title: Clinical-Scale Rapid Autologous BK Virus-Specific T Cell Line Generation From Kidney Transplant Recipients With Active Viremia for Adoptive Immunotherapy.
abstract: Polyomavirus-associated nephropathy (PVAN) after BK virus reactivation in kidney transplant recipients (KTR) can compromise graft survival. Lowering immunosuppression is the only established approach to prevent or treat PVAN but nonspecifically increasing host immune competence also augments rejection risk. Ex vivo T cell stimulation/expansion offers the possibility to generate BK-specific T cell lines for adoptive immunotherapy. The objective of this study was to develop and characterize a clinical-scale protocol to generate BK-specific T cell lines from viremic KTR.
  
  
  
  Peripheral blood mononuclear cells from healthy controls and viremic KTR were stimulated using BK virus peptide libraries loaded or not on monocytes-derived dendritic cells. Cell counts, flow cytometry, and next-generation sequencing were used to assess T cell expansion, differentiation, and clonal diversity. Enzyme-linked immunospots, cytotoxicity assays as well as adoptive transfer in NOD/SCID/IL2Rγ mice were used to assess for pathogen-specificity and evidence of nonspecific alloreactivity.
  
  
  
  T cell lines from KTR and healthy control showed similar characteristics, implying that ongoing immunosuppression and chronic virus exposure do not compromise the differentiation, specificity, or clonal diversity of T cell lines after ex vivo production. Using antigen-loaded dendritic cells improved T cell expansion and favored central memory T cell differentiation. The T cell lines were antigen-specific and showed no nonspecific alloreactivity in vitro and in vivo.
  
  
  
  Using a rapid, clinically compliant culture system, we show that autologous BK virus-specific T cell lines can be reliably generated from viremic KTR. Our results pave the way for the treatment or prevention of PVAN with adoptive immunotherapy.
authors.affiliation: 1 Centre de Recherche de l'Hôpital Maisonneuve-Rosemont (CRHMR), Université de Montréal, Montreal, Canada. 2 Division of Nephrology, Department of Medicine, Hôpital Maisonneuve-Rosemont (HMR), Université de Montréal, Montreal, Canada. 3 Division of Hematology, Department of Medicine, Hôpital Maisonneuve-Rosemont (HMR), Université de Montréal, Montreal, Canada.
authors.forename: Caroline
authors.initials: C
authors.lastname: Lamarche
authors.forename: Julie
authors.initials: J
authors.lastname: Orio
authors.forename: Victoria
authors.initials: V
authors.lastname: Georges-Tobar
authors.forename: Thomas
authors.initials: T
authors.lastname: Pincez
authors.forename: Mathieu
authors.initials: M
authors.lastname: Goupil
authors.forename: Amina
authors.initials: A
authors.lastname: Dahmani
authors.forename: Cedric
authors.initials: C
authors.lastname: Carli
authors.forename: Ann
authors.initials: A
authors.lastname: Brasey
authors.forename: Lambert
authors.initials: L
authors.lastname: Busque
authors.forename: Jean-Sébastien
authors.initials: JS
authors.lastname: Delisle
mesh_terms: D019264:Adoptive Transfer / Q000379:methods*; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D001739:BK Virus / Q000276:immunology*; D016022:Case-Control Studies; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology; D006801:Humans; D016867:Immunocompromised Host; D007156:Immunologic Memory; D007166:Immunosuppressive Agents / Q000009:adverse effects; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000235:genetics / Q000276:immunology; D016030:Kidney Transplantation / Q000009:adverse effects*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D008213:Lymphocyte Activation; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D010641:Phenotype; D027601:Polyomavirus Infections / Q000276:immunology / Q000628:therapy* / Q000821:virology; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation* / Q000821:virology; D013997:Time Factors; D014412:Tumor Virus Infections / Q000276:immunology / Q000628:therapy* / Q000821:virology; D014775:Virus Activation
chemical_list: D000956:Antigens, Viral; C507942:Il2rg protein, mouse; D007166:Immunosuppressive Agents; D053631:Interleukin Receptor Common gamma Subunit