35 results on '"Claudya Tenca"'
Search Results
2. Expression of Immunoglobulin Receptors with Distinctive Features Indicating Antigen Selection by Marginal Zone B Cells from Human Spleen
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Monica Colombo, Giovanna Cutrona, Daniele Reverberi, Silvia Bruno, Fabio Ghiotto, Claudya Tenca, Kostas Stamatopoulos, Anastasia Hadzidimitriou, Jenny Ceccarelli, Sandra Salvi, Simona Boccardo, Maria Grazia Calevo, Amleto De Santanna, Mauro Truini, Franco Fais, and Manlio Ferrarini
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Antigen Selection ,Intraclonal Diversification ,Recurrent Sequence ,Activation-induced Cytidine Deaminase (AID) ,Follicular Mantle (FM) ,Therapeutics. Pharmacology ,RM1-950 ,Biochemistry ,QD415-436 - Abstract
Abstract Marginal zone (MZ) B cells, identified as surface (s)IgMhighsIgDlowCD23low/−CD21+CD38− B cells, were purified from human spleens, and the features of their V(D)J gene rearrangements were investigated and compared with those of germinal center (GC), follicular mantle (FM) and switched memory (SM) B cells. Most MZ B cells were CD27+ and exhibited somatic hypermutations (SHM), although to a lower extent than SM B cells. Moreover, among MZ B-cell rearrangements, recurrent sequences were observed, some of which displayed intraclonal diversification. The same diversifying sequences were detected in very low numbers in GC and FM B cells and only when a highly sensitive, gene-specific polymerase chain reaction was used. This result indicates that MZ B cells could expand and diversify in situ and also suggested the presence of a number of activation-induced cytidine deaminase (AID)-expressing B cells in the MZ. The notion of antigen-driven expansion/selection in situ is further supported by the VH CDR3 features of MZ B cells with highly conserved amino acids at specific positions and by the finding of shared (“stereotyped”) sequences in two different spleens. Collectively, the data are consistent with the notion that MZ B cells are a special subset selected by in situ antigenic stimuli.
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- 2013
- Full Text
- View/download PDF
3. Dependence of Immunoglobulin Class Switch Recombination in B Cells on Vesicular Release of ATP and CD73 Ectonucleotidase Activity
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Francesca Schena, Stefano Volpi, Caterina Elisa Faliti, Federica Penco, Spartaco Santi, Michele Proietti, Ursula Schenk, Gianluca Damonte, Annalisa Salis, Marta Bellotti, Franco Fais, Claudya Tenca, Marco Gattorno, Hermann Eibel, Marta Rizzi, Klaus Warnatz, Marco Idzko, Cemil Korcan Ayata, Mirzokhid Rakhmanov, Thierry Galli, Alberto Martini, Marco Canossa, Fabio Grassi, and Elisabetta Traggiai
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Biology (General) ,QH301-705.5 - Abstract
Immunoglobulin (Ig) isotype diversification by class switch recombination (CSR) is an essential process for mounting a protective humoral immune response. Ig CSR deficiencies in humans can result from an intrinsic B cell defect; however, most of these deficiencies are still molecularly undefined and diagnosed as common variable immunodeficiency (CVID). Here, we show that extracellular adenosine critically contributes to CSR in human naive and IgM memory B cells. In these cells, coordinate stimulation of B cell receptor and toll-like receptors results in the release of ATP stored in Ca2+-sensitive secretory vesicles. Plasma membrane ectonucleoside triphosphate diphosphohydrolase 1 CD39 and ecto-5′-nucleotidase CD73 hydrolyze ATP to adenosine, which induces CSR in B cells in an autonomous fashion. Notably, CVID patients with impaired class-switched antibody responses are selectively deficient in CD73 expression in B cells, suggesting that CD73-dependent adenosine generation contributes to the pathogenesis of this disease.
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- 2013
- Full Text
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4. A soluble form of CTLA-4 is present in paediatric patients with acute lymphoblastic leukaemia and correlates with CD1d+ expression.
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Rita Simone, Claudya Tenca, Franco Fais, Matteo Luciani, Giulio De Rossi, Giampaola Pesce, Marcello Bagnasco, and Daniele Saverino
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Medicine ,Science - Abstract
CTLA-4 is a key factor in regulating and maintaining self tolerance, providing a negative signal to the T cell and thus limiting immune responses. Several polymorphisms within the CTLA-4 gene have been associated with an increased risk of developing autoimmune diseases and, very recently, with susceptibility to human cancer. Acute lymphoblastic leukemia is a clonal disorder of lymphoid progenitors representing the most frequent malignancy of childhood. Here, we show the presence at significantly elevated levels of a circulating soluble form of CTLA-4 in 70% of B-ALL pediatric patients with active disease, the positive correlation between the percentage of leukemic B lymphocytes and the amount of serum sCTLA-4, and the expression of sCTLA-4 transcript by B cells in patients. Finally, a correlation between CD1d expression (a negative prognostic marker) and the sCTLA-4 in B-ALL patients was observed. This suggests a possible role of this soluble molecule as a marker of progression or severity of the neoplastic disease.
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- 2012
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5. Correction: CXCR1 Is Expressed by Human B Lymphocytes and Mediates CXCL1 Driven Chemotaxis of Tonsil Centrocytes.
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Anna Corcione, Elisa Ferretti, Maria Bertolotto, Franco Fais, Lizzia Raffaghello, Andrea Gregorio, Claudya Tenca, Luciano Ottonello, Claudio Gambini, Glaucia Furtado, Sergio Lira, and Vito Pistoia
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Medicine ,Science - Published
- 2010
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6. CX3CR1 is expressed by human B lymphocytes and mediates [corrected] CX3CL1 driven chemotaxis of tonsil centrocytes.
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Anna Corcione, Elisa Ferretti, Maria Bertolotto, Franco Fais, Lizzia Raffaghello, Andrea Gregorio, Claudya Tenca, Luciano Ottonello, Claudio Gambini, Glaucia Furtado, Sergio Lira, and Vito Pistoia
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Medicine ,Science - Abstract
BACKGROUND:Fractalkine/CX(3)CL1, a surface chemokine, binds to CX(3)CR1 expressed by different lymphocyte subsets. Since CX(3)CL1 has been detected in the germinal centres of secondary lymphoid tissue, in this study we have investigated CX(3)CR1 expression and function in human naïve, germinal centre and memory B cells isolated from tonsil or peripheral blood. METHODOLOGY/PRINCIPAL FINDINGS:We demonstrate unambiguously that highly purified human B cells from tonsil and peripheral blood expressed CX(3)CR1 at mRNA and protein levels as assessed by quantitative PCR, flow cytometry and competition binding assays. In particular, naïve, germinal centre and memory B cells expressed CX(3)CR1 but only germinal centre B cells were attracted by soluble CX(3)CL1 in a transwell assay. CX(3)CL1 signalling in germinal centre B cells involved PI3K, Erk1/2, p38, and Src phosphorylation, as assessed by Western blot experiments. CX(3)CR1(+) germinal centre B cells were devoid of centroblasts and enriched for centrocytes that migrated to soluble CX(3)CL1. ELISA assay showed that soluble CX(3)CL1 was secreted constitutively by follicular dendritic cells and T follicular helper cells, two cell populations homing in the germinal centre light zone as centrocytes. At variance with that observed in humans, soluble CX(3)CL1 did not attract spleen B cells from wild type mice. OVA immunized CX(3)CR1(-/-) or CX(3)CL1(-/-) mice showed significantly decreased specific IgG production compared to wild type mice. CONCLUSION/SIGNIFICANCE:We propose a model whereby human follicular dendritic cells and T follicular helper cells release in the light zone of germinal centre soluble CX(3)CL1 that attracts centrocytes. The functional implications of these results warrant further investigation.
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- 2009
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7. Adoptive immunotherapy mediated by ex vivo expanded natural killer T cells against CD1d-expressing lymphoid neoplasms
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Davide Bagnara, Adalberto Ibatici, Mirko Corselli, Nadia Sessarego, Claudya Tenca, Amleto De Santanna, Andrea Mazzarello, Antonio Daga, Renzo Corvò, Giulio De Rossi, Francesco Frassoni, Ermanno Ciccone, and Franco Fais
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Diseases of the blood and blood-forming organs ,RC633-647.5 - Abstract
Background CD1d is a monomorphic antigen presentation molecule expressed in several hematologic malignancies. Alpha-galactosylceramide (α-GalCer) is a glycolipid that can be presented to cytotoxic CD1d-restricted T cells. These reagents represent a potentially powerful tool for cell mediated immunotherapy.Design and Methods We set up an experimental model to evaluate the use of adoptively transferred cytotoxic CD1d-restricted T cells and α-GalCer in the treatment of mice engrafted with CD1d+ lymphoid neoplastic cells. To this end the C1R cell line was transfected with CD1c or CD1d molecules. In addition, upon retroviral infection firefly luciferase was expressed on C1R transfected cell lines allowing the evaluation of tumor growth in xenografted immunodeficient NOD/SCID mice.Results The C1R-CD1d cell line was highly susceptible to specific CD1d-restricted T cell cytotoxicity in the presence α-GalCer in vitro. After adoptive transfer of CD1d-restricted T cells and α-GalCer to mice engrafted with both C1R-CD1c and C1R-CD1d, a reduction in tumor growth was observed only in CD1d+ masses. In addition, CD1d-restricted T-cell treatment plus α-GalCer eradicated small C1R-CD1d+ nodules. Immunohistochemical analysis revealed that infiltrating NKT cells were mainly observed in CD1d nodules.Conclusions Our results indicate that ex vivo expanded cytotoxic CD1d-restricted T cells and α-GalCer may represent a new immunotherapeutic tool for treatment of CD1d+ hematologic malignancies.
