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CD40L modulates transcriptional signatures of neutrophils in the bone marrow associated with development and trafficking
Tábata Takahashi França, Ashraf Al-Sbiei, Ghada Bashir, Yassir Awad Mohamed, Ranieri Coelho Salgado, Lucila Akune Barreiros, Sarah Maria da Silva Napoleão, Cristina Worm Weber, Janaíra Fernandes Severo Ferreira, Carolina Sanchez Aranda, Carolina Prando, Mayra B. de Barros Dorna, Igor Jurisica, Maria J. Fernandez-Cabezudo, Hans D. Ochs, Antonio Condino-Neto, Basel K. Al-Ramadi, Otavio Cabral-Marques
Tábata Takahashi França, Ashraf Al-Sbiei, Ghada Bashir, Yassir Awad Mohamed, Ranieri Coelho Salgado, Lucila Akune Barreiros, Sarah Maria da Silva Napoleão, Cristina Worm Weber, Janaíra Fernandes Severo Ferreira, Carolina Sanchez Aranda, Carolina Prando, Mayra B. de Barros Dorna, Igor Jurisica, Maria J. Fernandez-Cabezudo, Hans D. Ochs, Antonio Condino-Neto, Basel K. Al-Ramadi, Otavio Cabral-Marques
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Resource and Technical Advance Cell biology Immunology

CD40L modulates transcriptional signatures of neutrophils in the bone marrow associated with development and trafficking

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Abstract

Neutrophils are produced in the BM in a process called granulopoiesis, in which progenitor cells sequentially develop into mature neutrophils. During the developmental process, which is finely regulated by distinct transcription factors, neutrophils acquire the ability to exit the BM, properly distribute throughout the body, and migrate to infection sites. Previous studies have demonstrated that CD40 ligand (CD40L) influences hematopoiesis and granulopoiesis. Here, we investigate the effect of CD40L on neutrophil development and trafficking by performing functional and transcriptome analyses. We found that CD40L signaling plays an essential role in the early stages of neutrophil generation and development in the BM. Moreover, CD40L modulates transcriptional signatures, indicating that this molecule enables neutrophils to traffic throughout the body and to migrate in response to inflammatory signals. Thus, our study provides insights into the complex relationships between CD40L signaling and granulopoiesis, and it suggests a potentially novel and nonredundant role of CD40L signaling in neutrophil development and function.

Authors

Tábata Takahashi França, Ashraf Al-Sbiei, Ghada Bashir, Yassir Awad Mohamed, Ranieri Coelho Salgado, Lucila Akune Barreiros, Sarah Maria da Silva Napoleão, Cristina Worm Weber, Janaíra Fernandes Severo Ferreira, Carolina Sanchez Aranda, Carolina Prando, Mayra B. de Barros Dorna, Igor Jurisica, Maria J. Fernandez-Cabezudo, Hans D. Ochs, Antonio Condino-Neto, Basel K. Al-Ramadi, Otavio Cabral-Marques

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Figure 2

CD40L modulates the transcriptional profile of BM-derived neutrophils.

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CD40L modulates the transcriptional profile of BM-derived neutrophils.
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(A) Schematic diagram of the neutrophil isolation protocol used for RNA isolation. (B) Principal component analysis (PCA) of transcriptome profile exhibited by CD40L–/– and WT mice (n = 9/group). (C) Volcano plot representing the expression changes of all genes. Significantly down- and upregulated genes (adjusted P ≤ 0.05) are colored blue and yellow, respectively. Genes that do not show significant expression changes are colored black. Random labeling was performed in some genes of each side. (D and E) Modular gene coexpression analysis of all genes (figure showing the most enriched Module, called here as M1). Network interaction highlighting gene nodes with the potential hubs labeled (D) and gene set enrichment analysis shows the enrichment of Module 1 (symbol color represents the normalized enrichment score [NES]; top) and overrepresentation analysis of the enriched pathways in module 1 (−log10 adjusted P value, bottom; E).

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