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3. Impact of treatment on cellular immunophenotype in MS: A cross-sectional study

Author

Cellerino, Maria, Ivaldi, Federico, Pardini, Matteo, Rotta, Gianluca, Vila, Gemma, Bäcker-Koduah, Priscilla, Berge, Tone, Laroni, Alice, Lapucci, Caterina, Novi, Giovanni, Boffa, Giacomo, Sbragia, Elvira, Palmeri, Serena, Asseyer, Susanna, Høgestøl, Einar, Campi, Cristina, Piana, Michele, Inglese, Matilde, Paul, Friedemann, Harbo, Hanne F., Villoslada, Pablo, Kerlero de Rosbo, Nicole, and Uccelli, Antonio

Published
2020
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5. Protocol for flow cytometry immunophenotyping of human antigen-specific T cells by activation-induced marker and Th1 cytokine detection

Author

Rotta, Gianluca, Achille, Valentina, Bornheimer, Scott J., Duenas, Melanie, Fernandez, Marco, Gatti, Arianna, Giron, Sergio Haro, Rio, Irene Martinez, Massanella, Marta, Jiménez, Diego J., Monserrat, Jorge, Pfeifer, Verena, Pichler, Josefa, Prietl, Barbara, Sourij, Harald, Uras, Chiara R.M., Vlah, Sara, and Fenoglio, Daniela

Published
2025
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6. Genetic Variants Regulating Immune Cell Levels in Health and Disease

Author

Orrù, Valeria, Steri, Maristella, Sole, Gabriella, Sidore, Carlo, Virdis, Francesca, Dei, Mariano, Lai, Sandra, Zoledziewska, Magdalena, Busonero, Fabio, Mulas, Antonella, Floris, Matteo, Mentzen, Wieslawa I., Urru, Silvana A.M., Olla, Stefania, Marongiu, Michele, Piras, Maria G., Lobina, Monia, Maschio, Andrea, Pitzalis, Maristella, Urru, Maria F., Marcelli, Marco, Cusano, Roberto, Deidda, Francesca, Serra, Valentina, Oppo, Manuela, Pilu, Rosella, Reinier, Frederic, Berutti, Riccardo, Pireddu, Luca, Zara, Ilenia, Porcu, Eleonora, Kwong, Alan, Brennan, Christine, Tarrier, Brendan, Lyons, Robert, Kang, Hyun M., Uzzau, Sergio, Atzeni, Rossano, Valentini, Maria, Firinu, Davide, Leoni, Lidia, Rotta, Gianluca, Naitza, Silvia, Angius, Andrea, Congia, Mauro, Whalen, Michael B., Jones, Chris M., Schlessinger, David, Abecasis, Gonçalo R., Fiorillo, Edoardo, Sanna, Serena, and Cucca, Francesco

Published
2013
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7. Workflow for high-dimensional flow cytometry analysis of T cells from tumor metastases

Author

Faccani, Cristina, primary, Rotta, Gianluca, additional, Clemente, Francesca, additional, Fedeli, Maya, additional, Abbati, Danilo, additional, Manfredi, Francesco, additional, Potenza, Alessia, additional, Anselmo, Achille, additional, Pedica, Federica, additional, Fiorentini, Guido, additional, Villa, Chiara, additional, Protti, Maria P, additional, Doglioni, Claudio, additional, Aldrighetti, Luca, additional, Bonini, Chiara, additional, Casorati, Giulia, additional, Dellabona, Paolo, additional, and de Lalla, Claudia, additional

Published
2022
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9. Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants

Author

Dondero, Alessandra, primary, Morini, Martina, additional, Cangelosi, Davide, additional, Mazzocco, Katia, additional, Serra, Martina, additional, Spaggiari, Grazia Maria, additional, Rotta, Gianluca, additional, Tondo, Annalisa, additional, Locatelli, Franco, additional, Castellano, Aurora, additional, Scuderi, Francesca, additional, Sementa, Angela Rita, additional, Eva, Alessandra, additional, Conte, Massimo, additional, Garaventa, Alberto, additional, Bottino, Cristina, additional, and Castriconi, Roberta, additional

Published
2021
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11. Single Cell Analysis of Circulating Endothelial Cells in Allogeneic Hematopoietic Stem Cell Transplant; To Whom Do They Belong: Host or Donor?

