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In vitro permissiveness of bovine neutrophils and monocyte derived macrophages to Leishmania donovani of Ethiopian isolate.

Tasew G, Gadisa E, Abera A, Zewude A, Chanyalew M, Aseffa A, Abebe M, Ritter U, van Zandbergen G, Laskay T, Tafess K - Parasit Vectors (2016)

Bottom Line: Microscopy and flow cytometry were used to measure the infection rate while PCR-RLFP was used to confirm the infecting parasite.Similarly, we were able to show that bovine MDM can be infected by L. donovani, which transformed to amastigote forms in the cells.The in vitro infection of bovine PMN and MDM by L. donovani further strengthens the possibility that cattle might serve as source of L. donovani infection for humans.

View Article: PubMed Central - PubMed

Affiliation: Ethiopia Public Health Institute, Leishmaniasis Research Laboratory, P.O. Box 1242, Addis Ababa, Ethiopia.

ABSTRACT

Background: Epidemiological studies in Ethiopia have documented that the risk of visceral leishmaniasis (VL, Kala-azar) is higher among people living with domestic animals. The recent report on isolation of Leishmania donovani complex DNA and the detected high prevalence of anti-leishmanial antibodies in the blood of domestic animals further strengthen the potential role of domestic animals in the epidemiology of VL in Ethiopia. In mammalian hosts polymorphonuclear cells (PMN) and macrophages are the key immune cells influencing susceptibility or control of Leishmania infection. Thus to substantiate the possible role of cattle in VL transmission we investigate the permissiveness of bovine PMN and monocyte derived macrophages (MDM) for Leishmania (L.) donovani infection.

Methods: Whole blood was collected from pure Zebu (Boss indicus) and their cross with Holstein Friesian cattle. L. donovani (MHOM/ET/67/HU3) wild and episomal green fluorescent protein (eGFP) labelled stationary stage promastigotes were co-incubated with whole blood and MDM to determine infection of these cells. Engulfment of promastigotes by the cells and their transformation to amastigote forms in MDM was studied with direct microscopy. Microscopy and flow cytometry were used to measure the infection rate while PCR-RLFP was used to confirm the infecting parasite.

Results: L. donovani infected bovine whole blood PMN in the presence of plasma factors and all cellular elements. Morphological examinations of stained cytospin smears revealed that PMN engulfed promastigotes. Similarly, we were able to show that bovine MDM can be infected by L. donovani, which transformed to amastigote forms in the cells.

Conclusions: The in vitro infection of bovine PMN and MDM by L. donovani further strengthens the possibility that cattle might serve as source of L. donovani infection for humans.

No MeSH data available.


Related in: MedlinePlus

Whole blood and stationary phase promastigotes of L. donovani co-incubated for 22 h. Erythrocytes were lysed, and remaining cells were washed twice in PBS (pH 7.2). Cytospin smears from cell suspensions were stained with 10 % Giemsa staining solution, and examined for PMN infection by microscopic observation and enumeration of amastigote like forms inside them. PMN from Zebu-Holstein crossed animals are shown in panels (a-c), and from Zebu animals in panels (d-f). Panel a, d: uninfected PMN control; panels b, c, d, f: PMN infected with L. donovani parasites, designated by the red arrows, at 22 h of culture. The original photomicrographs were taken at 100× magnification
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Fig1: Whole blood and stationary phase promastigotes of L. donovani co-incubated for 22 h. Erythrocytes were lysed, and remaining cells were washed twice in PBS (pH 7.2). Cytospin smears from cell suspensions were stained with 10 % Giemsa staining solution, and examined for PMN infection by microscopic observation and enumeration of amastigote like forms inside them. PMN from Zebu-Holstein crossed animals are shown in panels (a-c), and from Zebu animals in panels (d-f). Panel a, d: uninfected PMN control; panels b, c, d, f: PMN infected with L. donovani parasites, designated by the red arrows, at 22 h of culture. The original photomicrographs were taken at 100× magnification

Mentions: Bovine PMN cells were observed to be infected by L. donovani after co-incubation for 22 h; as well amastigote like forms without flagella were observed. Such forms within PMN were observed more frequently at 22 h after infection compared with earlier time points. Not all PMN were equally infected, some PMN harboring many parasites per cell, others had 2–4 parasites per cell and still others with no observable parasites. The various morphology of the PMN is shown in Fig. 1.Fig. 1


In vitro permissiveness of bovine neutrophils and monocyte derived macrophages to Leishmania donovani of Ethiopian isolate.

