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Oleylphosphocholine (OlPC) arrests Cryptosporidium parvum growth in vitro and prevents lethal infection in interferon gamma receptor knock-out mice.

Sonzogni-Desautels K, Renteria AE, Camargo FV, Di Lenardo TZ, Mikhail A, Arrowood MJ, Fortin A, Ndao M - Front Microbiol (2015)

Bottom Line: Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30.Doses of 30 and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction.Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

View Article: PubMed Central - PubMed

Affiliation: National Reference Centre for Parasitology, Research Institute of the McGill University Health Centre, Montreal QC, Canada ; Institute of Parasitology, Macdonald Campus, McGill University, Montreal QC, Canada.

ABSTRACT
Cryptosporidium parvum is a species of protozoa that causes cryptosporidiosis, an intestinal disease affecting many mammals including humans. Typically, in healthy individuals, cryptosporidiosis is a self-limiting disease. However, C. parvum can cause a severe and persistent infection that can be life-threatening for immunocompromised individuals, such as AIDS patients. As there are no available treatments for these patients that can cure the disease, there is an urgent need to identify treatment options. We tested the anti-parasitic activity of the alkylphosphocholine oleylphosphocholine (OlPC), an analog of miltefosine, against C. parvum in in vitro and in vivo studies. In vitro experiments using C. parvum infected human ileocecal adenocarcinoma cells (HCT-8 cells) showed that OlPC has an EC50 of 18.84 nM. Moreover, no cell toxicity has been seen at concentrations ≤50 μM. C57BL/6 interferon gamma receptor knock-out mice, were infected by gavage with 4000 C. parvum oocysts on Day 0. Oral treatments, with OlPC, miltefosine, paromomycin or PBS, began on Day 3 post-infection for 10 days. Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30. Doses of 30 and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction. Mice treated with 10 mg/kg/day of miltefosine resulted in 50% survival at Day 30. In contrast, control mice, treated with PBS or 100 mg/kg/day of paromomycin, died or had to be euthanized between Days 6 and 13 due to severe illness. Results of parasite burden were obtained by qPCR and cross-validated by both flow cytometry of stool oocysts and histological sections of the ileum. Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

No MeSH data available.


Related in: MedlinePlus

Day 30 histological sections of the ileum of control and 20 mg/kg/day OlPC-treated C57BL/6 IFNγR-KO mice infected with C. parvum. Representative transverse sections of the ileum stained with H&E showing disease progression. (A) Ileum of mice treated with 10 mg/kg/day of miltefosine shows severe epithelial damage and abundance of oocysts. (B) Ileum of mice treated with 20 mg/kg/day of OlPC shows some epithelial damage and moderate level of oocysts. Each group is represented at 100 and 400x magnifications. Red arrows indicate C. parvum oocysts.
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Figure 7: Day 30 histological sections of the ileum of control and 20 mg/kg/day OlPC-treated C57BL/6 IFNγR-KO mice infected with C. parvum. Representative transverse sections of the ileum stained with H&E showing disease progression. (A) Ileum of mice treated with 10 mg/kg/day of miltefosine shows severe epithelial damage and abundance of oocysts. (B) Ileum of mice treated with 20 mg/kg/day of OlPC shows some epithelial damage and moderate level of oocysts. Each group is represented at 100 and 400x magnifications. Red arrows indicate C. parvum oocysts.

Mentions: At Day 30, the surviving mice treated with 10 mg/kg/day of miltefosine revealed an abundant presence of C. parvum oocysts and severe damage to the gut epithelium (Figure 7A). At the lowest concentration of OlPC, mice displayed a moderate presence of C. parvum oocysts and only mild damage to the intestinal epithelium (Figure 7B). At 30 and 40 mg/kg/day of OlPC, mice showed a complete absence of C. parvum oocysts and a healthy ileum in comparison with controls (Figures 8A,B). Sections of ileum, liver and spleen (data not shown) were also processed in uninfected (control) mice treated with 40 mg/kg/day for 10 days and no inflammation or toxicity was noticeable.


