Video summary
The Molecular Microbiology and Biotechnology Laboratory at the University of Siena is dedicated to developing safe and effective vaccines to address global health needs, with a specific focus on pathogens that pose pandemic risks or primarily affect low-income countries. Researchers in this department utilize preclinical and clinical studies alongside innovative technologies to investigate how the immune system responds in both healthy individuals and vulnerable populations. A central goal of their work is not only to measure antibody production but also to study immunological memory, which ensures the body can actively protect itself against subsequent infections. This research extends to serious diseases such as Ebola, invasive non-typhoidal salmonella affecting sub-Saharan Africa, and respiratory viruses like influenza and coronaviruses, aiming to create solutions for neglected diseases where economic incentives are often lacking.
To achieve these goals, the laboratory employs advanced techniques such as spectral flow cytometry and RNA sequencing to analyze complex immune responses in detail. Spectral flow cytometry allows scientists to measure molecules on cell surfaces and distinguish between different types of immune cells, such as those that immediately produce antibodies versus memory cells that provide long-term protection. This technology is crucial for understanding how vaccines perform in patients with compromised immune systems, such as those living with HIV or suffering from cystic fibrosis, where standard responses may differ significantly. By integrating biological data with computational approaches known as Systems Biology, the team can identify specific biomarkers that predict vaccine efficacy and tailor vaccination strategies to individual pathologies, ensuring broader protection across diverse populations.
Beyond human clinical trials, the research group relies heavily on preclinical models, including 3D organoids grown in vitro, to test new vaccines and reduce the need for animal testing. These miniature organs, which mimic the structure and function of human tissues like the nasal epithelium or intestinal mucosa, allow researchers to study infections caused by bacteria, viruses, and fungi under controlled conditions. The laboratory has successfully used these models to demonstrate that innovative vaccine candidates against salmonella can induce high-quality antibody responses and memory B cells capable of protecting subjects years after vaccination. This rigorous preclinical validation has accelerated the development of new vaccines, moving several candidates into advanced human clinical trials and contributing to coordinated European efforts through initiatives like the European Vaccine Hub for Pandemic Readiness.
In addition to scientific discovery, the University of Siena is committed to training the next generation of researchers to tackle future pandemics through a national doctoral program in infection diagnosis and prevention. This multidisciplinary initiative brings together students from various countries and backgrounds to collaborate on controlling infections at risk of becoming epidemics or pandemics. The laboratory also fosters partnerships with local businesses and international research centers, making its state-of-the-art equipment available for broader scientific collaboration. Ultimately, the combined efforts of basic research, cutting-edge technology application, and educational programs aim to build a robust network capable of responding rapidly and effectively to emerging global health challenges.
Read the full video transcript
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The
Molecular Microbiology and Biotechnology Laboratory of the
Department of Medical Biotechnology
at the University of Siena involves
researchers actively
engaged in vaccine development. and
in the study of the immune response
to vaccination and infection
through the application of
preclinical studies, clinical studies and
highly innovative technologies. The
goal of our research is very
concrete: to develop increasingly
effective and safe vaccines to
meet
global health needs and to study how
the immune system responds in
healthy and vulnerable individuals. We
study not only the
antibody response, but also, above all,
immunological memory, which is the memory
of the vaccine or microorganism that
is induced and enables us to
actively respond, protecting ourselves in the
event of subsequent infection. An
important part of our
research activity is the development of vaccines for
pathogens at risk of epidemic pandemics and
for pathogens that mainly affect
low-income countries.
For example, we
actively participated in two European projects
aimed at characterizing the
immune response to the Ebola vaccine,
a very serious disease that induces a
haemorrhagic fever with high mortality.
We conducted transcriptomic analysis studies
to identify
biomarkers of the immune response
in the vaccine that was the first vaccine
approved for human use against
this devastating disease.
We are also involved in studying
vaccines such as vaccines against
Scygella, invasive non-
typhoidal salmonella. Well, in particular, this is a
disease that mainly affects
sub-Saharan Africa. and it is
particularly serious for children and
immunocompromised individuals. This is a
vaccine developed with an
innovative technology from the GSK Vaccine Institute for
Global Health here in Siena, and we have
contributed and are contributing with the
analyses in preclinical studies and the
immunological studies in
clinical trials. This vaccine
is currently in an advanced stage of
clinical trials. Obviously
our research activity is
also focused on the study of
vaccines against the coronavirus. Well, in
particular, since the beginning of the
pandemic we have started clinical studies
with various departments of the
university hospital, including the
infectious diseases department, where we
are studying the
immunological response to different vaccines, to
different combinations of prime boosts, both
in healthy subjects and in
vulnerable subjects. We are
also conducting similar studies on
vaccines against BK pox and
influenza.
