Expansion of NK cells from PBMCs using immobilized 4-1BBL and interleukin-21

Li, Xiaomei; He, Chenhui; Liu, Changzhen; Ma, Juan; Ma, Pan; Cui, Honglian; Tao, Hua; Gao, Bin

doi:10.3892/ijo.2015.3005

Cited by 26 publications

(26 citation statements)

References 42 publications

(42 reference statements)

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“…Interestingly, chemokines including CCL19 that is known to attract DCs and central memory T cells (45) were also increased. Any LOAd-infected DCs could stimulate proliferation of antigenspecific T cells but LOAd703-infected DCs expanded NK cells as well, which is likely due to 4-1BBL as it is a known potent stimulator of NK-cell expansion (21,22). Similarly, an article by Galivo and colleagues demonstrated that adenoviruses with CD40L can stimulate T-cell responses against tumor antigens, and not only to adenoviral antigens which unfortunately was the case when using a vesicular stomatitis virus (46).…”

Section: Discussionmentioning

confidence: 99%

“…19) and full-length human 4-1BB ligand . While CD40L has documented antitumor activities including CD40-mediated tumor cell apoptosis (18), and adaptive immune activation (15,20), 4-1BBL is known to enhance immunologic memory and expand natural killer (NK) cells (21,22). Because of the actions by the virus per se and the CD40 pathway combined with 4-1BB signaling, we hypothesize that the LOAd703 virus has a broad mechanism of action by stimulating several arms of the immune system simultaneously to affecting the biological reality of the tumor and its stroma.…”

Section: Introductionmentioning

confidence: 99%

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Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus

Eriksson

Milenova

Wenthe

et al. 2017

Clinical Cancer Research

111

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Pancreatic cancer is a severe indication with short expected survival despite surgery and/or combination chemotherapeutics. Checkpoint blockade antibodies are approved for several cancer indications, but pancreatic cancer has remained refractory. However, there are clinical data suggesting that stimulation of the CD40 pathway may be of interest for these patients. Oncolytic viruses armed with immunostimulatory genes represent an interesting approach. Herein, we present LOAd703, a designed adenovirus armed with trimerized and that activates the CD40 and 4-1BB pathways, respectively. As many cells in the tumor stroma, including stellate cells and the infiltrating immune cells, express CD40 and some 4-1BB, we hypothesize that LOAd703 activates immunity and simultaneously modulates the biology of the tumor stroma. Tumor, stellate, endothelial, and immune cells were infected by LOAd703 and investigated by flow cytometry, proteomics, and functional analyses. LOAd703-infected pancreatic cell lines were killed by oncolysis, and the virus was more effective than standard-of-care gemcitabine. In xenograft models, LOAd703 efficiently reduced established tumors and could be combined with gemcitabine for additional effect. Infected stellate and tumor cells reduced factors that promote tumor growth (Spp-1, Gal-3, HGF, TGFβ and collagen type I), while chemokines were increased. Molecules involved in lymphocyte migration were upregulated on infected endothelial cells. Dendritic cells were robustly stimulated by LOAd703 to produce costimulators, cytokines and chemokines, and such DCs potently expanded both antigen-specific T cells and NK cells. LOAd703 is a potent immune activator that modulates the stroma to support antitumor responses. .

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus

Eriksson

Milenova

Wenthe

et al. 2017

Clinical Cancer Research

111

View full text Add to dashboard Cite

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“…In fact, the effects of IL-21 in NK cell proliferation are controversial. Immobilized or membrane-bound IL-21 has a positive effect on NK cell expansion (25, 61). Also, the longer survival of human NK cells following combined IL-21 and IL-2 stimulation has been reported (18).…”

