A resposta de pacientes com leishmaniose ao tratamento antimonial pode variar em função de fatores como a cepa do parasito envolvido, estado imunológico do paciente e a forma clínica da doença. Os esquemas terapêuticos com antimônio pentavalente (Sb5+) têm sido frequentemente modificados quanto à dose e duração da terapia. Os efeitos colaterais observados com o uso de 20 mg/kg/dia de Sb5+ durante quatro semanas (dose recomendada para o tratamento da forma mucosa) são mais acentuados em pacientes idosos, os mais afetados pela forma mucosa (espundia) da doença. Nossa experiência demonstra que a forma cutânea ocorrendo no Rio de Janeiro responde muito bem ao tratamento com a dose de 5 mg/kg/dia de Sb5+ durante 30 ou 45 dias. Neste estudo foi obtida elevada eficácia (91,4%) com o mesmo regime terapêutico em 36 pacientes com espundia (que evoluem, contudo, com lesão crônica menos destrutiva do que o observado em outras áreas endêmicas) procedentes desta região. Os efeitos colaterais foram reduzidos, não havendo também qualquer resistência ao emprego de doses maiores nos casos que não responderam ao tratamento inicial.

Response to treatment with antimonial drugs varies considerably depending on the parasite strain involved, immune status of the patient and clinical form of the disease. Therapeutic regimens with this first line drug have been frequently modified both, in dose and duration of therapy. A regimen of 20 mg/kg/day of pentavalent antimony (Sb5+) during four weeks without an upper limit on the daily dose is currently recommended for mucosal disease ("espundia"). Side-effects with this dose are more marked in elderly patients, more commonly affected by this form of leishmaniasis. According to our experience, leishmaniasis in Rio de Janeiro responds well to antimony and, in cutaneous disease, high cure rates are obtained with 5 mg/kg/day of Sb5+ during 30 to 45-days. In this study a high rate of cure (91.4%) employing this dose was achieved in 36 patients with mild disease in this same geographic region. Side-effects were reduced and no antimony refractoriness was noted with subsequent use of larger dose in patients that failed to respond to initial schedule.

A new parasite species of Leishmania is described, L. (Leishmania) forattinii sp. n., which was isolated from a pooled triturate of liver and spleen of a opossum (Didelphis marsupialis aurita) and from skin samples from a rodent (Proechmys iheringi denigratus), captured in primary forest on the Atlantic Cost of Brazil. Our results on the basis of biological and molecular criteria indicate that this taxonomically distinct parasite ias a new species of the L. mexicana complex, but closely related to L. (L.) aristidesi Laison & shaw, 1979, as revelated by phenetic and phylogenetic numerical analyses of the enzyme data. L. forattinii was clearly distinguishable from other Leishmania species of the genus usisng enzyme electrophoresis, monoclonal antibodies, molecular karyotypes, analysis of restriction enzyme digestion patterns of kinetoplast DNA (kDNA), as well as the use of kDNA hybridization procedures.

The authors report a case of culture-proven disseminated American muco-cutaneous leishmaniasis caused by Leishmania brasiliensis brasiliensis in an HIV positive patient. Lesions began in the oropharynx and nasal mucosa eventually spreading to much of the skin surface. The response to a short course of glucantime therapy was good.

The molecular karyotypes for 20 reference strais of species complexes of Leishmania were determined by contour-clamped homogeneous eletric field (CHEF) electrosphoresis. Determination of number/position of chromosome-sized bands and chromosomal DNA locations of house-keeping genes were the two criteria used for differentiating and classifying the Leishmania species. We have established two gel running conditions of optimal separation of chromosomes, wich resolved DNA molecules as large as 2,500 kilobase pairs (kb). Chromosomes were polymorphic in number (22-30) and size (200-2,500 kb) of bands among members of five complexes of Leishmania. Although each stock had a distinct karyotype, in general the differences found between strains and/or species within each complex were not clear enough for parasite identification. However, each group showed a specific number of size-concordant DNA molecules, wich allowed distinction among the Leishmania complex parasites. Clear differences between the Old and New world groups of parasites or among some New World Leishmania species were also apparent in relation to the chromosome locations of beta-tubulin genes. Based on these results as well as data from other published studies the potencial of using DNA karyotype for identifying and classifying leishmanial field isolates is discussed.

Characterization is given of a new parasite, Leishmania equatoriensis sp.n. wich was isolated from the viscera of a sloth (Choloepus hoffmanni) and a squirrel (Sciurus granatensis), captured in humid tropical forest onthe Pacific Coast of Ecuador. Data based on biological and molecular criteria, as well as numerical zymotaxonomical analysis, indicate that this parasite is a new species of the L. brasiliensis complex. L. equatoriensis is cleary distinguishable form all other known species within this complex, using the following molecular criteria: reactivity patterns with specific monoclonal antibodies, isoenzyme electrophoresis, and restriction-endonuclease fragment patterns of kinetoplast DNA (k-DNA).

