title: Tracking genetically engineered lymphocytes long-term reveals the dynamics of T cell immunological memory.
abstract: Long-lasting immune protection from pathogens and cancer requires the generation of memory T cells able to survive long-term. To unravel the immunological requirements for long-term persistence of human memory T cells, we characterized and traced, over several years, T lymphocytes genetically modified to express the thymidine kinase (TK) suicide gene that were infused in 10 patients after haploidentical hematopoietic stem cell transplantation (HSCT). At 2 to 14 years after infusion and in the presence of a broad and resting immune system, we could still detect effectors/effector memory (TEM/EFF), central memory (TCM), and stem memory (TSCM) TK(+) cells, circulating at low but stable levels in all patients. Longitudinal analysis of cytomegalovirus (CMV)- and Flu-specific TK(+) cells indicated that antigen recognition was dominant in driving in vivo expansion and persistence at detectable levels. The amount of infused TSCM cells positively correlated with early expansion and with the absolute counts of long-term persisting gene-marked cells. By combining T cell sorting with sequencing of integration (IS), TCRα and TCRβ clonal markers, we showed that T cells retrieved long-term were enriched in clones originally shared in different memory T cell subsets, whereas dominant long-term clonotypes appeared to preferentially originate from infused TSCM and TCM clones. Together, these results indicate that long-term persistence of gene-modified memory T cells after haploidentical HSCT is influenced by antigen exposure and by the original phenotype of infused cells. Cancer adoptive immunotherapy might thus benefit from cellular products enriched in lymphocytes with an early-differentiated phenotype.
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy. Vita-Salute San Raffaele University, Milan 20132, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Oliveira
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy. Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany.
authors.forename: Eliana
authors.initials: E
authors.lastname: Ruggiero
authors.affiliation: Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Maria Teresa Lupo
authors.initials: MT
authors.lastname: Stanghellini
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Cieri
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy. Vita-Salute San Raffaele University, Milan 20132, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: D'Agostino
authors.forename: Mattio
authors.initials: M
authors.lastname: D'Agostino
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Fronza
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Lulay
authors.affiliation: Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Dionisio
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy. Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Mastaglio
authors.affiliation: Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Greco
authors.affiliation: Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Jacopo
authors.initials: J
authors.lastname: Peccatori
authors.affiliation: Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Aiuti
authors.affiliation: Vita-Salute San Raffaele University, Milan 20132, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ambrosi
authors.affiliation: Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Biasco
authors.affiliation: Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy. Innovative Immunotherapies Unit, Division of Immunology, Transplantation and Infectious Diseases, PIBIC, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Attilio
authors.initials: A
authors.lastname: Bondanza
authors.affiliation: MolMed S.p.A, Milan 20132, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lambiase
authors.affiliation: MolMed S.p.A, Milan 20132, Italy.
authors.forename: Catia
authors.initials: C
authors.lastname: Traversari
authors.affiliation: Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy. Unit of Immunogenetics, Leukemia Genomics and Immunobiology, Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Vago
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Vita-Salute San Raffaele University, Milan 20132, Italy. MolMed S.p.A, Milan 20132, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Bordignon
authors.affiliation: Vita-Salute San Raffaele University, Milan 20132, Italy. Hematology and Bone Marrow Transplantation Unit, San Raffaele Scientific Institute, Milan 20132, Italy. Unit of Immunogenetics, Leukemia Genomics and Immunobiology, Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Ciceri
authors.affiliation: Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program in Immunology and Bio-Immunotherapy of Cancer (PIBIC), San Raffaele Scientific Institute, Milan 20132, Italy. Vita-Salute San Raffaele University, Milan 20132, Italy. bonini.chiara@hsr.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bonini
mesh_terms: D000328:Adult; D000368:Aged; D000941:Antigens / Q000276:immunology; D049109:Cell Proliferation; D058948:Cell Tracking; D002999:Clone Cells; D005260:Female; D041722:Genes, Transgenic, Suicide; D005818:Genetic Engineering; D015316:Genetic Therapy; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007156:Immunologic Memory; D018655:Lymphocyte Count; D008212:Lymphocyte Depletion; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D013601:T-Lymphocytes / Q000276:immunology*; D013937:Thymidine Kinase / Q000378:metabolism; D013997:Time Factors; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D000941:Antigens; D013937:Thymidine Kinase