated the high index of cysticercosis contamination of pork. The purpose of the present study was for measuring the prevalence of human taeniasis on the Pedra Branca population. For this it was used the Kato method, modificated by Katz in order to be easy in use and to supply the preparations quasi permanent. In this study were analyzed 831 samples of stool of persons of both sexes between 2 and 70 year, with 54 (6.5%) of positives. The results show prevalent of different types of helminths, where Ascaris lumbricoides 19 (2.30%) has the biggest index, second was ancilostomideos 12 (1.44%) and third Taenia sp 10 (1.20%). The prevalence of taeniasis was significant considering that the used method was not specific for it diagnostic.

Anti-inflammatory drugs block cytokine mRNA accumulation in the skin and improve the clinical condition of reactional leprosy patients.

The aim of this study was to investigate in what ways in vivo anti-inflammatory treatment affects cytokine mRNA expression in situ in both erythema nodosum leprosum and reversal reaction patients. Serial biopsies were collected from the patients undergoing leprosy reactions before and during pentoxifylline (n = 7) or thalidomide (n = 3) treatment for erythema nodosum leprosum and prednisone (n = 3) for reversal reaction. Clinical evolution of the skin lesion was assessed during the study and semiquantitative reverse transcription-polymerase chain reaction was used to investigate cytokine mRNA expression at the lesion site. Results showed expression of interferon-gamma, interleukin-6, interleukin-10, interleukin-12 p40, and tumor necrosis factor-alpha in all patients tested at the onset of reactional episodes, but interleukin-4 mRNA was rarely detected in the lesions (n = 4). Follow-up analysis showed that, irrespective of the drugs used, tumor necrosis factor-alpha mRNA was diminished in 10 of the 13 patients tested. A concomitant decrease of mRNA accumulation was also observed for interferon-gamma (nine of 11 patients), interleukin-6 (nine of 11), and interleukin-12 p40 (six of eight). An inhibitory effect on interleukin-10 mRNA was likewise seen after thalidomide and pentoxifylline, but not subsequent to prednisone treatment. The data also demonstrated that cytokine mRNA inhibition correlates to the resolution of the inflammatory response in situ (n = 10), whereas the persistence/enhancement of cytokine message expression after treatment was associated with worsening of the skin condition, as seen in three erythema nodosum leprosum patients whose maintenance of local inflammation was accompanied by the appearance/persistence of interleukin-4 gene expression in situ subsequent to anti-inflammatory treatment. In summary, the participation of cytokines in leprosy inflammatory episodes seems to be directly associated with the patients' clinical evolution following therapy for reaction.

[Expectations of hospitalized patients as to their relationship with the nursing team]. / Expectativas dos clientes hospitalizados frente ao relacionamento com a equipe de enfermagem.

This research has been made in infectious-parasitary disease sector of a public university hospital in Rio de Janeiro on the first semester of 1995. The logbook research has been introduced to 5 adult clients, from both sexes, who had been interned in the construction of two categories, which are: a) Category I: Feelings and expectations of the client and; b) Category II: Communication and relationship between client and nurse. The results found in the first category were: 1) Lack of affection; 2) Affection, fear and anxiety; 3) Loneliness and depression; 4) Reflection; 5) Insecurity; 6) Worry about the equipment, to be cut, to be impersonally treated, lose self-control and physical dependence. The results found in the second category were: 1) Doubts and interests in information about their disease, its evolution and how long they will stay in hospital: 2) Opportunity to express feelings and ideas; 3) Comprehension of the message given by the nurse. We emphasize the increase of interchange of information, ideas, beliefs, feelings and acts in nurse-client interaction to develop the therapeutic relationship aiming at helping the client to fulfill his basic needs.