Author

Almici, Camillo, primary, Fontana, Francesca, additional, Piovani, Giovanna, additional, Balzarini, Piera, additional, Neva, Arabella, additional, Bruno, Benedetto, additional, Skert, Cristina, additional, Rotta, GianLuca, additional, Fisogni, Simona, additional, Verardi, Rosanna, additional, Bianchetti, Andrea, additional, Braga, Simona, additional, Malagola, Michele, additional, Manaresi, Nicolò, additional, Facchetti, Fabio, additional, Marini, Mirella, additional, and Russo, Domenico, additional

Published
2019
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13. Additional file 2: Table S1. of High-throughput immunophenotypic characterization of bone marrow- and cord blood-derived mesenchymal stromal cells reveals common and differentially expressed markers: identification of angiotensin-converting enzyme (CD143) as a marker differentially expressed between adult and perinatal tissue sources

Author

Amati, Eliana, Perbellini, Omar, Rotta, Gianluca, Bernardi, Martina, Chieregato, Katia, Sella, Sabrina, Rodeghiero, Francesco, Ruggeri, Marco, and Astori, Giuseppe

Abstract

Mean and CV values of log2 FI for each of 246 investigated markers. (DOCX 22 kb)

Published
2018
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14. Additional file 3: Table S2. of High-throughput immunophenotypic characterization of bone marrow- and cord blood-derived mesenchymal stromal cells reveals common and differentially expressed markers: identification of angiotensin-converting enzyme (CD143) as a marker differentially expressed between adult and perinatal tissue sources

Author

Amati, Eliana, Perbellini, Omar, Rotta, Gianluca, Bernardi, Martina, Chieregato, Katia, Sella, Sabrina, Rodeghiero, Francesco, Ruggeri, Marco, and Astori, Giuseppe

Abstract

List of the lowest variable markers (n = 59, CV cut-off of log2 FI ≤ 0.5) sorted by function and cluster expression. Markers with no expression were assigned to cluster 1, low-expressed markers to cluster 2, and intermediate markers to cluster 3, while a total of 18 markers clustering as very strong were assigned to cluster 4. (PPTX 50 kb)

Published
2018
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16. NOTCH1 Is Aberrantly Activated in Chronic Lymphocytic Leukemia Hematopoietic Stem Cells

Author

Di Ianni, Mauro, primary, Baldoni, Stefano, additional, Del Papa, Beatrice, additional, Aureli, Patrizia, additional, Dorillo, Erica, additional, De Falco, Filomena, additional, Albi, Elisa, additional, Varasano, Emanuela, additional, Di Tommaso, Ambra, additional, Giancola, Raffaella, additional, Accorsi, Patrizia, additional, Rotta, Gianluca, additional, Rompietti, Chiara, additional, Silva Barcelos, Estevão Carlos, additional, Campese, Antonio Francesco, additional, Di Bartolomeo, Paolo, additional, Screpanti, Isabella, additional, Rosati, Emanuela, additional, Falzetti, Franca, additional, and Sportoletti, Paolo, additional

Published
2018
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17. A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges

Author

Lanuti, Paola, primary, Simeone, Pasquale, additional, Rotta, Gianluca, additional, Almici, Camillo, additional, Avvisati, Giuseppe, additional, Azzaro, Rosa, additional, Bologna, Giuseppina, additional, Budillon, Alfredo, additional, Di Cerbo, Melania, additional, Di Gennaro, Elena, additional, Di Martino, Maria Luisa, additional, Diodato, Annamaria, additional, Doretto, Paolo, additional, Ercolino, Eva, additional, Falda, Alessandra, additional, Gregorj, Chiara, additional, Leone, Alessandra, additional, Losa, Francesca, additional, Malara, Natalia, additional, Marini, Mirella, additional, Mastroroberto, Pasquale, additional, Mollace, Vincenzo, additional, Morelli, Michele, additional, Muggianu, Emma, additional, Musolino, Giuseppe, additional, Neva, Arabella, additional, Pierdomenico, Laura, additional, Pinna, Silvia, additional, Piovani, Giovanna, additional, Roca, Maria Serena, additional, Russo, Domenico, additional, Scotti, Lorenza, additional, Tirindelli, Maria Cristina, additional, Trunzo, Valentina, additional, Venturella, Roberta, additional, Vitagliano, Carlo, additional, Zullo, Fulvio, additional, Marchisio, Marco, additional, and Miscia, Sebastiano, additional

