Targeting Parasite-Produced Macrophage Migration Inhibitory Factor as an Antivirulence Strategy With Antibiotic–Antibody Combination to Reduce Tissue Damage Biologia Molecular- Su
Similar a Targeting Parasite-Produced Macrophage Migration Inhibitory Factor as an Antivirulence Strategy With Antibiotic–Antibody Combination to Reduce Tissue Damage Biologia Molecular- Su
Similar a Targeting Parasite-Produced Macrophage Migration Inhibitory Factor as an Antivirulence Strategy With Antibiotic–Antibody Combination to Reduce Tissue Damage Biologia Molecular- Su (20)
Targeting Parasite-Produced Macrophage Migration Inhibitory Factor as an Antivirulence Strategy With Antibiotic–Antibody Combination to Reduce Tissue Damage Biologia Molecular- Su
2. INTRODUCTION
M.O
Microscopic organisms, commonly
known as microorganisms or microbes,
are found all around us and even inside
the bodies.
Several medically important protozoan
parasites such as Plasmodium,
Entamoeba, Toxoplasma, and Leishmania
secrete an inflammatory macrophage
migration inhibitory factor (MIF)
MIF
cytokine homolog, a virulence
factor linked to severe disease.
combining antiparasite MIF-blocking
antibodies with current standard-of-
care antibiotics might improve
outcomes in severe proto- zoan
infections.
3. INTRODUCTION
ANTI VIRULENT
STRATEGY
Its directed to virulence
factors by inhibiting
specific mechanisms
that promote tissue
damage and disease
symptoms
"
MIF antibodies as an
add-on therapy to
antibiotics in severe
disease using E.
histolytica as the model
organism. We found
that blocking the
virulence factor E.
histolytica MIF (Eh-MIF)
with neutralizing
antibodies combined
with antibiotics resulted
in improved
inflammatory outcomes
and less host damage
in severe infection
4. OBJETIVE
The aim of this study was
to investigate the
effectiveness of targeting
parasite-produced MIF as
combination therapy with
standard antibiotics to
reduce disease severity.
The aim of this preclinical
study was to investigate
the benefit of neutralizing
antiprotozoan MIF
antibodies as an add-on
therapy to antibiotics in
severe disease using E.
histolytica as the model
organism
5. MATERIALES Y METODOS
ELISA
INMUNOTRANSFERENCIA RATONES
QUIMIOLUMINISCENCIA
ELISA
Prueba en la que se identifica
un anticuerpo o un antigeno
utilizando un antigeno o un
anticuerpo absorbido a una
superficie o fase solida
Se utiliza un conjugado anti gama
globulina humana al que se ha unido
una enzima y la enzima produce una
solucion coloreada, la intensidad de
color con espectrofotometro revela la
cantidad de anticuerpos o antigenos
Se preparo tejido intestinal para
ELISA
6. MATERIALES Y METODOS
Quimioluminiscencia
Identificar las reacciones de
un Antigeno - Anticuerpo
Para poder detectar la reaccion se
le debe añadir un indicador con el
cual se logra evidenciar y
cuantificar cuanta reaccion hay.
Utilizo un reactivo
Quimiolunicente para poder
marcar la reaccion
7. MATERIALES Y METODOS
RATONES
Mide la virulencia e infectividad,
detecta parasitos
Entamoeba histolytica que persistieron
en un intestino inflamado durante al
menos 5 días se usaron para
experimentos de colitis severa en
ratones.
Los ratones fueron tratados con factor
estimulante de colonias de granulocitos
por vía subcutánea dos veces al día
durante 3 días. El día 4, los animales
fueron anestesiados, laparotomizados e
infectados intracecalmente con
trofozoítos de E. histolytica.
8. MATERIALES Y METODOS
INMUNOBLOT
Técnica mediante la cual se separan
proteínas por electroforesis, donde
son detectadas con anticuerpos que
reconocen los antígenos que hay en
ellas.
Separación por tamaño, transferencia a
un soporte sólido y, finalmente,
visualización mediante la marcación de
proteínas con el uso de anticuerpos
primarios o secundarios apropiados.
Las membranas se incubaron durante la noche con
El anticuerpo anti- Eh- MIF de ratón purificado por
afinidad por la inmunoglobulina G antimousa G (IgG),
peroxidasa de rábano picante
conjugado (Sigma) anticuerpo secundario.
9. RESULTADOS
La similitud estructural de las
proteinas es un fuerte predictor de la
similitud funcional.
Por lo tanto, la baja homología de
secuencia facilita la generación
anticuerpos contra Eh- MIF que no
reaccionan de forma cruzada con
humanos.
10. RESULTADOS
Se encuentra que antiEhMIF
eran altamente específicos
para la proteína del parásito y
no reacciona con la proteína
del huésped.
.
11. RESULTADOS
El daño tisular causado por la infección
por E. histolytica resulta en la pérdida de
la barrera de permeabilidad intestinal
aumento de la infiltración de neutrófilos,
correlacionada con daño intestinal.
12. Los anticuerpos contra el factor de
virulencia amebiano MIF pueden
proporcionar
beneficios adicionales sobre los
antibióticos, evitando el daño tisular
y notable mejoria en epitelio
13. DISCUSION
AUTHOR CONCEPT YES OR NOT
Medzhitov During an infection, host tissue
destruction occurs by direct
damage by the pathogen and
inflammatory-mediated damage
(immunopathology)
YES
Dickey sw Another benefit of antivirulence
strategies may be lack of direct
effects on beneficial host
commensals, which are unlikely
to harbor virulence factors
YES
Shirley D.A Anti–Eh- MIF does not impair
amebic growth or have
amebicidal activity, meaning
that as antibody levels wane
with time, if the parasite has not
been killed, there could be a risk
of recurrence
NOT
14. CONCLUTION
2
CONCLUTION 1 CONCLUTION 2
Eh-MIF will be developed as a
strategy to minimize
inflammation and Induced
tissue damage, combined with
antibiotic therapy, thus seeks
to facilitate the ability to
perform this correctly and in a
timely manner.
The anti– Eh-MIF offers an
adequate and rational
mechanism to develop new
ways to reduce tissue damage
adequately and avoid
resistance to antibiotics.