Tasew G, Gadisa E, Abera A, Zewude A, Chanyalew M, Aseffa A, Abebe M, Ritter U, van Zandbergen G, Laskay T, Tafess K - Parasit Vectors (2016)

Whole blood and stationary phase promastigotes of L. donovani co-incubated for 22 h. Erythrocytes were lysed, and remaining cells were washed twice in PBS (pH 7.2). Cytospin smears from cell suspensions were stained with 10 % Giemsa staining solution, and examined for PMN infection by microscopic observation and enumeration of amastigote like forms inside them. PMN from Zebu-Holstein crossed animals are shown in panels (a-c), and from Zebu animals in panels (d-f). Panel a, d: uninfected PMN control; panels b, c, d, f: PMN infected with L. donovani parasites, designated by the red arrows, at 22 h of culture. The original photomicrographs were taken at 100× magnification
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4836163&req=5

Fig1: Whole blood and stationary phase promastigotes of L. donovani co-incubated for 22 h. Erythrocytes were lysed, and remaining cells were washed twice in PBS (pH 7.2). Cytospin smears from cell suspensions were stained with 10 % Giemsa staining solution, and examined for PMN infection by microscopic observation and enumeration of amastigote like forms inside them. PMN from Zebu-Holstein crossed animals are shown in panels (a-c), and from Zebu animals in panels (d-f). Panel a, d: uninfected PMN control; panels b, c, d, f: PMN infected with L. donovani parasites, designated by the red arrows, at 22 h of culture. The original photomicrographs were taken at 100× magnification
Mentions: Bovine PMN cells were observed to be infected by L. donovani after co-incubation for 22 h; as well amastigote like forms without flagella were observed. Such forms within PMN were observed more frequently at 22 h after infection compared with earlier time points. Not all PMN were equally infected, some PMN harboring many parasites per cell, others had 2–4 parasites per cell and still others with no observable parasites. The various morphology of the PMN is shown in Fig. 1.Fig. 1

Bottom Line: Microscopy and flow cytometry were used to measure the infection rate while PCR-RLFP was used to confirm the infecting parasite.Similarly, we were able to show that bovine MDM can be infected by L. donovani, which transformed to amastigote forms in the cells.The in vitro infection of bovine PMN and MDM by L. donovani further strengthens the possibility that cattle might serve as source of L. donovani infection for humans.

View Article: PubMed Central - PubMed

Affiliation: Ethiopia Public Health Institute, Leishmaniasis Research Laboratory, P.O. Box 1242, Addis Ababa, Ethiopia.

ABSTRACT

Background: Epidemiological studies in Ethiopia have documented that the risk of visceral leishmaniasis (VL, Kala-azar) is higher among people living with domestic animals. The recent report on isolation of Leishmania donovani complex DNA and the detected high prevalence of anti-leishmanial antibodies in the blood of domestic animals further strengthen the potential role of domestic animals in the epidemiology of VL in Ethiopia. In mammalian hosts polymorphonuclear cells (PMN) and macrophages are the key immune cells influencing susceptibility or control of Leishmania infection. Thus to substantiate the possible role of cattle in VL transmission we investigate the permissiveness of bovine PMN and monocyte derived macrophages (MDM) for Leishmania (L.) donovani infection.

Methods: Whole blood was collected from pure Zebu (Boss indicus) and their cross with Holstein Friesian cattle. L. donovani (MHOM/ET/67/HU3) wild and episomal green fluorescent protein (eGFP) labelled stationary stage promastigotes were co-incubated with whole blood and MDM to determine infection of these cells. Engulfment of promastigotes by the cells and their transformation to amastigote forms in MDM was studied with direct microscopy. Microscopy and flow cytometry were used to measure the infection rate while PCR-RLFP was used to confirm the infecting parasite.

Results: L. donovani infected bovine whole blood PMN in the presence of plasma factors and all cellular elements. Morphological examinations of stained cytospin smears revealed that PMN engulfed promastigotes. Similarly, we were able to show that bovine MDM can be infected by L. donovani, which transformed to amastigote forms in the cells.

Conclusions: The in vitro infection of bovine PMN and MDM by L. donovani further strengthens the possibility that cattle might serve as source of L. donovani infection for humans.

No MeSH data available.


Related in: MedlinePlus