Immunoblot analysis was used to investigate antigenic differences among clinical isolates of Leishmania amazonensis and their role in the etiology of the diseases. Western blots of promastigote homogenates were analyzed with either monoclonal antibodies (MAbs) specific for the L. mexicana complex (M-4, M-6, M-9 and M-11) or polyclonal sera from L. amazonensis infected patients with the various forms of clinical disease. In the case of the MAbs, no significant variation was observed among the strains of L. amazonensis, isolated from cases of cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL), diffuse cutaneous leishmaniasis (DCL), visceral leishmaniasis (VL) or post kala-azar dermal leishmaniasis (PKDL), in either the relative morbility (Mr) or the quantitative amount (intensity) of the antigenic determinats. In the case of the sera of the infected patients, the patterns of antigenic reactivity of these strains revealed that, despite showing the presence of shared antigens, differences were observed between some of the antigenic components of the various isolates of L. amazonensis that were recognized by a single serum. Differences were also demonstrated between the antigenic determinants of a single isolate of L. amazonensis that were recognized by the different patient's sera. No apparent association was consistently found, however, between the Mr components identified in these isolates and clinical form of the disease or the geographical area of isolation. In addition, the spectrum of antigens recognized by the sera from patients with the same clinical form were not identical; although in some instances, similar Mr antigens were shared. These results indicate that isolates of L. amazonensis are not antigenically identical (homogeneous) and that the immune responses (antibodies) observed among infected patients are heterogeneous.

Epidermal changes from 32 cutaneous and 3 mucosal American leishmaniasis (ACL) active lesions were studied for HLA-DR, -DP expression, Lanerhans cells and lymphocyte infiltration. In addition to a DR and DQ positivity at the surface of the cells of the inflammatory infiltrate, a strong reaction for DR antigens was detected on keratinocytes. Hyperplasia of Langerhans cells was present in al cutaneous lesions and epidermis was infiltrated by T lymphocytes. When healed lesions of 14 of these subjects were re-biopsied 1 to 12 months after the end of pentavalent antimonial therapy, MHC class antigens could no longer be seen on keratinocytes. Our data represrn evidence for hhe reversibility of the abnormal HLA-DR expression by keratinocytes in ACL after Glucantime therapy or spontaneous scar formation, demonstrating that this expresion is restricted to the period of active lesions. The present findings can be regarded as an indirect evidence that keratinocytes may be involved in the immunopathology of ACL.

De julho de 1984 a setembro de 1986, 105 casos de leishmaniose cutânea foram estudados numa localidade situada na imediata vizinhança da área urbanizada da região metropolitana da cidade do Rio de Janeiro. A ocupação do sítio deu-se aproximadamente há 20 anos, mas os primeiros casos foram registrados somente seis meses antes do início de nosso estudo. Os casos eram quase que exclusivamente cutâneos, da forma clínica ulcerada com um e seis meses de evolução. O teste de Montenegro foi positivo em todos os casos e anticorpos antileishmania foram detectados por imunofluorescência indireta em 74,3% dos pacientes. A demonstração do parasito foi obtida em 69,5%. Animais domésticos infectados foram facilmente encontrados: 32% dos cães examinados e 30,8% dos equinos mostravam presença de leishmânia em lesões ulceradas. Parasitos isolados, tanto de casos humanos como de cães e equinos, foram imulogicamente caracterizados e identificados com L. b. braziliensis. Da população de flebotomíneos encontrados 73% eram de Lutzomyia intermedia capturados principalmente com iscas humanas e de animais domésticos. Nossas observações sugerem que esta é uma área de estabelecimento recente da infecção por L. b. braziliensis e que a transmissão ocorre provavelmente tanto no peridomicílio como no interior das habitações.

From July 1984 to September 1986, 105 cases of American cutaneous leishmaniasis were studied in a locality closely situated to an urbanized area of the city of Rio de Janeiro, Brazil. Settement in this area was established at least 20 years ago but the first cases were noted six months prior to the beginning of this study. Cases were almost exlusively cutaneous and ulcerated, with one to six months of evolution. Montenegro's skin tests were positive in all cases and anti-Leishmania antibodies were detected by indirect immunofluorescence test in 74.3% of the patients. Parasites were demonstrated in 69.5% of cases. Domestic animals were easily found infected; 32% of the examined dogs and 30.8% of the examined equines were positive to the presence of Leishmania in cutaneous ulcerated lesions. Parasite isolates from human, dog andequines were immunologically characterized and identified as L. b. braziliensis. 73,0% of the sandfly population were Lutzomyia intermedia mainly caught on human baits and on domestic animals. Our observations suggest that this is an area of recent established L. b. braziliensis infection and that transmission probably occurs indoors or outdoors close to the houses.

