A leishmaniose visceral no Brasil estava inicialmente associada a áreas rurais, mas devido às diversas alterações no ambiente como, desmatamentos, urbanização e intenso processo migratório, ocorreu a expansão das áreas endêmicas, levando à urbanização da doença, principalmente nas regiões Sudeste e Centro Oeste do país. No município de Montes Claros, situado ao norte de Minas Gerais, foi feito um estudo para verificação da situação da LV. No ano de 2002 foi realizado inquérito sorológico canino e no período de setembro de 2002 a agosto de 2003 foi feito levantamento entomológico, utilizando armadilhas luminosas de CDC. A prevalência da LV canina apresentou taxa média de infecção em torno de 5%. A fauna de flebotomíneos estimada foi de 16 espécies, totalizando 1043 exemplares. Lutzomyia longipalpis foi a espécie predominante com 74%, o que sugere a sua participação na transmissão de LV em Montes Claros.

Visceral leishmaniasis in Brazil was initially associated with rural areas. However, due to several environmental modifications such as deforestation, urbanization and intense migratory processes, there has been an expansion of endemic areas, leading to urbanization of the disease, mainly in the central and northeastern regions of Brazil. In the municipality of Montes Claros, located in the north of the state of Minas Gerais, an epidemiological survey on VL was carried out. A canine serological inquiry was carried out in 2002 and an entomological survey, using luminous CDC traps, was performed from September 2002 to August 2003. Canine VL prevalence showed an average infection rate of approximately 5%. An estimated 16 species comprised the phlebotomine sand fly fauna, based on a total of 1043 specimens. The predominant species was Lutzomyia longipalpis with a rate of 74%, suggesting its participation in the transmission of VL in the municipality of Montes Claros.

A study of the phlebotomine sand fly fauna was carried out in an endemic area of American visceral leishmaniasis (AVL) in the municipality of Porteirinha, in the Brazilian state of Minas Gerais. Captures were performed with CDC light traps in 7 districts, 5 days per month, during 2 consecutive years (January 2000 to December 2001). A total of 3240 sand flies were captured and identified. Sixteen species were found, among which 15 belonged to the genus Lutzomyia and one to the genus Brumptomyia. Lutzomyia longipalpis, a proven vector of AVL, was the predominant species (71.85%) throughout the time period. The interference of climatic factors (temperature, humidity, and rainfall) over the populational dynamics of the sand flies was determined. Statistical analysis of the data showed a significant correlation among the number of phlebotomine sand flies collected, rainfall, and humidity, whereas the effect of temperature was negligible, in that particular region. The amount of collected phlebotomine, the number of human cases, and the prevalence of canine AVL in the districts of Porteirinha are discussed.

Proteínas imunogênicas da vacina polivalente de promastigotas mortas de leishmanias (Leishvacin®) produzida pela Biobrás Bioquímica do Brasil, Montes Claros, Minas Gerais, Brasil foram identificadas e purificadas por eletroforese em gel de poliacrilamida e eletroeluição. Camundongos C57BL/10 foram vacinados com proteínas de pesos moleculares estimados em 42, 46, 63, 66, 73, 87, 97 e 160kDa em três doses de 30µg de cada proteína combinada com 250µg de Corynebacteriumparvum em intervalos de 15 dias e desafiados com uma infecção desafio de 10(5) promastigotas infectantes de Leishmania (Leishmania) amazonensis na base da cauda. Foram avaliadas a habilidade dessas proteínas em induzir resposta imune e proteção dos animais vacinados após a infecção desafio. Nenhuma diferença estatística foi observada nos níveis de IFN-g nos grupos vacinados em comparação ao grupo controle. Proteção de 28,57; 42,86; 57,14; 42,86; 42,86, 57,14; 42,86; 57,14% foi demonstrado para cada proteína.

Immunogenic proteins from nonliving promastigote polyvalent Leishmania vaccine against American tegumentary leishmaniasis (Leishvacin®), produced by Biobrás (Biochemistry of Brazil ), Montes Claros, State of Minas Gerais, Brazil, were identified and purified by polyacrylamide electrophoresis gel and electroelution. C57BL/10 mice were vaccinated with proteins with estimated molecular weights of 42, 46, 63, 66, 73, 87, 97, and 160kDa in three doses of 30µg of each protein at 15-day intervals combined with 250µg of Corynebacterium parvum followed by a challenge infection with 10(5) infective promastigotes from Leishmania (Leishmania) amazonensis. The ability of these proteins to induce immune response and protection was analyzed. No statistical difference was observed in the level of IFN-g induced by proteins in vaccinated groups in comparison with control groups. Six months after challenge infection, protection levels of 28.57; 42.86; 57.14; 42.86; 42.86, 57.14; 42.86 and 57.14% were demonstrated for each purified protein.