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- 2009
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8. A reversible carnitine palmitoyltransferase (CPT1) inhibitor offsets the proliferation of chronic lymphocytic leukemia cells
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Davide Bagnara, Fabio Ghiotto, Franco Fais, Claudya Tenca, Daniele Reverberi, Andrea Nicola Mazzarello, Ermanno Ciccone, Elena Gugiatti, Adalberto Ibatici, Marina Fabbi, Silvia Ravera, Giovanna Cutrona, Daniela Zarcone, Silvia Bruno, and Zbigniew Darzynkiewicz
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0301 basic medicine ,Chronic lymphocytic leukemia ,Apoptosis ,Apoptosis, Cell Therapy and Immunotherapy, Chronic Lymphocytic Leukemia ,03 medical and health sciences ,medicine ,Humans ,Chronic Lymphocytic Leukemia ,Carnitine O-palmitoyltransferase ,Carnitine ,Enzyme Inhibitors ,Online Only Articles ,Cell Proliferation ,A549 cell ,Carnitine O-Palmitoyltransferase ,Cell growth ,Chemistry ,Hematology ,HCT116 Cells ,medicine.disease ,Cell Therapy and Immunotherapy ,Leukemia, Lymphocytic, Chronic, B-Cell ,Neoplasm Proteins ,Leukemia ,030104 developmental biology ,A549 Cells ,PC-3 Cells ,MCF-7 Cells ,Cancer research ,medicine.drug - Published
- 2018
9. Functional Activation of Osteoclast Commitment in Chronic Lymphocytic Leukaemia: A Possible Role for RANK/RANKL Pathway
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Davide Bagnara, Adalberto Ibatici, Daniela de Totero, Michele Pennone, Paolo Giannoni, Alessandro Bellini, Anna Maria Orengo, Michele Piana, Maurizio Miglino, Hülya Efetürk, Francesca La Rosa, Elena Gugiatti, Michele Cilli, Francesco Fiz, Cecilia Marini, Alberto Nieri, Franco Fais, Valerio Brucato, Laura Emionite, Giovanna Cutrona, Elisabetta Tedone, Cristina Campi, Carlo Emanuele Neumaier, Anna Maria Massone, Silvia Bruno, Serena Matis, Anna Borra, Roberta Piva, Gianmario Sambuceti, Matteo Bauckneht, Claudya Tenca, Paolo Bruzzi, Marini, C., Bruno, S., Fiz, F., Campi, C., Piva, R., Cutrona, G., Matis, S., Nieri, A., Miglino, M., Ibatici, A., Maria Orengo, A., Maria Massone, A., Neumaier, C., Totero, D., Giannoni, P., Bauckneht, M., Pennone, M., Tenca, C., Gugiatti, E., Bellini, A., Borra, A., Tedone, E., Efetã¼rk, H., Rosa, F., Emionite, L., Cilli, M., Bagnara, D., Brucato, V., Bruzzi, P., Piana, M., Fais, F., and Sambuceti, G.
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Male ,0301 basic medicine ,Chronic lymphocytic leukaemia ,Clone (cell biology) ,Osteoclasts ,lcsh:Medicine ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,Bone Marrow ,Positron Emission Tomography Computed Tomography ,hemic and lymphatic diseases ,80 and over ,Prospective Studies ,Chronic ,lcsh:Science ,Aged, 80 and over ,Settore ING-IND/24 - Principi Di Ingegneria Chimica ,Leukemia ,Multidisciplinary ,Bone Density Conservation Agents ,Receptor Activator of Nuclear Factor-kappa B ,biology ,Middle Aged ,Lymphocytic ,medicine.anatomical_structure ,Denosumab ,RANKL ,030220 oncology & carcinogenesis ,Female ,medicine.drug ,Adult ,Stromal cell ,Article ,03 medical and health sciences ,Osteoclast ,medicine ,Animals ,Humans ,Aged ,RANK/RANKL Pathway ,business.industry ,RANK Ligand ,lcsh:R ,B-Cell ,Diagnostic markers ,Glucose ,Leukemia, Lymphocytic, Chronic, B-Cell ,Xenograft Model Antitumor Assays ,medicine.disease ,030104 developmental biology ,Chronic Lymphocytic Leukaemia ,biology.protein ,Cancer research ,Inbred NOD ,lcsh:Q ,Bone marrow ,business - Abstract
Skeletal erosion has been found to represent an independent prognostic indicator in patients with advanced stages of chronic lymphocytic leukaemia (CLL). Whether this phenomenon also occurs in early CLL phases and its underlying mechanisms have yet to be fully elucidated. In this study, we prospectively enrolled 36 consecutive treatment-naïve patients to analyse skeletal structure and bone marrow distribution using a computational approach to PET/CT images. This evaluation was combined with the analysis of RANK/RANKL loop activation in the leukemic clone, given recent reports on its role in CLL progression. Bone erosion was particularly evident in long bone shafts, progressively increased from Binet stage A to Binet stage C, and was correlated with both local expansion of metabolically active bone marrow documented by FDG uptake and with the number of RANKL + cells present in the circulating blood. In immune-deficient NOD/Shi-scid, γcnull (NSG) mice, administration of CLL cells caused an appreciable compact bone erosion that was prevented by Denosumab. CLL cell proliferation in vitro correlated with RANK expression and was impaired by Denosumab-mediated disruption of the RANK/RANKL loop. This study suggests an interaction between CLL cells and stromal elements able to simultaneously impair bone structure and increase proliferating potential of leukemic clone.
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- 2017
10. Expression of Immunoglobulin Receptors with Distinctive Features Indicating Antigen Selection by Marginal Zone B Cells from Human Spleen
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Franco Fais, Jenny Ceccarelli, Amleto De Santanna, Giovanna Cutrona, Monica Colombo, Maria Grazia Calevo, Claudya Tenca, Manlio Ferrarini, Fabio Ghiotto, Daniele Reverberi, Mauro Truini, Kostas Stamatopoulos, Sandra Salvi, Simona Boccardo, Silvia Bruno, and Anastasia Hadzidimitriou
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Palatine Tonsil ,Somatic hypermutation ,Spleen ,Antigen ,Genetics ,medicine ,Humans ,Child ,Gene Rearrangement, B-Lymphocyte ,Molecular Biology ,Cells, Cultured ,Genetics (clinical) ,B-Lymphocytes ,biology ,Germinal center ,Sequence Analysis, DNA ,Articles ,Cytidine deaminase ,Gene rearrangement ,Germinal Center ,Marginal zone ,Molecular biology ,medicine.anatomical_structure ,Child, Preschool ,biology.protein ,Molecular Medicine ,Somatic Hypermutation, Immunoglobulin ,Antibody - Abstract
Marginal zone (MZ) B cells, identified as surface (s)IgM(high)sIgD(low)CD23(low/-)CD21(+)CD38(-) B cells, were purified from human spleens, and the features of their V(D)J gene rearrangements were investigated and compared with those of germinal center (GC), follicular mantle (FM) and switched memory (SM) B cells. Most MZ B cells were CD27(+) and exhibited somatic hypermutations (SHM), although to a lower extent than SM B cells. Moreover, among MZ B-cell rearrangements, recurrent sequences were observed, some of which displayed intraclonal diversification. The same diversifying sequences were detected in very low numbers in GC and FM B cells and only when a highly sensitive, gene-specific polymerase chain reaction was used. This result indicates that MZ B cells could expand and diversify in situ and also suggested the presence of a number of activation-induced cytidine deaminase (AID)-expressing B cells in the MZ. The notion of antigen-driven expansion/selection in situ is further supported by the VH CDR3 features of MZ B cells with highly conserved amino acids at specific positions and by the finding of shared ("stereotyped") sequences in two different spleens. Collectively, the data are consistent with the notion that MZ B cells are a special subset selected by in situ antigenic stimuli.
- Published
- 2013
11. Dependence of Immunoglobulin Class Switch Recombination in B Cells on Vesicular Release of ATP and CD73 Ectonucleotidase Activity
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Marta Rizzi, Marco Canossa, Stefano Volpi, Annalisa Salis, Thierry Galli, Marta Bellotti, Mirzokhid Rakhmanov, Gianluca Damonte, Klaus Warnatz, Marco Idzko, Claudya Tenca, Alberto Martini, Francesca Schena, Federica Penco, Elisabetta Traggiai, Franco Fais, Cemil Korcan Ayata, Fabio Grassi, Spartaco Santi, Michele Proietti, Ursula Schenk, Caterina Elisa Faliti, Marco Gattorno, Hermann Eibel, Schena, Francesca, Volpi, Stefano, Faliti, Caterinaelisa, Penco, Federica, Santi, Spartaco, Proietti, Michele, Schenk, Ursula, Damonte, Gianluca, Salis, Annalisa, Bellotti, Marta, Fais, Franco, Tenca, Claudya, Gattorno, Marco, Eibel, Hermann, Rizzi, Marta, Warnatz, Klau, Idzko, Marco, Ayata, Cemil Korcan, Rakhmanov, Mirzokhid, Galli, Thierry, Martini, Alberto, Canossa, Marco, Grassi, Fabio, and Traggiai, Elisabetta
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B-cell receptor ,B-Lymphocyte Subsets ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Immunoglobulin Class Switch Recombination ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Adenosine Triphosphate ,Antigens, CD ,medicine ,Animals ,Humans ,lcsh:QH301-705.5 ,5'-Nucleotidase ,030304 developmental biology ,B-Lymphocyte Subset ,Recombination, Genetic ,0303 health sciences ,B-Lymphocytes ,Biochemistry, Genetics and Molecular Biology (all) ,Animal ,Common variable immunodeficiency ,Apyrase ,B-Lymphocyte ,medicine.disease ,Isotype ,Adenosine ,Immunoglobulin Class Switching ,3. Good health ,Cell biology ,Common Variable Immunodeficiency ,Biochemistry ,Immunoglobulin class switching ,lcsh:Biology (General) ,Ectonucleoside Triphosphate Diphosphohydrolase 1 ,Antibody Formation ,biology.protein ,Antibody ,030215 immunology ,medicine.drug ,Human - Abstract
Immunoglobulin (Ig) isotype diversification by class switch recombination (CSR) is an essential process for mounting a protective humoral immune response. Ig CSR deficiencies in humans can result from an intrinsic B cell defect; however, most of these deficiencies are still molecularly undefined and diagnosed as common variable immunodeficiency (CVID). Here, we show that extracellular adenosine critically contributes to CSR in human naive and IgM memory B cells. In these cells, coordinate stimulation of B cell receptor and toll-like receptors results in the release of ATP stored in Ca2+-sensitive secretory vesicles. Plasma membrane ectonucleoside triphosphate diphosphohydrolase 1 CD39 and ecto-5′-nucleotidase CD73 hydrolyze ATP to adenosine, which induces CSR in B cells in an autonomous fashion. Notably, CVID patients with impaired class-switched antibody responses are selectively deficient in CD73 expression in B cells, suggesting that CD73-dependent adenosine generation contributes to the pathogenesis of this disease.
