The Leukocyte Composition of Peripheral Blood Buffy Coat

Teetson, Winfred; Cartwright, C A; Dreiling, Bernard J.; Steinberg, Martin H.

doi:10.1093/ajcp/79.4.500

Cited by 11 publications

(8 citation statements)

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“…PBMCs are a heterogeneous population of innate immune cells consisting of T-cells, B- cells, monocytes, NK cells and dendritic cells [44]. Human blood dendritic cells are classified as plasmacytoid dendritic cells (pDCs), myeloid BDCA1 + dendritic cells (mDC1s) and myeloid BDCA3 + dendritic cells (mDC2s) [45].…”

Section: Resultsmentioning

confidence: 99%

Induction of Type I and Type III Interferons by Borrelia burgdorferi Correlates with Pathogenesis and Requires Linear Plasmid 36

et al. 2014

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The capacity for Borrelia burgdorferi to cause disseminated infection in humans or mice is associated with the genotype of the infecting strain. The cytokine profiles elicited by B. burgdorferi clinical isolates of different genotype (ribosomal spacer type) groups were assessed in a human PBMC co-incubation model. RST1 isolates, which are more frequently associated with disseminated Lyme disease in humans and mice, induced significantly higher levels of IFN-α and IFN-λ1/IL29 relative to RST3 isolates, which are less frequently associated with disseminated infection. No differences in the protein concentrations of IFN-γ, IL-1β, IL-6, IL-8, IL-10 or TNF-α were observed between isolates of differing genotype. The ability of B. burgdorferi to induce type I and type III IFNs was completely dependent on the presence of linear plasmid (lp) 36. An lp36-deficient B. burgdorferi mutant adhered to, and was internalized by, PBMCs and specific dendritic cell (DC) subsets less efficiently than its isogenic B31 parent strain. The association defect with mDC1s and pDCs could be restored by complementation of the mutant with the complete lp36. The RST1 clinical isolates studied were found to contain a 2.5-kB region, located in the distal one-third of lp36, which was not present in any of the RST3 isolates tested. This divergent region of lp36 may encode one or more factors required for optimal spirochetal recognition and the production of type I and type III IFNs by human DCs, thus suggesting a potential role for DCs in the pathogenesis of B. burgdorferi infection.

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Section: Resultsmentioning

confidence: 99%

Induction of Type I and Type III Interferons by Borrelia burgdorferi Correlates with Pathogenesis and Requires Linear Plasmid 36

et al. 2014

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“…Factors in uencing cell-type composition can be false-positively identi ed as differentially expressed genes. Peripheral leukocytes are composed of PBMCs that include B, T lymphocytes, and monocytes 31 . PMNLs are considered as the granulocyte population composed of neutrophils, eosinophils, and basophils.…”

Section: Leukocyte Decomposition Between Peripheral Blood Mononuclear...mentioning

confidence: 99%

“…The ratio between normalized whole blood pChRO data and PBMC/PMNL data in signature genes will re ect the relative cell-type fraction. While PMNLs normally compose 65% of the leukocyte population 31 , whole blood pChRO is closer to PBMC pro le since PBMCs are more transcriptionally active than PMNLs. Our estimation from the data is that PBMCs are on average ~ 4.1 fold transcriptionally more active than PMNLs.…”

Section: Leukocyte Decomposition Between Peripheral Blood Mononuclear...mentioning

confidence: 99%

Integrative nascent RNA methods to reveal cell-type specific transcription programs in peripheral blood and its derivative cells

Kwak

Kimm

Kim

et al. 2022

Preprint

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Nascent RNA sequencing is a powerful method to measure transcription with high resolution, sensitivity, and directional information. We present an integrated package of nascent RNA-seq methods composed of ultrafast Precision Run On (uPRO) combined with deep learning and hierarchical co-expression network analysis to discover enhancers, promoters, and transcription factor networks that define cell-type specific transcription programs. We generated nascent transcription profiles using one-day uPRO in human blood-derived cell lines and raw whole blood of ~ 1 ml. Transcription sites at enhancers and genes were conserved in positions but varied in expression levels across cell types. Transcription factors such as TCF-3 and OCT1 were pivotally associated with enhancer-gene networks. Intriguingly, transcription factors related to cell stress and inflammation are associated with inter-individual variation of leukocyte transcription in whole blood. Our integration of experimental and computational nascent RNA methods will provide an efficient strategy to identify specific transcriptional programs with minimal sample requirements.

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“…Gene ontology analysis of the signature genes were performed using the PANTHER gene ontology analysis 35 . Cell type fractions were calculated from the relative expression levels of each signature gene subset relative to the reference data, assuming reference peripheral leukocyte compositions of normal healthy adult individual: PBMC = 0.35, PMNL = 0.65, B = 0.07, T = 0.21, and CD4 = 0.06 25 .…”

Section: Hierarchical Coexpression Network Analysismentioning

confidence: 99%

Integrative nascent RNA methods to reveal cell-type specific transcription programs in peripheral blood and its derivative cells

Kim

Dziubek

Lee

et al. 2019

Preprint

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Nuclear Run-On sequencing is a powerful method to measure transcription with high resolution, sensitivity, and directional information, which provides alternative perspective from existing methods such as chromatin immunoprecipitation or mRNA sequencing. Current form of Nuclear Run-On assays such as Precision Run-On sequencing (PRO-seq) involves multiple RNA chemistry steps including RNA end repairs and ligations. These have limited the widespread use of PRO-seq by requiring robust RNA handling skills and multiple days of effort. To solve this, we developed an ultrashort PRO-seq (uPRO) method that requires minimal steps. In uPRO, the requirement of only two reactions -RNA adaptor ligation and template switch reverse transcriptionreduced the procedure into less than a single day. Using uPRO, we generated genome-wide transcription profiles of human haploid cell lines (HAP1) and peripheral blood samples combined with Chromatin Run-On sequencing (pChRO). Blood cell handling procedure is dramatically reduced using pChRO directly on crude chromatin preparations, and enables utilizing archived specimens. As a result, we identified individual differences in the transcriptional profiles of human whole blood from a small volume (~1 ml). We also generated blood cell type specific transcription data, and deconvoluted the nucleated blood cell compositions by modeling to the reference datasets. Overall, uPRO and pChRO provided a powerful platform to identify differentially expressed genes between individuals with minimal sample requirements. Fig 1. Schematics of the uPRO procedure A. Comparison between conventional PRO-seq and uPRO procedures. Adapted from Mahat et al 12

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The Leukocyte Composition of Peripheral Blood Buffy Coat

Cited by 11 publications

References 0 publications

Induction of Type I and Type III Interferons by Borrelia burgdorferi Correlates with Pathogenesis and Requires Linear Plasmid 36

Induction of Type I and Type III Interferons by Borrelia burgdorferi Correlates with Pathogenesis and Requires Linear Plasmid 36

Integrative nascent RNA methods to reveal cell-type specific transcription programs in peripheral blood and its derivative cells

Integrative nascent RNA methods to reveal cell-type specific transcription programs in peripheral blood and its derivative cells

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