Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

SUMMARYThe kinetics of growth of Leishmania performed in vitro after internalization of the promastigote form in the cell and the occurrence of the transformation of the parasite into the amastigote form have been described by several authors. They used explants of macrophages in hamster spleen cell culture or in a human macrophage lineage cell, the U937. Using microscopy, the description of morphologic inter-relationship and the analysis of the production of specific molecules, it has been possible to define some of the peculiarities of the biology of the parasite. The present study shows the growth cycle of Leishmania chagasi during the observation of kinetic analysis undertaken with a McCoy cell lineage that lasted for a period of 144 hours. During the process, the morphologic transformation was revealed by indirect immunofluorescence (IF) and the molecules liberated in the extra cellular medium were observed by SDS-PAGE at 24-hour intervals during the whole 144-hour period. It was observed that in the first 72 hours the promastigote form of L. chagasi adhered to the cell membranes and assumed a rounded (amastigote-like) form. At 96 hours the infected cells showed morphologic alterations; at 120 hours the cells had liberated soluble fluorescent antigens into the extra cellular medium. At 144 hours, new elongated forms of the parasites, similar to promastigotes, were observed. In the SDS-PAGE, specific molecular weight proteins were observed at each point of the kinetic analysis showing that the McCoy cell imitates the macrophage and may be considered a useful model for the study of the infection of the Leishmania/ cell binomial.

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“…Promastigotes of Leishmania: produced in conditioned growth medium of McCoy cells culture 19 , according to the methodology described 22 .…”

Section: Methodsmentioning

confidence: 99%

“…Growth kinetic analyses made after the promastigotes internalization in the cells and their transformation into amastigotes have been made by several authors 6,11,13,15,22 who have also demonstrated the host-parasite relationship and so contributed to the comprehension of the biology of these protozoa.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of growth of Leishmania (Leishmania) chagasi cycle in McCoy cell culture

Nogueira

Nakamura

Galati

2006

Rev. Inst. Med. trop. S. Paulo

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Anti-leishmanial activity of the antimicrobial peptide DRS 01 observed in Leishmania infantum (syn. Leishmania chagasi) cells

Eaton¹,

Bittencourt²,

Silva³

et al. 2014

Nanomedicine: Nanotechnology, Biology and Medicine

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A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques

Nogueira

Odorizzi

Nakamura

2007

Rev. Inst. Med. trop. S. Paulo

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SUMMARYThe Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.

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Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Cited by 4 publications

References 17 publications

Kinetics of growth of Leishmania (Leishmania) chagasi cycle in McCoy cell culture

Kinetics of growth of Leishmania (Leishmania) chagasi cycle in McCoy cell culture

Anti-leishmanial activity of the antimicrobial peptide DRS 01 observed in Leishmania infantum (syn. Leishmania chagasi) cells

A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques

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