The Viral Immunopathology Laboratory of the Complex Structure of Infectious Diseases and Hepatology deals with the study of the pathogenetic mechanisms of organ damage and immunological control of hepatitis virus infections (HBV, HCV and HDV), SARS-CoV2, HIV and hepatocarcinoma with a view to the development of innovative diagnostic and therapeutic strategies tailored to the individual host response profile to the individual infection or neoplastic pathology.
A diagnostic section also operates within the laboratory in which immunological, virological and molecular tests are performed for the evaluation of respiratory virus infections (SARS-CoV2, Influenza and RSV) CMV, HIV and mycobacterium tuberculosis.
The integration of research, diagnostic and clinical activities is aimed at maintaining the high quality level of care provided by the Complex Structure of Infectious Diseases and Hepatology, through the development of innovative diagnostic technologies and continuous collaboration with industries involved in the development of new therapies for emerging respiratory viral infections (e.g., SARS-CoV-2), so as to enable the clinical use of emerging therapeutic strategies in the early stages of their development.
Main lines of research:
1. Hepatitis B virus (HBV) and Delta virus (HDV) infection.
Analysis of phenotype, function, and gene expression profiles of virus-specific T- and B-lymphocytes and NK cells isolated from peripheral blood and liver of patients with different ability to control infection (acute infection, chronic infection, spontaneously cured subjects, or following therapy), with the aim of:
- characterize the condition of functional depletion of virus-specific T and B lymphocytes to identify new molecular targets for the development of novel anti-HBV and anti-HDV therapeutic strategies based on restoring protective immune responses by correcting deregulated intracellular metabolic and signaling pathways;
- identify immunologic and molecular parameters predictive of response to immunomodulatory therapies in chronically infected patients;
- develop laboratory-based tests of cell-mediated immunity to diagnose occult serum-negative HBV infection in order to reduce the risk of viral reactivation in patients undergoing immunosuppressive or biologic drug therapies.
2. Hepatitis C virus (HCV) infection.
Characterization of phenotype, metabolic and anti-viral function of HCV-specific and circulating NK lymphocyte T cell populations in the blood of chronically infected subjects undergoing treatment with direct antiviral action (DAA) drugs with the aim of:
- to define immunologic parameters predictive of nonresponse to therapy, to identify patients who need enhanced anti-viral therapies and for whom immunomodulatory treatment might be prospective;
- to define strategies to correct impaired immune activities, to be prospective as immunotherapies for chronic C infection.
3. SARS-CoV-2 infection.
- Defining the immunological correlates of infection control and organ damage by SARS-CoV-2, through a qualitative-quantitative characterization of the anti-viral effector and regulatory activities expressed by different lymphocyte (B, T, NK) and monocyte/dendritic populations isolated not only from circulating blood, but also from target organ (lung) and through a fine dissection of the transcriptional, genomic, metabolic and intracellular signaling profiles of these cell populations.
- Identification of novel therapeutic molecular targets and outcome predictors of SARS-CoV-2 infection, based on humoral and cell-mediated immunity parameters, and lymphocyte transcriptional profiles, in order to develop new personalized therapies profiled on infected host characteristics and disease severity.
4. Human immunodeficiency virus (HIV) infection.
Identification of the presence of circulating HIV-specific lymphocyte T populations in the peripheral blood of high risk, uninfected, pre-exposure prophylactically treated HIV-exposed subjects and their characterization by:
- analysis of the frequency of CD8, HIV-specific T lymphocytes and evaluation of the phenotypic set-up and in-vitro functional capacity of both CD8 and CD4 virus-specific T cells;
- identification of serum-immunological parameters specifically related to protection from infection;
- analysis of phenotypic and functional characteristics of non-antigen-specific lympho-monocyte populations (T-lymphocytes, B-lymphocytes, and NK cells) in the case of occurrence of sexually transmitted diseases in relation also to their clinical severity.
5. Hepatocarcinoma (HCC).
Analysis of genomic and proteomic expression profiles and phenotypic/functional characterization of CD8 and NK T lymphocytes circulating and infiltrating hepatic neoplastic tissue with the aim of:
- identify the molecular and cellular determinants of immunosuppression in hepatocarcinoma, with a focus on the role of the tumor microenvironment in inducing lymphocyte T dysfunction;
- characterize the condition of functional depletion and plasticity of T and NK lymphocytes for the identification of lymphocyte T subsets with predictive/prognostic value of response to immunomodulatory therapies in patients with hepatocarcinoma;
- identify molecular targets for innovative therapeutic strategies for the treatment of hepatocarcinoma based on functional lymphocyte restoration, with a focus on the immunometabolism of these cell populations.
Techniques in use
- Cell Biology: cell cultures and three-dimensional liver models, cell phenotyping, lymphocyte metabolism and function assays, Elispot
- Immunofluorescence: cytofluorimetry and Fluorospot
- Molecular Biology: from basic techniques (RT-PCR, qPCR, etc.) to transcriptomics in "bulk" and "single cell."
Main equipment provided
The Viral Immunopathology Laboratory is fully equipped for immunology and molecular Biology studies, with laminar flow hoods, refrigerated centrifuges, CO2 incubators, direct and inverted transmission microscopes, -20°C and -80°C freezers, thermocyclers, gel electrophoresis apparatus, and high-speed centrifuges. Specifically, the laboratory is equipped with:
- two cytofluorometers: FACSLyric (3 lasers) and LSRFortessa (4 lasers) - BD Biosciences
- a cell sorter, FACSAria III - BD Biosciences
- a 3-color Elispot and Fluorospot reader - C.T.L.
- two platforms for the identification and isolation of rare cells for single-cell transcriptomics (scRNAseq) experiments:
- DepArray - Silicon Biosystems-Menarini Group
- Rhapsody HT Xpress System - BD Biosciences
- an analyzer for the study of cell metabolism Seahorse - Agilent Technologies