- Published
- 2013
- Full Text
- View/download PDF
12. Deciphering KRAS and NRAS mutated clone dynamics in MLL-AF4 paediatric leukaemia by ultra deep sequencing analysis
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Franco Fais, Valentina Serafin, Geertruy te Kronnie, Giovanni Cazzaniga, Giuseppe Basso, Claudya Tenca, Emanuela Giarin, Maria Grazia Valsecchi, Michela Bardini, Benedetta Accordi, Paola De Lorenzo, Luca Trentin, Silvia Bresolin, Trentin, L, Bresolin, S, Giarin, E, Bardini, M, Serafin, V, Accordi, B, Fais, F, Tenca, C, De Lorenzo, P, Valsecchi, M, Cazzaniga, G, Te Kronnie, G, and Basso, G
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0301 basic medicine ,Neuroblastoma RAS viral oncogene homolog ,Male ,Oncogene Proteins, Fusion ,Clone (cell biology) ,Biology ,medicine.disease_cause ,Deep sequencing ,Article ,GTP Phosphohydrolases ,Proto-Oncogene Proteins p21(ras) ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,hemic and lymphatic diseases ,medicine ,Humans ,Fusion ,Sanger sequencing ,Oncogene Proteins ,Mutation ,Multidisciplinary ,Leukemia ,Membrane Proteins ,High-Throughput Nucleotide Sequencing ,Amplicon ,Molecular biology ,Female ,Myeloid-Lymphoid Leukemia Protein ,030104 developmental biology ,030220 oncology & carcinogenesis ,symbols ,KRAS - Abstract
To induce and sustain the leukaemogenic process, MLL-AF4+ leukaemia seems to require very few genetic alterations in addition to the fusion gene itself. Studies of infant and paediatric patients with MLL-AF4+ B cell precursor acute lymphoblastic leukaemia (BCP-ALL) have reported mutations in KRAS and NRAS with incidences ranging from 25 to 50%. Whereas previous studies employed Sanger sequencing, here we used next generation amplicon deep sequencing for in depth evaluation of RAS mutations in 36 paediatric patients at diagnosis of MLL-AF4+ leukaemia. RAS mutations including those in small sub-clones were detected in 63.9% of patients. Furthermore, the mutational analysis of 17 paired samples at diagnosis and relapse revealed complex RAS clone dynamics and showed that the mutated clones present at relapse were almost all originated from clones that were already detectable at diagnosis and survived to the initial therapy. Finally, we showed that mutated patients were indeed characterized by a RAS related signature at both transcriptional and protein levels and that the targeting of the RAS pathway could be of beneficial for treatment of MLL-AF4+ BCP-ALL clones carrying somatic RAS mutations.
- Published
- 2016
13. N-(4-hydroxyphenyl)retinamide promotes apoptosis of resting and proliferating B-cell chronic lymphocytic leukemia cells and potentiates fludarabine and ABT-737 cytotoxicity
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Paola Luzzi, Franco Fais, Fortunato Morabito, Anna Grazia Recchia, Claudya Tenca, Silvia Bruno, Maria Bono, Andrea Nicola Mazzarello, Salvatore Casciaro, Fabio Ghiotto, and Andrea Decensi
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Cancer Research ,Fenretinide ,Chronic lymphocytic leukemia ,Antineoplastic Agents ,Apoptosis ,Protein degradation ,Piperazines ,Nitrophenols ,chemistry.chemical_compound ,Downregulation and upregulation ,immune system diseases ,hemic and lymphatic diseases ,medicine ,Humans ,Cytotoxic T cell ,Cell Proliferation ,Sulfonamides ,CD40 ,biology ,Biphenyl Compounds ,Drug Synergism ,Hematology ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Fludarabine ,Leukemia ,Proto-Oncogene Proteins c-bcl-2 ,Oncology ,chemistry ,Immunology ,biology.protein ,Cancer research ,Myeloid Cell Leukemia Sequence 1 Protein ,Reactive Oxygen Species ,Vidarabine ,medicine.drug - Abstract
The in vitro effects of the synthetic retinoid N-(4-hydroxyphenyl)retinamide (4HPR, fenretinide) on primary B-cell chronic lymphocytic leukemia (CLL) cells from previously untreated CLL patients were investigated. 4HPR promoted the intrinsic apoptotic pathway by reactive oxygen species (ROS) generation and was accompanied by drop of Mcl-1 protein expression. The latter was not attributable to transcriptional downregulation but to protein degradation mediated by jun N-terminal kinase activation, and likely by NF-kB downregulation and Noxa upregulation. CLL cells stimulated in vitro with CD40L did not increase 4HPR chemoresistance if activation was accompanied by proliferation. Intra-patient analysis confirmed that the proliferating pool of CLL cells was more sensitive to the cytotoxic action of 4HPR than the activated but resting CLL subpopulation. The different 4HPR susceptibility of the two subpopulations was associated with higher Noxa expression in proliferating CLLs. Combination experiments revealed that 4HPR strongly potentiated ABT-737 cytotoxicity, especially in proliferating CLL cells that displayed amplified chemoresistance to ABT-737 alone. Synergic cytotoxicity was also demonstrated in combination with fludarabine, in both resting and stimulated CLL samples. This study entitles 4HPR to be assayed as a chemotherapeutic adjuvant for the treatment of CLL.
- Published
- 2012
14. Adoptive immunotherapy mediated by ex vivo expanded natural killer T cells against CD1d-expressing lymphoid neoplasms
- Author
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Adalberto Ibatici, Francesco Frassoni, Franco Fais, Davide Bagnara, Ermanno Ciccone, Andrea Nicola Mazzarello, Antonio Daga, Amleto De Santanna, Mirko Corselli, Giulio De Rossi, Nadia Sessarego, Claudya Tenca, and Renzo Corvò
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Adoptive cell transfer ,T-Lymphocytes ,medicine.medical_treatment ,Antigen presentation ,chemical and pharmacologic phenomena ,Mice, SCID ,Immunotherapy, Adoptive ,Mice ,Interleukin 21 ,Mice, Inbred NOD ,parasitic diseases ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Antigen-presenting cell ,biology ,hemic and immune systems ,Hematology ,Immunotherapy ,Natural killer T cell ,Immunohistochemistry ,Killer Cells, Natural ,carbohydrates (lipids) ,Hematologic Neoplasms ,Immune System ,CD1D ,Immunology ,biology.protein ,Cancer research ,Natural Killer T-Cells ,Original Article ,lipids (amino acids, peptides, and proteins) ,Antigens, CD1d ,Neoplasm Transplantation - Abstract
Background CD1d is a monomorphic antigen presentation molecule expressed in several hematologic malignancies. Alpha-galactosylceramide (α-GalCer) is a glycolipid that can be presented to cytotoxic CD1d-restricted T cells. These reagents represent a potentially powerful tool for cell mediated immunotherapy.Design and Methods We set up an experimental model to evaluate the use of adoptively transferred cytotoxic CD1d-restricted T cells and α-GalCer in the treatment of mice engrafted with CD1d+ lymphoid neoplastic cells. To this end the C1R cell line was transfected with CD1c or CD1d molecules. In addition, upon retroviral infection firefly luciferase was expressed on C1R transfected cell lines allowing the evaluation of tumor growth in xenografted immunodeficient NOD/SCID mice.Results The C1R-CD1d cell line was highly susceptible to specific CD1d-restricted T cell cytotoxicity in the presence α-GalCer in vitro. After adoptive transfer of CD1d-restricted T cells and α-GalCer to mice engrafted with both C1R-CD1c and C1R-CD1d, a reduction in tumor growth was observed only in CD1d+ masses. In addition, CD1d-restricted T-cell treatment plus α-GalCer eradicated small C1R-CD1d+ nodules. Immunohistochemical analysis revealed that infiltrating NKT cells were mainly observed in CD1d nodules.Conclusions Our results indicate that ex vivo expanded cytotoxic CD1d-restricted T cells and α-GalCer may represent a new immunotherapeutic tool for treatment of CD1d+ hematologic malignancies.
- Published
- 2009
15. Inhibitory Receptors CD85j, LAIR-1, and CD152 Down-Regulate Immunoglobulin and Cytokine Production by Human B Lymphocytes
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Claudya Tenca, Lorenzo Battini, Franco Fais, Ermanno Ciccone, Andrea Merlo, Carlo E. Grossi, and Daniele Saverino
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CD4-Positive T-Lymphocytes ,Lipopolysaccharides ,Microbiology (medical) ,medicine.drug_class ,medicine.medical_treatment ,CD40 Ligand ,Lymphocyte Cooperation ,Clinical Biochemistry ,Immunology ,Down-Regulation ,Antibodies and Mediators of Immunity ,Monoclonal antibody ,Immunoglobulin E ,Antibodies ,Cell Line ,Leukocyte Immunoglobulin-like Receptor B1 ,Antigen ,Antigens, CD ,medicine ,Humans ,Immunology and Allergy ,CTLA-4 Antigen ,Receptors, Immunologic ,Cells, Cultured ,B-Lymphocytes ,CD40 ,biology ,Tumor Necrosis Factor-alpha ,Chemistry ,Interleukin-8 ,Antibodies, Monoclonal ,Antigens, Differentiation ,Immunoglobulin Class Switching ,Interleukin-10 ,Cytokine ,Immunoglobulin M ,Immunoglobulin class switching ,Immunoglobulin G ,biology.protein ,Interleukin-4 ,Antibody - Abstract
Class switching consists in the substitution of the heavy-chain constant region of immunoglobulin M (IgM) with that of IgG, IgA, or IgE. This enables antibodies to acquire new effector functions that are crucial to combat invading pathogens. Class switching usually requires engagement of CD40 on B cells by CD40 ligand (CD40L) on antigen-activated CD4 + T cells and the production of cytokines. The process must be regulated tightly because abnormal IgG and IgA production favors the onset of autoimmunity, whereas increased switching to IgE leads to atopy. These inflammatory disorders can be triggered or exacerbated by costimulatory signals. Although thoroughly investigated on T cells, the roles of the inhibitory receptors CD85j, LAIR-1, and CD152 on B-cell functions have not been fully elucidated. In this study we show that cross-linking of the B-cell inhibitory receptors by specific monoclonal antibodies inhibits IgG and IgE production, reduces the percentage of IgG- and IgE-expressing B cells, and down-regulates interleukin 8 (IL-8), IL-10, and tumor necrosis factor alpha production. These effects were demonstrated using different B-cell stimulatory pathways (recall antigens, CD40L-transfected cells plus IL-4, and lipopolysaccharide plus IL-4). It thus appears that CD85j, LAIR-1, and CD152 play a central role for the control of IL-4-driven isotype switching.
