Gene Expression Analysis of Ex Vivo Expanded and Freshly Isolated NK Cells From Cancer Patients

Park, Kyoung Un; Jin, Ping; Sabatino, Marianna; Ji, Feng; Civini, Sara; Khuu, Hanh; Berg, Maria; Childs, Richard W.; Stroncek, David F.

doi:10.1097/cji.0b013e3181f71b81

Cited by 23 publications

(23 citation statements)

References 30 publications

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“…Indeed, gene expression profiles were shown to undergo substantial changes during NK-cell in vitro expansion. 51 However, comparative functional experiments with non-expanded versus expanded NK cells from some healthy donors and CML patients confirmed the results obtained with expanded NK cells and argue against a major effect of in vitro expansion on NK-cell functionality.…”

Section: Discussionmentioning

confidence: 69%

“…No significant correlation of CD107a secretion with disease response at the time of analysis was found (Figure 4c). In contrast to the reduced degranulation responses, direct cytolysis of K562 by 51 Cr release was unaffected at various effector to target ratios in patients on imatinib treatment (Figure 4d, Supplementary Figure 2). Again, presence or absence of 1 mM imatinib in the culture medium during NK-cell expansion did not affect results ( Figure 4d, center panel), and non-expanded NK cells from healthy donors and patients were also equally effective (Figure 4d, right panel).…”

Section: Nk Cells In CML Patients Are Functionally Impaired Both At Dmentioning

confidence: 95%

“…For 51 Cr release assays, various numbers of NK cells were coincubated in triplicate with 2500 target cells labeled with 100 mCi 51 Cr/1 Â 10 6 cells (phycoerythrin Applied Biosystems) in a total volume of 200 ml in a V-bottomed 96-well plate at 37 1C and 5% CO 2 . Supernatants were harvested and radioactivity was counted in a gamma counter.…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

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NK cells are dysfunctional in human chronic myelogenous leukemia before and on imatinib treatment and in BCR–ABL-positive mice

et al. 2011

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Although BCR-ABL þ stem cells in chronic myeloid leukemia (CML) resist elimination by targeted pharmacotherapy in most patients, immunological graft-versus-leukemia effects can cure the disease. Besides cytotoxic T cells, natural killer (NK) cells may have a role in immune control of CML. Here, we explored the functionality of NK cells in CML patients and in a transgenic inducible BCR-ABL mouse model. Compared with controls, NK-cell proportions among lymphocytes were decreased at diagnosis of CML and did not recover during imatinib-induced remission for 10-34 months. Functional experiments revealed limited in vitro expansion of NK cells from CML patients and a reduced degranulation response to K562 target cells both at diagnosis and during imatinib therapy. Consistent with the results in human CML, relative numbers of NK1.1 þ NK cells were reduced following induction of BCR-ABL expression in mice, and the defects persisted after BCR-ABL reversion. Moreover, target-induced degranulation by expanded BCR-ABL þ NK cells was compromised. We conclude that CML is associated with quantitative and functional defects within the NK-cell compartment, which is reproduced by induced BCR-ABL expression in mice. Further work will aim at identifying the mechanisms of NK-cell deficiency in CML and at developing strategies to exploit NK cells for immunotherapy.

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Section: Discussionmentioning

confidence: 69%

Section: Nk Cells In CML Patients Are Functionally Impaired Both At Dmentioning

confidence: 95%

Section: Cytotoxicity Assaysmentioning

confidence: 99%

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NK cells are dysfunctional in human chronic myelogenous leukemia before and on imatinib treatment and in BCR–ABL-positive mice

et al. 2011

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“…The adoptive infusion of NK cells is a promising immunotherapy for patients with advanced malignancies [5]. Using gene and microRNA expression microarrays, Park et al provided distinct expression profiles of ex vivo expanded NK cells and freshly isolated NK cells from cancer patients [17]. Among approximately 25 100 genes evaluated, the expanded NK cells overexpressed 1098 genes and 28 microRNAs when compared with freshly isolated human NK cells [17].…”

Section: Using Nk Cells For Antitumor Therapymentioning

confidence: 99%

Genomic expression profiling of NK cells in health and disease

2014

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NK cells are important components of innate and adaptive immunity. Functionally, they play key roles in host defense against tumors and infectious pathogens. Within the past few years, genomic-scale experiments have provided us with a plethora of gene expression data that reveal an extensive molecular and biological map underlying gene expression programs. In order to better explore and take advantage of existing datasets, we review here the genomic expression profiles of NK cells and their subpopulations in resting or stimulated states, in diseases, and in different organs; moreover, we contrast these expression data to those of other lymphocytes. We have also compiled a comprehensive list of genomic profiling studies of both human and murine NK cells in this review.

