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Updated: Mar 1, 2026
Isolation and Characterization of Neutrophils with Anti-Tumor Properties
Published on: June 19, 2015
Gene silencing and a novel monoallelic expression pattern in distinct CD177 neutrophil subsets
Claudia Eulenberg-Gustavus1, Sylvia Bähring1, Philipp G Maass2,3
1Experimental and Clinical Research Center, a joint cooperation between the Charité Medical Faculty and the Max-Delbrück Center for Molecular Medicine at the Charité, Berlin, Germany.
Insights
Researchers discovered epigenetic mechanisms controlling CD177 expression in neutrophils. Hematopoietic stem cells silence one CD177 allele during differentiation, creating distinct CD177-positive and CD177-negative neutrophil subsets.
Area of Science:
- Immunology
- Epigenetics
- Cell Biology
Background:
- CD177 is expressed on neutrophils and implicated in autoimmune and alloimmune diseases.
- Individuals exhibit distinct CD177-deficient, CD177-negative (CD177neg), and CD177-positive (CD177pos) neutrophil subsets.
- Mechanisms governing this subset-restricted CD177 expression remain largely unknown.
Purpose of the Study:
- To elucidate the epigenetic mechanisms controlling CD177 expression in bimodal neutrophil subsets.
- To investigate the unique monoallelic expression pattern of the CD177 gene.
- To understand how hematopoietic stem cells regulate CD177 allele silencing during neutrophil differentiation.
Main Methods:
- Analysis of CD177 protein and mRNA expression in CD177pos and CD177neg neutrophils.
- Haplotype analysis to determine CD177 allele transcription patterns.
- Chromatin immunoprecipitation (ChIP) and reporter assays in HeLa cells.
- DNA methylation analysis and transcription factor binding assays (c-Jun, c-Fos) in human neutrophils.
Main Results:
- CD177 protein and mRNA were exclusively produced by CD177pos neutrophils.
- A novel monoallelic CD177 expression pattern was identified, with offspring stably transcribing either the maternal or paternal allele.
- Hematopoietic stem cells expressed both CD177 alleles but silenced one during neutrophil differentiation.
- Epigenetic modifications, specifically DNA methylation, regulated CD177 reporter activity.
- Demethylation promoted biallelic CD177 expression, while methylation reduced it.
- Transcription factors c-Jun and c-Fos increased CD177 mRNA levels.
- CD177pos neutrophils exhibited an unmethylated, euchromatic CD177 promoter with bound c-Jun and c-Fos, unlike CD177neg neutrophils.
Conclusions:
- Epigenetic mechanisms, including DNA methylation and transcription factor binding (c-Jun, c-Fos), explain the distinct CD177pos and CD177neg neutrophil subsets.
- A novel form of monoallelic CD177 expression, distinct from classical random monoallelic expression or imprinting, governs allele silencing during neutrophil differentiation.
- These findings provide a molecular basis for understanding CD177 neutrophil heterogeneity in health and disease.
Abstract:
CD177 presents antigens in allo- and autoimmune diseases on the neutrophil surface. Individuals can be either CD177-deficient or harbor distinct CD177neg and CD177pos neutrophil subsets. We studied mechanisms controlling subset-restricted CD177 expression in bimodal individuals. CD177pos, but not CD177neg neutrophils, produced CD177 protein and mRNA. Haplotype analysis indicated a unique monoallelic CD177 expression pattern, where the offspring stably transcribed either the maternal or paternal allele. Hematopoietic stem cells expressed both CD177 alleles and silenced one copy during neutrophil differentiation. ChIP and reporter assays in HeLa cells with monoallelic CD177 expression showed that methylation reduced reporter activity, whereas demethylation caused biallelic CD177 expression. HeLa cell transfection with c-Jun and c-Fos increased CD177 mRNA. Importantly, CD177pos human neutrophils, but not CD177neg neutrophils, showed a euchromatic CD177 promoter, unmethylated CpGs, and c-Jun and c-Fos binding. We describe epigenetic mechanisms explaining the two distinct CD177 neutrophil subsets and a novel monoallelic CD177 expression pattern that does not follow classical random monoallelic expression or imprinting.