- Published
- 2005
16. Death of T cell precursors in the human thymus: a role for CD38
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Carlo Pesce, Silvia Deaglio, Andrea Merlo, Daniele Saverino, Claudya Tenca, Carlo E. Grossi, Fabio Malavasi, Marina Fabbi, Amleto De Santanna, Silvia Bruno, Ermanno Ciccone, Dunia Ramarli, and Daniela Zarcone
- Subjects
CD4-Positive T-Lymphocytes ,CD3 Complex ,Cell Survival ,T-Lymphocytes ,T cell ,Immunology ,Receptors, Antigen, T-Cell ,Apoptosis ,Thymus Gland ,CD8-Positive T-Lymphocytes ,In Vitro Techniques ,Biology ,Mice ,CD28 Antigens ,Antigens, CD ,medicine ,Animals ,Humans ,Immunology and Allergy ,ADP-ribosyl Cyclase ,Membrane Glycoproteins ,T-cell receptor ,Models, Immunological ,Antibodies, Monoclonal ,CD28 ,General Medicine ,Transfection ,T lymphocyte ,Flow Cytometry ,ADP-ribosyl Cyclase 1 ,Cell biology ,Platelet Endothelial Cell Adhesion Molecule-1 ,Haematopoiesis ,Thymocyte ,medicine.anatomical_structure ,B7-1 Antigen ,Signal Transduction - Abstract
Thymic T cell maturation depends on interactions between thymocytes and cells of epithelial and hematopoietic lineages that control a selective process whereby developing T cells with inappropriate or self-reactive receptors die. Molecules involved in this process are the TCR expressed on thymocytes together with the CD3 complex and MHC-peptide on accessory cells. However, other molecules may favor or prevent death of thymocytes, thus playing a role in selection. CD38 is expressed by the majority of human thymocytes, mainly at the double-positive (DP) stage. In contrast, CD38 is not found on subcapsular double-negative (DN) thymocytes and on a proportion of medullary single-positive (SP) thymocytes. CD38 enhances death of thymocytes when it is cross-linked by goat anti-mouse (GAM) antiserum or by one of its ligands, CD31, expressed by thymic epithelial cells or transfected into murine fibroblasts (L cells). As most thymocytes are at an intermediate (DP) stage of development, it is likely that these cells are most vulnerable to death mediated via MHC-peptide-TCR interactions that is increased by CD38 cross-linking. DN and SP thymocytes are refractory to CD38-induced apoptosis. Accessory molecules, e.g. CD38, are expressed during thymic cell maturation and their presence is relevant for the survival or death of DP T cells in the course of selection. Based on our data, CD38 enhances thymocyte death by interacting with CD31 expressed by accessory cells. In addition, CD28 expression on developing thymocytes also appears to play a role for their selection and it synergizes with CD38 to induce apoptosis of DP thymocytes.
- Published
- 2003
17. CD85/LIR-1/ILT2 and CD152 (Cytotoxic T Lymphocyte Antigen 4) Inhibitory Molecules Down-Regulate the Cytolytic Activity of Human CD4+T-Cell Clones Specific forMycobacterium tuberculosis
- Author
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Silvia Bruno, Andrea Merlo, Daniele Saverino, Carlo E. Grossi, Claudya Tenca, and Ermanno Ciccone
- Subjects
CD4-Positive T-Lymphocytes ,Interleukin 2 ,Immunoconjugates ,CD3 Complex ,Immunology ,Down-Regulation ,Leukocyte Immunoglobulin-like Receptor B1 ,Biology ,Lymphocyte Activation ,Tuberculin ,Major histocompatibility complex ,Microbiology ,Abatacept ,Interferon-gamma ,Antigen ,Antigens, CD ,medicine ,Humans ,Cytotoxic T cell ,CTLA-4 Antigen ,Interferon gamma ,Receptors, Immunologic ,Autologous Monocytes ,Mycobacterium tuberculosis ,T lymphocyte ,Cytotoxicity Tests, Immunologic ,Antigens, Differentiation ,Clone Cells ,Infectious Diseases ,Microbial Immunity and Vaccines ,biology.protein ,Interleukin-2 ,Parasitology ,Cell Division ,medicine.drug - Abstract
Antigen-specific cytolytic CD4+T lymphocytes controlMycobacterium tuberculosisinfection by secreting cytokines and by killing macrophages that have phagocytosed the pathogen. However, lysis of the latter cells promotes microbial dissemination, and other macrophages engulf the released bacteria. Subsequently, CD4+T-cell-mediated killing of macrophages goes on, and this persistent process may hamper control of infection, unless regulatory mechanisms maintain a subtle balance between lysis of macrophages by cytolytic CD4+cells and activation of cytolytic CD4+cells by infected macrophages. We asked whether inhibitory molecules expressed by CD4+cytolytic T lymphocytes could play a role in such a balance. To this end, human CD4+T-cell clones specific forM. tuberculosiswere produced that displayed an autologous major histocompatibility complex class II-restricted lytic ability against purified protein derivative (PPD)-pulsed antigen-presenting cells. All T-cell clones expressed CD152 (cytotoxic T-lymphocyte antigen 4 [CTLA-4]) and CD85/leukocyte immunoglobulin-like receptor 1 (LIR-1)/immunoglobulin-like transcript 2 (ILT2) inhibitory receptors, but not CD94 and the killer inhibitory receptor (or killer immunoglobulin-like receptor [KIR]) p58.2. CD3-mediated activation of the clones was inhibited in a redirected killing assay in which CD152 and CD85/LIR-1/ILT2 were cross-linked. Specific antigen-mediated proliferation of the clones was also sharply reduced when CD152 and CD85/LIR-1/ILT2 were cross-linked by specific monoclonal antibody (MAb) followed by goat anti-mouse antiserum. In contrast, blockade of the receptors by specific MAb only increased their proliferation. Production of interleukin 2 (IL-2) and gamma interferon (IFN-γ) by the T-cell clones was also strongly reduced when CD152 and CD85/LIR-1/ILT2 were cross-linked. The lytic activity of the T-cell clones against PPD-pulsed autologous monocytes or Epstein-Barr virus-activated B cells was increased by blockade and decreased by cross-linking of the receptors. These results indicate that CD152 and CD85/LIR-1/ILT2 play a role in the regulation of the antigen-specific activity of CD4+cytolytic T lymphocytes against PPD-presenting cells.
- Published
- 2001
18. Igs expressed by chronic lymphocytic leukemia B cells show limited binding-site structure variability
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Fortunato Morabito, Franco Fais, Nicholas Chiorazzi, Anna Tramontano, Claudya Tenca, Monica Colombo, Paolo Marcatili, Manlio Ferrarini, Davide Bagnara, Silvia Bruno, Fabio Ghiotto, Emilia Albesiano, Giovanna Cutrona, Anna Chailyan, Xiao-Jie Yan, and Andrea Nicola Mazzarello
- Subjects
Models, Molecular ,Protein Conformation ,Chronic lymphocytic leukemia ,Immunology ,Cell ,Immunoglobulin Variable Region ,Gene Expression ,Immunoglobulins ,Receptors, Antigen, B-Cell ,Biology ,medicine.disease_cause ,Pathogenesis ,Antigen ,medicine ,Immunology and Allergy ,Cluster Analysis ,Humans ,Binding site ,Antigens ,Receptor ,Genetics ,Mutation ,Binding Sites ,Structural classification ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,medicine.anatomical_structure ,Immunoglobulin Heavy Chains ,Hydrophobic and Hydrophilic Interactions ,Protein Binding - Abstract
Ag selection has been suggested to play a role in chronic lymphocytic leukemia (CLL) pathogenesis, but no large-scale analysis has been performed so far on the structure of the Ag-binding sites (ABSs) of leukemic cell Igs. We sequenced both H and L chain V(D)J rearrangements from 366 CLL patients and modeled their three-dimensional structures. The resulting ABS structures were clustered into a small number of discrete sets, each containing ABSs with similar shapes and physicochemical properties. This structural classification correlates well with other known prognostic factors such as Ig mutation status and recurrent (stereotyped) receptors, but it shows a better prognostic value, at least in the case of one structural cluster for which clinical data were available. These findings suggest, for the first time, to our knowledge, on the basis of a structural analysis of the Ab-binding sites, that selection by a finite quota of antigenic structures operates on most CLL cases, whether mutated or unmutated.
- Published
- 2013
19. MOESM1 of Expression of Immunoglobulin Receptors with Distinctive Features Indicating Antigen Selection by Marginal Zone B Cells from Human Spleen
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Colombo, Monica, Cutrona, Giovanna, Reverberi, Daniele, Bruno, Silvia, Ghiotto, Fabio, Claudya Tenca, Stamatopoulos, Kostas, Hadzidimitriou, Anastasia, Ceccarelli, Jenny, Salvi, Sandra, Boccardo, Simona, Calevo, Maria, Santanna, Amleto, Truini, Mauro, Fais, Franco, and Ferrarini, Manlio
- Abstract
Supplementary material, approximately 2.13 MB.