Oleylphosphocholine (OlPC) arrests Cryptosporidium parvum growth in vitro and prevents lethal infection in interferon gamma receptor knock-out mice.

Sonzogni-Desautels K, Renteria AE, Camargo FV, Di Lenardo TZ, Mikhail A, Arrowood MJ, Fortin A, Ndao M - Front Microbiol (2015)

Day 30 histological sections of the ileum of control and 20 mg/kg/day OlPC-treated C57BL/6 IFNγR-KO mice infected with C. parvum. Representative transverse sections of the ileum stained with H&E showing disease progression. (A) Ileum of mice treated with 10 mg/kg/day of miltefosine shows severe epithelial damage and abundance of oocysts. (B) Ileum of mice treated with 20 mg/kg/day of OlPC shows some epithelial damage and moderate level of oocysts. Each group is represented at 100 and 400x magnifications. Red arrows indicate C. parvum oocysts.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585137&req=5

Figure 7: Day 30 histological sections of the ileum of control and 20 mg/kg/day OlPC-treated C57BL/6 IFNγR-KO mice infected with C. parvum. Representative transverse sections of the ileum stained with H&E showing disease progression. (A) Ileum of mice treated with 10 mg/kg/day of miltefosine shows severe epithelial damage and abundance of oocysts. (B) Ileum of mice treated with 20 mg/kg/day of OlPC shows some epithelial damage and moderate level of oocysts. Each group is represented at 100 and 400x magnifications. Red arrows indicate C. parvum oocysts.
Mentions: At Day 30, the surviving mice treated with 10 mg/kg/day of miltefosine revealed an abundant presence of C. parvum oocysts and severe damage to the gut epithelium (Figure 7A). At the lowest concentration of OlPC, mice displayed a moderate presence of C. parvum oocysts and only mild damage to the intestinal epithelium (Figure 7B). At 30 and 40 mg/kg/day of OlPC, mice showed a complete absence of C. parvum oocysts and a healthy ileum in comparison with controls (Figures 8A,B). Sections of ileum, liver and spleen (data not shown) were also processed in uninfected (control) mice treated with 40 mg/kg/day for 10 days and no inflammation or toxicity was noticeable.

Bottom Line: Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30.Doses of 30 and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction.Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

View Article: PubMed Central - PubMed

Affiliation: National Reference Centre for Parasitology, Research Institute of the McGill University Health Centre, Montreal QC, Canada ; Institute of Parasitology, Macdonald Campus, McGill University, Montreal QC, Canada.

ABSTRACT
Cryptosporidium parvum is a species of protozoa that causes cryptosporidiosis, an intestinal disease affecting many mammals including humans. Typically, in healthy individuals, cryptosporidiosis is a self-limiting disease. However, C. parvum can cause a severe and persistent infection that can be life-threatening for immunocompromised individuals, such as AIDS patients. As there are no available treatments for these patients that can cure the disease, there is an urgent need to identify treatment options. We tested the anti-parasitic activity of the alkylphosphocholine oleylphosphocholine (OlPC), an analog of miltefosine, against C. parvum in in vitro and in vivo studies. In vitro experiments using C. parvum infected human ileocecal adenocarcinoma cells (HCT-8 cells) showed that OlPC has an EC50 of 18.84 nM. Moreover, no cell toxicity has been seen at concentrations ≤50 μM. C57BL/6 interferon gamma receptor knock-out mice, were infected by gavage with 4000 C. parvum oocysts on Day 0. Oral treatments, with OlPC, miltefosine, paromomycin or PBS, began on Day 3 post-infection for 10 days. Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30. Doses of 30 and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction. Mice treated with 10 mg/kg/day of miltefosine resulted in 50% survival at Day 30. In contrast, control mice, treated with PBS or 100 mg/kg/day of paromomycin, died or had to be euthanized between Days 6 and 13 due to severe illness. Results of parasite burden were obtained by qPCR and cross-validated by both flow cytometry of stool oocysts and histological sections of the ileum. Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

No MeSH data available.


Related in: MedlinePlus