Sensitive intranuclear flow cytometric quantification of IRF4 protein in multiple myeloma and normal human hematopoietic cells

Vora, Ashni A.; Mondala, Phoebe; Costello, Caitlin; MacLeod, A. Robert; Crews, Leslie

doi:10.1016/j.xpro.2021.100565

Cited by 5 publications

(1 citation statement)

References 21 publications

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“…IRF8 apparently lacks physical interaction with STAT2, which may explain its nuclear localization. IRF1,2,4 are also well-known nuclear proteins (Schaper et al, 1998; Vora et al, 2021).…”

Section: Resultsmentioning

confidence: 99%

The Co-Evolution of Jawed Vertebrates and Interferon Regulatory Factor 5 Generates Unique Inflammation and Innate Immunity

Hubing,

Marquis,

Ziemann

et al. 2024

Preprint

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The emergence of jaws in early vertebrates introduced a novel feeding apparatus and potent oral defenses but also increased the risk of physical injury and pathogen exposure. Innate immunity and inflammation constitute the body’s first line of defense against invading microbes and tissue damage, aiming to eliminate threats and restore internal homeostasis. Interferon regulatory factor 5 (IRF5) plays a critical role in orchestrating innate immunity and inflammation by regulating the transcription of genes that encode type I interferons (IFNs) and pro-inflammatory cytokines. Despite this, the evolution of IRF5 has remained poorly understood. We have identified the IRF5 and IRF6 genes in cartilaginous fish, including sharks. As cartilaginous fish represent one of the oldest surviving jawed vertebrate lineages, the presence of these genes suggests the genes have ancient origins potentially dating back hundreds of millions of years to early jawed vertebrates. Furthermore, our analysis shows that IRF5 has conserved nuclear export sequences and phosphorylation sites for activation throughout evolution from cartilaginous fish to humans, indicating these regulatory elements evolved early and have been maintained across jawed vertebrates. Additionally, the shift in subcellular localization of IRF5 from nucleus to cytosol, and of other interferon related IRFs, aligns with functional enhancements of IRFs in innate immunity and the emergence of IFNs across jawed vertebrates. This analysis implies that the evolution of jaws may have driven the emergence of new IRF members, the expansion of their functions, and the development of a unique inflammation and innate immune system.

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“…IRF8 apparently lacks physical interaction with STAT2, which may explain its nuclear localization. IRF1,2,4 are also well-known nuclear proteins (Schaper et al, 1998; Vora et al, 2021).…”

Section: Resultsmentioning

confidence: 99%

The Co-Evolution of Jawed Vertebrates and Interferon Regulatory Factor 5 Generates Unique Inflammation and Innate Immunity

Hubing,

Marquis,

Ziemann

et al. 2024

Preprint

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Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi‐pass acquisition

Fahlberg,

Forward,

Assita

et al. 2024

Cytometry Pt A

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The fixation and permeabilization of cells are essential for labeling intracellular biomarkers in flow cytometry. However, these chemical treatments often alter fragile targets, such as cell surface and fluorescent proteins (FPs), and can destroy chemically‐sensitive fluorescent labels. This reduces measurement accuracy and introduces compromises into sample workflows, leading to losses in data quality. Here, we demonstrate a novel multi‐pass flow cytometry approach to address this long‐standing problem. Our technique utilizes individual cell barcoding with laser particles, enabling sequential analysis of the same cells with single‐cell resolution maintained. Chemically‐fragile protein markers and their fluorochrome conjugates are measured prior to destructive sample processing and adjoined to subsequent measurements of intracellular markers after fixation and permeabilization. We demonstrate the effectiveness of our technique in accurately measuring intracellular FPs and methanol‐sensitive antigens and fluorophores, along with various surface and intracellular markers. This approach significantly enhances assay flexibility, enabling accurate and comprehensive cellular analysis without the constraints of conventional one‐time measurement flow cytometry. This innovation paves new avenues in flow cytometry for a wide range of applications in immuno‐oncology, stem cell research, and cell biology.

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Nonlinear hybridization chain reaction-based flow cytometric immunoassay for the detection of prostate specific antigen

Zeng

Zhou

Sun³

et al. 2022

Analytica Chimica Acta

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Sensitive intranuclear flow cytometric quantification of IRF4 protein in multiple myeloma and normal human hematopoietic cells

Cited by 5 publications

References 21 publications

The Co-Evolution of Jawed Vertebrates and Interferon Regulatory Factor 5 Generates Unique Inflammation and Innate Immunity

The Co-Evolution of Jawed Vertebrates and Interferon Regulatory Factor 5 Generates Unique Inflammation and Innate Immunity

Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi‐pass acquisition

Nonlinear hybridization chain reaction-based flow cytometric immunoassay for the detection of prostate specific antigen

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