A high throughput method for enrichment of natural killer cells and lymphocytes and assessment of in vitro cytotoxicity

So, Edward; Sallin, Michelle; Zhang, Xiaoyu; Chan, Siaw-Lin; Sahni, Lepakshi; Schulze, Dan H.; Dávila, Eduardo; Strome, Scott E.; Jain, Ajay

doi:10.1016/j.jim.2013.05.001

Cited by 12 publications

(9 citation statements)

References 21 publications

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“…After obtaining informed consent, peripheral blood was obtained from a 25-year-old Caucasian male (patient hB53), and PBMCs were isolated by density gradient centrifugation (So et al., 2013). To induce reprogramming and increase the infection efficiency of PBMCs, cells were infected with lentiviruses expressing OCT4, KLF4, SOX2, and c-MYC from a polycistronic cassette (pHAGE2-TetOminiCMV-hSTEMCCA) (Sommer et al., 2010; Staerk et al., 2010).…”

Section: Methodsmentioning

confidence: 99%

Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

et al. 2014

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SummaryAdvances in induced pluripotent stem cell (iPSC) technology have set the stage for routine derivation of patient- and disease-specific human iPSC-cardiomyocyte (CM) models for preclinical drug screening and personalized medicine approaches. Peripheral blood mononuclear cells (PBMCs) are an advantageous source of somatic cells because they are easily obtained and readily amenable to transduction. Here, we report that the electrophysiological properties and pharmacological responses of PBMC-derived iPSC CM are generally similar to those of iPSC CM derived from other somatic cells, using patch-clamp, calcium transient, and multielectrode array (MEA) analyses. Distinct iPSC lines derived from a single patient display similar electrophysiological features and pharmacological responses. Finally, we demonstrate that human iPSC CMs undergo acute changes in calcium-handling properties and gene expression in response to rapid electrical stimulation, laying the foundation for an in-vitro-tachypacing model system for the study of human tachyarrhythmias.

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Section: Methodsmentioning

confidence: 99%

Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

et al. 2014

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“…Cytolytic activity of patients' peripheral blood mononuclear cells (PBMCs) 19 and healthy donors' CD56 + -isolated NK cells 20 were tested in a 2 h BATDA Europium release assay against K562 (MHC class I-negative erythroleukemia cell line) and A673 (EWS cell line). In healthy donors, the following target cell lines were tested additionally A204, TE671 and AX-OH-1 (rhabdomyosarcoma), GROH-1 and KNOH-1 (primitive neuroectodermal tumor (PNET)), SWOH-1 and PL377 (rhabdoid tumor).…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

Favorable NK cell activity after haploidentical hematopoietic stem cell transplantation in stage IV relapsed Ewing’s sarcoma patients

Schlegel

Feuchtinger

Nitschke-Gérard

et al. 2015

Bone Marrow Transplant

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Natural killer (NK) cell activity has been shown to have potential activity against Ewing's sarcoma (EWS) especially in tumors with low HLA I expression and high NKG2D expression. Two patients with metastatic relapsed and primary metastatic stage IV EWS who had received two courses of high dose chemotherapy with autologous stem cell rescue were transplanted from a haploidentical parental stem cell donor. Patients are alive in ongoing CR for 10.2 and 3.4 years now. Post transplant local second and first relapses were treated successfully in both patients. In vivo IL-2 stimulation not only increased the number and activity of effector cells in one patient but was also associated with severe GvHD. In vitro studies demonstrated high NK cell activity against K562 and relevant activity against EWS cell line A673 post transplant. NK activity was enhanced by cytokine prestimulation as well as by EWS targeting anti-GD2 Ab. Haploidentical hematopoietic stem cell transplantation (HSCT) might contribute to long-term survival by NK cellmediated effect exerted by donor-derived NK cells. Local tumor recurrence was manageable in both high-risk patients indicating systemic immune control preventing subsequent metastasizing. The efficacy of haploidentical HSCT, cytokine application and tumor targeting antibodies for the use of Ab-dependent cellular cytotoxicity needs evaluation in clinical trials.

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“…Human PBMCs were prepared from healthy donors' blood using Ficoll density centrifugation as described previously. 17 T cells were then isolated from the PBMCs using an EasySep Human CD3 Positive Selection Kit (STEMCELL Technologies, Inc., Vancouver, Canada) according to the manufacturer's instructions. The isolated T cells were cultured in complete RPMI 1640 with 10% FBS and 1% penicillin/streptomycin at 37°C in a 5% CO 2 humidified incubator before assays.…”

Section: Methodsmentioning

confidence: 99%

BiHC, a T-Cell–Engaging Bispecific Recombinant Antibody, Has Potent Cytotoxic Activity Against Her2 Tumor Cells

Xing

Lin

et al. 2017

Translational Oncology

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Among different cancer immunotherapy approaches, bispecific antibodies (BsAbs) are of great interest due to their ability to recruit immune cells to kill tumor cells directly. Various BsAbs against Her2 tumor cells have been proposed with potent cytotoxic activities. However, most of these formats require extensive processing to obtain heterodimeric bispecific antibodies. In this study, we describe a bispecific antibody, BiHC (bispecific Her2-CD3 antibody), constructed with a single-domain anti-Her2 and a single-chain Fv (variable fragment) of anti-CD3 in an IgG-like format. In contrast to most IgG-like BsAbs, the two arms in BiHC have different molecular weights, making it easier to separate hetero- or homodimers. BiHC can be expressed in Escherichia coli and purified via Protein A affinity chromatography. The purified BiHC can recruit T cells and induce specific cytotoxicity of Her2-expressing tumor cells in vitro. The BiHC can also efficiently inhibit the tumor growth in vivo. Thus, BiHC is a promising candidate for the treatment of Her2-positive cancers.

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A high throughput method for enrichment of natural killer cells and lymphocytes and assessment of in vitro cytotoxicity

Cited by 12 publications

References 21 publications

Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

Favorable NK cell activity after haploidentical hematopoietic stem cell transplantation in stage IV relapsed Ewing’s sarcoma patients

BiHC, a T-Cell–Engaging Bispecific Recombinant Antibody, Has Potent Cytotoxic Activity Against Her2 Tumor Cells

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