0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
tral nervous system, host immune response, and host reparative response.
This will be achieved by analysis of oligoclonal total IgG and EBV-specific
bands by isoelectric focusing and capillary immunoblotting; CXCL13, lymphotoxin-a,
Bcl-2, soluble CD8, soluble CD4, perforin, human Granzyme B,
and protein tau by ELISA; Nogo by Western blot.
U.O.15 – In the attempt to dissect the pathogenetic events characterizing
the early stage of MS, this unit will determine: 1) a complete profile of
the intrathecal humoral immune response, IgM index included; 2) EBV
specific CSF IgM; 3) EBV seropositivity; 4) EBV DNA and antibodies
against early lytic cycle viral proteins in the CSF; 5) the number of Cortical
Inflammatory Lesions by Double Inversion Recovery (DIR), in order to
demonstrate a possible association between the presence of an altered
intrathecal immune response, as well as a specific response to EBV (EBV
specific IgM, etc.), and cortical pathology, at the clinical onset of MS (pediatric
patients, Clinically Isolated Syndromes and possible MS) and in
patients with the primary progressive form of the disease. Standard analysis
and IgG and IgM index will be performed in CSF samples. In addition,
IgM will be characterized for their specificity against the main EBV epitopes
by immunoblotting. EBV DNA sequences will be searched both in
serum and CSF by RT-PCR.
GENERAL TRANSFERIBILITY AND POTENTIAL IMPACT OF RESULTS
Recent advances in the study of MS pathogenesis represent the basis for
the identification of novel biomarkers of disease. We will approach the study
of the immune response toward EBV and its correlation with both molecular
and cellular markers and disease course by analyzing blood and CFS from MS
patients. Specifically, we will perform experiments aimed at understanding
the following issues:
1) Frequency and activation state of EBV-specific lymphocyte populations
in the various phases of the disease.
2) Alterations in the signaling pathways of Toll-like receptors and in the
IFN-signature, which may both be indicators of the dysregulation of the
immune response towards EBV or of the development into a chronic phase of
the disease.
3) Gene expression profiling of cells of the immune system with highthroughput
screening methodologies.
4) Analysis of oligoclonal bands (total and EBV-specific) and of soluble
immune molecules in the CSF of MS patients.
5) Serum alterations of biochemical parameters measuring oxidative
stress.
The identification of novel biomarkers is crucial for monitoring the disease
course and for assessment of therapeutic efficacy. With advanced technologies
it will be possible to identify, in the blood and CSF of MS patients,
molecular and functional indicators of the different phases of the disease,
642 2009