Section: Discussionmentioning

confidence: 99%

A Two-Phase Expansion Protocol Combining Interleukin (IL)-15 and IL-21 Improves Natural Killer Cell Proliferation and Cytotoxicity against Rhabdomyosarcoma

et al. 2017

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Rhabdomyosarcoma (RMS) is the most common soft tissue malignancy in children. Despite intensive research in recent decades the prognosis for patients with metastatic or relapsed diseases has hardly improved. New therapeutic concepts in anti-tumor therapy aim to modulate the patient’s immune system to increase its aggressiveness or targeted effects toward tumor cells. Besides surgery, radiotherapy and chemotherapy, immune activation by direct application of cytokines, antibodies or adoptive cell therapy are promising approaches. In the last years, adoptive transfer of natural killer (NK) cells came into the focus of translational medicine, because of their high cytotoxic potential against transformed malignant cells. A main challenge of NK cell therapy is that it requires a high amount of functional NK cells. Therefore, ex vivo NK cell expansion protocols are currently being developed. Many culturing strategies are based on the addition of feeder or accessory cells, which need to be removed prior to the clinical application of the final NK cell product. In this study, we addressed feeder cell-free expansion methods using common γ-chain cytokines, especially IL-15 and IL-21. Our results demonstrated high potential of IL-15 for NK cell expansion, while IL-21 triggered NK cell maturation and functionality. Hence, we established a two-phase expansion protocol with IL-15 to induce an early NK cell expansion, followed by short exposure to IL-21 that boosted the cytotoxic activity of NK cells against RMS cells. Further functional analyses revealed enhanced degranulation and secretion of pro-inflammatory cytokines such as interferon-γ and tumor necrosis factor-α. In a proof of concept in vivo study, we also observed a therapeutic effect of adoptively transferred IL-15 expanded and IL-21 boosted NK cells in combination with image guided high precision radiation therapy using a luciferase-transduced RMS xenograft model. In summary, this two-phased feeder cell-free ex vivo culturing protocol combined efficient expansion and high cytolytic functionality of NK cells for treatment of radiation-resistant RMS.

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“…In collaboration with CytoSen Therapeutics (recently acquired by Kiadis Pharma NV), we re‐derived and validated clone CSTX002 and its MCB, which is now being shared academically and is supporting NK cell propagation for several clinical trials. Similar versions have been developed and published by others, and feeder cells with soluble IL‐21 have also been used, but appear much less effective …”

Section: Development Of Il‐21 Feeder Cellsmentioning

confidence: 99%

“…Similar versions have been developed and published by others, [77][78][79] and feeder cells with soluble IL-21 have also been used, but appear much less effective. [80][81][82][83][84][85][86][87]…”

Section: De Velopment Of Il-21 Feeder Cell Smentioning

confidence: 99%

Cellular therapy: Adoptive immunotherapy with expanded natural killer cells

Lee

2019

Immunological Reviews

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Adoptive immunotherapy with natural killer cells was pioneered 30 years ago in human clinical trials with the development of cytokine‐induced killer cells—unfractionated peripheral blood mononuclear cell (PBMC) populations activated overnight with IL‐2. Higher doses were subsequently made possible through the advent of steady‐state apheresis, allowing the collection of PBMC numbers equivalent to an entire adult blood volume, and increased purity made feasible through magnetic CD3‐depletion and/or CD56‐selection methods. Still, these approaches rarely achieved clinical dosing above a single infusion of 108 NK cells/kg, except with substantial donor‐recipient size mismatch (eg, parents donating cells to children). To address this shortcoming, leukemia cell lines with NK cell‐like function or ex vivo expansion approaches centered on the homeostatic cytokine IL‐15 were developed. Here, we describe the development of an ex vivo expansion system based on a feeder cell expressing membrane‐bound IL‐21 that enables log‐phase growth of primary NK cells for many weeks without inducing senescence, and describe the biology, correlative science, and translation to clinical trials for patients with leukemia, brain tumors, and solid tumors.

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Expansion of NK cells from PBMCs using immobilized 4-1BBL and interleukin-21

Cited by 26 publications

References 42 publications

Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus

Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus

A Two-Phase Expansion Protocol Combining Interleukin (IL)-15 and IL-21 Improves Natural Killer Cell Proliferation and Cytotoxicity against Rhabdomyosarcoma

Cellular therapy: Adoptive immunotherapy with expanded natural killer cells

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