Published
2018
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18. A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges

Author

Lanuti, P, Simeone, P, Rotta, G, Almici, C, Avvisati, G, Azzaro, R, Bologna, G, Budillon, A, Di Cerbo, M, Di Gennaro, E, Di Martino, M, Diodato, A, Doretto, P, Ercolino, E, Falda, A, Gregorj, C, Leone, A, Losa, F, Malara, N, Marini, M, Mastroroberto, P, Mollace, V, Morelli, M, Muggianu, E, Musolino, G, Neva, A, Pierdomenico, L, Pinna, S, Piovani, G, Roca, M, Russo, D, Scotti, L, Tirindelli, M, Trunzo, V, Venturella, R, Vitagliano, C, Zullo, F, Marchisio, M, Miscia, S, Lanuti, Paola, Simeone, Pasquale, Rotta, Gianluca, Almici, Camillo, Avvisati, Giuseppe, Azzaro, Rosa, Bologna, Giuseppina, Budillon, Alfredo, Di Cerbo, Melania, Di Gennaro, Elena, Di Martino, Maria Luisa, Diodato, Annamaria, Doretto, Paolo, Ercolino, Eva, Falda, Alessandra, Gregorj, Chiara, Leone, Alessandra, Losa, Francesca, Malara, Natalia, Marini, Mirella, Mastroroberto, Pasquale, Mollace, Vincenzo, Morelli, Michele, Muggianu, Emma, Musolino, Giuseppe, Neva, Arabella, Pierdomenico, Laura, Pinna, Silvia, Piovani, Giovanna, Roca, Maria Serena, Russo, Domenico, Scotti, Lorenza, Tirindelli, Maria Cristina, Trunzo, Valentina, Venturella, Roberta, Vitagliano, Carlo, Zullo, Fulvio, Marchisio, Marco, Miscia, Sebastiano, Lanuti, P, Simeone, P, Rotta, G, Almici, C, Avvisati, G, Azzaro, R, Bologna, G, Budillon, A, Di Cerbo, M, Di Gennaro, E, Di Martino, M, Diodato, A, Doretto, P, Ercolino, E, Falda, A, Gregorj, C, Leone, A, Losa, F, Malara, N, Marini, M, Mastroroberto, P, Mollace, V, Morelli, M, Muggianu, E, Musolino, G, Neva, A, Pierdomenico, L, Pinna, S, Piovani, G, Roca, M, Russo, D, Scotti, L, Tirindelli, M, Trunzo, V, Venturella, R, Vitagliano, C, Zullo, F, Marchisio, M, Miscia, S, Lanuti, Paola, Simeone, Pasquale, Rotta, Gianluca, Almici, Camillo, Avvisati, Giuseppe, Azzaro, Rosa, Bologna, Giuseppina, Budillon, Alfredo, Di Cerbo, Melania, Di Gennaro, Elena, Di Martino, Maria Luisa, Diodato, Annamaria, Doretto, Paolo, Ercolino, Eva, Falda, Alessandra, Gregorj, Chiara, Leone, Alessandra, Losa, Francesca, Malara, Natalia, Marini, Mirella, Mastroroberto, Pasquale, Mollace, Vincenzo, Morelli, Michele, Muggianu, Emma, Musolino, Giuseppe, Neva, Arabella, Pierdomenico, Laura, Pinna, Silvia, Piovani, Giovanna, Roca, Maria Serena, Russo, Domenico, Scotti, Lorenza, Tirindelli, Maria Cristina, Trunzo, Valentina, Venturella, Roberta, Vitagliano, Carlo, Zullo, Fulvio, Marchisio, Marco, and Miscia, Sebastiano

