Electrical lysis of biological cells on a microfluidic platform has evoked significant interest because of its applications in rapid recovery of intracellular contents such as nucleic acids or proteins without introducing lytic agents. Applying a direct current (DC) field for cell lysis typically requires field strength of 1-10 kV/cm, which is dependent on the cell type: prokaryotes or eukaryotes. Bubble generation and Joule heating can often be induced under such high field strengths. In this study we fabricated a simple microfluidic device using low-cost soft lithography with channel widths considerably larger than the cell size to avoid clogging and ensure stable performance, which was able to lyse green fluorescent protein (GFP)-expressing Escherichia coli cells under continuous DC voltage while cells were flowing through the channels. The cell lysis only happened in a defined section of a microfluidic channel because of local field amplification by geometric modification. The geometric modification also effectively decreased the required voltage for lysis severalfold. The cell lysis was verified by plate count on nutrient agar plates and by fluorescence spectroscopy.
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http://dx.doi.org/10.1007/978-1-59745-426-1_3 | DOI Listing |
Sci Adv
January 2025
Texas Children's Cancer Center, Texas Children's Hospital, Baylor College of Medicine, Houston, TX 77030, USA.
Chimeric antigen receptor T cells (CART) targeting CD19 through CD28.ζ signaling induce rapid lysis of leukemic blasts, contrasting with persistent tumor control exhibited by 4-1BB.ζ-CART.
View Article and Find Full Text PDFJ Virol
January 2025
University of Central Florida, College of Medicine, Orlando, Florida, USA.
Unlabelled: Persistent viral infections can be an important medical problem, with persistently infected (PI) cells extending viral shedding, maintaining inflammation, and providing potential sources for new viral variants. Given that PI cells can acquire resistance to some innate immune pathways, we tested the hypothesis that complement (C')-mediated lysis of parainfluenza virus 5 (PIV5)-infected cells would differ between acute-infected and PI cells. Biochemical and real-time cell viability assays showed effective C'-mediated lysis of A549 lung cells acutely infected with PIV5, through pathways that depended on C3 and C5, but largely independent of C6.
View Article and Find Full Text PDFCells
December 2024
BIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité-Universitätsmedizin Berlin, 13353 Berlin, Germany.
Antibody-dependent cell-mediated cytotoxicity (ADCC) by NK cells is a key mechanism in anti-cancer therapies with monoclonal antibodies, including cetuximab (EGFR-targeting) and avelumab (PDL1-targeting). Fc gamma receptor IIIa (FcγRIIIa) polymorphisms impact ADCC, yet their clinical relevance in NK cell functionality remains debated. We developed two complementary flow cytometry assays: one to predict the FcγRIIIa-V158F polymorphism using a machine learning model, and a 15-color flow cytometry panel to assess antibody-induced NK cell functionality and cancer-immune cell interactions.
View Article and Find Full Text PDFSTAR Protoc
January 2025
Division of Immunology and Infectious Disease Biology, INtegrative GENomics of HOst-PathogEn (INGEN-HOPE) Laboratory, CSIR-Institute of Genomics and Integrative Biology (CSIR-IGIB), Mall Road, Delhi 110007, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India. Electronic address:
Intracellular microorganisms like viruses and bacteria impact immune cell function. However, detection of these microbes is challenging as the majority exist in a non-culturable state. This protocol presents detailed steps to investigate intracellular microbial diversity using single-cell RNA sequencing (scRNA-seq) in immune-cells of SARS-CoV-2-positive and recovered patients.
View Article and Find Full Text PDFClin Kidney J
January 2025
Department of Medicine, Universidad Autonoma de Madrid and IIS-Fundacion Jimenez Diaz, Madrid, Spain.
Chimeric antigen receptor T (CAR-T) cell therapy, an emerging personalized immunotherapy for various haematologic malignancies, autoimmune diseases and other conditions, involves the modification of patients' T cells to express a chimeric antigen receptor that recognizes tumour or autoimmune cell antigens, allowing CAR-T cells to destroy cancerous and other target cells selectively. Despite remarkable clinical improvements in patients, multiple adverse effects have been associated with CAR-T cell therapy. Among the most recognized adverse effects are cytokine release syndrome, immune effector cell-associated neurotoxicity syndrome and tumour lysis syndrome.
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