title: Characterization of cord blood natural killer and lymphokine activated killer lymphocytes following ex vivo cellular engineering.
abstract: Cord blood (CB) natural killer (NK) and lymphokine-activated killer (LAK) cytotoxic cells are poorly characterized but might be used to treat minimal residual and/or recurrent malignant disease. Currently, there is no mechanism to use CB for adoptive cancer cellular immunotherapy after CB transplantation (CBT). Recognizing this as a deficiency, we hypothesized that CB aliquots could be engineered ex vivo for potential donor lymphocyte infusion after CBT. Cryopreserved CB aliquots were thawed, depleted of monocytes, and cultured in serum-free medium alone or serum-free medium with anti-CD3 and interleukins 2, 7, and 12 combined with antibody/cytokines for 48 hours. Immunophenotyping, cytotoxicity, and proliferation were evaluated. A significant expansion of CD3+ was seen, in addition to increases in lymphocyte subsets of CD8+, CD8+/CD25+, and CD3+/45RO+ versus medium alone. A significant enhancement of CD3 proliferation (P<.001), NK cytotoxicity, NK subset expansion, LAK cytotoxicity, and T-helper 1 subset expansion was also demonstrated. Significant enrichment was seen in NK CD16+/CD56+bright, CD16+/CD56+dim, CD56+bright and CD56+dim/KIR3DL1+, CD56+bright and CD56+dim/KIR2DL1+, CD56+bright and CD56+dim/KIR2DL2+ and CD94+/NKG2a+ subsets. These increases in CB NK subsets were in part secondary to augmentation of cell survival. Further, survival of NOD-SCID mice xenografted with human K562 cells and treated with CB cells expanded with antibody/cytokines was significantly higher than that in animals that received no treatment (phosphate buffered saline) and those that were treated with CB ex vivo expanded in medium alone (P<.005, respectively). These data suggest that cryopreserved CB cells could be ex vivo engineered for potential use as adoptive cancer cellular immunotherapy for donor lymphocyte infusion after CBT.
authors.affiliation: Department of Pediatrics, Columbia University, New York, New York 10032, USA.
authors.forename: Janet
authors.initials: J
authors.lastname: Ayello
authors.forename: Carmella
authors.initials: C
authors.lastname: van de Ven
authors.forename: Weiwei
authors.initials: W
authors.lastname: Fortino
authors.forename: Cheryl
authors.initials: C
authors.lastname: Wade-Harris
authors.forename: Prakash
authors.initials: P
authors.lastname: Satwani
authors.forename: Laxmi
authors.initials: L
authors.lastname: Baxi
authors.forename: Lynn L
authors.initials: LL
authors.lastname: Simpson
authors.forename: Warren
authors.initials: W
authors.lastname: Sanger
authors.forename: Diana
authors.initials: D
authors.lastname: Pickering
authors.forename: Joanne
authors.initials: J
authors.lastname: Kurtzberg
authors.forename: Mitchell S
authors.initials: MS
authors.lastname: Cairo
mesh_terms: D000818:Animals; D002455:Cell Division; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D020014:K562 Cells; D015979:Killer Cells, Lymphokine-Activated / Q000166:cytology* / Q000276:immunology*; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology*; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D023822:Tissue Engineering / Q000379:methods; D014183:Transplantation, Heterologous