Abstract

Circulating endothelial cells (CEC) represent a restricted peripheral blood (PB) cell subpopulation with high potential diagnostic value in many endothelium-involving diseases. However, whereas the interest in CEC studies has grown, the standardization level of their detection has not. Here, we undertook the task to align CEC phenotypes and counts, by standardizing a novel flow cytometry approach, within a network of six laboratories. CEC were identified as alive/nucleated/CD45negative/CD34bright/CD146positive events and enumerated in 269 healthy PB samples. Standardization was demonstrated by the achievement of low inter-laboratory Coefficients of Variation (CVL), calculated on the basis of Median Fluorescence Intensity values of the most stable antigens that allowed CEC identification and count (CVL of CD34bright on CEC ∼ 30%; CVL of CD45 on Lymphocytes ∼ 20%). By aggregating data acquired from all sites, CEC numbers in the healthy population were captured (medianfemale = 9.31 CEC/mL; medianmale = 11.55 CEC/mL). CEC count biological variability and method specificity were finally assessed. Results, obtained on a large population of donors, demonstrate that the established procedure might be adopted as standardized method for CEC analysis in clinical and in research settings, providing a CEC physiological baseline range, useful as starting point for their clinical monitoring in endothelial dysfunctions.

Published
2018

20. High-throughput immunophenotypic characterization of bone marrow- and cord blood-derived mesenchymal stromal cells reveals common and differentially expressed markers: identification of angiotensin-converting enzyme (CD143) as a marker differentially expressed between adult and perinatal tissue sources

Author

Amati, Eliana, primary, Perbellini, Omar, additional, Rotta, Gianluca, additional, Bernardi, Martina, additional, Chieregato, Katia, additional, Sella, Sabrina, additional, Rodeghiero, Francesco, additional, Ruggeri, Marco, additional, and Astori, Giuseppe, additional

Published
2018
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21. Endothelial progenitor cells, defined by the simultaneous surface expression of VEGFR2 and CD133, are not detectable in healthy peripheral and cord blood

Author

Lanuti, Paola, primary, Rotta, Gianluca, additional, Almici, Camillo, additional, Avvisati, Giuseppe, additional, Budillon, Alfredo, additional, Doretto, Paolo, additional, Malara, Natalia, additional, Marini, Mirella, additional, Neva, Arabella, additional, Simeone, Pasquale, additional, Di Gennaro, Elena, additional, Leone, Alessandra, additional, Falda, Alessandra, additional, Tozzoli, Renato, additional, Gregorj, Chiara, additional, Di Cerbo, Melania, additional, Trunzo, Valentina, additional, Mollace, Vincenzo, additional, Marchisio, Marco, additional, and Miscia, Sebastiano, additional

Published
2015
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23. Endothelial progenitor cells, defined by the simultaneous surface expression of VEGFR2 and CD133, are not detectable in healthy peripheral and cord blood.

Author

Lanuti, Paola, Rotta, Gianluca, Almici, Camillo, Avvisati, Giuseppe, Budillon, Alfredo, Doretto, Paolo, Malara, Natalia, Marini, Mirella, Neva, Arabella, Simeone, Pasquale, Di Gennaro, Elena, Leone, Alessandra, Falda, Alessandra, Tozzoli, Renato, Gregorj, Chiara, Di Cerbo, Melania, Trunzo, Valentina, Mollace, Vincenzo, Marchisio, Marco, and Miscia, Sebastiano

