title: 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.
abstract: 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.
authors.affiliation: Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Anna K
authors.initials: AK
authors.lastname: Thomas
authors.forename: Marcela V
authors.initials: MV
authors.lastname: Maus
authors.forename: Waleed S
authors.initials: WS
authors.lastname: Shalaby
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: James L
authors.initials: JL
authors.lastname: Riley
mesh_terms: D000906:Antibodies / Q000276:immunology; D000938:Antigen-Presenting Cells; D015703:Antigens, CD; D060887:B7 Antigens; D018122:B7-1 Antigen / Q000235:genetics; D060890:B7-H1 Antigen; D001798:Blood Proteins; D018106:CD28 Antigens / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002455:Cell Division; D002460:Cell Line; D018920:Coculture Techniques; D004274:DNA, Recombinant; D018274:Electroporation; D006801:Humans; D019409:Interleukin-15 / Q000235:genetics; D007376:Interleukin-2 / Q000378:metabolism; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins; D010455:Peptides; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D060887:B7 Antigens; D018122:B7-1 Antigen; D060890:B7-H1 Antigen; D001798:Blood Proteins; C423236:CD274 protein, human; C423237:CD276 protein, human; D018106:CD28 Antigens; D017252:CD3 Complex; D004274:DNA, Recombinant; D019409:Interleukin-15; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D010455:Peptides; D012333:RNA, Messenger; D011971:Receptors, Immunologic