ArticleCells2020
Flow Cytometric Methods for the Detection of Intracellular Signaling Proteins and Transcription Factors Reveal Heterogeneity in Differentiating Human B Cell Subsets.
Article in Cells, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 26 citations in OpenAlex.
- From Selection to Use: Aptamers as Targeting Reagents in Hematology.Biomedicines · 2026Review
- Sex Differences in Lung B Cell Responses in a Murine Model of Asthma.International archives of allergy and immunology · 2026Article
- Lymphocytes and monocytes undergo swift suppression of IL-10R, IL-6R, and IL-2Rβγ signaling under high concentrations of different cytokines.bioRxiv : the preprint server for biology · 2025Article
- Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi-pass acquisition.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2024Article
- Shedding Light on Intracellular Proteins using Flow Cytometry.Cell biochemistry and biophysics · 2024Review
- Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi-pass acquisition.bioRxiv : the preprint server for biology · 2024Article
- Tertiary lymphoid structure-related immune infiltrates in NSCLC tumor lesions correlate with low tumor-reactivity of TIL products.Oncoimmunology · 2024Article
- Article
- B-cell targeting with anti-CD38 daratumumab: implications for differentiation and memory responses.Life science alliance · 2023Article
- B Cell Signaling and Activation in Autoimmunity.Cells · 2023Article
- A synthetic human 3DiScience · 2023Article
- Review
- Termination of CD40L co-stimulation promotes human B cell differentiation into antibody-secreting cells.European journal of immunology · 2022Article
- Oxygen level is a critical regulator of human B cell differentiation and IgG class switch recombination.Frontiers in immunology · 2022Article
- Optimized Protocols forFrontiers in immunology · 2022Article
- Human influenza virus challenge identifies cellular correlates of protection for oral vaccination.Cell host & microbe · 2021Article
- Minimalistic In Vitro Culture to Drive Human Naive B Cell Differentiation into Antibody-Secreting Cells.Cells · 2021Article
- G-CSF as a suitable alternative to GM-CSF to boost dinutuximab-mediated neutrophil cytotoxicity in neuroblastoma treatment.Journal for immunotherapy of cancer · 2021Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The flow cytometric detection of intracellular (IC) signaling proteins and transcription factors (TFs) will help to elucidate the regulation of B cell survival, proliferation and differentiation. However, the simultaneous detection of signaling proteins or TFs with membrane markers (MMs) can be challenging, as the required fixation and permeabilization procedures can affect the functionality of conjugated antibodies. Here, a phosphoflow method is presented for the detection of activated NF-κB p65 and phosphorylated STAT1, STAT3, STAT5 and STAT6, together with the B cell differentiation MMs CD19, CD27 and CD38. Additionally, a TF-flow method is presented that allows the detection of the B cell TFs PAX5, c-MYC, BCL6 and AID and antibody-secreting cell (ASC) TFs BLIMP1 and XBP-1s, together with MMs. Applying these methods on in vitro-induced human B cell differentiation cultures showed significantly different steady-state levels, and responses to stimulation, of phosphorylated signaling proteins in CD27-expressing B cell and ASC populations. The TF-flow protocol and Uniform Manifold Approximation and Projection (UMAP) analysis revealed heterogeneity in TF expression within stimulated CD27- or CD38-expressing B cell subsets. The methods presented here allow for the sensitive analysis of STAT, NF-κB p65 signaling and TFs, together with B cell differentiation MMs, at single-cell resolution. This will aid the further investigation of B cell responses in both health and disease.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.