A indução de imunidade no homem contra a leishmaniose cutânea tem sido estudada por vários pesquisadores usando uma grande variedade de preparações antigênicas, como: promastigotas vivas ou atenuadas, extratos de promastigotas, antígenos parcialmente purificados e proteínas puras. Neste trabalho foram isoladas 11 proteínas de L. (L.) amazonensis com pesos moleculares variando de 13.5 a 97 kDa por eletroforese em gel de poliacrilamida e por eletroeluição. Estas proteínas foram combinadas em diferentes preparações vacinais com gp63 e BCG. As vacinas foram avaliadas in vitro quanto à capacidade de estimular linfócitos de pessoas vacinadas com Leishvacin<FONT FACE="Symbol">â</font> a produzirem IFN-<FONT FACE="Symbol">g</font> e a estimularem a proliferação de linfócitos de camundongos vacinados. Assim, camundongos C57BL/10 foram vacinados em intervalos de 15 dias com três doses de cada vacina contendo 30 <FONT FACE="Symbol">m</font>g de cada proteína. 100 <FONT FACE="Symbol">m</font>g de BCG foram usados somente na primeira dose. Sete dias após a última dose os animais receberam a primeira infecção desafiado com 105 promastigotas infectantes e um segundo desafio foi administrado 143 dias após, com o mesmo número de parasitas. Sessenta dias após o segundo desafio, proteções de 42,86% foram obtidas com as vacinas constituídas de gp63+46+22kDa, gp63+13.5+25+42kDa, gp63+46+42kDa, gp63+66kDa e gp63+97kDa; 57,14% de proteção foi obtido com a vacina gp63+46+97kDa, gp63+46+97+13.5kDa, gp63+46+33kDa, e 71,43% com a vacina constituída de gp63 mais todas as proteínas. Em contraste, a vacina gp63+46+33kDa não induziu proteção nos camundongos vacinados, indicando que possivelmente a proteína de 40kDa induziu a uma atividade imunossupressora da resposta imunoprotetora. Estes resultados sugerem que uma futura vacina contra a leishmaniose cutânea deverá conter, excluindo-se a proteína de 40kDa, um coquetel de proteínas imunogênicas indutoras de proteção de camundongos contra a leishmaniose cutânea.

In the past few years, induction of protective immunity to cutaneous leishmaniasis has been attempted by many researchers using a variety of antigenic preparations, such as living promastigotes or promastigote extracts, partially purified, or defined proteins. In this study, eleven proteins from Leishmania (Leishmania) amazonensis (LLa) with estimated molecular mass ranging from 97 to 13.5kDa were isolated by polyacrylamide gel electrophoresis and electro-elution. The proteins were associated as vaccine in different preparations with gp63 and BCG (Bacilli Calmette-Guérin). The antigenicity of these vaccines was measured by their ability to induce the production of IFN-<FONT FACE="Symbol">g</font> by lymphocyte from subjects vaccinated with Leishvacin<FONT FACE="Symbol">â</font> . The immunogenicity was evaluated in vaccinated mice. C57BL/10 mice were vaccinated with three doses of each vaccine consisting of 30 <FONT FACE="Symbol">m</font>g of each protein at 15 days interval. One hundred <FONT FACE="Symbol">m</font>g of live BCG was only used in the first dose. Seven days after the last dose, they received a first challenge infection with 105 infective promastigotes and four months later, a second challenge was done. Two months after the second challenge, 42.86% of protection was obtained in the group of mice vaccinated with association of proteins of gp63+46+22kDa, gp63+13.5+25+42kDa, gp63+46+42kDa, gp63+66kDa, and gp63+97kDa; 57.14% of protection was demonstrated with gp63+46+97+13.5kDa, gp63+46+97kDa, gp63+46+33kDa, and 71.43% protection for gp63 plus all proteins. The vaccine of gp63+46+40kDa that did not protect the mice, despite the good specific stimulation of lymphocytes (LSI = 7.60) and 10.77UI/ml of IFN-<FONT FACE="Symbol">g</font> production. When crude extract of L. (L.) amazonensis was used with BCG a 57.14% of protection was found after the first challenge and 28.57% after the second, the same result was observed for gp63. The data obtained with the vaccines can suggest that the future vaccine probably have to contain, except the 40kDa, a cocktail of proteins that would protect mice against cutaneous leishmaniasis.

A comparative study was undertaken on the immunogenic properties of 63kDa glycoproteins obtained from five different strains/species of Leishmania and assessed in C57BL/10 mice. The humoral immune response was assessed by ELISA against the five different antigens of the immunized animals. The cellular immune response was derived from Leishmania. The response was found to be species-specific in all of determined by means of the cytokine profiles secreted by the spleen cells of immunized animals. The presence of ³-IFN and IL-2, and the absence of IL-4 in the supernatants of cells stimulated by L. amazonensis antigen established that the cellular response is of Th1 type. The five glycoproteins tested were equally effective in protecting C57BL/10 mice against challenge by L. amazonensis. About 50% of the immunized animals were protected for six months.