Frozen cord blood hematopoietic stem cells differentiate into higher numbers of functional natural killer cells in vitro than mobilized hematopoietic stem cells or freshly isolated cord blood hematopoietic stem cells.
Adoptive natural killer (NK) cell therapy relies on the acquisition of large numbers of NK cells that are cytotoxic but not exhausted. NK cell differentiation from hematopoietic stem cells (HSC) has become an alluring option for NK cell therapy, with umbilical cord blood (UCB) and mobilized peripheral blood (PBCD34(+)) being the most accessible HSC sources as collection procedures are less invasive. In this study we compared the capacity of frozen or freshly isolated UCB hematopoietic stem cells (CBCD34(+)) and frozen PBCD34(+) to generate NK cells in vitro. By modifying a previously published protocol, we showed that frozen CBCD34(+) cultures generated higher NK cell numbers without loss of function compared to fresh CBCD34(+) cultures. NK cells generated from CBCD34(+) and PBCD34(+) expressed low levels of killer-cell immunoglobulin-like receptors but high levels of activating receptors and of the myeloid marker CD33. However, blocking studies showed that CD33 expression did not impact on the functions of the generated cells. CBCD34(+)-NK cells exhibited increased capacity to secrete IFN-γ and kill K562 in vitro and in vivo as compared to PBCD34(+)-NK cells. Moreover, K562 killing by the generated NK cells could be further enhanced by IL-12 stimulation. Our data indicate that the use of frozen CBCD34(+) for the production of NK cells in vitro results in higher cell numbers than PBCD34(+), without jeopardizing their functionality, rendering them suitable for NK cell immunotherapy. The results presented here provide an optimal strategy to generate NK cells in vitro for immunotherapy that exhibit enhanced effector function when compared to alternate sources of HSC.
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Martha
M
Luevano
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Anna
A
Domogala
Centre for Immunodeficiency, Molecular Immunology Unit, UCL Institute of Child Health, London, United Kingdom.
Michael
M
Blundell
University College London, Cancer Institute, London, United Kingdom.
Nicola
N
Jackson
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Isabela
I
Pedroza-Pacheco
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Sophie
S
Derniame
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Michelle
M
Escobedo-Cousin
Programa Concordia Banc de Sang i Teixits, Barcelona, Spain.
Sergio
S
Querol
Centre for Immunodeficiency, Molecular Immunology Unit, UCL Institute of Child Health, London, United Kingdom.
Adrian
A
Thrasher
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Alejandro
A
Madrigal
University College London, Cancer Institute, London, United Kingdom ; Anthony Nolan Research Institute, London, United Kingdom.
Aurore
A
Saudemont
D019264:Adoptive Transfer; D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D015925:Cryopreservation; D003602:Cytotoxicity, Immunologic; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D015870:Gene Expression; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000378:metabolism; D018664:Interleukin-12 / Q000494:pharmacology; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology* / Q000187:drug effects / Q000276:immunology / Q000637:transplantation; D051379:Mice; D016688:Mice, Inbred NOD; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000235:genetics / Q000276:immunology
D018952:Antigens, CD34; D015415:Biomarkers; D063268:Sialic Acid Binding Ig-like Lectin 3; D018664:Interleukin-12; D007371:Interferon-gamma