A cell-based artificial antigen-presenting cell coated with anti-CD3 and CD28 antibodies enables rapid expansion and long-term growth of CD4 T lymphocytes

Clin Immunol. 2002 Dec;105(3):259-72. doi: 10.1006/clim.2002.5277.


We compared the ability of two genetically modified myeloid cells, K562 and U937, to serve as artificial antigen-presenting cells (aAPC). Both aAPC were stably transfected with the low-affinity Fcgamma receptor CD32 (K32/U32 cells). K32 cells loaded with anti-CD3 and anti-CD28 Ab (K32/CD3/28) induced more rapid CD4 T-cell expansion than CD3/28-coated beads. In contrast, U32/CD3/28 induced high levels of CD4 T-cell thymidine uptake but were unable to sustain long-term T-cell expansion. K32 cells, but not U32 cells, loaded with anti-CD3 alone also stimulated CD4 T-cell growth and IL-2 secretion, indicating the expression of additional costimulatory molecules on K32 cells. We found constitutive expression of B7-H3 and a strong upregulation of mRNA encoding for IL-15, PD-L1, and PD-L2 after coculture with CD4 T cells activated by K32/CD3/28 but not U32/CD3/28. We conclude that K32 aAPCs are a robust system for clinical scale ex vivo expansion of CD4 T cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies / immunology
  • Antigen-Presenting Cells*
  • Antigens, CD
  • B7 Antigens
  • B7-1 Antigen / genetics
  • B7-H1 Antigen
  • Blood Proteins*
  • CD28 Antigens / immunology*
  • CD3 Complex / immunology*
  • CD4-Positive T-Lymphocytes / immunology*
  • Cell Division
  • Cell Line
  • Coculture Techniques
  • DNA, Recombinant
  • Electroporation
  • Humans
  • Interleukin-15 / genetics
  • Interleukin-2 / metabolism
  • Lymphocyte Activation
  • Membrane Glycoproteins
  • Peptides*
  • RNA, Messenger
  • Receptors, Immunologic
  • Time Factors
  • Up-Regulation


  • Antibodies
  • Antigens, CD
  • B7 Antigens
  • B7-1 Antigen
  • B7-H1 Antigen
  • Blood Proteins
  • CD274 protein, human
  • CD276 protein, human
  • CD28 Antigens
  • CD3 Complex
  • DNA, Recombinant
  • Interleukin-15
  • Interleukin-2
  • Membrane Glycoproteins
  • Peptides
  • RNA, Messenger
  • Receptors, Immunologic