Staff: Dr. M. Rossi, Dr. C. Gianfrani, Dr. G. Mazzarella, Dr. F. Maurano

Collaborators from other National Institutions: Prof. R. De Luna, Prof R.Troncone, Dr. P Bergamo
Fellows: Dr. D. Luongo, Dr. S. Senger, Dr. R. Stefanile, Dr. R. D' Arienzo,
Students: M Gaita, A. Ferraro, F. Blasi

The Unit of Immunobiology (UIB) focuses on the interaction between food and immune system, particularly on the gut associated lymphoid tissue (GALT) as it functions in the maintainance of health and as it malfunctions in the production of food-related diseases. Current research is directed both at understanding the mechanisms that lead to the development of food intolerances and to generate an expanded knowledge base of the GALT physiology. Fundamental aims are then seen in the detection and production of molecular and cell tools useful to recover oral tolerance.

Research activities.

• Food intolerances:
  • Studies of murine models of gluten hypersensitivity and development of immunomodulatory strategies. An animal model of celiac disease (CD) is still lacking; to address this issue studies have been done in transgenic mice expressing the CD-linked HLA-DQ8 molecule by using several approaches to break the tolerance to dietary gluten. Immunomodulation protocols for down-regulating the gliadin-specific immune response in this model are also under investigation [1-4].
  • Identification of T cell epitopes of gliadin. A recombinant _-gliadin has been produced to identify, in the DQ8 mouse model, the immunodominant epitope/s; this study will give the opportunity to gain further insight on the peptides responsible for gut inflammation in CD.
  • Relationship between gluten and type I diabetes in NOD mice. This work is aimed to analyse the role of gluten in the onset of this autoimmune disease, and to look for the development of possible related enteropathy signs by using a known murine model of IDDM.
  • Analysis of antigen specificity, phenotype, and biological function of CD8+ T lymphocytes infiltrating normal and celiac intestinal mucosa. CD8+ T lymphocytes are hypothesized to have a key role in the induction of gluten-sensitive enteropathy, but little is known about their specificity [5].
  • Identification and characterization of food antigen-specific regulatory T cells and cytokines from small intestinal biopsies. We have data indicating the presence, in the intestinal mucosa of CD patients, of T regulatory cells which are able to suppress the gliadin-reactive inflammatory T cells in vitro. The role of IL-10 as a regulatory cytokine, has been also investigated by using in vitro organ cultures and intestinal T cell lines from CD patients [6-8].
  • Evaluation of gliadin peptides toxicity by using the in vitro organ culture system. A series of inflammation markers have been identified by imunohistochemistry, following culture of jejunal biopsies from CD patients in the presence of gliadin; these markers are useful to assess the toxicity of untested peptides from wheat or other cereals [9-16]
  • Analysis of the intracellular signalling pathway and transcription factor involved in the gliadin- induced mucosal inflammation. This work aims to identify the molecules that have a key role in the activation of mucosa inflammation in CD [17].
• GALT physiology:
  • Spore probiotics: an alternative to antibiotics. The overall aim of this project is to study the suppressive effect of Bacillus spores upon a mouse specific enteropathogen, Citrobacter rodentium and to analyse the mucosal immune responses to the pathogen.
  • Safety assessment in food and environment of genetically modified plants (SAFE). The project is focused on the development of in vivo protocols in animal models, useful to evaluate the ability of genetically modified foods to induce immunological tolerance.
  • Effect of conjugated linoleic acid (CLA) on the immune system. Most research into how fat in the diet may influence the functioning of the immune system has essentially focused on specific types of fatty acids. Among these ones, CLA has been extensively studied but contrasting effects were reported. To more directly address the question of a specific effect of CLA on immune cells, we have analysed the activity of CLA on the Jurkat cell line, a classical in vitro model of human T lymphocyte [18].