Our commitment is also reflected
in our active participation in
the European Vaccine Hub for
Pandemic Readiness, a highly
ambitious program funded by the
European Commission
that involves the
leading European centers for
vaccine development and
pandemic response. It is coordinated right here in
Siena, with the University of Siena playing a key role
and the
Siena Biotechnopole Foundation playing a central role.
This
initiative aims to coordinate
the main European centers to
enable a coordinated,
rapid, and effective response to
possible future infections at risk of
epidemic or pandemic. Our
research activity therefore integrates basic research
with innovative research and the
application of
cutting-edge technologies,
conducting clinical studies,
collaborating with
local companies and
European and international research centers with
the common goal of contributing to the
development of increasingly safe and
effective vaccines to meet
health challenges. global.
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This is the cytoflorimetry center,
a technology that allows us to measure
the expression of molecules on the
surface of a cell resuspended in
a fluid sample, for example in
blood. And in the field
of vaccination response,
studying the response to vaccination,
cytoflorimetry is a very
important technique, and in
recent years, in the context of the pandemic,
we have used this instrumentation
to characterize the response to
vaccination, also using new
vaccination technologies, therefore the
ARNA vaccines, Pfizer, Moderna, which
were administered as
mass vaccination for the first time
in this context. And the response that
emerges from
cytopronometric analyses is not only
quantitative, but also qualitative. That is,
what type of cells can I
stimulate with this
vaccine formulation? They are cells, for
example, capable of immediately producing
antibodies or rather
memory cells that persist over time and
provide protection to the subject.
These studies were conducted not only
on healthy volunteers, but also on
patients. Patients with various
pathologies that somehow alter
the capacity of their
immune system, making them particularly
vulnerable to infection, but it is
also important to understand how they
respond to a vaccination precisely
because of these immunological deficiencies.
And this study was
conducted in collaboration with
Professor Montagnani of the
Infectious Diseases Unit, with the
participation of many clinics at
Siena University Hospital.
They were interested in understanding how
patients with different diseases
might respond, and our
results have effectively
highlighted that different pathologies can have
different responses to the
same vaccination. So this convinced us
that it's still very important to
continue conducting these types of
studies, to understand, even beyond the
Covid vaccination,
how these subjects can
generate a
protective response and thus
also help guide, um, schedules,
vaccination programs
specifically designed for each
individual pathology.
And this instrument you see here is a
tool that combines the technology of
cytoflorimetry, which
allows you to highlight and identify
individual cell populations in a sample
with the possibility of
recovering them. So it's called a sorter
precisely because it allows us to
collect purified populations
and then use these cells
to perform downstream assays, for example,
to characterize their
hygienic expression or their functionality. This
entire laboratory
you see here is a laboratory that has been
built over the years thanks to
funding from the university,
the Tuscany Region, national funds, and the
European Community. And this is the latest
instrument, a cytoflorimeter that
implements
spectral technology, which is somewhat of an evolution
compared to
conventional cytoflorimetry. It was purchased
by a group of researchers from
the University of Siena, some from
different departments,
because we were all interested,
let's say, in purchasing this
instrument, which was also obtained
thanks to funding from the PNRR.
And this technology, this center is
available for collaborations, it's open
to all university researchers who
are interested in conducting these studies.
Furthermore, we also offer services to
local businesses that may
not be able to afford all the
equipment, but are interested in conducting
these analyses. This has allowed us
over the years to create
collaborations with researchers and
local businesses, which have
generated a scientific and technical network that is
important for the quality of our
research. [music]
In this laboratory I
work on sequencing techniques and
in particular to study the
immune response and the response to
infections, I use a technique
called RNA sequencing. This technique
allows us to characterize the
expression levels of thousands of
vaccination or infection genes. In this
way, we analyze the
expression levels of all these genes and then
try to understand the
overall function of these genes
in the response to vaccination.
We do this essentially
early after the immune response
because we then want to try to
correlate the type of response
we observe with RNA sequencing to the
immune response that
typically occurs 2 to 6 weeks
after vaccination.
