title: Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells.
abstract: OBJECTIVE
  Cytomegalovirus (CMV) disease constitutes a serious complication after stem cell transplantation and has been treated by adoptive transfer of donor-derived CMV-specific CD8(+) T cells. CMV-specific CD8(+) T cells were selected by multimers, and the technologies may alter the function of these T cells. Therefore, here we evaluated the impact of multimer reagents on the function of CD8(+) T lymphocytes.
  
  
  METHODS
  CMV-specific CD8(+) T cells were purified from the peripheral blood of donors using tetra- and streptamer technologies. The functional status of purified CMV-specific CD8(+) T cells was assessed by multiparametric immunophenotyping and carboxyfluorescein succinimidyl ester proliferation assays as well as by enzyme-linked immunospot assays.
  
  
  RESULTS
  A similar percentage of CMV-specific CD8(+) T cells could be purified by both tetra- (90%) and streptamer (92%) technologies. That constitutes a 30- to 50-fold concentration of CMV-specific CD8(+)CD45RA(+)CCR7-effector T cells. Selected cells secreted interferon-gamma and granzyme B upon stimulation with CMVpp65 peptide, thus demonstrating their functionality.
  
  
  CONCLUSIONS
  Our study demonstrated that both tetra- and streptamer technologies can be used to purify CMV-specific cytotoxic CD8(+) T cells for adoptive T-cell transfer. Both multimer technologies did not have any negative influence on the proliferation of selected T cells. Importantly, streptamer technology is available at good manufacturing practice level.
authors.affiliation: Department of Hematology, The Affiliated Zhongda Hospital of Southeast University, Nanjing, China.
authors.forename: Xin-Chao
authors.initials: XC
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Pang
authors.forename: Xun
authors.initials: X
authors.lastname: Xu
authors.forename: Anita
authors.initials: A
authors.lastname: Schmitt
authors.forename: Mathias
authors.initials: M
authors.lastname: Freund
authors.forename: Michael
authors.initials: M
authors.lastname: Schmitt
authors.forename: Bao-An
authors.initials: BA
authors.lastname: Chen
mesh_terms: D019264:Adoptive Transfer; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D003587:Cytomegalovirus / Q000276:immunology*; D053804:Granzymes / Q000032:analysis; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D018189:Immunomagnetic Separation; D007371:Interferon-gamma / Q000032:analysis; D008213:Lymphocyte Activation
chemical_list: D015789:HLA-A2 Antigen; D007371:Interferon-gamma; D053804:Granzymes