- Published
- 2013
- Full Text
- View/download PDF
20. Targeting the Bcl-2 family in B-cell chronic lymphocytic leukemia
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Claudya Tenca, Franco Fais, Fabio Ghiotto, and Silvia Bruno
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Programmed cell death ,business.industry ,Chronic lymphocytic leukemia ,Cell ,Bcl-2 family ,Hematology ,medicine.disease ,Subclass ,Pathogenesis ,Leukemia ,medicine.anatomical_structure ,Oncology ,Apoptosis ,hemic and lymphatic diseases ,Immunology ,Medicine ,Pharmacology (medical) ,business - Abstract
SUMMARY B-cell chronic lymphocytic leukemia (CLL) is the most common leukemia in human adults of the western world and no definitive cure is yet available. One key factor in CLL pathogenesis and disease progression is misbalanced Bcl-2 cell death machinery that is shifted towards protection from apoptosis. Thus, strategies to counteract the antiapoptotic action of the Bcl-2 family in CLL cells are being explored. The Bcl-2 family is composed of a growing number of proteins related to Bcl-2 by sequence homology and their interactions regulate the cell’s decision to die. The features of one particular subclass, the BH3-only proteins, are being studied and exploited for the development of therapeutic anticancer approaches that specifically target antiapoptotic Bcl-2 proteins overexpressed in tumors, including CLL. Preclinical and clinical efficacy and toxicity of the most effective among these ‘BH3 mimetics’ are presented, together with a model that accounts for the differential sensitivity of CLL and normal cells to Bcl-2 neutralization.
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- 2013
21. BH3-Only Proteins in Cancer and Apoptosis
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Franco Fais, Fabio Ghiotto, Claudya Tenca, and Silvia Bruno
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Programmed cell death ,Multicellular organism ,medicine.anatomical_structure ,Apoptosis ,Bcl-2 family ,Cell ,medicine ,Biology ,Inner mitochondrial membrane ,Tissue homeostasis ,Intracellular ,Cell biology - Abstract
In multicellular organisms, an enormous number of cells die every day, because of stress, injury, infection, or natural turnover necessary for proper tissue homeostasis. In most of these events, cell death is orchestrated by the dying cell itself, a sort of cellular suicide called “apoptosis”. Malfunctioning of this death program at any stage leads to severe human diseases, including cancer and autoimmune disorders. Apoptosis is elicited by intracellular or extracellular stimuli that activate a common cell death machinery, culminating in permeabilization of the mitochondrial membrane. In this chapter we describe the state of the art of the knowledge of how cells execute “mitochondrial” or “intrinsic” apoptosis. A fundamental and crucial role as regulators of the cell’s commitment to apoptosis is assumed by BH3-only proteins, a class of small molecules belonging to the Bcl-2 family. They are both sensors of death signals and vectors of information to the core apoptotic machinery, exerting their activity by hierarchical and finely tuned interactions with the other Bcl-2 family members. We discuss the groundbreaking research from several laboratories that have contributed to disclosing the complex activity of BH3-only proteins and the efforts made to translate these results into novel tools for cancer therapy.
- Published
- 2013
22. A Soluble Form of CTLA-4 Is Present in Paediatric Patients with Acute Lymphoblastic Leukaemia and Correlates with CD1d+ Expression
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Franco Fais, Daniele Saverino, Matteo Luciani, Giampaola Pesce, Giulio Rossi, Claudya Tenca, Marcello Bagnasco, and Rita Simone
- Subjects
Male ,B Cells ,Messenger ,Gene Expression ,Pediatrics ,Hematologic Cancers and Related Disorders ,Antigens, CD1d ,B-Lymphocytes ,Biomarkers, Tumor ,CTLA-4 Antigen ,Case-Control Studies ,Child ,Child, Preschool ,Female ,Humans ,Immunophenotyping ,Lymphocyte Count ,Polymorphism, Genetic ,Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ,RNA, Messenger ,Solubility ,Pathology ,Cytotoxic T cell ,Multidisciplinary ,Tumor ,biology ,Hematology ,Acute Lymphoblastic Leukemia ,Leukemia ,medicine.anatomical_structure ,Oncology ,CD1D ,Medicine ,Research Article ,Science ,T cell ,Immune Cells ,Immunology ,Malignancy ,CD1d ,Immunomodulation ,Immune system ,Genetic ,Diagnostic Medicine ,Leukemias ,medicine ,Pediatric Hematology ,Antigens ,Polymorphism ,Preschool ,Biology ,business.industry ,Cancers and Neoplasms ,medicine.disease ,CTLA-4 ,Pediatric Oncology ,biology.protein ,RNA ,Clinical Immunology ,business ,Biomarkers ,General Pathology - Abstract
CTLA-4 is a key factor in regulating and maintaining self tolerance, providing a negative signal to the T cell and thus limiting immune responses. Several polymorphisms within the CTLA-4 gene have been associated with an increased risk of developing autoimmune diseases and, very recently, with susceptibility to human cancer. Acute lymphoblastic leukemia is a clonal disorder of lymphoid progenitors representing the most frequent malignancy of childhood. Here, we show the presence at significantly elevated levels of a circulating soluble form of CTLA-4 in 70% of B-ALL pediatric patients with active disease, the positive correlation between the percentage of leukemic B lymphocytes and the amount of serum sCTLA-4, and the expression of sCTLA-4 transcript by B cells in patients. Finally, a correlation between CD1d expression (a negative prognostic marker) and the sCTLA-4 in B-ALL patients was observed. This suggests a possible role of this soluble molecule as a marker of progression or severity of the neoplastic disease.
- Published
- 2012
23. Adhesion Molecule Expression on B-Cells from Acute and Chronic Lymphoid Leukemias
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G. Cerruti, Luciana Annino, G. De Rossi, Francesca Romana Mauro, Daniela Zarcone, Antonio Tabilio, C. E. Grossit, Claudya Tenca, and Anna Favre
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Male ,Cancer Research ,Stromal cell ,Fluorescent Antibody Technique ,Cell–cell interaction ,Humans ,Lymphocyte homing receptor ,Cell adhesion ,B-Lymphocytes ,CD40 ,biology ,Cell adhesion molecule ,Infant, Newborn ,Hematology ,Middle Aged ,Flow Cytometry ,Burkitt Lymphoma ,Leukemia, Lymphocytic, Chronic, B-Cell ,Cell biology ,Phenotype ,Oncology ,biology.protein ,Female ,Neural cell adhesion molecule ,Cell Adhesion Molecules ,Homing (hematopoietic) - Abstract
Adhesion molecule expression on acute and chronic lymphoid leukemia cells of B lineage (B-ALL and B-CLL) may subserve several functions. Adhesion of leukemic cells to endothelial cells and to extracellular matrix components is relevant to homing, trafficking and spread of the malignant cells, and thus to clinical presentation, course and disease prognosis. Adhesive interactions between malignant cells and accessory cells, particularly stromal cells in the bone marrow environment, may support growth of the malignant cells via cytokine-delivered messages. They may also deliver signals that prevent or trigger programmed cell death of tumor cells. Here we review data on the adhesive phenotype of leukemic blasts from pro-B (CALLA +) ALL and of cells from B-CLL cases. We show that expression of certain adhesion molecules may help define disease subsets with distinctive clinical and prognostic features. One adhesion molecule, the lymphocyte homing receptor CD44, allows definition of two groups of B-CLL patients with significantly different survival.
- Published
- 1994
24. High frequency of development of B cell lymphoproliferation and diffuse large B cell lymphoma in Dbl knock-in mice
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Franco Fais, Claudya Tenca, Emilio Hirsch, Manuela Fedele, Fiorella Altruda, Alessandra Eva, Luca Mastracci, Federica Sabatini, Laura Emionite, Barbara Salani, Simonetta Astigiano, Amleto De Santanna, Marzia Ognibene, Cristina Vanni, Roberta Resaz, Luigi Varesio, and Ottavia Barbieri
- Subjects
Male ,rho GTP-Binding Proteins ,Dbl ,Lymphoma, B-Cell ,Mutant ,Blotting, Western ,CDC42 ,Diffuse large B cell lymphoma ,Biology ,Exon ,Mice ,Gene knockin ,Drug Discovery ,medicine ,In Situ Nick-End Labeling ,Knock-in mice ,Animals ,Guanine Nucleotide Exchange Factors ,Humans ,Genetics (clinical) ,B cell ,Genetics ,Lung tumors ,Small lymphocytic lymphoma ,Gene targeting ,medicine.disease ,Molecular biology ,Phenotype ,Immunohistochemistry ,medicine.anatomical_structure ,NIH 3T3 Cells ,Molecular Medicine ,Lymphoma, Large B-Cell, Diffuse ,Diffuse large B-cell lymphoma - Abstract
Dbl is the prototype of a large family of GDP–GTP exchange factors for small GTPases of the Rho family. In vitro, Dbl is known to activate Rho, Rac, and Cdc42 and to induce a transformed phenotype in murine fibroblasts. We previously reported that Dbl-null mice are viable and fertile but display defective dendrite elongation of distinct subpopulations of cortical neurons, suggesting a role of Dbl in controlling dendritic growth. To gain deeper insights into the role of Dbl in development and disease, we attempted a knock-in approach to create an endogenous allele that encodes a missense-mutation-mediated loss of function in the DH domain. We generated, by gene targeting technology, a mutant mouse strain by inserting a mutagenized human proto-Dbl cDNA clone expressing only the Dbl N terminus regulatory sequence at the starting codon of murine exon 1. Animals were monitored over a 21-month period, and necropsy specimens were collected for histological examination and immunohistochemistry analysis. Dbl knock-in mice are viable and did not manifest either decreased reproductive performances or gross developmental phenotype but revealed a reduced lifespan compared to wild-type (w.t.) mice and showed, with aging, a B cell lymphoproliferation that often has features of a frank diffuse large B cell lymphoma. Moreover, Dbl knock-in male mice displayed an increased incidence of lung adenoma compared to w.t. mice. These data indicate that Dbl is a tumor susceptibility gene in mice and that loss of function of Dbl DH domain by genetic missense mutations is responsible for induction of diffuse large B cell lymphoma.
