| Autor: |
Chayé MAM; Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands., Tontini C; Allergy Unit, Department of Internal Medicine, University Hospital of Ancona, Ancona, Italy., Ozir-Fazalalikhan A; Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands., Voskamp AL; Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands., Smits HH; Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. h.h.smits@lumc.nl. |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2270, pp. 235-261. |
| DOI: |
10.1007/978-1-0716-1237-8_13 |
| Abstrakt: |
Toll-like receptors (TLRs) are pattern recognition receptors (PRRs), which constitute key components in the recognition of pathogens, thereby initiating innate immune responses and promoting adaptive immune responses. In B cells, TLR ligation is important for their activation and, together with CD40, for their differentiation. TLR ligands are also strong promoters of regulatory B (Breg)-cell development, by enhancing the production of IL-10 and their capacity to induce tolerance. In inflammatory diseases, such as autoimmunity or allergies, Breg-cell function is often impaired, while in chronic infections, such as with helminths, or cancer, Breg-cell function is boosted. Following pathogen exposure, B cells can respond directly by producing cytokines and/or IgM (innate response) and develop into various memory B (Bmem)-cell subsets with class-switched immunoglobulin receptors. Depending on the disease state or chronic infection conditions, various Breg subsets can be recognized as well. Currently, a large array of surface markers is known to distinguish between these large range of B-cell subsets. In recent years, the development of mass cytometers and spectral flow cytometry has allowed for high-dimensional detection of up to 48 markers, including both surface and intracellular/intranuclear markers. Therefore, this novel technology is highly suitable to provide a comprehensive overview of Bmem/Breg-cell subsets in different disease states and/or in clinical intervention trials. Here, we provide detailed instructions of the steps necessary to obtain high-quality data for high-dimensional analysis of multiple human Breg-cell subsets using various TLR ligands. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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