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An Experimental Study Comparing the Expansion of Peripheral Blood Natural Killer (NK) Cells Cultured with Artificial Antigen-Presenting Cells, in the Presence or Absence of Bone Marrow Mesenchymal Stem Cells (MSCs)

Abstract

NK cells have been seen as potential agents in adoptive immunotherapy for cancer. The main challenge for the success of this approach is to obtain a great quantity of activated NK cells for adoptive transfer. The present study had aimed to evaluate the effect of a feeder layer of irradiated MSCs in the in vitro expansion of NK cells. MSCs were obtained from the bone marrow (BM) cells remaining in the bag and filter used in the transplantation of hematopoietic stem cells. NK cells were obtained from peripheral blood (PB) of healthy volunteers. NK expansion and activation were stimulated by culture with artificial antigen-presenting cells (aAPCs) and IL-2, in the presence or absence of BM-MSCs. NK cell proliferation, phenotypic expression and cytotoxic activity were evaluated. Both culture conditions showed high NK purity with predominance of NK CD56brightCD16+ subset post expansion. However, cultures without the presence of MSCs showed higher NK proliferation, expression of activation markers (CD16 and NKG2D) and related cytotoxic activity. In this experimental study, the presence of a feeder layer of irradiated BM-MSCs interfered negatively in the expansion of PB-NKs, limiting their growth and activation. Further investigation is needed to understand the mechanisms of NK-MSC interaction and its implications.

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Data Availability

The datasets supporting the conclusions of this article are included within the article.

Abbreviations

NK:

Natural killer

PB:

Peripheral blood

KIR:

Killer cell immunoglobulin-like receptor

HLA-I:

Human leukocyte antigen class I

ADCC:

Antibody-dependent cellular cytotoxicity

GVL:

Graft versus leukemia

GVHD:

Graft versus host disease

HSCT:

Hematopoietic stem cell transplantation

WBC:

White blood cells

aAPCs:

Artificial antigen-presenting cells

mbIL21:

Membrane-bound IL-21

MSCs:

Mesenchymal stem cells

BM:

Bone marrow

FBS:

Fetal bovine serum

PB-MNCs:

Peripheral blood mononuclear cells

CD3dep PB-MNCs:

CD3-depleted PB-MNCs

MFI:

Median fluorescence intensity

CB:

Cord blood

UC:

Umbilical cord

IDO:

Indoleamine 2,3-dioxygenase

PGE2:

Prostaglandin E2

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Acknowledgements

The authors thank the volunteers for their donation of blood.

Funding

This work was financially supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Financiadora de Estudos e Projetos (Finep) and Fundo de Incentivo à Pesquisa e Eventos do Hospital de Clínicas de Porto Alegre (FIPE-HCPA).

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Authors and Affiliations

Authors

Contributions

JFP, VSV, JMF and LMRS designed the study. JFP coordinated the collection of blood samples, performed the BM-MSCs generation and characterization, isolation of MNCs from blood, NK enrichment and culture expansion. FS and ADG performed the flow cytometry. JFP and GL performed the cytotoxicity assay. JFP and FSP analyzed the flow cytometry data. JFP conduct statistical analyses. JFP and LMRS drafted this manuscript.

Corresponding author

Correspondence to Lucia Mariano da Rocha Silla.

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Conflict of interest

The authors declare that they have no competing interests.

Ethics Approval

Informed consent was obtained from all volunteers. The study was performed according to the declaration of Helsinki and was approved by the Research Ethics Committee of Hospital de Clínicas de Porto Alegre.

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Pedroso, J.F., de Souza Valim, V., Pezzi, A. et al. An Experimental Study Comparing the Expansion of Peripheral Blood Natural Killer (NK) Cells Cultured with Artificial Antigen-Presenting Cells, in the Presence or Absence of Bone Marrow Mesenchymal Stem Cells (MSCs). Mol Biotechnol 62, 306–315 (2020). https://doi.org/10.1007/s12033-020-00250-2

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