- Published
- 2011
25. Mutation Pattern of Paired Immunoglobulin Heavy and Light Variable Domains in Chronic Lymphocytic Leukemia B Cells
- Author
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Franco Fais, Claudya Tenca, Davide Bagnara, Manlio Ferrarini, Nicholas Chiorazzi, Fabio Ghiotto, Charles C. Chu, Fortunato Morabito, Emilia Albesiano, Anna Tramontano, Giovanna Cutrona, Xiao-Jie Yan, Monica Colombo, Silvia Bruno, Paolo Marcatili, and Maria Grazia Calevo
- Subjects
Mutation rate ,mutations ,antibody ,cll ,Chronic lymphocytic leukemia ,DNA Mutational Analysis ,Immunoglobulin Variable Region ,Somatic hypermutation ,Complementarity determining region ,Biology ,medicine.disease_cause ,Mutation Rate ,Genetics ,medicine ,Humans ,Mutation frequency ,Molecular Biology ,Genetics (clinical) ,B-Lymphocytes ,Mutation ,Articles ,medicine.disease ,Complementarity Determining Regions ,Leukemia, Lymphocytic, Chronic, B-Cell ,Isotype ,Molecular biology ,Protein Structure, Tertiary ,Molecular Medicine ,Immunoglobulin Light Chains ,Immunoglobulin Heavy Chains ,IGHV@ - Abstract
B-cell chronic lymphocytic leukemia (CLL) patients display leukemic clones bearing either germline or somatically mutated immunoglobulin heavy variable (IGHV ) genes. Most information on CLL immunoglobulins (Igs), such as the definition of stereotyped B-cell receptors (BCRs), was derived from germline unmutated Igs. In particular, detailed studies on the distribution and nature of mutations in paired heavy- and light-chain domains of CLL clones bearing mutated Igs are lacking. To address the somatic hyper-mutation dynamics of CLL Igs, we analyzed the mutation pattern of paired IGHV-diversity-joining (IGHV-D-J ) and immunoglobulin kappa/lambda variable-joining (IGK/LV-J ) rearrangements of 193 leukemic clones that displayed ≥ 2% mutations in at least one of the two immunoglobulin variable (IGV ) genes (IGHV and/or IGK/LV ). The relationship between the mutation frequency in IGHV and IGK/LV complementarity determining regions (CDRs) and framework regions (FRs) was evaluated by correlation analysis. Replacement (R) mutation frequency within IGK/LV chain CDRs correlated significantly with mutation frequency of paired IGHV CDRs in λ but not κ isotype CLL clones. CDRs of IGKV-J rearrangements displayed a lower percentage of R mutations than IGHVs. The frequency/pattern of mutations in kappa CLL Igs differed also from that in κ-expressing normal B cells described in the literature. Instead, the mutation frequency within the FRs of IGHV and either IGKV or IGLV was correlated. Notably, the amount of diversity introduced by replaced amino acids was comparable between IGHVs and IGKVs. The data indicate a different mutation pattern between κ and λ isotype CLL clones and suggest an antigenic selection that, in κ samples, operates against CDR variation.
- Published
- 2011
26. Correction: CX3CR1 Is Expressed by Human B Lymphocytes and Mediates CX3CL1 Driven Chemotaxis of Tonsil Centrocytes
- Author
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Maria Bertolotto, Andrea Gregorio, Lizzia Raffaghello, Claudya Tenca, Claudio Gambini, Luciano Ottonello, Glaucia C. Furtado, Vito Pistoia, Franco Fais, Elisa Ferretti, Anna Corcione, and Sergio A. Lira
- Subjects
Multidisciplinary ,business.industry ,Science ,Correction ,Chemotaxis ,medicine.anatomical_structure ,Tonsil ,CX3CR1 ,Immunology ,Medicine ,CX3CL1 ,business - Abstract
The word "Mediates" was misspelled in the article title. The correct title is: CX₃CR1 Is Expressed by Human B Lymphocytes and Mediates CX₃CL1 Driven Chemotaxis of Tonsil Centrocytes. The correct citation is: Corcione A, Ferretti E, Bertolotto M, Fais F, Raffaghello L, et al. (2009) CX₃CR1 Is Expressed by Human B Lymphocytes and Mediates CX₃CL1 Driven Chemotaxis of Tonsil Centrocytes. PLoS ONE 4(12): e8485. doi:10.1371/journal.pone.0008485
- Published
- 2010
27. CD1d expression on B-precursor acute lymphoblastic leukemia subset with poor prognosis
- Author
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Davide Bagnara, Giuseppe Cimino, Carlo E. Grossi, Sabrina Zanardi, G. De Rossi, Valentina Coletti, Daniela Zarcone, Ermanno Ciccone, Claudya Tenca, Franco Fais, and Daniele Saverino
- Subjects
Cancer Research ,Cell Communication ,Acute lymphoblastic leukemia ,Antigens, CD1 ,Immunophenotyping ,Cytotoxic T cell ,Killer Cells ,Child ,Acute leukemia ,α-galactosylceramide ,B-Lymphocytes ,Tumor ,hemic and immune systems ,Hematology ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Natural killer T cell ,Prognosis ,Killer Cells, Natural ,Survival Rate ,Leukemia ,medicine.anatomical_structure ,Oncology ,Natural ,Natural killer T cells ,lipids (amino acids, peptides, and proteins) ,chemical and pharmacologic phenomena ,Galactosylceramides ,CD1d ,Cell Line ,Predictive Value of Tests ,Acute lymphocytic leukemia ,parasitic diseases ,medicine ,Biomarkers, Tumor ,Humans ,Antigens ,CD1d-restricted T cells ,Antigens, CD1d ,Hematopoietic Stem Cells ,Infant ,Anesthesiology and Pain Medicine ,CD1 ,business.industry ,Gene rearrangement ,medicine.disease ,carbohydrates (lipids) ,Immunology ,Bone marrow ,business ,Biomarkers - Abstract
Acute lymphoblastic leukemia (ALL) is the most frequent malignancy of childhood. Although therapeutical advances have been achieved, some ALL subgroups still fare poorly. CD1d is a monomorphic molecule that provides a suitable target for immunotherapy in view of the characterization of a glycolipid, alpha-galactosylceramide (alpha-GalCer), capable of being presented to CD1d-restricted T cells with cytotoxic potential. We investigated CD1d expression in 80 pediatric B-cell precursor (BCP) ALL cases defined according to immunophenotype, cytogenetic features and age at onset. CD1d was detected on ALL cells in 15% of the patients. CD1d+ ALLs were significantly associated with infant leukemia, pro-B phenotype and mixed-lineage leukemia (MLL)/AF4 gene rearrangement. Accordingly, overall survival of patients with CD1d+ ALL was significantly shorter. CD1d+ leukemic blasts were able to present alpha-GalCer via CD1d to cytotoxic CD1d-restricted T cells, which induced apoptosis of ALL cells that was inhibited by mAb to CD1d. CD1d+ blasts loaded with alpha-GalCer elicited cytokine secretion by CD1d-restricted T cells. Analysis of bone marrow (BM) cells derived from normal donors revealed that CD19+/CD1d+ cells were mostly mature B lymphocytes. However, a minority of BCPs expressed CD1d. Thus, expression of CD1d in ALL cases heralds an adverse prognosis but may provide a therapeutic tool.
- Published
- 2005
28. Apoptosis of squamous cells at different stages of carcinogenesis following 4-HPR treatment
- Author
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Ermanno Ciccone, Silvia Bruno, Daniele Saverino, Claudya Tenca, and Carlo E. Grossi
- Subjects
Keratinocytes ,Cancer Research ,Programmed cell death ,Pathology ,medicine.medical_specialty ,Fas Ligand Protein ,Time Factors ,Fenretinide ,Cell ,Antineoplastic Agents ,Apoptosis ,medicine.disease_cause ,Caspase 8 ,Cell Line ,Tumor Cells, Cultured ,medicine ,Humans ,Neoplasms, Squamous Cell ,fas Receptor ,Caspase ,Membrane Glycoproteins ,biology ,Cell growth ,General Medicine ,Flow Cytometry ,Mitochondria ,medicine.anatomical_structure ,Epidermoid carcinoma ,Caspases ,Cancer research ,biology.protein ,Carcinogenesis ,Cell Division - Abstract
Squamous cell carcinoma (SCC) is the end product of a multistep process characterized by a progression from normal epithelial cells through metaplastic or dysplastic intraepithelial changes that evolve into invasive cancer. Since retinamides have shown promising in vivo anti-tumoral activity, we studied effects and effector mechanisms of the synthetic retinoid N-(4-hydroxyphenyl)retinamide (4-HPR) on squamous cells at progressing stages of tumorigenesis. To this end, an in vitro model of squamous carcinogenesis consisting of normal human keratinocytes, human papilloma virus (HPV)-immortalized keratinocytes (UP) and tumorigenic HPV-immortalized/v-Ha-ras transfected keratinocytes (UPR) was used. 4-HPR treatment affected cell growth at doses higher than 1.5 microM. Flow cytometric measurements of DNA content and annexin V revealed that cell growth decrease was mainly due to apoptosis at 4-HPR concentrations of or below 15 microM, and necrosis at higher concentrations. The effects were similar in the three cell types of the in vitro model, as well as in three SCC cell lines, suggesting that sensitivity to 4-HPR is independent of the degree of squamous cell tumorigenesis in the in vitro model. We further investigated whether mitochondrial damage was involved in the course of 4-HPR-induced apoptosis. Treatment of squamous cells with the antioxidant L-ascorbic acid inhibited apoptosis, indicating that 4-HPR increases production of free radicals. Measures of mitochondrial membrane potentials showed that 4-HPR induced membrane permeability transition (MPT), and that MPT-inhibitors were able to reduce apoptosis. This indicates that MPT is involved in apoptosis signalling by 4-HPR. Finally, we studied the role of caspases. We found that caspases 8, 9 and 3 participate in 4-HPR-mediated apoptosis of squamous cells, and that MPT is an upstream event that regulates caspase activity. Caspase 8 was activated independently of the Fas-Fas ligand pathway.
