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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

MDM infection with eGFP labelled L. donovani was analyzed using the BD FACS Calibur. a A representative forward scatter (FSC) and sideward scatter (SSC) plot shows gating for cultured macrophages. b A representative infection assay result for eGFP L. donovani in bovine MDM at two time points of infection. GFP fluorescence was evaluated among gated macrophages from either cross-breed MDM (i) or Zebu cattle (ii). Grey colored plots represent MDM infected with wild type parasite infections, whereas green colored plots depict MDM infected with eGFP-labelled L. donovani. Cells were evaluated after 24 h of infection (upper panels) or 48 h of infection (lower panels)
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Fig4: MDM infection with eGFP labelled L. donovani was analyzed using the BD FACS Calibur. a A representative forward scatter (FSC) and sideward scatter (SSC) plot shows gating for cultured macrophages. b A representative infection assay result for eGFP L. donovani in bovine MDM at two time points of infection. GFP fluorescence was evaluated among gated macrophages from either cross-breed MDM (i) or Zebu cattle (ii). Grey colored plots represent MDM infected with wild type parasite infections, whereas green colored plots depict MDM infected with eGFP-labelled L. donovani. Cells were evaluated after 24 h of infection (upper panels) or 48 h of infection (lower panels)

Mentions: Quantitation of eGFP labelled L. donovani infection rates by flow cytometry revealed more infected MDM in the cross-breeds, 73.9 % at 24 h, and 77.8 % at 48 h, compared to the MDMs from pure Zebu, 47.16 % at 24 h and 49.55 % at 48 h (Fig. 4).Fig. 4


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)

MDM infection with eGFP labelled L. donovani was analyzed using the BD FACS Calibur. a A representative forward scatter (FSC) and sideward scatter (SSC) plot shows gating for cultured macrophages. b A representative infection assay result for eGFP L. donovani in bovine MDM at two time points of infection. GFP fluorescence was evaluated among gated macrophages from either cross-breed MDM (i) or Zebu cattle (ii). Grey colored plots represent MDM infected with wild type parasite infections, whereas green colored plots depict MDM infected with eGFP-labelled L. donovani. Cells were evaluated after 24 h of infection (upper panels) or 48 h of infection (lower panels)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig4: MDM infection with eGFP labelled L. donovani was analyzed using the BD FACS Calibur. a A representative forward scatter (FSC) and sideward scatter (SSC) plot shows gating for cultured macrophages. b A representative infection assay result for eGFP L. donovani in bovine MDM at two time points of infection. GFP fluorescence was evaluated among gated macrophages from either cross-breed MDM (i) or Zebu cattle (ii). Grey colored plots represent MDM infected with wild type parasite infections, whereas green colored plots depict MDM infected with eGFP-labelled L. donovani. Cells were evaluated after 24 h of infection (upper panels) or 48 h of infection (lower panels)
Mentions: Quantitation of eGFP labelled L. donovani infection rates by flow cytometry revealed more infected MDM in the cross-breeds, 73.9 % at 24 h, and 77.8 % at 48 h, compared to the MDMs from pure Zebu, 47.16 % at 24 h and 49.55 % at 48 h (Fig. 4).Fig. 4

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