Technical skills - Please describe technical and methodological skills that you have acquired during your research

projects or internships. *

During my Masters in USA, I gained a lot of experience in technical skills since I was leader for two
projects and also my own research for my master thesis. For these two projects, on the first on we
worked with the S1P receptor and its antagonist, which gained me enough experience to mater the
usage of conjugated antibodies and their administration to lab animals in my case mice. Furthermore, I
learned how to use Flow Cytometry and during that time I helped the department of Microbiology and
other PhD students from Department of Biochemistry in their work with the Flow Cytometry Analysis,
which this experience increased my knowledge about data analyzing and also troubleshooting if needed.
Besides Flow Cytometry, I learned PCR and qPCR Technique during our experiments which remains
unpublished about the long-term infection of spleen and the clusters of macrophages that interact with
Chlamydia muridarum. Parallelly, in this project I had the chance to learn Immunoflurohistochemistry
(IFM) respectively Immunohistochemistry and immunostaining, from which I visualized in fluorescent
microscope the images I took from spleen samples that I sliced using Cryostat, which is another
technique that I possess. Besides this, in another project I worked with depletion of FoxP3, CD4 and CD8
T cells. During this I had a chance to learn about administration of substances in IV, IP and SC way.
During this experiment, we removed spleen as well and it was a survival surgery, so I learned how to
perform it under anesthesia which gained me with experience and practical skill to work in that
methodology under stress of time and potential infection of animals itself. In my own thesis and in
another publication that I have, I developed and learned the technique of Western Blot, ELISA and
Vaccine development. During this time which lasted 1 year, we had to work periodically every 3 days
with administration of treatments and then every week to perform experiments and analysis to gather
enough data to move to another stage of experiment. Besides this, during my masters and bachelors I
was caretaker of vivarium as well and have training from Animal Facilities commissions which made me
ready to work with animals and to already master the technique of handling and performing
experiments with them in ethic conditions.
What are the most interesting scientific questions for you within the research areas of infectious diseases and
immunity? Please also explain your choice! (max. 1500 characters)

The most interesting areas of studies that I consider personally in infectious diseases and immunity are the systemic
infections and host pathogen interactions. One of them that I would to further study is how gut microbiota interacts
with other bacteria and how other bacteria survive or find death, the way how this persists. Furthermore, for some
bacteria studies already show that gut is a really hidden cover for them and that the bacteria (the pathogens) are safe
from the immunity itself. I want to explore further how these bacteria use the systemic infection and spread to that
far distances in bodies. Besides this, the immunity of gut is fascinating since its still unexplored at all. In our
previous publication we shoved that gut is responsible for producing IgA which was transferred to the vaginal tract
of mice and helped from secondary infection with Chlamydia. But, my question still remains unclear, like which
cells are interaction and HOW this transport is being done. Are there any receptors involved that we can inhibit or
stop to find their role or is there any secret way that we are not knowing yet. So far, only by reviewing other studies
I somehow have a lead upon VCAM-1 receptors but I haven’t tried it experimentally yet, so that is one of the
questions that I find interesting.

Besides that, I think I should highly consider it mentioning that I am specifically focused on choroidal blood-
cerebrospinal fluid barrier. This is one of the paths that I wanted to follow but didn’t have enough time and
resources during Masters since my supervisor had other priorities in projects. My main objective was studying the
way how Streptococcus bacteria crosses this barrier during Meningitis and the immune cell trafficking interfering
into this. So far there are studies that have identified some proteins playing a role into this but none of them is
focused into immune cells if possibly playing a hidden role into this. Besides this, NK cells are other my lead that I
want to follow for future research. My main focus is to deplete NK Cells respectively to stop production of IL-5 and
to investigate the effect that this would have on macrophages/dendritic cells and the fate of systemic infection itself.
It would be my pleasure to share more about this if required.