- Published
- 2002
29. 'Increased serum levels of soluble CD44 standard, but not of variant isoforms v5 and v6, in B cell chronic lymphocytic leukemia'
- Author
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M Paganuzzi, Francesca Romana Mauro, P Marroni, Stefano Molica, G. De Rossi, Andrea Velardi, Carlo E. Grossi, Daniela Zarcone, and Claudya Tenca
- Subjects
Adult ,Male ,Cancer Research ,viruses ,Genetic enhancement ,Chronic lymphocytic leukemia ,Response element ,B-CLL, adhesion molecules, CD44 ,Biology ,chemistry.chemical_compound ,Antigens, Neoplasm ,medicine ,Biomarkers, Tumor ,Humans ,adhesion molecules ,CD44 ,Aged ,Aged, 80 and over ,Oligonucleotide ,RNA ,Hematology ,B-CLL ,Middle Aged ,medicine.disease ,Virology ,Molecular biology ,Leukemia, Lymphocytic, Chronic, B-Cell ,Hyaluronan Receptors ,Oncology ,Viral replication ,chemistry ,RNA splicing ,Female ,DNA - Abstract
The CD44 cell surface proteoglycan participates in a variety of functions including lymphohematopoiesis, lymphocyte homing and tumor metastasis. In addition to the standard form (CD44st), a large family of variant isoforms (CD44v) is generated by alternative splicing of a single gene. Certain CD44v (v5 and V6) are upregulated in the course of neoplastic progression and reflect the metastatic potential of tumor cells. CD44 v6 is expressed in high-grade non-Hodgkin's lymphoma cells and is released in the serum, thus providing a soluble marker that reflects tumor burden, disease progression and treatment response. Here we show that serum CD44st is elevated in approximately half of B-CLL patients. In contrast, CD44v5 and v6 are detected at normal levels in the large majority of the cases. CD44st serum levels correlate significantly with the number of circulating leukemic B cells and with the levels of another soluble B-CLL marker, beta2-microglobulin. Immunoprecipitation analyses of B-CLL sera allow detection of several high molecular weight bands and of a 78 kDa band that represents a soluble form of CD44st and is 4 kDa lower than a similar band (82 kDa) detected in B-CLL cell lysates. Elevated serum CD44st associates with a number of unfavorable prognostic factors such as high peripheral blood lymphocytosis, splenomegaly, advanced disease stage and therapy requirement. A follow-up study indicates that serum levels of CD44st are related to disease status, thus reinforcing our veiw that this molecule may represent a reliable tumor marker in B-CLL.
- Published
- 1997
30. Antibodies to adhesion molecules inhibit the lytic function of MHC-unrestricted cytotoxic cells by preventing their activation
- Author
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O Viale, Claudya Tenca, Walter Malorni, Daniela Zarcone, Carlo E. Grossi, Francesca Iosi, Ricciarda Galandrini, Giannamaria Cerruti, Andrea Velardi, Alessandro Moretta, and Giuseppe Arancia
- Subjects
Antigens, Differentiation, T-Lymphocyte ,Cytotoxicity, Immunologic ,CD3 Complex ,Immunology ,Lymphocyte Cooperation ,CD2 Antigens ,Receptors, Antigen, T-Cell ,Fluorescent Antibody Technique ,Biology ,In Vitro Techniques ,Lymphocyte Activation ,Natural killer cell ,Major Histocompatibility Complex ,Cell–cell interaction ,Antigens, CD ,Tubulin ,medicine ,Cell Adhesion ,Cytotoxic T cell ,Humans ,Receptors, Immunologic ,Killer Cells, Lymphokine-Activated ,Cells, Cultured ,Cytoskeleton ,Lymphokine-activated killer cell ,Cluster of differentiation ,Effector ,Cell adhesion molecule ,Antibodies, Monoclonal ,Actins ,CD56 Antigen ,Lymphocyte Function-Associated Antigen-1 ,Lymphocyte Subsets ,Cell biology ,Clone Cells ,Killer Cells, Natural ,Cytolysis ,medicine.anatomical_structure ,CD18 Antigens ,Microscopy, Electron, Scanning ,Cell Adhesion Molecules - Abstract
We evaluated the effect of the antibodies to adhesion molecules CD2, CD11a/CD18 (LFA-1), and CD56 (N-CAM) on MHC-unrestricted cytotoxicity mediated by polyclonal NK cells and LAK cells or by CD3 + or CD3 − cytolytic cell clones against a panel of tumor cell targets selected according to expression or absence of the corresponding ligands. We show that (i) antibodies to CD11a/CD18 and, to a lesser extent, antibodies to CD2 inhibit target cell lysis, whereas anti-CD56 antibodies exert little if any effect; (ii) in a model system using polyclonal NK/LAK cells as effectors and K562 or HL60-R (NK-resistant) cells as targets, inhibition of cytotoxicity occurs without a significant impairment of effector to target cell binding; (iii) the cytotoxic function of CD3 + or CD3 − cytotoxic cell clones is inhibited differentially by antibodies to adhesion molecules; (iv) conjugates formed in the presence of antibodies which inhibit target cell lysis display a significant reduction of target to effector cell contact surface; and (v) this may lead to defective activation of effector cells, as indicated by lack of redistribution of the microtubular apparatus. We conclude that (i) MHC-unrestricted cytotoxicity is regulated by a number of molecular interactions that span far beyond our present knowledge and that it is strictly dependent on the surface phenotype of the effector cell and of the target cell; (ii) in certain types of effector/target cell interactions, antibodies to adhesion molecules do not prevent conjugate formation but reduce the extent of cell-to-cell surface contact which, in turn, leads to defective activation of the effector cell and, therefore, to inhibition of target cell lysis.
- Published
- 1992
31. CX3CR1 Is Expressed by Human B Lymphocytes and Meditates CX3CL1 Driven Chemotaxis of Tonsil Centrocytes
- Author
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Franco Fais, Maria Bertolotto, Claudya Tenca, Claudio Gambini, Andrea Gregorio, Elisa Ferretti, Glaucia C. Furtado, Anna Corcione, Vito Pistoia, Sergio A. Lira, Luciano Ottonello, and Lizzia Raffaghello
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Chemokine ,Ovalbumin ,Palatine Tonsil ,Immunology ,CX3C Chemokine Receptor 1 ,lcsh:Medicine ,Spleen ,Cell Biology/Cell Signaling ,Immunoglobulin G ,Immunophenotyping ,Mice ,Antibody Specificity ,medicine ,Centroblasts ,Animals ,Humans ,Cell Biology/Leukocyte Signaling and Gene Expression ,lcsh:Science ,B-Lymphocytes ,Multidisciplinary ,CD40 ,Follicular dendritic cells ,biology ,Chemokine CX3CL1 ,Chemotaxis ,lcsh:R ,Germinal center ,Germinal Center ,Molecular biology ,Lymphocyte Subsets ,medicine.anatomical_structure ,Child, Preschool ,biology.protein ,Immunization ,Receptors, Chemokine ,lcsh:Q ,Signal Transduction ,Research Article - Abstract
BACKGROUND:Fractalkine/CX(3)CL1, a surface chemokine, binds to CX(3)CR1 expressed by different lymphocyte subsets. Since CX(3)CL1 has been detected in the germinal centres of secondary lymphoid tissue, in this study we have investigated CX(3)CR1 expression and function in human naïve, germinal centre and memory B cells isolated from tonsil or peripheral blood. METHODOLOGY/PRINCIPAL FINDINGS:We demonstrate unambiguously that highly purified human B cells from tonsil and peripheral blood expressed CX(3)CR1 at mRNA and protein levels as assessed by quantitative PCR, flow cytometry and competition binding assays. In particular, naïve, germinal centre and memory B cells expressed CX(3)CR1 but only germinal centre B cells were attracted by soluble CX(3)CL1 in a transwell assay. CX(3)CL1 signalling in germinal centre B cells involved PI3K, Erk1/2, p38, and Src phosphorylation, as assessed by Western blot experiments. CX(3)CR1(+) germinal centre B cells were devoid of centroblasts and enriched for centrocytes that migrated to soluble CX(3)CL1. ELISA assay showed that soluble CX(3)CL1 was secreted constitutively by follicular dendritic cells and T follicular helper cells, two cell populations homing in the germinal centre light zone as centrocytes. At variance with that observed in humans, soluble CX(3)CL1 did not attract spleen B cells from wild type mice. OVA immunized CX(3)CR1(-/-) or CX(3)CL1(-/-) mice showed significantly decreased specific IgG production compared to wild type mice. CONCLUSION/SIGNIFICANCE:We propose a model whereby human follicular dendritic cells and T follicular helper cells release in the light zone of germinal centre soluble CX(3)CL1 that attracts centrocytes. The functional implications of these results warrant further investigation.
