Accuracy of mucocutaneous leishmaniasis diagnosis using polymerase chain reaction: systematic literature review and meta-analysis.

The diagnosis of mucocutaneous leishmaniasis (MCL) is hampered by the absence of a gold standard. An accurate diagnosis is essential because of the high toxicity of the medications for the disease. This study aimed to assess the ability of polymerase chain reaction (PCR) to identify MCL and to compare these results with clinical research recently published by the authors. A systematic literature review based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses: the PRISMA Statement was performed using comprehensive search criteria and communication with the authors. A meta-analysis considering the estimates of the univariate and bivariate models was performed. Specificity near 100% was common among the papers. The primary reason for accuracy differences was sensitivity. The meta-analysis, which was only possible for PCR samples of lesion fragments, revealed a sensitivity of 71% [95% confidence interval (CI) = 0.59; 0.81] and a specificity of 93% (95% CI = 0.83; 0.98) in the bivariate model. The search for measures that could increase the sensitivity of PCR should be encouraged. The quality of the collected material and the optimisation of the amplification of genetic material should be prioritised.

LACK-specific CD4(+) T cells that induce gamma interferon production in patients with localized cutaneous leishmaniasis during an early stage of infection.

The profile of cytokines induced by soluble leishmania antigen (SLA) and the Leishmania homologue of the mammalian receptor for activated C kinase (LACK), a candidate vaccine against leishmaniasis, and the cellular source of the cytokines produced in response to these antigens were analyzed in patients infected with Leishmania guyanensis. Gamma interferon (IFN-gamma) and interleukin-10 (IL-10) were produced in response to LACK. Although LACK-specific CD4(+) cells producing IFN-gamma were isolated only during the early phase of infection (less than 30 days following the onset of infection), cells producing IL-10 in response to LACK were detected in all patients. CD4(+) T cells producing IFN-gamma and IL-13 were produced in response to SLA in all patients. SLA- and LACK-specific T cells are effector memory cells, as they are CD45RA(-) CCR7(-) CD4(+) T cells. CD4(+) T cells producing IFN-gamma are CD62L(-), and CD4(+) T cells producing IL-10 are CD62L(+), indicating that these cells have different tissue-homing capacities. These findings show that SLA and LACK induce both type 1 (IFN-gamma) and type 2 (IL-10 or IL-13) cell responses.

Diagnóstico laboratorial das formas mucosa e muco-cutânea da leishmaniose tegumentar americana

Neste estudo, avaliamos a eficiência dos métodos mais comumente utilizados na rotina do diagnóstico da leishmaniose mucosa (LM) e muco-cutânea (LMC). O grupo de (LM) foi composto por 19 pacientes, sendo 13(68,4 por cento)homens e 6(31,6 por cento)mulheres. A média de idade foi de 56,9 com mediana de 58 anos (min=29 e máx=95). O tempo médio da lesão foi de 65 meses. O grupo de (LMC) foi composto por 17 pacientes, sendo 12(70,6 por cento) homens e 5(29,4 por cento) mulheres. A idade média do grupo LMC foi de 46,7 com mediana de 49(min=18 e máx=79), com tempo médio de lesão de 148 meses. Foram realizadas hemoculturas antes do diagnóstico, aos 30, 90 e 180 dias após o tratamento, sendo os resultados sempre negativos. As culturas de fragmento de mucosa também foram negativas. A impressão por aposição do fragmento de biópsia foi positivo em um paciente do grupo LM e um grupo LMC. A intradermo reação de Montenegro foi realizada em 36 pacientes, conferindo sensibilidade de 95,2 por cento sendo de 100 por cento para o grupo de LM e 83,3 por cento do grupo LMC. A sensibilidade da PCR de fragmento de biópsia foi de 84,6 por cento e 100 por cento para os grupos de LM e LMC, respectivamente. A especificidade da PRC foi de 93,3 por cento. Antes do tratamento, detectou-se por PRC parasitário ou DNA do parasito circulante em um paciente do grupo LM. Após o tratamento, a PRC foi positiva em dois pacientes do grupo LM(aos 90 dias e, aos 180 dias) e um paciente do grupo LMC(aos 30 dias). A pesquisa de anticorpos foi avaliada através dos testes ELISA (IgA, IgG, total, IgG1, IgG2, IgG3 e IgG4) e RIFI(IgG total) utilizando antígenos de L.braziliensis e de L.amazonensis. Não foram detectados anticorpos IgA, IgG2 e IgG4. A sensibilidade da RIFI variou entre 100 por cento(Lb)e 89,5 por cento(La).

