Evaluation of parasitological and immunological parameters of Leishmania chagasi infection in BALB/c mice using different doses and routes of inoculation of parasites.

Experimental vaccines to protect against visceral leishmaniasis (VL) have been developed by using BALB/c mice infected with a large (107 to 108) inoculum of parasites. Remarkably, prior literature has reported that the poor protection observed is mainly due to the high susceptibility of this strain. To determine factors inherent to mice that might abrogate vaccine-induced efficacy, the present research sought to investigate the impact of the administration of different infective inoculums of Leishmania chagasi (syn. L. infantum) in BALB/c mice, evaluating subcutaneous and intravenous routes of administration as well as parasitological and immunological parameters over different periods of time. This study shows that the injection of a highly infective inoculum in mice, through both subcutaneous and intravenous routes, results in a sustained infection. The mice developed a high parasite load in the liver; however, these values diminished over time. This result did not corroborate with the parasite load in the bone marrow and brain and proved to be expressively different in the spleen and draining lymph nodes, where the values increased over time. Mice infected with a low dose of parasites (10³) showed a certain resistance against infection, based mainly on the IFN-γ and oxide nitric production. Considering all the elements, it could be concluded that the models employing high doses (107) of L. chagasi in BALB/c mice can bring about an imbalance in the animals' immune response, thus allowing for the development of the disease at the expense of efficacy within the vaccine candidates.

Antiproliferative activity of Actinobacillus (Haemophilus) actinomycetemcomitans and Fusobacterium nucleatum in peripheral blood mononuclear cells.

Several studies indicate Actinobacillus (Haemophilus) actinomycetemcomitans and Fusobacterium nucleatum as etiologic agents of periodontal disease. Immunosuppressive factors produced by microorganisms probably contribute to the initiation and evolution of this disease. This study evaluated the antiproliferative activity of ammonium precipitate fractions of A. (H.) actinomycetemcomitans and F. nucleatum isolates from humans and marmosets both with and without periodontal disease. All A. (H.) actinomycetemcomitans and most F. nucleatum strains inhibited PBMC proliferation in a dose-dependent manner. The degree of cell proliferative inhibition of each bacterial species differed among the strains and was independent of host clinical status. The in vitro inhibition of stimulated lymphocyte proliferation induced by different A. (H.) actinomycetemcomitans and F. nucleatum isolates demonstrated the importance of this phenomenon in bacterial virulence, playing a possible suppressor role in host defense mechanisms in vivo. Moreover, our findings pointed out a marked difference between A. (H.) actinomycetemcomitans and F. nucleatum cytoplasmic extracts in their antiproliferative activity, regarding the antigen concentration required for maximum inhibition and their vulnerability to heating and proteolytic treatment.

Molecular diagnosis of leishmaniasis.

This review describes the worldwide situation of visceral and tegumentary leishmaniasis with an emphasis on diagnosis, including methods for the detection of antibodies, antigens, parasite DNA and of skin testing. The advantages and problems of each method are discussed and the need for a rapid, sensitive and low-cost diagnostic method for use in field conditions is highlighted. Recent advances in Leishmania genome sequencing, the use of DNA microarrays and protein microarray methodologies and their potential use for leishmaniasis diagnosis are presented.

Diagnosis of American visceral leishmaniasis in humans and dogs using the recombinant Leishmania donovani A2 antigen.

A2 proteins are expressed in the amastigote stage of Leishmania donovani and are composed predominantely by a conserved repetitive element. Here, we have investigated the presence of anti-A2 antibodies in a panel of American Visceral Leishmaniasis (VL) sera. Anti-A2 antibodies were detected by ELISA, using a recombinant A2 protein containing a tag of six histidine residues (A2-HIS), in 77% of patients sera with symptomatic VL and in 87% of sera from dogs that tested positive in Leishmania immunofluorescent-antibody test (IFAT) or in the parasitological evaluation. Anti-A2 antibodies were also detected in 14 out of 15 symptomatic and in 10 out of 13 asymptomatic dogs. In addition, among the asymptomatic/anti-A2 positive animals, 9 were also positive for the presence of parasites. No significant cross reactivity was observed with sera of animals with other common canine diseases. Our findings suggest that A2 protein is a potential tool for the diagnosis of VL in the New World, and will be particularly useful for diagnosis of dogs.

Shistosoma mansoni: Acquired resistance in mice by implantation of young irradiated worms into the portal system

Em dois experimentos distintos, vermes imaturos de Schistosoma mansoni, com 20 dias de idade, obtidos do sistema porta de camundongos albinos, foram irradiados com 14 e 4 Krad e posteriormente inoculados diretamente na veia porta de camundongos normais. Cada animal, de cada experimento específico, recebeu 20 vermes irradiados. Decorridos 50 dias de inoculaçäo, os camundongos com os vermes irradiados com 4 e 14 Krad e seus respectivos controles foram infectados pela via transcutânea, com cercárias da cepa LE Schistosoma mansoni. No 20§ dia após esta infecçäo desafio, os camndongos foram sacrificados e perfundidos para as contagens dos vermes maduros irradiados (90 dias de idade) e imaturos (20 dias de idade). A análise dos resultados mostrou que ocorreu proteçäo, estatísticamente signifciativa, contra cercárias nos grupos previamente inoculados com vermes irradiados com as doses de 4 e 14 Krad

Schistosoma mansoni: implante de vermes na cavidade peritoneal visando imunoproteçäo / Schistosoma mansoni: implantation of worms in to the peritoneal cavity aiming immunoprotection

Tentou-se obter imunoproteçäo contra infecçäo cercariana pela inoculaçäo de vermes vivos na cavidade peritoneal de camundongos. Embora os vermes sobrevivessem bem nestas condiçöes e näo ocorresse postura de ovos, näo foi possível obter imunoproteçäo. Também a inoculaçäo de ovos viáveis por via sanguínea e peritoneal näo propiciou o aparecimento de imunoproteçäo nos camundongos com vermes na cavidade peritoneal