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Short-Term Exercise and Immune Function
Published in Ronald R. Watson, Marianne Eisinger, Exercise and Disease, 2020
The more recent use of fluorescence-activated cell sorting allows differentiation of 10,000 or more cells per sample, which intuitively is more accurate than counting 100 or 200 cells under a microscope. Accordingly, bouts of maximal or submaximal short-term exercise appear to result in a decrease in the percentage of B-cells without a concomitant change in absolute numbers. Therefore, it appears that B-cells are not mobilized during short-term exercise.
Detection of Lysosomal Membrane Permeabilization
Published in Bruno Gasnier, Michael X. Zhu, Ion and Molecule Transport in Lysosomes, 2020
Anne-Marie Ellegaard, Line Groth-Pedersen, Marja Jäättelä
This protocol applies to adherent cells but can easily be modified to cells in suspension, in which case the analysis is performed on a fluorescence-activated cell sorter (FACS). Depending on the treatment of interest, it can be added to the cells before or after loading with AO. If treatment is applied before AO loading, one must check that the loading of AO is not significantly affected by the treatment.
T Cells
Published in Miroslav Holub, Immunology of Nude Mice, 2020
Since the earliest days of the discovery of “athymia”, or, more exactly, thymic dysgenesis in the nude mouse, it has been known that the animal is not entirely devoid of the T-lineage cells. Raff’s studies have revealed small but significant numbers of Thy-1-positive cells in nude mouse spleens and lymph nodes;1,2 3 to 20% of Thy-1-positive cells were demonstrated by a variety of techniques and their immaturity confirmed by the finding of TL expression which is normally confined to the thymus.3-5 In a pioneer study with fluorescence-activated cell sorting, Cantor et al.6 found two populations of Thy-1.2-positive cells: “bright T cells” (with high Thy 1 expression), preferentially homing to the spleen and present in normal mice (BALB/c) in constant numbers from birth to 30 weeks of age; and “dull T cells”, homing more to lymph nodes, increasing in numbers with age, not affected by adult thymectomy and rapidly recirculating. In nude mice, the dull T cells are much less reduced than the bright T cells which amount here to about 20% of the dull T cells.6
Simultaneous affinity maturation and developability enhancement using natural liability-free CDRs
Published in mAbs, 2022
Andre A. R. Teixeira, Sara D’Angelo, M. Frank Erasmus, Camila Leal-Lopes, Fortunato Ferrara, Laura P. Spector, Leslie Naranjo, Esteban Molina, Tamara Max, Ashley DeAguero, Katherine Perea, Shaun Stewart, Rebecca A. Buonpane, Horacio G. Nastri, Andrew R. M. Bradbury
Given the size of each of these libraries, we performed first and second rounds of selection using magnetic-assisted cell sorting (MACS) at antigen concentrations of 10 nM and 1 nM, respectively. This allowed us to label and sort a larger number of cells than would have been practical using a flow cytometer. For subsequent rounds, we used fluorescence-activated cell sorting (FACS) to enable more precise sorting of the cells of interest. After the first three rounds of equilibrium sorting, we performed two rounds of kinetic sorting with 4 hours of competition for the L3 and H1H2 libraries. Only one 4 h kinetic sort was performed for the L1L2 library, followed by a negative sort, where the population was only incubated with secondary reagents and negative cells were sorted, which was done out of concern that enrichment of polyreactive antibodies may occur in light of the weak positivity observed in the absence of antigen
Nanoliposomes as drug delivery systems: safety concerns
Published in Journal of Drug Targeting, 2022
Yu. A. Tereshkina, T. I. Torkhovskaya, E. G. Tikhonova, L. V. Kostryukova, M. A. Sanzhakov, E. I. Korotkevich, Yu. Yu. Khudoklinova, N. A. Orlova, E. F. Kolesanova
Even though nanoparticle characterisation is often complicated by polydispersity, the safety plan considers it sufficient to obtain a summary estimate using the methods described in Table 1 [67], dynamic light scattering, transmission electron microscopy, scanning electron microscopy, atomic force microscopy, or nanoparticle tracking (track) analysis [6,135]. The latter is based on the observation of the Brownian motion of separate nanoparticles, which speed depends on their properties and media viscosity [135]. This property affects the nanoliposome motion in the extracellular matrix. Among the methods recommended (Table 1), fluorescent-activated cell sorting (FACS), a method commonly used for cells, is also considered an essential one. This is a variant of flow cytometry wherein the fluorescent dye is attached to nanoparticles as opposed to cells, which allows detection of nanoparticle aggregates, including nanoliposomes, in the flow [67].
Combinatorial effect of radium-223 and irreversible electroporation on prostate cancer bone metastasis in mice
Published in International Journal of Hyperthermia, 2021
Raniv D. Rojo, Joy Vanessa D. Perez, Jossana A. Damasco, Guoyu Yu, Song-Chang Lin, Francisco M. Heralde, Nora M. Novone, Elmer B. Santos, Sue-Hwa Lin, Marites P. Melancon
To generate C4-2B cells expressing BMP4 and a luciferase reporter (LT), cDNAs encoding human BMP4 were inserted into the bicistronic retroviral vector pBMN-I-Neo was used to generate retroviruses containing BMP4. The resulting retroviruses were used to infect C4-2B–LT cells. The cells were then selected by G418 (Corning 30-234-CR). BMP4-expressing Myc-CaP and TRAMP-C2 cells were produced by transfection with a bicistronic retroviral vector containing the mouse BMP4 cDNA and floxed-green fluorescent protein (GFP) through internal ribosome entry sites (IRES). Cells were selected by fluorescence-activated cell sorting through GFP. GFP was then removed using an adenoviral vector containing cre recombinase.