Abstract

Circulating endothelial cells (CEC) and their progenitors (EPC) are restricted subpopulations of peripheral blood (PB), cord blood (CB), and bone marrow (BM) cells, involved in the endothelial homeostasis maintenance. Both CEC and EPC are thought to represent potential biomarkers in several clinical conditions involving endothelial turnover/remodeling. Although different flow cytometry methods for CEC and EPC characterization have been published so far, none of them have reached consistent conclusions, therefore consensus guidelines with respect to CEC and EPC identification and quantification need to be established. Here, we have carried out an in depth investigation of CEC and EPC phenotypes in healthy PB, CB and BM samples, by optimizing a reliable polychromatic flow cytometry (PFC) panel. Results showed that the brightness of CD34 expression on healthy PB and CB circulating cells represents a key benchmark for the identification of CEC (CD45neg/CD34bright/CD146pos) respect to the hematopoietic stem cell (HSC) compartment (CD45dim/CD34pos/CD146neg). This approach, combined with a dual-platform counting technique, allowed a sharp CEC enumeration in healthy PB ( n = 38), and resulting in consistent CEC counts with previously reported data (median = 11.7 cells/ml). In parallel, by using rigorous PFC conditions, CD34pos/CD45dim/CD133pos/VEGFR2pos EPC were not found in any healthy PB or CB sample, since VEGFR2 expression was never detectable on the surface of CD34pos/CD45dim/CD133pos cells. Notably, the putative EPC phenotype was observed in all analyzed BM samples ( n = 12), and the expression of CD146 and VEGFR2, on BM cells, was not restricted to the CD34bright compartment, but also appeared on the HSC surface. Altogether, our findings suggest that the previously reported EPC antigen profile, defined by the simultaneous expression of VEGFR2 and CD133 on the surface of CD45dim/CD34pos cells, should be carefully re-evaluated and further studies should be conducted to redefine EPC features in order to translate CEC and EPC characterization into clinical practice. © 2015 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

Published
2016
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29. Workflow for high-dimensional flow cytometry analysis of T cells from tumor metastases

Author

Cristina Faccani, Gianluca Rotta, Francesca Clemente, Maya Fedeli, Danilo Abbati, Francesco Manfredi, Alessia Potenza, Achille Anselmo, Federica Pedica, Guido Fiorentini, Chiara Villa, Maria P Protti, Claudio Doglioni, Luca Aldrighetti, Chiara Bonini, Giulia Casorati, Paolo Dellabona, Claudia de Lalla, Faccani, Cristina, Rotta, Gianluca, Clemente, Francesca, Fedeli, Maya, Abbati, Danilo, Manfredi, Francesco, Potenza, Alessia, Anselmo, Achille, Pedica, Federica, Fiorentini, Guido, Villa, Chiara, Protti, Maria P, Doglioni, Claudio, Aldrighetti, Luca, Bonini, Chiara, Casorati, Giulia, Dellabona, Paolo, and de Lalla, Claudia

Subjects
Ecology, Liver, T-Lymphocyte Subsets, Health, Toxicology and Mutagenesis, Plant Science, Flow Cytometry, Biochemistry, Genetics and Molecular Biology (miscellaneous), Workflow
Abstract

We describe a multi-step high-dimensional (HD) flow cytometry workflow for the deep phenotypic characterization of T cells infiltrating metastatic tumor lesions in the liver, particularly derived from colorectal cancer (CRC-LM). First, we applied a novel flow cytometer setting approach based on single positive cells rather than fluorescent beads, resulting in optimal sensitivity when compared with previously published protocols. Second, we set up a 26-color based antibody panel designed to assess the functional state of both conventional T-cell subsets and unconventional invariant natural killer T, mucosal associated invariant T, and gamma delta T (γδT)-cell populations, which are abundant in the liver. Third, the dissociation of the CRC-LM samples was accurately tuned to preserve both the viability and antigenic integrity of the stained cells. This combined procedure permitted the optimal capturing of the phenotypic complexity of T cells infiltrating CRC-LM. Hence, this study provides a robust tool for high-dimensional flow cytometry analysis of complex T-cell populations, which could be adapted to characterize other relevant pathological tissues.