A especificidade foi avaliada em pacientes portadores de Doença de Chagas, Malária, Sífilis e indivíduos não-infectados de área endêmica, sendo encontrada especificidade entre 65 por cento e 100 por cento. No ELISA, a sensibilidade variou de 64 por cento a 100 por cento e a especificidade de 40 por cento a 100 por cento. No acompanhamento pós-tratamento, não foi observada queda dos níveis de anticorpos pela técnica de RIFI. Pelo ELISA, observou-se queda dos anticorpos IgG totais anti-L braziliensis a partir de 90 dias pós-tratamento para o grupo LM(sendo a IgG3 a principal responsável pela queda) a aos 180 dias pós-tratamento para o grupo LMC. Para IgG1, a queda dos níveis de anticorpos só foi observada aos 180 dias pós-tratamento para o grupo LM nos dois antígenos estudados. A queda dos níveis de IgG3 só aconteceu utilizando a L.braziliensis, no grupo LM, a partir dos 90 dias pós-tratamento.

Diagnóstico laboratorial das formas mucosa e muco-cutânea da leishmaniose tegumentar americana / Laboratory diagnosis of mucous membrane forms the skin and muco-cutaneous leishmaniasis

Neste estudo, avaliamos a eficiência dos métodos mais comumente utilizados na rotina do diagnóstico da leishmaniose mucosa (LM) e muco-cutânea (LMC). O grupo de (LM) foi composto por 19 pacientes, sendo 13 (68,4 por cento) homens e 6 (31,6 por cento) mulheres. A média de idade foi de 56,9 com mediana de 58 anos (min=29 e máx=95). O tempo médio da lesão foi de 65 meses. O grupo de (LMC) foi composto por 17 pacientes, sendo 12 (70,6 por cento) homens e 5 (29,4 por cento) mulheres. A idade média do grupo LMC foi de 46,7 com mediana de 49 (min=18 e máx=79), com tempo médio de lesão de 148 meses. Foram realizadas hemoculturas antes do diagnóstico, aos 30, 90 e 180 dias após o tratamento, sendo os resultados sempre negativos. As culturas de fragmento de mucosa também foram negativas. A impressão por aposição do fragmento de biópsia foi positivo em um paciente do grupo LM e um grupo LMC. A intradermo reação de Montenegro foi realizada em 36 pacientes, conferindo sensibilidade de 95,2 por cento sendo de 100 por cento para o grupo de LM e 83,3 por cento do grupo LMC. A sensibilidade da PCR de fragmento de biópsia foi de 84,6 por cento e 100 por cento para os grupos de LM e LMC, respectivamente. A especificidade da PRC foi de 93,3 por cento. Antes do tratamento, detectou-se por PRC parasitário ou DNA do parasito circulante em um paciente do grupo LM. Após o tratamento, a PRC foi positiva em dois pacientes do grupo LM (aos 90 dias e, aos 180 dias) e um paciente do grupo LMC (aos 30 dias). A pesquisa de anticorpos foi avaliada através dos testes ELISA (IgA, IgG, total, IgG1, IgG2, IgG3 e IgG4) e RIFI (IgG total) utilizando antígenos de L.braziliensis e de L.amazonensis. Não foram detectados anticorpos IgA, IgG2 e IgG4. A sensibilidade da RIFI variou entre 100 por cento (Lb)e 89,5 por cento (La). A especificidade foi avaliada em pacientes portadores de Doença de Chagas, Malária, Sífilis e indivíduos não-infectados de área endêmica, sendo encontrada especificidade entre 65 por cento e 100 por cento. No ELISA, a sensibilidade variou de 64 por cento a 100 por cento e a especificidade de 40 por cento a 100 por cento. No acompanhamento pós-tratamento, não foi observada queda dos níveis de anticorpos pela técnica de RIFI. Pelo ELISA, observou-se queda dos anticorpos IgG totais anti-L braziliensis a partir de 90 dias pós-tratamento para o grupo LM (sendo a IgG3 a principal responsável pela queda) a aos 180 dias pós-tratamento para o grupo LMC. Para IgG1, a queda dos níveis de anticorpos só foi observada aos 180 dias pós-tratamento para o grupo LM nos dois antígenos estudados. A queda dos níveis de IgG3 só aconteceu utilizando a L. braziliensis, no grupo LM, a partir dos 90 dias pós-tratamento.