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“…Indeed, EBV-LCL stimulation has been reported as a method to expand and culture NK cells for both experimental and clinical use [347][348][349][350]. In contrast, the final drug product generated cell drug product; which, has been shown to be preferred for ACT immunotherapy [353][354][355].…”

Section: Discussionmentioning

confidence: 99%

Strategies for improving T-cell manufacturing for adoptive immunotherapies

Corvino¹

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Immunotherapy is a novel approach to disease treatment and has shown numerous clinical successes. In brief, immunotherapy exploits the principles of immunology to generate an anti-viral or anti-cancer response. One of the myriad methods that exist to stimulate an immunotherapeutic response is adoptive T-cell transfer (ACT). ACT is an immunotherapy approach whereby patient-derived antigen-specific T-cells are expanded in vitro and infused into the patient. Thus, a central factor of ACT immunotherapy is the in vitro T-cell stimulation and expansion step. In recent years, T-cell stimulation and expansion methods have focused on maximising cell yield. These expansion methods rely on the use of high-dose and often repeated cycles of stimulation. While effective in deriving large cell yields, this Tcell expansion strategy appears to compromise T-cell quality. Furthermore, various antigen sources are used to stimulate and expand antigen-specific T-cells. The impact these differing antigen sources have on T-cell quality is poorly understood. An aspect of this project aims to describe the consequences of antigen stimulation dose on T-cell quality. This project also evaluated common antigen stimulation sources, including Epstein-Barr Virus (EBV) transformed lymphoblastoid cell lines (EBV-LCLs), adenoviral vector-infected antigen presenting cells (APCs), and peptide-pulsed APCs, to determine their impact on T-cell quality. Of particular interest is the impact of EBV-LCLbased stimulation as this method is widely used for the manufacture of ACT drug products. To this end, we hypothesised that the therapeutic potential of an ACT drug product is heavily influenced by the stimulation and expansion method used. Furthermore, this project aimed to demonstrate the utility of comprehensive ACT drug product assessment. Indeed, our findings revealed that T-cell cultures stimulated with varying stimulation doses exhibit different phenotypic, transcriptional, and T cell receptor (TCR) repertoire profiles that reflect functional differences. This analysis can potentially be used to distinguish between ACT drug products of superior quality. The comprehensive analysis approach used also revealed that the dose of EBV-LCL-based stimulation had a profound impact on the overall ACT drug product composition. Specifically, it was observed that a high-dose EBV-LCL stimulation promoted a significant increase in natural killer cells but limited expansion of CD4 + T-cells. In contrast, varying stimulation dose of other antigen stimulation sourcesadenoviral vector-infected APCs and peptide-pulsed APCs-had less influence on culture composition. Overall, these two stimulation approaches resulted in ACT drug products that predominately contained T-cells. Additionally, T-cells derived from a high-dose EBV-LCL stimulation showed hallmarks of T-cell exhaustion, via the co-expression of multiple I would like to thank everyone who has helped me directly or indirectly throughout my Ph.D degree. First and foremost, I must express my sincere gratitude to Prof. R...

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Gene Expression Analysis of Ex Vivo Expanded and Freshly Isolated NK Cells From Cancer Patients

Cited by 23 publications

References 30 publications

NK cells are dysfunctional in human chronic myelogenous leukemia before and on imatinib treatment and in BCR–ABL-positive mice

NK cells are dysfunctional in human chronic myelogenous leukemia before and on imatinib treatment and in BCR–ABL-positive mice

Genomic expression profiling of NK cells in health and disease

Strategies for improving T-cell manufacturing for adoptive immunotherapies

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