Published
2021

30. A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges

Author

Gianluca Rotta, Paola Lanuti, Elena Di Gennaro, Natalia Malara, Alfredo Budillon, Mirella Marini, Pasquale Mastroroberto, Valentina Trunzo, Francesca Losa, Silvia Pinna, Maria Cristina Tirindelli, Marco Marchisio, Paolo Doretto, Alessandra Falda, Emma Muggianu, Lorenza Scotti, Camillo Almici, Giuseppe Musolino, Annamaria Diodato, Carlo Vitagliano, Laura Pierdomenico, Fulvio Zullo, Rosa Azzaro, Sebastiano Miscia, María Roca, Melania Di Cerbo, Domenico Russo, Chiara Gregorj, Michele Morelli, Vincenzo Mollace, Arabella Neva, Giovanna Piovani, Pasquale Simeone, Giuseppe Avvisati, Eva Ercolino, Alessandra Leone, Roberta Venturella, Maria Luisa Di Martino, Giuseppina Bologna, Lanuti, P, Simeone, P, Rotta, G, Almici, C, Avvisati, G, Azzaro, R, Bologna, G, Budillon, A, Di Cerbo, M, Di Gennaro, E, Di Martino, M, Diodato, A, Doretto, P, Ercolino, E, Falda, A, Gregorj, C, Leone, A, Losa, F, Malara, N, Marini, M, Mastroroberto, P, Mollace, V, Morelli, M, Muggianu, E, Musolino, G, Neva, A, Pierdomenico, L, Pinna, S, Piovani, G, Roca, M, Russo, D, Scotti, L, Tirindelli, M, Trunzo, V, Venturella, R, Vitagliano, C, Zullo, F, Marchisio, M, Miscia, S, Lanuti, Paola, Simeone, Pasquale, Rotta, Gianluca, Almici, Camillo, Avvisati, Giuseppe, Azzaro, Rosa, Bologna, Giuseppina, Budillon, Alfredo, Di Cerbo, Melania, Di Gennaro, Elena, Di Martino, Maria Luisa, Diodato, Annamaria, Doretto, Paolo, Ercolino, Eva, Falda, Alessandra, Gregorj, Chiara, Leone, Alessandra, Losa, Francesca, Malara, Natalia, Marini, Mirella, Mastroroberto, Pasquale, Mollace, Vincenzo, Morelli, Michele, Muggianu, Emma, Musolino, Giuseppe, Neva, Arabella, Pierdomenico, Laura, Pinna, Silvia, Piovani, Giovanna, Roca, Maria Serena, Russo, Domenico, Scotti, Lorenza, Tirindelli, Maria Cristina, Trunzo, Valentina, Venturella, Roberta, Vitagliano, Carlo, Zullo, Fulvio, Marchisio, Marco, and Miscia, Sebastiano

Subjects
Adult, Male, 0301 basic medicine, Median Fluorescence Intensity, Circulating endothelial cell, Science, Large population, Cell Separation, Biology, Sensitivity and Specificity, Article, Flow cytometry, Andrology, Young Adult, 03 medical and health sciences, Reference Values, medicine, Humans, High potential, Multidisciplinary, medicine.diagnostic_test, Healthy population, Endothelial Cells, Hematology, Middle Aged, Flow Cytometry, Healthy Volunteers, Peripheral blood, Blood Cell Count, 030104 developmental biology, Biological Variation, Population, cardiovascular system, Feasibility Studies, Medicine, Female, Endothelium, Vascular, Laboratories, Biological variability
Abstract

Circulating endothelial cells (CEC) represent a restricted peripheral blood (PB) cell subpopulation with high potential diagnostic value in many endothelium-involving diseases. However, whereas the interest in CEC studies has grown, the standardization level of their detection has not. Here, we undertook the task to align CEC phenotypes and counts, by standardizing a novel flow cytometry approach, within a network of six laboratories. CEC were identified as alive/nucleated/CD45negative/CD34bright/CD146positive events and enumerated in 269 healthy PB samples. Standardization was demonstrated by the achievement of low inter-laboratory Coefficients of Variation (CVL), calculated on the basis of Median Fluorescence Intensity values of the most stable antigens that allowed CEC identification and count (CVL of CD34bright on CEC ~ 30%; CVL of CD45 on Lymphocytes ~ 20%). By aggregating data acquired from all sites, CEC numbers in the healthy population were captured (medianfemale = 9.31 CEC/mL; medianmale = 11.55 CEC/mL). CEC count biological variability and method specificity were finally assessed. Results, obtained on a large population of donors, demonstrate that the established procedure might be adopted as standardized method for CEC analysis in clinical and in research settings, providing a CEC physiological baseline range, useful as starting point for their clinical monitoring in endothelial dysfunctions.

Published
2018
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