Persistence of Leishmania antigen in C57Bl/6j inbred mice infected with Leishmania (Leishmania) amazonensis.

PURPOSE: To develop an animal model for studying mucocutaneous leishmaniasis. METHODS: The hind footpad of C57Bl/6j inbred mice was experimentally infected with 10(7) Leishmania (Leishmania) amazonensis promastigote and the skin was studied through light and electron transmission microscopy and immunohistochemistry (PAP) techniques. RESULTS: There were morphological evidences of cellular immune mechanisms and hypersensitivity reaction after eight weeks of infection and metastasis and well shaped parasites at ultrastructural level by fifty-one weeks post infection. Relapse of infection with mucosa lesions occurred around the 50th week after inoculation. CONCLUSION: The use of this animal model in long term follow up could be an useful experimental model for human mucocutaneous leishmaniasis.

Um caso de leishmaniose cutâneo-mucosa tratado com sucesso com baixa dose de antimonial pentavalente / A case of mucocutaneous leishmaniasis treated with success with a low dose of pentavalent antimonial

Os autores relatam um caso de leishmaniose cutâneo-mucosa em uma paciente de 89 anos, diabética e hipertensa, tratada inicialmente com alopurinol por 10 meses não havendo cicatrização das lesões. Posteriormente, recebeu antimoniato de N-metil glucamina (glucantime) por 4 dias, na dose total de 2.380mg do Sbv, mas desenvolveu cardiotoxicidade e hipocalemia, sendo suspenso o tratamento, entretanto, evoluiu com regressão clínica total das lesões, apesar de ter recebido pequena dose desta medicação.

The authors report a case of a 89 years-old woman with mucocutaneous leishmaniasis and previous diabetes mellitus and high blood pressure, who had been treated with allopurinol for 10 months without healing of lesions. Afterwards, she has been treated with meglumine antimonate, [quot ]glucantime[quot ] for 4 days, with a total dose 2,380 mg of Sbv, but developed cardiac side effects and hypokalemia, hence the treatment was withdrawn. However, this patient developed total clinical regression of lesions, in spite of she has been received low dose of this drug.

Impaired production of cytokines in a case of human leishmaniasis.

A patient presented with the unique clinical picture of diffuse cutaneous and mucosal leishmaniasis caused by Leishmania tropica. Elevated serum levels of several cytokines including interleukin (IL) 2, interferon gamma (IFN-gamma), and tumor necrosis factor alpha were found. All cytokine levels returned to normal during therapy. No IL-10 or IL-4 levels were detectable. In whole blood cultures, induction of IFN-gamma by lipopolysaccharide (LPS) was completely negative, even after therapy. Concanavalin A (Con A)-induced release of IFN-gamma, like Con A-induced release of the other cytokines, was only initially impaired but returned to normal during therapy. Induction of the other cytokines by LPS was never impaired. The low expression of human leukocyte antigen DR on monocytes increased during IFN-gamma therapy but dropped when IFN-gamma treatment was ceased. We conclude that in this patient one or more of the routes of IFN-gamma production was impaired, thus resulting in insufficient IFN-gamma production in the infected lesions (although IFN-gamma was systemically present).

Análisis de la Intradermoreacción de montenegro e inmunoflourescencia indirecta en pacientes con leishmaniasis cutánea y mucocutánea de zonas endémicas del Paraguay / Analysis of the intradermo reaction of montenegro and indirect inmunoflourescence in patients with cutaneous and mucocutaneous leishmaniasis in endemic zones of Paraguay

Estudio prospectivo, de corte transversal donde fueron incluídos 102 pacientes con diagnóstico clínico y serológico de Leishmania Cutánea (LC) y Mucocutánea (LMC), que concurrieron al Laboratorio Central de Salud Pública, durante los años 1996 y 97, para la determinación del título de anticuerpos IgG anti-leismania se utilizó la Inmunoflourescencia Indirecta (IFI) y la Intradermoreacción de Montenegro (IDRM)

Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (viannia) braziliensis

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100 positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100 positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100 positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95 of Kala-azar and 35 of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15 of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.