Um estudo sobre o grau de maturação das células do Sistema Fagocítico Mononuclear foi realizado durante a infecção in vivo e in vitro com a Leishmania mexicana amazonensis. A caracterização da diferenciação das células fagocíticas foi obtida com a localização ultraestrutural de dois marcadores enzimáticos bam conhecidos: a enzima 5'-Nucleotidase marcadora de membrana plasmática de células maduras e a enzima peroxidase, presente em grânulos, marcadora de células imaturas. A atividade da enzima 5'-Nucleotidase foi encontrada apenas em alguns macrófagos, presentes no foco inflamatório, em projeções da membrana plasmática e em algumas vesículas citoplasmáticas. Macrófagos peritoneais de camundongo apresentaram a mesma reatividade para este marcador. Contudo a análise da atividade peroxidásica demonstrou a predominância da presença de fagócitos mononucleares imaturos nas lesões crônicas induzidas neste sistema por Leishmania mexicana amazonensis.

The degree of maturation of cells of the Mononuclear Phagocyte System (MPS), during in vivo and in vitro infection by Leishmania mexicana amazonenesis, was evaluated in this study. The macrophages' differentiation was assayed by cytochemical characterization at the ultrastrctural level, using two well-established markers: 5'-nucleotidase enzyme activity, for revealing the mature cells, and the peroxidase activity present in the cell granules to demonstrate immature mononuclear phagocytes. only a few mcrophages, demonstrating 5'-nucleotidase positive reaction in both the plasma membrane and within their cytoplasmic vesicles, were found scattered in the chronic inflammation at the L. m. amazonensis lesions in albino mice. However, by the peroxidase activity analysis, we were also able to demonstrate the presence of immature MPS cells, which predominate, together with parasitized vacuolated macrophages, in chronic lesions induced in this systemby L. m. amazonensis. The implications of these results on the pathogenesis of murine cutaneous leishmaniasis are discussed.

Inhibição do crescimento de um subespécie de Leishmania por exometabólitos de outra subespécie, um fenômeno ainda não notificado, é sugerido em nossas recentes observações em experimentos de clonagem celular com Leishmania mexicana mexicana e Leishmania mexicana amazonensis. Os clones foram identificados usando a técnica de análise de esquizodemas. O fenômeno observado é claramente relevante em estudos de isolamento parasitário, metabolismo, imunidade cruzada e quimioterapia.

Inhibition of one Leishmania subspecies by exometabolites of another subspecies, a phenomenon not previously reported, is suggested by our recent observations in cell cloning experiments with Leishmania mexicana mexicana and Leishmania mexicana amazonensis. Clones were identified using the technique of schizodeme analysis. The phenomenon observed is clearly relevant to studies of parasite isolation, leishmanial metabolism, cross-immunity and chemotherapy.

Em lesões cutâneas, de um caso humano e de um cão, procedente de área endêmica de leishmaniose tegumentar no Rio de Janeiro, foi isolada L.d. chagasi. Ambas as culturas foram identificadas por caracterização molecular e immunológica do parasito utilizando três diferentes métodos: mobilidade eletroforética de isoenzimas, análise do kDNA e anticorpos monoclonais. Este parece ser o primeiro caso humano bem documentado, no Novo Mundo, de uma Leishmania "viscerotrópica" induzindo lesões cutâneas e demonstra que o diagnóstico do agente etiológico baseado somente na observação clínica e dados epidemiológicos pode levar a conclusões errôneas.

L.d. chagasi was isolated from active cutaneous leishmaniasis in both human and canine infections in an endemic area in Rio de Janeiro, Brazil. Both isolates were identified by molecular and immunological characterization of the parasite using three different methods: electrophoretic mobility of isoenzymes; restriction endonuclease fragment analysis of kDNA and serodeme analysis using monoclonal antibodies. This seems to be the first well documented case in the New World of a "viscerotropic" Leishmania inducing a case of cutaneous leishmaniasis. This observation emphasizes that the diagnosis of the etiologic agent of human or canine visceral leishmaniasis based solely upon clinical and epidemiological critwria may lead to erroneous conclusions.

Os autores lograram infectar flebótomos em um caso humano de leishmaniose tegumentar americana fazendo fêmeas de Lutzomyia longipalpis se alimentarem em paciente com lesões devida à Leishmania mexicana amazonensis.

The authors were able to infect phlebotomine sandflies on a human case of American Cutaneous Leishmaniasis by feeding females of Lutzomyia longipalpis on a patient with a lesion due to Leishmania mexicana amazonensis.