Simultaneous Isolation of CD8+ and CD4+ T Cells Specific for Multiple Viruses for Broad Antiviral Immune Reconstitution After Allogeneic Stem Cell Transplantation

• DOI: 10.1097/cji.0b013e318213cb90
• Publication Date: 2011-03-09T11:16:17Z
• Authors: Maarten L. Zandvliet; Ellis van Liempt; Inge Jedema; Simone Kruithof; Michel G.D. Kester; Henk-Jan Guchelaar; J.H. Frederik Fal...; Pauline Meij
• Keywords: CD4-Positive T-Lymphocytes; 0301 basic medicine; Herpesvirus 4, Human; Immunomagnetic Separation; Adenoviruses, Human; Molecular Sequence Data; allogeneic stem cell transplantation adoptive immunotherapy virus-specific T cells interferon gamma production CD137 expression epstein-barr-virus cytomegalovirus-specific cd4(+) adoptive immunotherapy lymphoproliferative disease adenovirus infection detailed analysis ebv reactivation antigen responses lymphocytes; Cytomegalovirus; Adaptive Immunity; CD8-Positive T-Lymphocytes; Opportunistic Infections; Lymphocyte Activation; Orthomyxoviridae; Adoptive Transfer; 3. Good health; Immunocompromised Host; Interferon-gamma; 03 medical and health sciences; Humans; Transplantation, Homologous; Amino Acid Sequence; Antigens, Viral; Immunologic Memory; Stem Cell Transplantation

Opportunistic viral infections can cause serious morbidity and mortality in immunocompromised patients after allogeneic stem cell transplantation. Clinical studies have shown that adoptive transfer of donor-derived T cells specific for cytomegalovirus (CMV), Epstein-Barr virus (EBV), or human adenovirus (HAdV) can be a safe and effective treatment of infections with these major viral pathogens. The aim of this study was to develop a method for the simultaneous isolation of coordinated CD8(+) and CD4(+) memory T-cell responses against a broad repertoire of viral epitopes. To ensure that the method was applicable to a wide variety of virus-specific T cells that may differ in phenotypic and functional properties, we focused on T cells specific for the persistent viruses, CMV and EBV, and T cells specific for HAdV and influenza (FLU), which are not repetitively activated in vivo after initial viral clearance. Following in vitro activation, nearly all T cells specific for these viruses produced interferon γ (IFN-γ) and tumor necrosis factor α, and expressed CD137, whereas the populations varied in the production of interleukin-2, degranulation, and expression of phenotypic markers. Different kinetics of IFN-γ production were observed in CMV/EBV-specific T cells and HAdV/FLU-specific T cells. However, after the stimulation of peripheral blood from seropositive donors with viral protein-spanning peptide pools, the activated virus-specific CD8(+) and CD4(+) T cells could be simultaneously isolated by either IFN-γ-based or CD137-based enrichment. This study provides an efficient and widely applicable strategy for the isolation of virus-specific T cells, which may be used for the reconstitution of virus-specific immunity in allogeneic stem cell transplantation recipients.