title: Characterization of migratory activity and cytokine profile of helper and cytotoxic CMV-specific T-cell lines expanded by a selective peptide library.
abstract: OBJECTIVE
  Reconstitution of cellular immunity by infusion of cytomegalovirus (CMV)-specific T lymphocytes is an attractive alternative to drugs currently used to control CMV reactivation in immunocompromised patients. For this purpose, we established a method for generating both anti-CMV CD4 and CD8 T cells following Good Manufacturing Practice indications, and we extensively characterized their immune functions.
  
  
  METHODS
  For generating CD4 and CD8 CMV-specific lymphocytes, T cells from 11 CMV-seropositive donors were stimulated three times with dendritic cells (DC) pulsed with a library of selected CMV peptides, recognized by >85% of the Caucasian population. At the end of the culture, T cells were analyzed for their specificity, cytotoxicity, chemotactic migration, proliferation, and cytokine production.
  
  
  RESULTS
  T cells were successfully expanded and enriched in CMV-specific subsets with an effector memory or an effector memory CD45 RA(+) phenotype. CMV-specific T-cell lines showed specific cytotoxicity (average lysis: 47%) against CMV peptides-pulsed DCs, and were depleted of auto- and alloreactivity. Moreover, the ability to proliferate following antigenic stimulation and the presence of functional CD4 lymphocytes producing Th1 and Th2 cytokines can ensure long-term antiviral immunity after in vivo injection. CMV-specific T lymphocytes also proved to be fully equipped to reach CMV-infected tissues, because they expressed CD49d and CCR1, CXCR3, CXCR4, necessary to recruit effector cells to inflamed sites. In accordance with this profile, they significantly migrated towards inflammatory chemokines and towards the supernatant collected from inflamed lung fibroblasts, frequently involved in CMV pathology.
  
  
  CONCLUSIONS
  This strategy allows expansion of effector T cells capable to exert CD8 and CD4-mediated immune functions and, thus, is suitable for clinical use.
authors.affiliation: Centro Ricerca "M. Tettamanti," Clinica Pediatrica Università Milano-Bicocca, Ospedale San Gerardo, Monza, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Dander
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Li Pira
authors.forename: Ettore
authors.initials: E
authors.lastname: Biagi
authors.forename: Paolo
authors.initials: P
authors.lastname: Perseghin
authors.forename: Giuliano
authors.initials: G
authors.lastname: Renoldi
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gaipa
authors.forename: Martino
authors.initials: M
authors.lastname: Introna
authors.forename: Virna
authors.initials: V
authors.lastname: Marin
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Manca
authors.forename: Andrea
authors.initials: A
authors.lastname: Biondi
authors.forename: Giovanna
authors.initials: G
authors.lastname: D'Amico
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement / Q000276:immunology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis*; D003587:Cytomegalovirus / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D007159:Immunologic Tests; D019151:Peptide Library; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000276:immunology*; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology
chemical_list: D016207:Cytokines; D019151:Peptide Library; D014764:Viral Proteins