Seminario - "The Role of LC3-Associated Phagocytosis Inhibits the Inflammatory Response in Aspergillus fumigatus Keratitis"

VIDEO: https://www.youtube.com/watch?v=FBeiDVBaIVM
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COMENTRAIO DE JUNEIDY: https://www.youtube.com/watch?v=6YFFVg488-0

1. Carlos A. Monsalve Landero
Juneidy J. Meneses Fandino
Pontifical Bolivarian University
Faculty of Medicine
Molecular Biology
Semester III

2. INTRODUCTION
Aspergillus fumigatus is a ubiquitous
filamentous hyaline fungus, which has short,
round conidiophores or pseudospherical
conidia. It is found worldwide and grows on
almost any substrate.
Its spores are widely distributed throughout the
environment, and it is thought that we inhale
several of them every day.
it poses a very high risk to immunosuppressed
people since they can cause major pathologies
in various systems.

3. The immune response to A. Fumigatus keratitis
starts with immune cells recognizing the fungus
through receptors like Dectin-1, triggering an
inflammatory response and attracting more
immune cells.
LC3-associated phagocytosis (LAP) is crucial in this
process, helps immune cells engulf and destroy
the fungus efficiently while regulating
inflammation to protect the cornea from damage.
INTRODUCTION

4. GENERAL
OBJECTIVES
The purpose of this study was to investigate
the role and mechanism of microtubule-
associated protein light chain-3 (LC3)-
associated phagocytosis (LAP) in the
immune response to Aspergillus fumigatus
(A. fumigatus) keratitis.

5. METHODS
1.
Polymerase Chain Reaction (PCR)
Allows for the amplification of specific
DNA sequences.
This method is based on the enzymatic
amplification of the target DNA through
repeated cycles of denaturation,
annealing, and extension, which
generates enough copies of a specific
sequence for further analysis.

6. Allows for the identification of the
presence and size of proteins, confirming
their expression and facilitating the study
of their function.
The process involves separating proteins
by gel electrophoresis, transferring them
to a membrane, and detecting them with
specific antibodies.
2.
METHODS
Western blot

7. METHODS
Immunofluorescence Assays
Method used in the laboratory to detect certain
antigens (markers) in a cell or tissue sample.
Immunofluorescence uses antibodies that have a
special dye attached to them, and they bind to the
antigen in the sample.
It was a method used to explore and determine the
expression of proteins in the laboratory in the
corneas of mice, after a fungal infection
3.

8. 1
4
Enzyme-Linked Immunosorbent Assay
ELISA tests can be used to detect everything from
antibodies, hormones and enzymes to viruses and
bacteria.
The principle behind ELISA is that antibodies will
bind to specific antigens.
It was used to observe which induced antibodies
could bind and their reaction to a fungal antigen,
in this case Declin-1 and plasmids overexpressed
with Declin-1 were used.
METHODS

9. Single membrane phagosomes, indicative of
(LAP), are observed in A. fumigatus-infected
corneas(B-C,E-F) but not in normal
corneas(A,D).
The mouse model shows increasing severity of
disease over time.
There is a significant influx of macrophages
into the cornea during A. fumigatus infection.
RESULTS
Human: Mouse:
1 2
3
2
1
3

10. RESULTS
Significantly and sustained increased
levels of these proteins over time is
strong evidence that key components of
LAP are positively regulated during A
keratitis.

11. RESULTS
In this case, protein expression in RAW 264
cells is analyzed after stimulation with the
fungus Aspergillus A.
It was shown that Rubicon, AGT-7 and Beclin
increase progressively over a period of 24
hours after infection, so the greater the
exposure to the infection, the greater the
amount of proteins.

12. RESULTS
The results seen are a decrease in the
expression of Rubicon and LCII
proteins, this is because these cells
were treated with si-rubicon, which
influenced the expression of messenger
RNA, in this case it could be said that si-
rubicon has an anti-inflammatory
effect.

13. DISCUSSION
AUTHOR STATEMENT AGREE/DISAGREE
Wang Y, et al
LAP is a noncanonical
autophagy pathway
Martinez, et al.
Demonstrated that Rubicon
functioned as a molecular
switch for LAP
Smeekens et al.
Found high recruitment
of LC3-II to Candida
albicans-containing
phagosomes

14. CONCLUSIONS
The LAB is essential
to mediate the
inflammatory
response at the ocular
level caused by an
infection by the
fungus Asperillus
fumigatus.
The antibody declin-1
can help in the
immune response by
decreasing
inflammation and
protein expression,
which is why it is
widely used in this
type of infection.
Molecular biology is very
important for the performance
of clinical trials by providing
the basis for performing tests
and understanding them. This
allows us to understand the
cellular functioning of each
organism and enhance its
functions to facilitate
responses to pathogens or
conditions that cannot be
solved externally.
1
2 3

15. Hecho por: Juneidy J. Meneses Fandino

16. Hecho por: Carlos A. Monsalve Landero