Selected Publications

• 1. Rossi M, Maurano M, Caputo N, Auricchio S, Sette A, Capparelli R, Troncone R. In mice, intravenous or intranasal administration of gliadin is able to down-regulate the specific immune response. Scand J Immunol 1999, 50:177-182.
• 2. Maurano F, Siciliano RA, De Giulio B, Luongo D, Mazzeo MF, Troncone R, Auricchio S, Rossi M. Intranasal administration of one alpha gliadin can down-regulate the immune response to whole gliadin in mice. Scand J Immunol 2001; 53: 1-7.
• 3. Rossi M, Maurano F, Luongo D, Fasano A, Uzzau S, Auricchio S, Troncone R. Zonula occludens toxin (zot) intereferes with the induction of nasal tolerance to gliadin. Immunol Letters 2002; 81, 217-221.
• 4. Senger S, Luongo D, Maurano F, Mazzeo MF, Sicialino RA, Gianfrani C, David C, Troncone R, Auricchio S, Rossi M. Intranasal administration of a recombinant _-gliadin down-regulates the immune response to wheat gliadin in DQ8 transgenic mice. Immunol Letters 2003; 88/2: 127-134.
• 5. Gianfrani C., Troncone R., Mugione P., Cosentini E., De Pascale M., Faruolo C., Senger S., Terrazzano G., Southwood S., Auricchio S., Sette A. Coeliac Disease association with CD8+ T cell responses: identification of a novel gliadin-derived HLA-A2 restricted epitope. J Immunol 170, 2719-2726, 2003.
• 6. Gianfrani C, Rossi M, Mazzarella G, Maurano F, Salvati V, Zanzi D, Auricchio S, Troncone R. Immunotherapy of coeliac disease: where do we stand? In "Primary prevention of coeliac disease. The utopia of new millenium? Perspective in Coeliac Disease". Catassi C. Fasano A. Corazza GR. (eds), Vol 1, 83-88.
• 7. F. Clot, C. Gianfrani, MC. Babron, F. Bouguerra, S. Southwood, MF. Kagnoff, R. Troncone, S. Percopo, JF. Eliaou, FC. Darpoux, A. Sette, and L. Greco. HLA-DR53 molecules are associated with susceptibility to Celiac Disease and selectively bind gliadin-derived peptides. Immunogenetics 1999, 49, 800-807.
• 8. R. Troncone, C. Gianfrani, G. Mazzarella, L. Greco, J. Guardiola, S. Auricchio and P. De Berardinis. The majority of gliadin-specific T cell clones from the coeliac small intestinal mucosa produce both interferon and IL4. Dig Dis Sci 1998, 43, 156-161.
• 9. R.Troncone, G.Mazzarella, A.Leone, M.Mayer, M.De Vincenzi, L.Greco, S.Auricchio."Gliadin Activates Mucosal Cell-Mediated Immunity In In Vitro Cultured Rectal Mucosa from Coeliac Patients and a Subset of Their Siblings" Gut 1998; 43: 484-489
• 10. Troncone R., Maurano F., Iovine G., Petrone E., Paparo F. Diagnostic criteria for coeliac disease. Changing Featurea of Coeliac Disease, edited by Lohiniemi S., Collin P., Maki M. (The Finnish Coeliac Society, Tampere), 1998, 7-12.
• 11. G. Mazzarella, VM Salvati, M Borrelli, F Riccio, C Gianfrani, G Iaquinto, S Auricchio, R Troncone. Cytokines in coeliac disease. J Dig Protection, 2000, 2, 49-56.
• 12. G.Mazzarella, F.Paparo, M.Maglio, R.Troncone "Organ Culture Of Rectal Mucosa" Methods In Molecular Medicine Vol 41, Coeliac Disease: Methods And Protocol Humana Press Inc. ( Edited By Dr. Marsh Michael ) 2000;Cap.12:163-173.
• 13. VM. Salvati, M Bajaj-Elliott, R Poulsom, G Mazzarella, K E A. Lundin, E M.Nilsen, R Troncone and T T. MacDonald: "Keratinocyte growth factor and coeliac disease". Gut 2001;49 176-181.
• 14. Mazzarella G, Maglio M, Paparo F, Nardone G, Stefanile R, Greco L, Van De Wal Y, Kooy Y, Koning F, Auricchio S, Troncone R. An immunodominant DQ8 restricted gliadin peptide activates small intestinal immune response in in vitro cultured mucosa from HLA-DQ8 positive but not HLA-DQ8 negative coeliac patients. Gut. 2003 Jan;52(1):57-62.
• 15. Troncone R, Franzese R, Mazzarella G, Paparo F, Auricchio R, Coto I, Mayer M, Greco L. Gluten sensitivity in a subset of children with insulin-dependent diabetes mellitus. Am J Gastroenterol 2003;98 (3):590-5.
• 16. Esposito C, Paparo F, Caputo I, Porta R, Salvati V.M., Mazzarella G, Auricchio S, Troncone R. Expression and Enzymatic Activity of Small Intestinal Tissue Transglutaminase in Coeliac Disease Am J Gastroenterol 2003 in press
• 17. G. Mazzarella , T.T. MacDonald, VM Salvati, P. Mulligan, L.Pasquale, R. Stefanile, P.Lionetti, R. Troncone, G. Monteleone. Activation of signal transducer and activator of transcription 1 in celiac disease. Am J Path 2003;162(6):1845-55.
• 18. P Bergamo, D Luongo, M Rossi. Conjugated linoleic acid - mediated apoptosis in Jurkat T cells involves the production of reactive oxygen species. Cell Physiol Biochem 2003, in press.

Research Grants 2002-2003:

• 2002. European Commission (contract QLK1-CT-1999): Evaluation of the prevalence of the coeliac disease and its genetic components in the European population, subcontractor.
• 2002-03. European Commission (contract QLK5-CT-2001-01729): Spore probiotics: an alternative to antibiotics", subcontractor.
• 2003-2005. MIUR (Italian Ministry of Education, University and Research) Safety Assessment in food and environment of genetically modified plants (SAFE), subcontractor of Metapontum Agrobios srl.