Serum levels of tumor necrosis factor in patients with American cutaneous leishmaniasis.

The purpose of this study was to evaluate serum levels of TNF-alpha in patients with either of the three clinical forms of American cutaneous leishmaniasis. The 86 patients examined were classified as having either the localized (LCL: 22 patients), mucocutaneous (MCL: 45 patients), or the rare diffuse (DCL: 19 patients) form of the disease. Our results show a significant increase in the mean level of TNF-alpha in the three groups of patients when compared to controls. MCL patients produce significantly higher levels of TNF-alpha than DCL patients. The proportion of individuals positive for serum TNF-alpha was significantly higher in both MCL and DCL patients than in the controls. No significant differences were found in the level of TNF-alpha when compared between before and after cure of 12 patients with MCL. There were no significant correlations between the level of TNF-alpha and the skin test diameter. The results will be discussed in terms of the pathogenesis of the disease in its different clinical forms.

Serum levels of tumor necrosis factor in patients with American cutaneous leishmaniasis

The purpose of this study was to evaluate serum levels of TNF-alpha in patients with either of the three clinical forms of American cutaneous leishmaniasis. The 86 patients examined were classified as having either the localized (LCL: 22 patients), mucocutaneous (MCL: 45 patients), or the rare diffuse (DCL: 19 patients) form of the disease. Our results show a significant increase in the mean level of TNF-alpha in the three groups of patients when compared to controls. MCL patients produce significantly higher levels of TNF-alpha than DCL patients. The proportion of individuals positive for serum TNF-alpha was significantly higher in both MCL and DCL patients than in the controls. No significant differences were found in the level of TNF-alpha when compared between before and after cure of 12 patients with MCL. There were no significant correlations between the level of TNF-alpha and the skin test diameter. The results will be discussed in terms of the pathogenesis of the disease in its different clinical forms

Serum interferon activity of patients with leishmaniasis

Serum from a significant proportion of 29 cutaneous and 12 mucocutaneous leishmaniasis patients exhibited interferon activity in a cytopathic assay: positive tests were obtained for 24.1% and 41.7% of the patients, respectively. Similar positive frequencies were observed in other parasitic diseases (schistosomiasis, 12.5%; toxoplasmosis, 20%; Chagas' disease, 16%; leprosy, 12.5%; tuberculosis, 30%). In contrast, none of the 44 serum samples from American visceral leishmaniasis (AVL) patients had interferon activity during the active stage of the disease. However, 13% of the samples obtained from patients recovered from AVL were positive

Blood groups of the ABO and Rh systems and muco-cutaneous leishmaniasis

Estudou-se a distribuiçäo dos grupos sangüíneos dos sistemas ABO e Rh em 203 pacientes com leishmaniose cutanea ou cutaneo-mucosa, a maioria deles provenientes da zona rural do município de Viana, Estado do Espirito Santo. Para controle foram utilizadas duas populaçöes: 183 pacientes sem leishmaniose, provenientes da mesma área e 5154 doadores voluntários do banco de sangue do Hospital Universitário Cassiano A.Moraes. A comparaçäo pelo método do Quiquadrado näo revelou diferenças significativas na freqüência dos grupos sangüineos estudados nos pacientes com leishmaniose em relaçäo às duas populaçöes controle utilizadas. Esta observaçäo sugere que os parasitas do genero Leishmania näo utilizam as glicoproteinas que sintetizam e que se assemelham a antígenos de eritrócitos, para evasäo dos mecanismos de defesa do hospedeiro

American mucocutaneous leishmaniasis. Culture of a Leishmania species from peripheral blood leukocytes.

A Leishmania species was recovered in culture from the buffy coat of peripheral blood of a patient who had American mucocutaneous leishmaniasis, although cultures of biopsy material from the skin lesions were unsuccessful. Blood culture has not previously been recommended in non-visceral leishmaniasis. The importance of recognizing this tropical disease is stressed. Blood culture may be useful in confirming the diagnosis.