- Published
- 2009
32. Cell activation via CD44 occurs in advanced stages of squamous cell carcinogenesis
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Barbara Santamaria, Claudya Tenca, Fabio Ghiotto, Silvia Bruno, Carlo E. Grossi, Marina Fabbi, Micaela Tiso, Daniele Saverino, Silvano Ferrini, Daniela Zarcone, and Ermanno Ciccone
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Cancer Research ,Pathology ,medicine.medical_specialty ,Cell ,Apoptosis ,medicine.disease_cause ,Cell–cell interaction ,medicine ,Tumor Cells, Cultured ,Humans ,Cell Line, Transformed ,DNA Primers ,biology ,Base Sequence ,Cell growth ,CD44 ,General Medicine ,medicine.anatomical_structure ,Cell Transformation, Neoplastic ,Hyaluronan Receptors ,Epidermoid carcinoma ,Cancer research ,biology.protein ,Carcinoma, Squamous Cell ,Calcium ,Carcinogenesis ,Cell activation ,Intracellular ,Signal Transduction - Abstract
Squamous cell carcinoma (SCC) derives from dysplastic or metaplastic stratified epithelia. The process of squamous cell carcinogenesis has been investigated for the potential role of the adhesion molecule CD44, whose standard form (CD44s) and isoforms generated by alternative splicing of variant exons are known to display altered expression during tumorigenesis in other systems. We have utilized an in vitro correlate of squamous cell carcinogenesis, in which progression stages from normal squamous epithelium to dysplastic lesions and to SCC are represented by primary cultures of normal keratinocytes, by human papilloma virus-immortalized keratinocytes (UP) and by HPVimmortalized/v-Ha-ras transfected tumorigenic keratinocytes (UPR). We investigated expression of CD44 and of variant isoforms, from mRNA to intracellular and surface protein levels, and found no relationship between expression of CD44 and stages of squamous cell carcinogenesis. However, when the function of CD44 was analyzed as Ca(2+) mobilization ability upon monoclonal antibody binding and crosslinking, signal transduction via CD44 was found only for the neoplastic stage (UPR cells). Ca(2+) mobilization was completely independent of density of surface CD44. We have performed similar analyses in an in vitro model of SCC in which four squamous tumor cell lines and UPR cells were sorted according to increasing resistance to external cytotoxic stimuli, i.e. starving conditions, treatment with the retinoid N-(4-hydroxyphenyl)retinamide and cytolytic activity of effector lymphokine-activated killer cells. No relationship between expression of CD44 and level of cell resistance against external cell death-inducing stimuli was found, while CD44-mediated Ca(2+) mobilization ability was restricted to the highly resistant tumor cell lines. Our results indicate that the role(s) of CD44 in squamous cell proliferative disorders can be evinced from the functional features of the molecule, rather than from its phenotypic repertoire.
33. The CD85/LIR-1/ILT2 inhibitory receptor is expressed by all human T lymphocytes and down-regulates their functions
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Marina Fabbi, Fabio Ghiotto, Daniele Saverino, Carlo E. Grossi, Daniela Zarcone, Claudya Tenca, Andrea Merlo, Giuseppe Santoro, Ermanno Ciccone, Silvia Bruno, Micaela Tiso, Giuseppe Anastasi, and David Cosman
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CD4-Positive T-Lymphocytes ,Cytotoxicity, Immunologic ,Intracellular Fluid ,Cytoplasm ,CD3 Complex ,CD3 ,T cell ,T-Lymphocytes ,Immunology ,Down-Regulation ,Epitopes, T-Lymphocyte ,Biology ,Lymphocyte Activation ,Epitope ,Leukocyte Immunoglobulin-like Receptor B1 ,Antigen ,Antigens, CD ,medicine ,Immunology and Allergy ,Humans ,Calcium Signaling ,RNA, Messenger ,Receptors, Immunologic ,Receptor ,Interphase ,Membrane Glycoproteins ,Antibodies, Monoclonal ,Cytotoxicity Tests, Immunologic ,Molecular biology ,Cell biology ,Clone Cells ,Cytolysis ,medicine.anatomical_structure ,Cell culture ,Receptor-CD3 Complex, Antigen, T-Cell ,biology.protein ,Immunologic Memory ,CD8 ,Immunosuppressive Agents ,T-Lymphocytes, Cytotoxic - Abstract
The inhibitory molecule CD85/LIR-1/ILT2 has been detected previously on the surface of a small proportion of T lymphocytes. In this study, evidence is provided that, although only a fraction of CD3+ cells are stained by mAb specific for CD85/LIR-1/ILT2 on their surface, this inhibitory receptor is present in the cytoplasm of all T lymphocytes, and that it is detectable on the surface of all T cell clones by the M402 mAb. Biochemical analyses further demonstrate that CD85/LIR-1/ILT2 is present in all T clones analyzed, and that the protein is tyrosine-phosphorylated. Expression of mRNA coding for CD85/LIR-1/ILT2 has been assessed by RT-PCR. Notably, in the NKL cell line and in one T cell clone, amplification of the messenger required 30 cycles only, whereas, in other T cell clones, an amplification product was detected by increasing the number of cycles. CD85/LIR-1/ILT2 inhibits CD3/TCR-mediated activation in both CD4+ and CD8+ clones, and it down-regulates Ag recognition by CD8+ cells in a clonally distributed fashion. Addition of anti-ILT2 HP-F1 mAb in the cytolytic assay enhances target cell lysis mediated by Ag-specific CTL. This could be due to interference of the mAb with receptor/ligand interactions. In contrast, HP-F1 mAb cross-linking triggers inhibitory signals that reduce cytotoxicity. CD85/LIR-1/ILT2 also controls responses to recall Ags and, in low responders, its engagement sharply increases T cell proliferation. The inhibitory function of the molecule is also confirmed by its ability to reduce CD3/TCR-induced intracellular Ca2+ mobilization.
34. Apoptosis of B-cell chronic lymphocytic leukemia cells induced by a novel BH3 peptidomimetic
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Fabio Ghiotto, Franco Fais, Claudya Tenca, Anna Mumot, Silvia Bruno, Gabriele Zoppoli, Valeria Tomati, Silvio Parodi, Ermanno Ciccone, Salvatore Casciaro, and Fortunato Morabito
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Cancer Research ,Peptidomimetic ,Chronic lymphocytic leukemia ,Cell Culture Techniques ,bcl-X Protein ,Down-Regulation ,Apoptosis ,Phosphatidylserines ,hemic and lymphatic diseases ,medicine ,Humans ,bcl-2-Associated X Protein ,Pharmacology ,CD20 ,biology ,Myeloid leukemia ,Flow Cytometry ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,In vitro ,Leukemia ,bcl-2 Homologous Antagonist-Killer Protein ,medicine.anatomical_structure ,Proto-Oncogene Proteins c-bcl-2 ,Oncology ,Immunology ,biology.protein ,Cancer research ,Myeloid Cell Leukemia Sequence 1 Protein ,Molecular Medicine ,Bone marrow ,Peptides ,Biomarkers - Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is the most common leukemia in human adults of the Western world and no definitive cure is yet available. The disease is characterized by accumulation of clonal malignant B lymphocytes resistant to apoptosis. Strategies to hit the anti-apoptotic drift of the Bcl-2 family in B-CLL cells are being explored. A novel peptidomimetic based on the BH3 domain of the pro-apoptotic protein Bim and recently shown to exert significant apoptotic activity on acute myeloid leukemia cells, both in vitro and in vivo, was assayed on ex-vivo derived leukemic cells from untreated B-CLL patients (n = 7). We found that this peptide, named 072RB, induced apoptosis of B-CLL samples at a concentration that does not affect viability of peripheral and bone marrow derived lymphocytes from healthy donors. Apoptosis was demonstrated by activation of Bak and Bax, externalization of plasma membranes phosphadydilserines, appearance of hypodiploid events in DNA flow cytometry histograms and was accompanied by dissipation of the mitochondrial transmembrane potential. Before the onset of marked apoptotic signs a progressive decline of the relevant anti-apoptotic proteins Bcl-X(L) and Mcl-1 could be observed. The negative control peptide 072RBL94A was ineffective for B-CLL cells, supporting the sequence specificity of 072RB activity. No relationship was found between responsiveness to 072RB and Mcl-1/Bcl-X(L) basal levels or decrease magnitude, possibly because of the limited sample size of the study. Altogether, we demonstrate that 072RB induces significant apoptosis of B-CLL cells subsequent to Bcl-X(L) and Mcl-1 downregulation.
35. Specific recognition of the viral protein UL18 by CD85j/LIR-1/ILT2 on CD8+ T cells mediates the non-MHC-restricted lysis of human cytomegalovirus-infected cells
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Amleto De Santanna, Lucia Baldi, Angela Bachi, Daniele Saverino, Fabio Ghiotto, Ermanno Ciccone, Andrea Merlo, Lorenzo Battini, Massimo E. Maffei, Carlo E. Grossi, Marzia Occhino, Claudya Tenca, Marina Fabbi, Silvia Bruno, and Stefano Pileri
- Subjects
Cytotoxicity, Immunologic ,Cytomegalovirus ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,CD8-Positive T-Lymphocytes/*immunology/metabolism ,Interleukin 21 ,Jurkat Cells ,Mice ,Leukocyte Immunoglobulin-like Receptor B1 ,Immunologic ,Receptors ,Leukocytes ,Immunology and Allergy ,Cytotoxic T cell ,Capsid Proteins/biosynthesis/*metabolism ,IL-2 receptor ,Receptors, Immunologic ,Cytotoxicity Tests ,CD/biosynthesis/*metabolism/physiology ,Tumor ,ZAP70 ,Interphase/immunology ,Cytomegalovirus Infections/immunology/virology ,Middle Aged ,medicine.anatomical_structure ,Cytomegalovirus Infections ,Immunologic/biosynthesis/*metabolism/physiology ,Cytomegalovirus/*immunology ,Fibroblasts/immunology/virology ,Adult ,T cell ,Immunology ,Biology ,Lymphocyte Activation/immunology ,Cell Line ,Antigen ,Antigens, CD ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Antigens ,Interphase ,Interleukin 3 ,Histocompatibility Antigens Class I ,Fibroblasts ,Cytotoxicity Tests, Immunologic ,Virology ,Hela Cells ,Cytotoxicity ,Histocompatibility Antigens Class I/*physiology ,Leukocytes, Mononuclear ,Capsid Proteins ,Mononuclear/cytology/immunology ,CD8 - Abstract
Immune evasion mechanisms of human CMV are known; however, the immune control of infection remains poorly elucidated. We show that interaction between the viral protein UL18 on infected cells and the invariant receptor CD85j/LIR-1/ILT2 expressed on CTL is relevant for the control of infection. Resting and activated CD8+ T cells lysed UL18 expressing cells, whereas cells infected with CMV defective for UL18 were not killed. Lysis was not dependent on CD8+ T cell Ag specificity, MHC-unrestricted and specifically blocked by anti-CD85j and anti-UL18 mAb. Moreover, soluble recombinant UL18Fc immunoprecipitated CD85j from T cells. Activation is mediated by CD85j and its pathway is unrelated to CD3/TCR engagement. UL18 is detected in immunocompromised patients with productive infection and the mechanism used in vivo by human CMV to ensure survival of the immunocompetent host may be mediated by activation signals delivered by infected cells to T lymphocytes via UL18/CD85j interactions.
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