The goal, therefore, is to correlate the
expression levels and
activated immunological patterns with the levels of
response to vaccination,
essentially antibody levels.
We have used this approach in
several types of vaccines within
national and international projects, for
example the Ebola virus vaccine
with a live attenuated vaccine platform
, the yellow fever vaccine
, the vesicle-based vaccine
against invasive non-typhoid salmonellosis
and the therapeutic vaccine against
cutaneous spaniosis. All these are
so-called neglected diseases, meaning there
is no economic interest
or public interest in
developing therapies or vaccinations for them.
When we use our RNA
sequencing data we are faced with enormous
data complexity, because we have
thousands of pieces of information to
put together and synthesize. For
this reason, we use not only
biological and immunological skills, but we
also talk about
computational skills such as those that
physical or mathematical engineers may have in
a comprehensive approach called
Systems Biology. The goal of
Instance Biology and the analysis of
this type of multidimensional data is
to simplify the model and
then search for a few biomarkers that
can explain the
immune response and possibly
correlate the response we see with an
outcome that could be effective
vaccination or protection from the
disease. The goal of using
these technologies to study
vaccines is to understand not only how
vaccines work,
but also why
some vaccines work better in
some subpopulations than others
and, if necessary,
to seek intervention approaches
in subpopulations for which
vaccines are less effective.
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In our laboratory we have been involved in
preclinical research
applied to the development of new
vaccines for many years. Well, preclinical research
allows us, before a vaccine is
studied in humans, to
first characterize its immunogenicity, that is, the
ability of a vaccine to induce an
immune response in the subject who
then receives it. But even more
importantly, what is a
unique tool that preclinical research offers us
is the ability to evaluate
protection, that is, the ability of a
subject who has received a
vaccination to be protected years
after vaccination when they
encounter the pathogen. Among the
many vaccines
we have tested and sampled in
preclinical research in recent years, there are two
innovative vaccine candidates
developed by a local company,
JSCK Vaxing for Global Health. These are
two vaccines based on an
innovative platform designed to protect against
invasive non-typhoidal salmonella infection
and cigella infection.
These two pathogens, in fact,
cause severe infections in some areas
of the world such as sub-Saharan Africa,
in certain specific
population groups, particularly children
and immunocompromised individuals.
Well, thanks to preclinical research
we have been able to demonstrate that both of
these vaccines are capable of not only
inducing an excellent antibody response.
We also characterized the
antibodies produced not only in quantity,
but also in terms of their quality and
their effector functions, that is, the
ability of the antibodies induced by
vaccination to kill the bacteria in the
event of an infection. The
study of these vaccines in
preclinical models has allowed us to demonstrate not
only the induction of a good
immune or moral response, but also
the induction of memory B cells, which
will be able, years after
vaccination, to protect the subject
from contracting the infection.
Data obtained from preclinical research
integrated with data obtained from the first
clinical studies have allowed a huge
step forward in research as
both of these vaccines are now in
an advanced phase of
clinical trials on humans and this represents
a very important result
for the development of new
vaccination strategies to combat new
emerging infections and
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in our research group and we
deal, among other things, also
with organs and organoids are
3D systems, therefore three-dimensional
of human tissues or organs that are
built in vitro, therefore in the
laboratory, starting from
undifferentiated human cells that, under
appropriate stimuli, self-organize to form
real miniature organs
that mimic both from a structural point of view
, therefore in terms of
three-dimensionality and of
cell typology and from a functional point of view
of human organs. And in
this way, in our laboratory
we have constructed, for example, a
human nasal epithelium, but we can also
create an intestinal mucosa, a
vaginal epithelium, and these are systems that
are very suitable for carrying out
pretonic studies, thus limiting
the use of
animal testing. And the interesting thing is that it is
possible to construct both
healthy organoids, for example a
normal respiratory vesicle with
cilia that beat normally or
cells that produce normal mucus,
and diseased organs that
therefore animate important climatic conditions
for study. Well,
for example, it is possible to replace
a healthy mucous with a diseased mucous, as
happens for example in patients with
cystic fibrosis, who have a
particularly thick and sticky mucus in
which bacteria get trapped and
are not eliminated by the
pulmonary thrush. And this is why
these patients suffer from
chronic infections practically
throughout their lives. And in conclusion, these
organs are
extremely ductile, extremely
plastic platforms, which can be used for studies
of infections, therefore with bacteria, viruses,
fungi, or they can be used to
test new drugs. We
are microbiologists here, so we taste
antimicrobials, we can taste
antibiotics, but also
more innovative drugs, such as
monoclonal antibodies. And if the system
is made more complex, for example,
by adding human immune cells
such as white blood cells, they can be
used to study the
immune and inflammatory response to
microbial infections.
