Induction of CMV-specific T-cell lines using Ag-presenting cells pulsed with CMV protein or peptide.
BACKGROUND
CMV disease is still associated with a high morbidity and mortality in recipients of a solid organ or stem cell graft, especially in patients undergoing allogenic stem cell transplantation. Reconstitution of CMV-specific CD4(+) and CD8(+) cytotoxic T cell responses are essential to control CMV infection following allogenic stem cell transplantation. The transfer of unselected populations of lymphocytes from the peripheral blood of a CMV-scropositive donor to a transplant recipient can be used to control CMV infection. However, such transfer of unselected donor lymphocytes is limited by potentially fatal complications that arise from alloreactive T cells, also present in the unselected donor lymphocytes. Thus to make infusion of donor T cells safe and also more effective in controlling CMV infection in the recipient of the T cell infusion, T cells are manipulated in vitro to deplete alloreactive T cells and to enrich for CMV-specific T cells.


METHODS
Using various antigen-presenting cells (monocytes/PBMNCs/dendritic cells) and different modes of antigen presentation (infected APCs, pulsing of protein or peptide antigen) different CMV-specific T cell populations can be generated and expanded.


RESULTS
Using protein-/or peptide-pulsed DCs CMV-specific CD8(+) cytoxic T cell lines (can be generated and expanded) in addition CMV-specific CD4(+) T cell lines can be generated when CMV-protein-pulsed DCs are used as antigen-presenting cells. When peripheral blood mononuclear cells were stimulated with CMV lysates predominantly CMV-specific CD4(+) T cells are generated and expanded ex vivo.


CONCLUSIONS
Depending on the APC used (monocytes versus DC) and the mode of antigen presentation (protein versus peptide pulsing) different CMV-specific T cell populations of varying purity can be generated which show preserved function when tested for specific proliferation, cytokine production and cytotoxicity.
Einsele
H
H
Medizinische Klinik und Poliklinik, Universität Tübingen, Germany.
Rauser
G
G
Grigoleit
U
U
Hebart
H
H
Sinzger
C
C
Riegler
S
S
Jahn
G
G
D017951:Antigen Presentation / Q000276:immunology*; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology; D005240:Feasibility Studies; D006801:Humans; D010455:Peptides / Q000276:immunology; D010750:Phosphoproteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000276:immunology
D000956:Antigens, Viral; D010455:Peptides; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa