0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
Subproject Mechanism
P26 – Fondazione Santa Lucia
Luca Battistini
Objectives
The objective is to accurately define the ab CD8+ and gd T cell subsets
that respond to EBV antigens in peripheral blood obtained from MS patients
in the different phases of the disease. The crucial aspect of this project is to
monitor these responses in a follow-up study, and to correlate them with the
activity of MS, measured both clinically and by MRI. Furthermore, these cell
subsets will be functionally characterized, in order to determine their cytotoxic
potential and/or their ability to release pro-inflammatory cytokines.
Description of work
The experimental plan will develop according to the following steps:
1) Selection of 50 MS patients willing to participate in the follow-up
study, which consists of an average of 4 blood drawings per year. These
patients, when possible, will also undergo imaging studies by MRI. The clinical
evaluation will be performed by Diego Centonze, Neurologist at FSL,
before each blood drawing.
2) Phenotypic characterization of EBV-specific T cell subsets will be performed
with polychromatic flow cytometry using pentamers containing EBVderived
peptides (from proteins specific for the lytic or the latent phase of disease).
These pentamers have been tested successfully and the most appropriate
combination of markers has already been established.
3) Functional characterization of EBV-specific T cell subsets will be performed
by analyzing their cytotoxic potential and their ability to secrete
proinflammatory cytokines.
4) CD8 ab and gd T cell clones obtained from the CSF of MS patients will
be tested for EBV-specificity.
Deliverables
1 – Identification and characterization of EBV-specific ab CD8+ and gd T
cells in MS patients (54 month).
2 – Correlation of the EBV-specific immune response with the clinical
course of the disease (60 month).
Milestones and expected result
1 – Knowledge on the phenotypical and functional characteristics of EBVspecific
T cells in MS patients in different phases of disease activity.
2 – Knowledge on the role of EBV-specific T cells in MS pathogenesis.
602 2009