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Alongside research, we are also
strongly committed to training the
new generation of
researchers involved in
pandemic preparedness. The University of Siena,
in fact, coordinates the national doctorate
in innovation in the diagnosis,
prevention, and treatment of infections at
risk of epidemic and pandemic.
This doctorate of national interest
involves 16 Italian universities, sees
the participation of
national and international research centers, and receives
significant support from the
Siena Biotecnopolo Foundation, the National Anti-Pandemic Center
. Every year we recruit over 30
PhD students. There
are currently 91 students from 12
different countries and with backgrounds in
various disciplinary fields. Our
goal is precisely to offer
multidisciplinary training
that is also based on close
collaboration and planning among
our doctoral students, so as to enable them
to contribute
to research for the control of
infections at risk of epidemic and pandemic.
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In my work, I study how our
immune system responds to vaccines,
particularly against respiratory viruses
like Covid-19.
Um, specifically I'm interested in understanding
what type of antibodies we produce and which
cells are involved in this
response. To do this I use a
very advanced technique based on
fluorescence, namely
spectral cytoflorimetry, which allows us to simultaneously analyze
many types of antibody cellular parameters
within the same
biological sample. By better understanding
how this response works, we can
help make vaccines
increasingly effective, not only for the
general population, but also for those who are more vulnerable
than those who are most vulnerable.
My research focuses on the
immune response to vaccines in
people living with HIV. Although
antiviral therapies are very
effective, their immune systems
can still behave
differently, affecting the
vaccination response.
Well, in particular I deal with the
study of B cells, responsible for
antibody production and the formation of
immune memory. Understanding these
responses is crucial to
assessing how
strong and long-lasting the vaccine protection is.
To do this, we use
very advanced technologies such as spectral flow cytometry,
which allows us to
simultaneously analyze many characteristics of
single immune cells and offers
very detailed insights into how
different populations of B cells
respond before and after vaccination.
And our goal is to
better understand these immunological mechanisms
and contribute to the development of
vaccination strategies for people
living with compromised immune systems
.
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My PhD project
focuses on the development,
characterization and analysis of
3D models of human respiratory epithelium at Air
Liquid Interface. This type of model
allows us to recreate in vitro a
differentiated and
pseudostratified epithelium physiologically
similar to the human one. Specifically,
my project focuses
on host-pathogen interactions with
particular interest in pathogens of
clinically relevant endocrine agents
such as Streptococcus pneumoniae, which is the
leading cause of bacterial pneumonia
worldwide and, despite commercial vaccines,
remains a priority pathogen
for the WHO.
Well, specifically I deal with the
study, analysis, adhesion,
internalization, and
inflammatory response of epithelial damage.
Furthermore, this project is
also expanding to include respiratory viruses of
clinical interest such as
respiratory syncytial virus, RSV, and
seasonal coronaviruses, seeking to
better understand the
underlying mechanisms of respiratory infection,
especially in cases of bacterial co-infection
.
The ultimate goal of the project is to
recreate a physiologically relevant 3D preclinical model
, not only for
studying the pathogenesis of
respiratory infections, but also,
above all, for analyzing and testing
new therapeutic strategies.
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Here in the laboratory I am involved in
characterizing the response of our
immune system to
vaccines and in particular for my
project and this response is
evaluated against vaccines that
are obtained using as an antigen
a sugar that is naturally present
on the membrane of bacteria. In
detail, in fact, through
spectral cytoflorimetry I am concerned with
characterizing the interaction between
B cells and these sugars. At the
same time, however, we are also developing
functional assays that allow us to
evaluate the antibodies
produced by these B cells in their
activity in fighting and preventing
infections caused by the same bacterium that
produces them.
I am part of a larger group
called the European Vaccine Hub, and
we are currently studying
preclinical models of
vaccination response. This is a topic that has
interested me a lot since high school,
so I'm really
happy to be able to explore it further here.
I believe that a great advantage of the
national doctorate is precisely the fact
that it fosters interactions and
collaborations between different
research centers and institutions. And
I think what
excites me most about this course right now is
the ability to
constantly interact with different experts from
different fields—a young,
dynamic, and extremely stimulating team.
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