This document presents research analyzing the effects of edelfosine, a phospholipase C inhibitor, on mRNA and protein expression of GRA1 and MIC3 in acute toxoplasmosis models. The study finds that edelfosine reduces both GRA1 and MIC3 mRNA and protein expression in a dose-dependent manner when administered at the time of infection or 24 hours later. This suggests edelfosine may be an effective treatment for toxoplasmosis by blocking gene expression of the parasite. The results provide insight into using edelfosine and investigating other substances' potential as treatments by examining their effects on pathogen gene expression.

1. Presented by: Andres Calle Meneses and Federico Buenahora Restrepo
3rd semester. Medicine. Universidad Pontificia Bolivariana. 2020

2. Toxoplasma gondii and Toxoplasmosis
Is an obligate
intracellular parasite.
It needs a host cell for its
survival.
The process of
adaptation with a new
environment requires
gene expression
changes
Many people is
Asymptomatic but its
symptoms are similar to
the ones of influenza.
It affects mostly the fetus
Its main reservoir is cats

3. EDELFOSINE
Antiparasitic Antineoplastic effects
Prototype of synthetic lipids
against cancer
Immunomodulatory
properties
Affects gene expression
Selective apoptosis

4. Objective
“Analyze the effects of edelfosine as a PI-PLC inhibitor on the levels of GRA1 and MIC3 mRNA and
protein expressions as the markers of dense granules and microneme organelles in acute toxoplasmosis
models.”

5. Materials and methods
● Female BALB/c mice
● 6 to 8 weeks old
● Specific pathogen free conditions
ANIMALS
PARASITE
● T. gondii was maintained on the mice by
serial intraperitoneal passages.
● The parasite suspension was injected into
the mice to prepare the fresh tachyzoites.

6. Materials and methods
A real time polymerase chain reaction was
made to evaluate the GRA1 and MIC3
mRNA expressions.
RT PCR

7. Materials and methods
● Kidney fibroblast vero cells were cultured and
prepared in culture plates.
● They prepared edelfosine and it was applied to
the tachyzoites which were divided in 2
subgroups.
● it was put on one group at the moment of the
infection, and to the other group 24h later.
● Different concentrations of edelfosine were
applied to see the effect on infected cells
IN VITRO

8. Materials and methods
● A proteases inhibitor was added to the
culture as a buffer
● The extracted cell proteins were submitted
to gel electrophoresis
● The sample was transferred to a
membrane, which was incubated with anti
GRA1, anti MIC3, and anti SAG1, they also
added IgG
● It was visualized with chemiluminescence
and the results were quantified
WESTERN BLOT

9. Results “Effect of edelfosine on the levels of GRA1 mRNA and protein
expressions”.

10. Results “Effect of edelfosine on the levels of GRA1 mRNA and protein
expressions”.

11. Results: “Effect of edelfosine on the levels on MIC3 mRNA and protein expressions”

12. Results “Effect of edelfosine on the levels on MIC3 mRNA and protein expressions”

13. DISCUSSION
AUTHOR STATEMENT DOES IT
MATCH?
Lovett et al, 2002.
McCoy et al 2013
“It has been shown that the PI-PLC path- way through IP3
and Ca2+ and subsequent calcium-dependent protein kinase
signaling cascade has an important role in microneme
exocytosis”
Oliveira, 1999 ; Lux y
col. 2000
“Studies have shown that both miltefosine and edelfosine
inhibit the PI-PLC, lipid metabolism and remodeling,
glycosylphosphatidylinositol biosynthesis, signal transduction
in a dose-dependent manner in the disease”
Azzouz et al, 2015 “The edelfosine, miltefosine, and ilmofosine can be effective
treatment options in parasitosis through direct action on the
parasite membranes”

14. Conclusions
The importance of this type of articles in medicine relays on the fact that based on
the results of the investigation, substances such as edelfosine can very useful on
the treatment of diseases like toxoplasmosis because it was shown that some
dosis of edelfosine had very significant effects by blocking the gene expression of
the disease whether it was at the moment of the infection or time later.
This can also be used as a model of investigation for other diseases and other
substances, so it’s a potential advance for medicine to find cures and treatments
for various pathologies.
Make known laboratory methods implemented in projects
to search for answers to scientific medical problems.

15. Conceptual map:
Federico Buenahora Restrepo

16. Conceptual map:
Andres Calle Meneses.