Genetic tool for specific tracking of mature neutrophils.
8 affiliations
- 1Department of Neurosurgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
- 2Shanghai Immune Therapy Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
- 3Joint Laboratory for Biomedical Research and Pharmaceutical Innovation, The Unit of Pathogenic Fungal Infection & Host Immunity, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
- 4Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A∗STAR), Singapore, Singapore.
- 5Institute of Experimental Pathology, Center for Molecular Biology of Inflammation, University Hospital Münster, University of Münster, Münster, Germany.
- 6Department of Allergy, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
- 7Department of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
- 8School of Medicine, Westlake University, Zhejiang, China.
Abstract
Tracking mature neutrophils remains challenging due to the lack of reliable cell surface markers. Although CD101 is a promising candidate as a marker for mature neutrophils, its stability under pathological conditions is unclear. Here, we developed a novel CD101-tdTomato reporter mouse model to specifically identify mature neutrophils. CD101 expression, tdTomato fluorescence, and neutrophil physiology were characterized by flow cytometry across peripheral tissues under homeostasis and by a transwell migration assay, in parallel with morphological features assessed by Wright-Giemsa staining. To assess marker stability, CD101-tdTomato and wild-type (WT) mice were treated with lipopolysaccharide (LPS) (200 ng, intraperitoneally, 2 h, n = 8), influenza virus (5 × 10³ PFU/mL, intratracheally, days 1 and 5, n = 6), or granulocyte colony-stimulating factor (G-CSF) (2.5 μg, intravenously, 2-6 h, n = 5), followed by flow cytometry analyses. CD101-tdTomato mice were crossed with Lysozyme-GFP mice to validate reporter specificity across myeloid subsets. We confirmed that CD101⁺ and tdTomato⁺ neutrophils display identical characteristics of mature neutrophils, including poly-segmented nuclei, cell size, and key functional properties under homeostasis. Under pathological challenges, the decrease in CD101 expression was not attributed to shedding or degradation of the protein, supporting its robustness under inflammatory conditions. Using CD101-tdTomato × Lysozyme-GFP mice, we established a dual-reporter system that enables precise distinction of mature neutrophils from total myeloid cells. Collectively, our findings support CD101 as a robust and reliable marker of neutrophil maturity, providing a foundation for its future applications in spatial transcriptomics and lineage tracing studies of the heterogeneity and functions of neutrophils.
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BibTeX
@article{cao2026genetic,
title = {{Genetic tool for specific tracking of mature neutrophils.}},
author = {Jiayu Cao and Hui Ping Yaw and Shilin Yi and Yanxi Zhou and Shifan Qin and Yuanyuan Wang and Raul da Costa and Lei Zhang and Dandan Wu and Changbin Chen and Melissa Ng and Immanuel Kwok and Oliver Soehnlein and Xiaoxiang Chen and Jieqing Wan and Lai Guan Ng},
journal = {HLife},
year = {2026},
month = may,
doi = {10.1016/j.hlife.2026.05.007},
url = {https://europepmc.org/article/MED/42903866},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Genetic tool for specific tracking of mature neutrophils.
AU - Jiayu Cao
AU - Hui Ping Yaw
AU - Shilin Yi
AU - Yanxi Zhou
AU - Shifan Qin
AU - Yuanyuan Wang
AU - Raul da Costa
AU - Lei Zhang
AU - Dandan Wu
AU - Changbin Chen
AU - Melissa Ng
AU - Immanuel Kwok
AU - Oliver Soehnlein
AU - Xiaoxiang Chen
AU - Jieqing Wan
AU - Lai Guan Ng
PY - 2026
DA - 2026/05/28
JO - HLife
DO - 10.1016/j.hlife.2026.05.007
UR - https://europepmc.org/article/MED/42903866
AB - Tracking mature neutrophils remains challenging due to the lack of reliable cell surface markers. Although CD101 is a promising candidate as a marker for mature neutrophils, its stability under pathological conditions is unclear. Here, we developed a novel CD101-tdTomato reporter mouse model to specifically identify mature neutrophils. CD101 expression, tdTomato fluorescence, and neutrophil physiology were characterized by flow cytometry across peripheral tissues under homeostasis and by a transwell migration assay, in parallel with morphological features assessed by Wright-Giemsa staining. To assess marker stability, CD101-tdTomato and wild-type (WT) mice were treated with lipopolysaccharide (LPS) (200 ng, intraperitoneally, 2 h, n = 8), influenza virus (5 × 10³ PFU/mL, intratracheally, days 1 and 5, n = 6), or granulocyte colony-stimulating factor (G-CSF) (2.5 μg, intravenously, 2-6 h, n = 5), followed by flow cytometry analyses. CD101-tdTomato mice were crossed with Lysozyme-GFP mice to validate reporter specificity across myeloid subsets. We confirmed that CD101⁺ and tdTomato⁺ neutrophils display identical characteristics of mature neutrophils, including poly-segmented nuclei, cell size, and key functional properties under homeostasis. Under pathological challenges, the decrease in CD101 expression was not attributed to shedding or degradation of the protein, supporting its robustness under inflammatory conditions. Using CD101-tdTomato × Lysozyme-GFP mice, we established a dual-reporter system that enables precise distinction of mature neutrophils from total myeloid cells. Collectively, our findings support CD101 as a robust and reliable marker of neutrophil maturity, providing a foundation for its future applications in spatial transcriptomics and lineage tracing studies of the heterogeneity and functions of neutrophils.
SN - 2949-9283
ER -