Comparative evaluation of phenol and thimerosal as preservatives for a candidate vaccine against American cutaneous leishmaniasis.

For decades thimerosal has been used as a preservative in the candidate vaccine for cutaneous leishmaniasis, which was developed by Mayrink et al. The use of thimerosal in humans has been banned due to its mercury content. This study addresses the standardization of phenol as a new candidate vaccine preservative. We have found that the proteolytic activity was abolished when the test was conducted using the candidate vaccine added to merthiolate (MtVac) as well as to phenol (PhVac). The Montenegro's skin test conversion rates induced by MtVac and by PhVac was 68.06% and 85.9%, respectively, and these values were statistically significant (p < 0.05). The proliferative response of peripheral mononuclear blood cells shows that the stimulation index of mice immunized with both candidate vaccines was higher than the one in control animals (p < 0.05). The ability of the candidate vaccines to induce protection in C57BL/10 mice against a challenge with infective Leishmania amazonensis promastigotes was tested and the mice immunized with PhVac developed smaller lesions than the mice immunized with MtVac. Electrophoresis of phenol-preserved antigen revealed a number of proteins, which were better preserved in PhVac. These results do in fact encourage the use of phenol for preserving the immunogenic and biochemical properties of the candidate vaccine for cutaneous leishmaniasis.

Comparative evaluation of phenol and thimerosal as preservatives for a candidate vaccine against American cutaneous leishmaniasis

For decades thimerosal has been used as a preservative in the candidate vaccine for cutaneous leishmaniasis, which was developed by Mayrink et al. The use of thimerosal in humans has been banned due to its mercury content. This study addresses the standardization of phenol as a new candidate vaccine preservative. We have found that the proteolytic activity was abolished when the test was conducted using the candidate vaccine added to merthiolate (MtVac) as well as to phenol (PhVac). The Montenegro's skin test conversion rates induced by MtVac and by PhVac was 68.06 percent and 85.9 percent, respectively, and these values were statistically significant (p < 0.05). The proliferative response of peripheral mononuclear blood cells shows that the stimulation index of mice immunized with both candidate vaccines was higher than the one in control animals (p < 0.05). The ability of the candidate vaccines to induce protection in C57BL/10 mice against a challenge with infective Leishmania amazonensis promastigotes was tested and the mice immunized with PhVac developed smaller lesions than the mice immunized with MtVac. Electrophoresis of phenol-preserved antigen revealed a number of proteins, which were better preserved in PhVac. These results do in fact encourage the use of phenol for preserving the immunogenic and biochemical properties of the candidate vaccine for cutaneous leishmaniasis.

Immunotherapy, immunochemotherapy and chemotherapy for American cutaneous leishmaniasis treatment.

The first choice of treatment for American cutaneous leishmaniasis is the pentavalent antimonial drug. Although it has been shown that this treatment is mostly effective and indicated, some disadvantages should be taken into account such as side effects, long term treatment inconveniences and counter-indication for patients suffering from cardiopathy, nephropathy; yet, aging, pregnancy and other conditions. With the advent of the vaccine anti-American cutaneous leishmaniasis as a prophylactic measure, studies on therapy using the vaccine associated or not with other drugs have been performed by many investigators and it is currently among the alternative treatments and prevention measures for American cutaneous leishmaniasis. In conclusion, the association between antimony and vaccine (immunochemotherapy) showed the same cure rate when compared with the standard treatment (100%) and it was also able to reduce the salt volume in 17.9% and treatment length from 87 to 62 days, decreasing side effects.

Immunotherapy, immunochemotherapy and chemotherapy for American cutaneous leishmaniasis treatment

O tratamento de primeira escolha para leishmaniose tegumentar americana é o antimonial pentavalente. Embora este tratamento seja na maioria das vezes efetivo e indicado, devem ser consideradas as desvantagens tais como efeitos colaterais, longa duracão do tratamento e contra-indicacão para cardiopatas, nefropatas, idosos, grávidas e outras condicões. Com o advento da vacina antileishmaniose tegumentar americana para fins profiláticos e terapêuticos, associando-a ou não a outros fármacos, muitas pesquisas têm sido desenvolvidas, sendo a vacina a principal entre os atuais recursos no tratamento e prevencão da leishmaniose tegumentar americana. Em conclusão, a associacão do antimônio com a vacina (imunoquimioterapia) apresentou o mesmo índice de cura em relacão ao tratamento padrão (100%), e ainda reduziu o volume do sal em 17,9% e o tempo de cura significativamente, de 87 para 62 dias; conseqüentemente, reduzindo os efeitos colaterais.

Vaccination of C57BL/10 mice against cutaneous leishmaniasis using killed promastigotes of different strains and species of Leishmania

Antigenic extracts from five Leishmania stocks were used to vaccinate C57BL/10 mice. The Leishvacin® and PH8 monovalent vaccine yielded the highest IFN-gamma levels in the supernatants of spleen cell culture from vaccinated animals. Each single strain immunized group showed evidence of protective immunity six months after the challenge with promastigotes of Leishmania (Leishmania) amazonensis. No differences were detected between the vaccinated groups. It can be concluded that vaccines composed of single Leishmania stocks can provide protection to C57BL/10 mice against L. (L.) amazonensis infection

Mílio Colóide do Adulto

Mílio colóide é doença degenerativa relativamente rara, de etiologia desconhecida. São descritos três tipos de degeneração colóide da pele: mílio colóide tipo adulto, mílio colóide juvenil e degeneração nodular colóide, Na variante do adulto a luz solar é fator importante, havendo casos desencadeados por bronzeamento artificial, na histologia observa-se lesão dérmica contendo material eosinofilico. Os autores relatam dois casos de mílio colóide tipo adulto e dazem uma revisão breve sobre a possível etiologia, a clínica e os achados histopatológicos, bem como sobre as modalidades terapêuticas para a doença

Hemorragia macular em uma paciente com calazar: relato de caso / Macular hemorrhage in a patient with visceral leishmaniasis case report

Os autores relatam um caso de leishmeniose visceral (calazar) com alteraçäo ocular. Descreve-se um caso de uma paciente de 17 anos, sexo feminino, com calazar, que apresentou hemorragia macular documentada através de retinopatia simples. A hemorragia macular desapareceu após 3 semanas de tratamento específico com Glucantime e controle hematológico. Os autores chamam atençäo para a possibilidade de encontro de hemorragia retiniana em pacientes com leishmaniose visceral e alertam para o surgimento de inúmeros casos diante de reemergência desta parasitose, principalmente nos grandes centros urbanos.

Protection of C57BL/10 mice by vaccination with association of purified proteins from Leishmania (Leishmania) amazonensis

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-gamma by lymphocyte from subjects vaccinated with Leishvacin. The immunogenicity was evaluated in vaccinated mice. C57BL/10 mice were vaccinated with three doses of each vaccine consisting of 30 mu/g of each protein at 15 days interval. One hundred mu/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 percent 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 percent of protection was demonstrated with gp63+46+97+13.5kDa, gp63+46+97kDa, gp63+46+33kDa, and 71.43 percent 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-gamma production. When crude extract of L. (L.) amazonensis was used with BCG a 57.14 percent of protection was found after the first challenge and 28.57 percent 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

Leishmaniose visceral canina estudo parasitológico e histopatológico em olhos de cäes: parte I / Canine visceral leishmaniosis parasitological and histopathological study in dog's eyes: part I

Vários autores têm relatado o envolvimento ocular externo em cäes naturalmente infectados com Leihmania infantum e Leishmania donovani. Entretanto, ainda näo foram realizados estudos dos olhos de cäes no nosso meio. Objetivando investigar a possibilidade de parasitismo ocular, quatro cäes controle e 12 cäes com leishmaniose visceral foram examinados, todos provinientesda regiäo metropolitana de Belo Horizonte, MG, Brasil.O diagnóstico foi realizado através de sorologia convencional (RIFI e ELISA) e exame parasilógico de aspirado de medula óssea. Para o isolamento do parasito foi realizada a inoculaçäo de aproximadamente 100 ml de humor aquoso e/ou corpo vítreo em meio de cultura de NNN/LIT, que foi mantido a ...

Leishmaniose visceral canina estudo imunológico em olhos de cäes: parte II / Canine visceral leishmaniasis immunological study in dog's eyes

O quadro clínico da leishmaniose visceral canina se assemelha muito ao quadro clínico da leishmaniose visceral humana. Embora estas alteraçöes sejam amplamente abordadas em praticamente todos os sistemas no cäo, o estudo das alteraçöes imunológicas näo têm sido profundamente estudados. Aproximadamente 20ml de humor aquoso e corpo vítreo foram utilizados para exames imunológicos através do ensaio imunoenzimático (ELISA) para detecçäo de anticorpos anti leishmania (IgM e IgG) e 10ml de humor aquoso foram utilizados para dosagem de proteína pela micro técnica de Lowry. Detectou se anticorpos anti leishmania (IgM e IgG) em níveis que variaram de 1:80 a 1:2560 no humor aquoso e/ou corpo vítreo dos animais infectados e menores que 1:5 nos animais controle...

Immune responses induced by a Leishmania (Leishmania) amazonensis recombinant antigen in mice and lymphocytes from vaccinated subjects

A resposta imune induzida por uma proteina recombinante de Leishmania (Leishmania) amazonensis de 33 kD (Larp33) foi avaliada em linfocitos de individuos vacinados com a Leishvacin e em camundongos atraves de vacinacao. Larp33 foi expressa em Escherichia coli apos clonagem de um fragmento genomico de L. (L.) amazonensis de 2,2 kb no vetor pDS56-6His. Larp33 foi reconhecida por anticorpos IgG presentes no soro de individuos vacinados com Leishvacin e induziu proliferacao em linfocitos desses individuos em niveis comparaveis ao antigeno total de Leishmania...

Assessment of immunity induced in mice by glycoproteins derived from different strains and species of Leishamnia

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 y-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 por cento of the immunized animals were protected for six months.

Tratamento da leishmaniose tegumentar americana utilizando vacina / Treatment of american cutaneous leishmaniasis by means of vaccine

Foi realizado o tratamento de 62 pacientes portadores de leishmaniose tegumentar utilizando como agente imunoterápico uma vacina constituída de promastigotas mortas. O esquema terapêutico utilizado mostrou ser uma alternativa adequada ao tratamento de leishmaniose tegumentar já que houve sucesso terapêutico em 75,8% dos casos, näo se observando efeitos colaterais. A eficiência terapêutica foi semelhante nos casos de lesöes cutâneas ou mucosas